»Where the Future Challenges in Pharmaceutical Technology Meet 30+ Years of Experience« 25th th-27 September 2025, Bled, Slovenia 1 1 Impressum The 15th Central European Symposium on Pharmaceutical Technology Where the Future Challenges in Pharmaceutical Technology Meet 30+ Years of Experience: Book of Abstracts Editors: Mirjana Gašperlin, Rok Dreu, Alenka Zvonar Pobirk, Barbara Sterle Zorec Reviewers: Zrinka Abramović, Petra Kocbek et al. Authors: Peter Kleinebudde, Adam Haimhoffer, Maja Kus et al. Prepress design: Barbara Sterle Zorec, Alenka Zvonar Pobirk, Anže Zidar Design: Sebastjan Jenko Cover photo: Lake Bled (Aleš Krivec, www.bled.si) Publisher: Slovensko farmacevtsko društvo in Univerza v Ljubljani, Fakulteta za farmacijo URL address: The book of abstracts will be pdf format, available at: https://www.sfd.si/cespt2025/programme/scientific-programme/ Ljubljana, 2025 Kataložni zapis o publikaciji (CIP) pripravili v Narodni in univerzitetni knjižnici v Ljubljani COBISS.SI-ID 249007107 ISBN 978-961-97122-0-7 (Slovensko farmacevtsko društvo, PDF) 2 Welcome letter Dear Colleagues, The 15th Central European Symposium on Pharmaceutical Technology (CESPT) returns to its original venue, Bled, Slovenia, where the symposium was first established in October 1995 under the patronage of EUFEPS. Since that time, CESPT has been successfully hosted in a number of cities across Europe, including Portorož (Slovenia, three times), Vienna and Graz (Austria), Ljubljana (Slovenia, twice), Siófok and Szeged (Hungary), Dubrovnik (Croatia), Belgrade (Serbia), Gdańsk (Poland), and Ohrid (North Macedonia). This year’s symposium carries special significance as we celebrate the 30th anniversary, so the title “Where the Future Challenges in Pharmaceutical Technology Meet 30+ Years of Experience,” reflects both our tradition and our forward-looking perspective. The program will address five major thematic areas: • Pharmaceutical Manufacturing and Engineering • Delivery of Biopharmaceutics and Nanomedicine • Strategies for Optimizing Pharmacotherapy: Addressing Clinical Outcomes and Patient-Friendly Drug Delivery • Current Trends in Pharmaceutical Analytics • Biorelevant Methods of Assessment Across these areas, key topics include the evolving landscape of drug manufacturing such as continuous manufacturing, 3D printing, process intensification and automation — the design and delivery of advanced biopharmaceuticals, including peptides, proteins, and nucleic acid-based therapeutics as well as innovative delivery systems, such as nanoparticulate carriers. Further focus will be placed on bridging science with clinical practice, exploring strategies to enhance outcomes while supporting patient adherence and quality of life. The final section highlights innovations in analytical sciences, focusing on biorelevant dissolution testing and bio-predictive testing. The scientific program comprises 5 plenary lectures, 10 invited lectures, 21 oral presentations, and over 120 poster presentations, providing a comprehensive overview of today’s pharmaceutical challenges and opportunities. With more than 250 participants from 20 countries, CESPT 2025 promises to be an inspiring international forum for scientific exchange, collaboration, and innovation. We extend our sincere thanks to all sponsoring and supporting organizations and societies whose contributions have strengthened the visibility of this symposium within the international scientific community. We look forward to welcoming you to Bled, 25–27 September 2025. Prof. dr. Mirjana Gašperlin Prof. dr. Rok Dreu President of the symposium Co-president of the symposium 3 OU UR KNOWLEDG GE FOR YOUR HEALTH. www.krka a.biz 4 Sartorius BIA Separ rations Strategic Location: %" & !'% "#"( " (&% #"!% & % !# business. Slovenia combines a rich trad dition in pharmacy with cutting-edge innovations, making it an n ideal location for biopharma partnerships. Monolithic chr Biopharmaceutical Excellen nce: oma t ogr raph y columns and bioproces ssing Sartorius BIA Separations leads in the n next-gen biopharmaceutical processes, collaborating with business s partners from the USA (80%) solutions for l i f production d i n, " "& " '# " !&"#" &#! ! ! ! !'&' " $ % ! %" $ % ! % % "&!% " " & (' %&$#" %! " ""% large biomolecules for g gene Innovative Campus: therapy and vaccines. The campus features state-of-the-art f facilities and highly skilled workforce with close to 30 years’ expe rience in gene therapy processes and products development to meet biopharma manufacturing needs. Advanced Therapies: ! "&! &%' ("&"!% "& !& & % ! $ "##% &'"!% !&" ( "#!&% (" human plasma fractionation and more, driving the present and the future of medicine. Our products and e expertise were utilized in the development and manufacture of gene therapy product Zolgensma www.biaseparations.com and many others—some not disclosed and many still in trials. 5 Partner organisations (Photo: Jošt Gantar, www.bled.si) 6 CONTENT Committees 8 General Information 9 Key Information 10 Social Events 10 Symposium Timetable 13 Conference Floor Plan 15 Scientific Programme 16 Poster session 20 Sponsors 21 Plenary lectures 25 Invited lectures 33 Oral presentations 52 Poster presentations 95 (Photo: Jošt Gantar, www.bled.si) 7 COMMITTEES Honorary Committee Organizing Committee • Dominique Duchêne, France • Katarina Bolko Seljak • Atilla Hincal, Turkey • Miha Homar • Franc Kozjek, Slovenia • Mojca Kerec Kos • Julijana Kristl, Slovenia • Petra Kocbek • Aleš Mrhar, Slovenia (president) • Peter Molek • Fulvio Rubessa, Italy • Odon Planinšek • Stanko Srčič, Slovenia • Barbara Sterle Zorec • Jelka Šmid Korbar, Slovenia • Jurij Trontelj • Natalija Škrbina Zajc International Scientific committee • Tomaž Vovk • Zrinka Abramovič, Slovenia • Anže Zidar • Ildikó Bácskay, Hungary • Carla Caddeo, Italy President of the Symposium • Ildikó Csóka, Hungary Mirjana Gašperlin • Jelena Filipović-Grčić, Croatia • Co-President of the Symposium Nikola Geskovski, N. Macedonia • Rok Dreu Svetlana Ibrić, Serbia • Albin Kristl, Slovenia General Secretary of the Symposium • Jasmina Lovrić, Croatia • Alenka Zvonar Pobirk Janina Lulek, Poland • Jelena Parojčić, Serbia Secretary of the Symposium • Luka Peternel, Slovenia Andrijana Tivadar • Beatrice Perissutti, Italy • Eva Roblegg, Austria (Photo on the right: • Maja Simonoska Crcarevska, N. Macedonia Statue of the Genius • Malgorzata Sznitowska, Poland (sculptured by Dane Zajc, • 1935) located above Zvone Simončič, Slovenia • the entrance to the Edina Vranić, Bosnia and Hercegovina University of Ljubljana Faculty of Pharmacy) The 1st CESPT Symposium took place in October 1995 in Bled, Slovenia, under the patronage of EUFEPS. Since then, the event has been successfully organized in several locations across different countries, including Portorož (Slovenia, twice), Vienna and Graz (Austria), Ljubljana (Slovenia), Siofok and Szeged (Hungary), Dubrovnik (Croatia), Belgrade (Serbia), Gdansk (Poland), and Ohrid (North Macedonia). After 30 years, 15 th CEPST Symposium is returning to its original venue – Bled. Over the past three decades, CESPT has been recognized as a well-established platform for promotion of new scientific, professional and business collaborations. Today it is a renowned international event regularly attracting more than 200 experts from industry, academia and regulatory authorities worldwide and in particular from Central and Eastern Europe. 8 GENERAL INFORMATION CONFERENCE VENUE: Rikli Balance Hotel, Cankarjeva cesta 4, SI–4260 Bled. The ARNOLD congress center, in the ground floor. PRE-EVENT WORKSHOP VENUE: University of Ljubljana, Faculty of Pharmacy, Aškerčeva cesta 7, SI–1000 Ljubljana Meeting point in the faculty lobby. THE EXHIBITION is located in SONCE – ZRAK congress center in the ground floor of Rikli Balance Hotel. REGISTRATION AND INFORMATION is located in SONCE – ZRAK congress center in the ground floor of Rikli Balance Hotel. Opening hours: Thursday, September 25, 12.00–18.30 Friday, September 26, 8.00–18.30 Saturday, September 27, 8.00–13.30 BADGES Each participant will receive a badge. All participants are required to carry the badges during the conference and social events. FOOD AND DRINKS Coffee breaks (coffee, tea and water & sweet and fruit snacks) will be served in the SONCE – ZRAK congress center during breaks. On Friday lunch will be served during lunch break in the dining hall of the Rikli Balance Hotel (included in the registration fee for participants). INTERNET A wireless internet connection is provided free of charge. (Photo: Faculty of Pharmacy, University of Ljubljana (left), and Arnold Hall, Rikli Balance Hotel Bled (right)) 9 KEY INFORMATION – SCIENTIFIC PRESENTATIONS If you are a session chairperson, you are kindly asked to arrive at the ARNOLD lecture hall at least 15 minutes before the start of the session to meet with the speakers. There are three types of lectures at the 15th CESPT: - plenary lectures – 40 minute and 5 minutes for discussion, - invited lectures – 25 minutes and 5 minutes for discussion, - oral presentations – 10 minutes and 5 minutes for discussion. The chairs are kindly requested to adhere strictly to the scheduled times of sessions. As a speaker, you are kindly asked to arrive in the ARNOLD lecture hall at least 15 minutes prior to the start of the session to meet with the chairpersons and the technician in charge of projection. The standard format for presentations is MS-PowerPoint. Please use the “embedded fonts” to avoid certain problems when projecting files prepared on a different computer. For all other types of video presentations, a laptop of the speaker must be used. Authors may keep their presentation on a USB flash drive. Files must be uploaded to the central computer in the ARNOLD lecture hall before the session begins. It is recommended that the presentation be uploaded before 8.45 a.m. for the morning sessions and during the lunch break for the afternoon sessions. Preview of presentations is available during breaks in the ARNOLD lecture hall. If you are a poster presenter, please refer to the poster list at the website https://www.sfd.si/cespt2025/programme/poster-session/ for the number. Authors are asked to hang their posters on the display boards on Thursday and remove them at the end of the Saturday session. Some mounting materials are available at the information desk. Posters will be on display throughout the 15th CESPT Symposium. The main poster session is scheduled for: Friday, September 26, 2025. SOCIAL PROGRAM All social events are included in the symposium fee. All participants will be asked to present their conference badge and admission ticket before being admitted. Welcome Reception Date: Thursday, September 25, 2025 Time: 19.00 Venue: Rikli Balance Hotel Lobby You will enjoy meeting colleagues and friends in the unique atmosphere of the lake Bled. Conference Dinner Date: Friday, September 26, 2025 Time: 19.30 Venue: Grand Hotel Toplice, Cesta svobode 12, Bled Program: Dinner and Musical entertaining program 10 T echn e nological Expertise Alway ys Available v for Y ou o HARKE Pharma GmbH +49 (0)208 3069-0 0 Business Unit Pharma Xantener Straße 1 pharma@harke.com m HARKE GROUP 45479 Mülheim an der Ruhr 11 Germany www.harke.com 11 12 12 SCIENTIFIC PROGRAMME 15th Central European Symposium on Pharmaceutical Technology Program with topics presented at the Symposium September 24–25, 2025 Pre-Event: CEEPUS Workshop for PhD Students: Introduction to Electrospinning University of Ljubljana, Faculty of Pharmacy, Ljubljana and Rikli Balance Hotel, Bled September 25, 2025 Opening of the 15 th CESPT 14.00 – 14.30 Session title: Pharmaceutical Manufacturing & Engineering Plenary lecture: Some learnings about roll compaction (Peter Kleinebudde) 14.30 – 15.15 Invited lectures 16.15 – 16.45 noon sessions Coffee break 16.10 – 16.45 After Invited lectures & oral presentations 16.45 – 18.45 Welcome reception 19.00 September 26, 2025 Session title: Delivery of Biopharmaceutics Plenary lecture: Understanding the formulation challenges of protein drugs 9.00 – 9.45 (Wolfgang Frieß) Invited lectures & oral presentations 9.45 – 11.00 Coffee break 11.00 – 11.30 ning sessions Session title: Nanomedicine Mor Plenary lecture: Nanomedicine for the treatment of childhood cancer 11.30 – 12.15 (María Blanco-Prieto) Invited lectures & oral presentations 12.15 – 13.45 Luncheon & Poster session 13.45 – 15.15 Session title: Strategies for Optimizing Pharmacotherapy Addressing Clinical Outcomes and Patient Friendly Drug Delivery Plenary lecture: Patient-centric drug product development acceptability across patient populations - Science and evidence 15.15 – 16.00 (Jörg Breitkreutz) noon sessions Invited lectures & oral presentations 16.00 – 17.00 After Coffee break 17.00 – 17.30 Invited lectures & oral presentations 17.00 – 18.30 Conference dinner 19.30 13 September 27, 2025 Session title: Strategies for Optimizing Pharmacotherapy Addressing Clinical Outcomes and Patient Friendly Drug Delivery (cont.) Invited lectures & oral presentations 9.00 – 10.30 Coffee break 10.30 – 11.15 Session title: Current Trends in Pharmaceutical Analytics and Biorelevant Methods of Assessment ning sessions Plenary lecture: Personalized Biorelevant Dissolution Testing: Addressing Mor the Impact of Population & Disease Variability on Oral Drug 11.15 – 12.00 Performance (Grzegorz Garbacz) Invited lectures & oral presentations 12.00 – 13.30 Closing of the Symposium 13.30 14 Confere ence floor plan Exhibitors 1. Krka 8. RED EYE M MONKEY 2. Asahi KASEI 9. AstrinexLa ab Arnold 3. MICRO+POLO 10. Hamilton 4. Anton Paar 11. ABL&E-JA ASCO 5. Lek 12. Mettler To oledo 6. LabTim 13. Novartis 7. Analysis Adria 14. Harke Pha arma P119 - Voda P150 P47 - P118 12 13 14 P0 – P46 11 10 1 9 2 Re Reg gi ist trat ti ion 8 3 7 6 5 4 Sonce Zrak CEEPUS Workshop for PhD Students September 24–28, 2025 Wednesday, September 24, 2025 (University of Ljubljana, Faculty of Pharmacy) 16:00 – 17.00 Registration for the CEEPUS Workshop 17:00 – 17.30 Welcome and Introduction to workshop Karin Kogermann, University of Tartu, Estonia 17:30 – 18.15 Nanofibers in Biomedicine with a Focus on Tissue Regeneration and Drug Delivery Špela Zupančič, University of Ljubljana, Slovenia 18:15 – 19.00 Nanofibers with Probiotics 19.00 Participant gathering and networking Thursday, September 25, 2025 (University of Ljubljana, Faculty of Pharmacy) Practical part of the workshop, including • Demonstration of nanofiber electrospinning using different devices 9:00 – 12.00 • Presentation of how selected parameters affect the process • Introduction to basic methods for characterizing the prepared samples Organised transport to Bled and participation in the 15 th CESPT 15th Central European Symposium on Pharmaceutical Technology Where the Future Challenges in Pharmaceutical Technology Meet 30+ Years of Experience Detailed Programme Thursday, September 25, 2025 (Rikli Balance Hotel, Bled) 12.00 – 18.00 th 15 CESPT Registration Opening ceremony 14:00 – 14.30 Chairpersons: Mirjana Gašperlin, Rok Dreu Aleš Mrhar, 30 years of CESPT Symposium (Opening lecture) Section 1 – Pharmaceutical Manufacturing & Engineering Chairpersons: Rok Dreu, Jelena Parojčić [PL] Peter Kleinebudde, Heinrich Heine University Düsseldorf, Germany 14:30 –15:15 Some learnings about roll compaction [IL] Adam Haimhoffer, University of Debrecen, Hungary 15:15 – 15:45 Development of hormone-containing vaginal film and applicator using QbD approach [IL] Maja Kus, Novartis d.o.o., Slovenia 15.45 – 16.15 Innovation in Pharma: Development and Implementation of Continuous Manufacturing for Commercial Production 16.15 – 16.45 Coffee break Plenary lecture (PL), Invited lectures (IL), Oral Presentations (OP), Poster Presentations (PP) 16 Section 1 – Pharmaceutical Manufacturing & Engineering (cont.) Chairpersons: Peter Kleinebudde, Malgorzata Sznitowska [IL] Klemen Korasa, Krka d.d., Novo Mesto, Slovenia 16.45 – 17.15 Process Analytical Technology: Cases and Insights from the Pharmaceutical Industry [OP] Teodora Tasevska, Ss. Cyril & Methodius University in Skopje, R North Macedonia 17.15 – 17.30 Predicting 3D printability of semi-solid extrusion formulations using rheological data and machine learning models [OP] Paola Šurina, Research Center Pharmaceutical Engineering & Graz University, Austria 17.30 – 17.45 From Powder to a Personalized Tablet: Optimizing Direct Powder Extrusion 3D Printing for Tablet Manufacturing [OP] Magdalena Urbanowicz, National medicine institute, Warsaw, Poland 17.45 – 18.00 Pre-formulation studies of SLS 3D printed tablets [OP] Hadi Shammout, University of Szeged, Hungary 18.00 – 18.15 Laser marking of tablets for anti-counterfeiting and its effect on coating integrity 18:15– 18.30 ® Dehydration and Purification of Microparticle Suspensions via Vibro [OP] Lukas Vergeiner, University of Graz, Austria TFF [OP] Karoliina Inno, University of Tartu, Estonia 18.30 - 18.45 The effect of tablet design and excipients on drug release from 3D printed tablets 19:00 – 22:00 Welcome reception at Rikli Balance Hotel Friday, September 26, 2025 08.00 – 18.00 th 15 CESPT Registration Section 2 – Delivery of Biopharmaceutics Chairpersons: María Blanco-Prieto, Eva Roblegg [PL] Wolfgang Frieß, Ludwig-Maximilians-University Munich, Germany 9.00 – 9.45 Understanding the formulation challenges of protein drugs Rok Sekirnik, Sartorius BIA Separations d.o.o, Slovenia 9.45 – 10.15 In-Process Analytics in Manufacturing of mRNA Therapeutics (promotion lecture) [OP] Monika Prašnikar, University of Ljubljana, Slovenia 10.15 – 10.30 Monoclonal antibody formulation optimization using novel viscosity-reducing and stabilizing excipients [OP] Martin Deak, University of Szeged, Hungary 10.30 – 10.45 Optimization of solid self-emulsifying drug delivery systems to achieve oral peptide therapy [OP] Ágnes Rusznyák, University of Debrecen, Hungary 10.45 – 11.00 Cyclodextrin polymers: Formulation and investigations of new RNA delivery systems 11.00 – 11.30 Coffee break Section 3 – Nanomedicine Chairpersons: Petra Kocbek, Karin Kogermann [PL] María Blanco-Prieto, University of Navarra, Spain 11.30 – 12.15 Nanomedicine for the treatment of childhood cancer Plenary lecture (PL), Invited lectures (IL), Oral Presentations (OP), Poster Presentations (PP) 17 [IL] Jasmina Lovrić, University of Zagreb, Croatia 12.15 – 12.45 Innovative aspects in the development of drug delivery platforms for the treatment of dry eye disease [OP] Luka Casula, University of Cagliari, Italy 12.45 – 13.00 Phospholipid vesicle-based topical formulation to unlock the potential of a biosynthetic melanin [OP] Ivana Ruseska, University of Graz, Austria 13.00 – 13.15 The role of protein corona in enhancing microRNA delivery via nanostructured lipid carriers [OP] Nina Katarina Grilc, University of Ljubljana, Slovenia 13.15 – 13.30 Sticky business: Investigating the role of nanofiber encapsulation in Lactobacilli surface adhesion and biofilm formation [OP] Imren Esenturk-Guzel, University of Health Sciences, Istanbul, Turkey 13.30 – 13.45 Development and in vivo evaluation of haemostatic silk fibroin-based nanofiber formulations 13.45 – 15.15 Luncheons / Poster session Section 4 – Strategies for Optimizing Pharmacotherapy Addressing Clinical Outcomes and Patient Friendly Drug Delivery Chairpersons: Odon Planinšek, Ildikó Bácskay [PL] Jörg Breitkreutz, University of Düsseldorf, Germany 15.15 – 16.00 Patient-centric drug product development acceptability across patient populations - Science and evidence [IL] Tomasz Osmałek, Poznan University of Medical Sciences, Poland 16.00 – 16.30 Tiny needles, big possibilities – microneedle systems in modern drug delivery [OP] Mercedes Vitek, University of Ljubljana, Slovenia 16.30 – 16.45 In situ forming liquid crystals for peptide drug delivery: From gelation dynamics and sustained release to biocompatibility assessment [OP] Rabia Ashfaq, University of Szeged, Hungary 16.45 – 17.00 Nanolipid gel-based local therapy for periodontal diseases: A dual-action anti-inflammatory and antibacterial strategy 17.00 – 17.30 Coffee Break Section 4 - Strategies for Optimizing Pharmacotherapy Addressing Clinical Outcomes and Patient Friendly Drug Delivery (cont.) Chairpersons: Jelena Filipović-Grčić, Maja Simonoska Crcarevska [IL] Odon Planinšek, University of Ljubljana, Slovenia Buccal films and Granules-in-straw 17.30 – 18.00 as patient friendly dosage forms [OP] Ilenia D’Abbrunzo, University of Trieste, Italy 18.00 – 18.15 Assembling three anthelmintic molecules in a single solid: A praziquantel/niclosamide/ acetic acid cocrystal solvate [OP] Ana Krese, Sandoz, Lek d.d., Ljubljana, Slovenia 18.15 – 18.30 Assessing the performance of ophthalmic gel: IVRT methodology 19.30 – 00.00 Conference dinner at Grand Hotel Toplice Plenary lecture (PL), Invited lectures (IL), Oral Presentations (OP), Poster Presentations (PP) 18 Saturday, September 27, 2025 Section 4 - Strategies for Optimizing Pharmacotherapy Addressing Clinical Outcomes and Patient Friendly Drug Delivery (cont.) Chairpersons: Tomaz Vovk, Grzegorz Garbacz [IL] Katarina Vučičević, University of Belgrade, Serbia 9.00 – 9.30 Optimizing Dosing Regimen of Vedolizumab in Patients with Inflammatory Bowel Disease: Choosing the Right Pharmacokinetic Model for Routine Therapeutic Drug Monitoring [IL] Lea Knez, University Clinic Golnik & University of Ljubljana, Faculty of Pharmacy, Slovenia 9.30 – 10.00 Immunocheckpoint Inhibitors in Cancer Immunotherapy: Optimizing Drug Therapy to Improve Treatment Outcomes [OP] Jadwiga Paszkowska, Physiolution Polska, Poland 10.00 – 10.15 From in vitro to in vivo: How reliable are our bioequivalence predictions? [OP] Jiri Zeman, Masaryk University, Czech Republic 10.15 – 10.30 Are original and generic extended-release metformin tablets truly interchangeable? 10.30 – 11.15 Coffee break Section 5 - Current Trends in Pharmaceutical Analytics and Biorelevant Methods of Assessment Chairpersons: Jurij Trontelj, TBC [PL] Grzegorz Garbacz, Physiolution, Wroclaw, Poland 11.15 – 12.00 Innovative methods and software tools for biopredictive testing of oral medicines [IL] Sandra Cvijić, University of Belgrade, Serbia 12.00 – 12.30 Personalized Biorelevant Dissolution Testing: Addressing the Impact of Population & Disease Variability on Oral Drug Performance [IL] Tomasz Bączek, Medical University of Gdańsk, Poland 12.30 – 13.00 Profiling of phytocannabinoids in medicinal cannabis plant material via solid-phase microextraction approach [OP] Andrej Grobin, University of Ljubljana Slovenia 13.00 – 13.15 A useful method for monitoring of antibiotics in wastewaters by SPE_LC_MS/MS [OP] Ewelina Baran, Pedagogical University of Cracow, Poland 13.15 – 13.30 In situ characterization of 3D printed pharmaceuticals using magnetic resonance imaging and relaxometry methods 13.30 Closing of the Symposium Plenary lecture (PL), Invited lectures (IL), Oral Presentations (OP), Poster Presentations (PP) 19 POSTER SESSION The list of posters with assigned numbers is available here: NOTES 20 SPONSORS PLATINUM SPONSORS KRKA At Krka, our dedication lies in producing high quality products, and keeping people healthy is what motivates us. From start to finish, everything we do revolves around our patients and our attempts to help them preserve and improve their health. Each day, more than 100 million patients in 70 markets are treated with our medicines, which are marketed under our own brand names. This is possible because of our innovative products, investments, and market expansion. Stable sales growth, multiple research projects, and sound business performance have seen us grow into a multinational pharmaceutical business group with almost 13,000 employees. SARTORIUS BIA SEPARATIONS Sartorius BIA Separations develops and manufactures leading CIM® monolithic chromatographic columns for purifying and analyzing large biomolecules like viruses, plasmids, and mRNA, used in cell and gene therapies. With over 25 years of experience, their Cornerstone® Biomanufacturing Development Services offer integrated process development solutions and novel technology to enhance the robustness and yield of biomolecule production while ensuring product safety. Their technology is pivotal in manufacturing-scale purification for advanced therapeutics and novel drug candidates in clinical trials. GOLD SPONSOR HARKE HARKE Pharma & Nutra is your technology driven distributor for innovative excipients. Our technical team provides R&D support and gives periodical online seminars. Our customers can count on a good quality, using special warehousing and transport partners following GDP guidelines. We are able to support our customers to be fully compliant with legal and regulatory requirements. We support with a comprehensive range of products for applications including coating, tableting, sustained-release dosage forms, solubility enhancement, granulation, pelletization, semi-solids and capsules. NOVARTIS In Novartis in Slovenia, we develop, manufacture, and market innovative medicines, supporting over half of Novartis’s priority treatments across various stages. Leveraging cutting-edge technology, we address diseases with unmet needs and play a key role in strengthening Slovenia’s healthcare. With locations in Ljubljana, Mengeš, and Slovenj Gradec, we employ over 3,800 skilled professionals. Our purpose is to reimagine medicine to improve lives, driven by research, innovation, and advanced manufacturing to enhance treatment accessibility … 21 21 SILVER SPONSOR LEK Lek, a Sandoz company, is the first and oldest pharmaceutical company in Slovenia. It develops, manufactures, and markets effective, safe and high-value generic and biosimilar medicines for patients around the world. Their work is guided by high ethical standards, innovative thinking, and breakthrough achievements of their top experts. With more than 3,700 employees, they have been actively contributing to the Slovenian, European, and global research and development landscape for decades. They foster a culture of open dialogue and proudly and responsibly hold the title of the most reputable employer in Slovenia. LABTIM Labtim provides complete solutions for analytical laboratories, with a strong focus on the pharmaceutical and biopharmaceutical industries. As an official distributor for leading manufacturersincluding Waters Corporation, Wyatt Technology, Sotax, TA Instruments, and Merck Millipore—we combine world-class technologies with expert local support. Our services span sales, application support, technical maintenance, education, and IT consultations. With expertise in LC, LCMS, SFC, tablet testing, sample prep, columns, water purification systems, and our own GMP lab, we ensure accuracy, compliance, and confidence at every step. BRONZE AND OTHER SPONSORS ASAHI KASEI EUROPE We are grateful to our sponsors who support pharmaceutical technology and drug delivery science and add value to the 15 th CEPST. Many of the symposium events are made possible only through the generosity of sponsors. 22 22 23 Are you ready for absolute answers in an uncertain world? GMP data analysis in Astra™ or Empower™ Achieve absolute molecular weight, size, and aggregation status Premier MaxPeak™ HPS for metal-sensitive Characterize mAbs, oligonucleotides, analytes ADCs, AAVs, PEGylated proteins Complete solutions Application Support, Technical Support, Educational Training, Sales, IT Consultations for analytical laboratories. and our in-house GMP Laboratory. 24 Peter Kleinebudde 1. ROLL COMPACTION/ DRY /*:.8 4/8 :7654:9=738 +7<:;8 43,8 )458 "=,9. 8 43,8 GRANULATION 649;<=4-854<46;9; :7;++=:=;39$8 ;43".=-; 8 /76;8 /=65-=+=;,8 >=<;:98:7654:9=7382>108=/89.;8/=65-;/986;9.7,8 455<74:.;/84<;84!4=-4(-;8".=:.84<;8!4-=,8*3,;<8 +7<85<7,*:=3)894(-;9/8'&%$8#7";!;< 8>18=/87+9;38 :;<94=38:73,=9=73/8'%$8.=/8+7<6/89.;8(4/=/8+7<8 3798 57//=(-;8 ,*;8 978 !4<=7*/8 =//*;/ 8 /*:.8 4/8 5<7:;//8 9<43/+;<8 43,8 /:4-=3)8 *58 =38 1$8 .;8 =3/*++=:=;398 +-7"4(=-=9 8 ";994(=-=9 8 6;4/*<;,8 /5;:=+=:8 :7654:9=738 +7<:;8 :438 (;8 94(-;94(=-=9 8 ,;6==3)8 7<8 ,*/98 5<7,*:9=73$88 <;-49;,89789.;85;485<;//*<; 8".=:.8=/8=657<94398 <43*-49=738=/8485<7:;//87+8=3:<;4/=3)854<9=:-;8 +7<8,;9;<6=3=3)89.;8/7-=,8+<4:9=7387+89.;8<=((73$ /=;8 ".;<;(8 /64--;<8 54<9=:-;/8 4<;8 :73!;<9;,8 =3978 -4<);<8 4))-76;<49;/$8 4<=7*/8 )<43*-49=738 7,;<38<7--8:7654:97 6;9.7,/84<;84!4=-4(-;89.498:4387!;<:76;8/76;8 (79.89.;8/5;:=+=:8:7654:9=738+7<:;843,89.;8)458 7<84--87+89.;84+7<;6;39=73;,8>185<7(-;6/$83;8 "=,9.8 /=6*-943;7*/-$8 ;!;<4-8 :739<7-8 :=<:*=9/8 6;9.7,89.498,7;/83798<;*=<;8(=3,=3)8-=*=,/8=/8 "7<8/=6*-943;7*/-843,8*=:-$8.=/8;34(-;/8 <7--8 :7654:9=73,<8 )<43*-49=73$8 38 9.=/8 9.;8 87+89.;8<=((738978(;8;598:73/9439$8;!;<4-8 6;9.7, 89.;8<4"8649;<=4-/84<;8+=455<74:.;/8.4!;8(;;385*(-=/.;,8+7<86;4/*<=3)8 <=((73/8(848<7--8:7654:97<843,89.;38)<43*-49;,8 9.;8<=((738 8=38<;4-89=6;$ =38 48 /;:73,8 /9;58 (8 )<=3,=3)$8 ,!4394);/8 =3:-*,;89.;84(/;3:;87+8-=*=,/ 8".=:.84!7=,/89.;8 .;8 6;438 /7-=,8 +<4:9=738 7+8 9.;8 <=((738 :438 (;8 3;;,8 +7<8 +*<9.;<8 ,<=3) 8 ;3!=<736;394-8 :739<7--;,8 7!;<8 9=6;$8 #7";!;< 8 9.;<;8 =/8 +<=;3,-=3;// 8;4/;87+8/:4-=3)8*5 843,89.;8+4:989.498 !4<=49=738 "=9.=38 9.;8 <=((73$8 55<74:.;/8 978 =98=/8438=3.;<;39-8:739=3*7*/85<7:;//89.498:438 =65<7!;89.;8.767);3;=987+89.;8 8"=9.=389.;8 ;4/=-8 (;8 =39;)<49;,8 =3978 48 :739=3*7*/8 <=((738.4!;8(;;385<757/;,$ 643*+4:9*<=3)8-=3;$8.;864=38,=/4,!4394);/84<;8 9.;8 .=).8 +=3;/8 :739;398 :7654<;,8 "=9.8 79.;<8 )<43*-49=738 5<7:;//;/8 43,8 9.;8 <;,*:;,8 3;87+89.;864=38,=/4,!4394);/8=/89.;854<9=4-8-7//8 94(-;94(=-=987+8/76;8649;<=4-/8'%$ 7+8 94(-;994(=-=98 +7--7"=3)8 1$8 .=/8 =/8 54<9=:*-4<-8 ;!=,;398 =38 5-4/9=:4--8 ,;+7<6=3)8 649;<=4-/$8 .=/8 5.;376;3738 .4/8 (;;38 ;9;3/=!;-8 /9*,=;,8 =38 9.;8 :739;98 7+8 7.43/738 '%8 5;<+7<6;,8 +*3,46;394-8 "7<8 6=:<7: <;)4<,=3)8 9.;8 *3,;<;-;!4398 978 6438 79.;<8 ;:=5=;39/8 43,8 /$8 :7654:9=73821085<7:;//8=389.;8&/$84/;,8 ;!;<4-8 <;4/73/8 .4!;8 (;;38 /*));/9;,8 +7<8 9.=/8 738 .=/8 "7< 8 3*6;<7*/8 67,=+=:49=73/8 ";<;8 54<9=4-8 -7//8 7+8 94(-;994(=-=9 8 "=9.8 54<9=:-;8 /=;8 5*(-=/.;,8 =38 9.;8 +7--7"=3)8 ,;:4,;/$8.;8 67/98 ;3-4<);6;398 43,8 )<43*-;8 .4<,;3=3)8 (;=3)8 9.;8 =657<94398=39;<6;,=49;8:<=9=:4-8*4-=98499<=(*9;8 67/98/=)3=+=:43986;:.43=/6/8'%$ =/8 9.;8 /7-=,8 +<4:9=738 7+8 9.;8 <=((73$8 .;8 /7-=,8 ;!;<4-8 455<74:.;/8 :438 .;-58 978 7!;<:76;8 9.;8 +<4:9=738 2 08 =/8 =657<94398 +7<8 9.;8 /*(/;*;398 54<9=4-8 -7//8 7+8 94(-;994(=-=9$8 /=3)8 /94<9=3)8 5<7:;//;/843,8,;9;<6=3;/89.;8<=((73/8/9<;3)9.8 649;<=4-/8 7+8 9.;8 /46;8 95; 8 (*98 "=9.8 /64--;<8 43,8)<=3,=3)8(;.4!=7*< 84/8";--84/89.;854<9=:-;8 =3=9=4-854<9=:-;8/=;/ 8:438:765;3/49;8+7<89.;8-7//8 /=;8 ,=/9<=(*9=73 8 +-7"4(=-=9 8 43,8 94(-;94(=-=98 =38 94(-;994(=-=9$88 /;:73,8 455<74:.8 =/8 978 */;8 7+89.;8)<43*-;/$8.;<;+7<; 848;87(;:9=!;8=/8978 /*=94(-;8 (=3,; :739<7-8 9.;8 8 43,8 ;;58 =98 4/8 :73/=/9;398 4/8 3*6;<7*/8 9=6;/$8 79.8 455<74:.;/8 :438 (;8 57//=(-;$ :76(=3;,$8 <;/;4<:.;5<757/;,8 9.498 48 -*(<=:4398 (;8 4,,;,8 5<=7<8 978 =39;<,;5;3,;3:;87+8;*=56;39854<46;9; 7<8649;<=4-/89.498/9=:89789.;8<7--/ 8=98.4/8(;;38 4/8<7--8,=46;9;<843,8"=,9. 85<7:;//854<46;9;5<;!;398 /9=:=3) 8 =98 4-/78 <;,*:;/8 9.;8 +<=:9=738 <7--=3)8 :765<;//=738 210$8 .=-;8 9.=/8 648 26 )('&%)($#"!#)( #"(#$(%)($#%# (#%# # (# )!( "%# )"%& )# "# "(# " )"# !#"(# )!(##%"#"(# $$%"%!#!# #&)% "# "(# !&"# $$($# #(# #"(#%%#"%(# $# %"(%"# )(# %!)" "# "!)# %# $("()%%#"(#!&"!( # %# (# &$()"!!$# !$()# ('&%("# %# % (#"!#!")!#"(#)!(#"(# (#&# $# )!(#") ()#%#( ## #(#%!)!) "($# %"!# !"%&!&# & "&)%# %(# !$(# )&( # (# ((# )!!($# "!# !()!(# "(# $% $ " (## 1. Leane, M. et al., A proposal for a drug product Manufacturing Classification System (MCS) for oral solid dosage forms. Pharmaceutical Development and Techology. 2015. 20: 12-21. 2. Kleinebudde, P. Roll compaction/dry granulation: pharmaceutical applications European Journal of Pharmaceutics and Biopharmaceutics. 2004. 58: 317-326. 3. Johanson, J.R. A Rolling Theory for Granular Solids. Journal of Applied Mechanics. 1965. 32: 842. 4. Kleinebudde, P. Improving Process Understanding in Roll Compaction. Journal of Pharmaceutical Sciences. 2022. 111: 552-558. 5. Sun, C.C. and Kleinebudde, P. Mini review: Mechanisms to the loss of tabletability by dry granulation. European Journal of Pharmaceutics and Biopharmaceutics. 2016. 106: 9-14. %# !)# !&$# !"# (# ((# !%(# %"!&"#"(#(# $#( (("#!#&()!&# # "&$("# !"$!"!) # )(( )()# $# )"()#"#%#!"#!%(#"!#("%!#"(# # %$%%$& ## #$((#) "(&#"!# #!#"(# ! !) "%!# %# (# "!# (() "%# (# !($(# 27 Wolfgang Friess #"! " " " ! " " " 1. INTRODUCTION c) Computational Models d) Bulk Freezing e) Stabilization by Lyophilization 1. Koepf E., Eisele S., Schroeder R., Brezesinski G., Friess W., Notorious but not understood: How liquid-air interfacial stress triggers protein aggregation. International Journal of Pharmaceutics, 2018. 537: 202-212. 2. Marschall C., Witt M., Hauptmeier B., Friess W., Powder suspensions in non- aqueous vehicles for delivery of therapeutic proteins. European Journal of Pharmaceutics and Biopharmaceutics, 2021. 161:37-49. 3. Deiringer N., Friess W., Proteins on the Rack: Mechanistic Studies on Protein Particle Formation During Peristaltic Pumping. Journal of Pharmaceutical Science, 2022. 111: 1370- 1378. 4. Isaev N., Lo Presti K., Friess W., Insights into Folding and Molecular Environment of Lyophilized Proteins Using Pulsed Electron Paramagnetic Resonance Spectroscopy. Molecular Pharmaceutics, 2025. 22: 424-432. 5. Binder J., Zalar M., Huelsmeyer M., Siedler M., a) Protein Self Interaction protein self-interaction simulations. European Curtis R., Friess W., Enhancing Martini 3 for b) Protein Interaction with various Journal of Pharmaceutical Sciences, 2025. 209: Interfaces 107068 28 NANOMEDICINE FOR PEDIATRIC CANCER THERAPY AND TISSUE REGENERATION "! "! " " 54321 0/.-,+/1 *3001 )+/2/(-1 -4/1 '/&/0%)$/(-1 %#1 .+3-3."01 $/'3."01 .4"00/(!/2 1 )/'3"-+3.1 ."(./+1 1. %'+!,/-Nogales C, et al., ACS Nano. 12 "'&"(./'1-4/+")/,-3.12-+"-/!3/21-%1"''+/221-*%1 -+/"-$/(-1"('1+"3(1-322,/1+/!/(/+"-3%( 2. (2018) 7482-7496. Lasa-Saracibar B, et al., Cancer Lett 334:2 54/1 #3+2-1 )"+-1 *3001 #%.,21 %(1 -4/1 '/23!(1 "('1 (2013) 302-310. 3. Aznar MA, et al., Int J Pharm, 454:2 (2013) "))03."-3%(1 %#1 03)3'1 ("(%)"+-3.0/21 0%"'/'1 *3-41 720-726 "(-3-,$%+1 "!/(-21 #%+1 -4/1 -+/"-$/(-1 %#1 4. %( 0/-/+( ('/11/-1"01"(./+1/-11 %2-/%2"+.%$"1 "1 43!401 "!!+/223&/1 )/'3"-+3.1 (2017) 262 268. ."(./+1 54/2/1 ("(%)"+-3.0/21 "+/1 2)/.3#3."001 5. %( 0/-/+( ('/11/-1"01"(./+1/--11 /(!3(//+/'1 #%+1 %+"01 "'$3(32-+"-3%(1 %##/+3(!1 "1 (2018) 193-200. (%(3(&"23&/1 )"-3/(-#+3/('01 "))+%".41 -4"-1 6. Garbayo E et al., Biomaterials, 110 (2016) 11-/(4"(./21 '+,!1 2-"303-1 "('1 3%"&"30"303-1 23. *430/1$3(3$33(!122-/$3.1-%3.3-1+/.03(3."01 7. Garbayo E et al., WIREs Nanomedicine and /##3.".13(1+/',.3(!1)+3$"+1-,$%+1,+'/(1"('1 10.1002/wnan.1637. 8. Torres-Ortega et al., Biomacromolecules, 629 )+/&/(-3(!1$/-"2-"-3.12)+/"'1)%23-3%(3(!1-4/$1 2-,'3/21 4"&/1 '/$%(2-+"-/'1 -4/3+1 +/$"+"0/1 Nanobiotechnology, (2020), (2022), 122356. "21 "1 )+%$323(!1 -4/+")/,-3.1 2-+"-/!1 #%+1 %2-/%2"+.%$"1 %+/%&/+1 -4/1 %+"01 '/03&/+1 $/-4%'1321"(-3.3)"-/'1-%123!(3#3."(-013$)+%&/1 )"-3/(-1.%$)03"(./1"1.+,.3"01#".-%+13(1)/'3"-+3.1 %(.%0%! 54/1 2/.%('1 )"+-1 %#1 -4/1 0/.-,+/1 *3001 /)0%+/1 "'&"(./$/(-213(1+"3(1-322,/1+/)"3+1*3-43(1-4/1 .%(-/-1%#1"+3(2%(21'32/"2/1)/.3#3."0013-1 *3001 '32.,221 4%*1 -4/1 '/&/0%)$/(-1 %#1 '/03&/+1 22-/$21 "2/'1 %(1 3%'/!+"'"0/1 )%0$/+21 .%$3(/'1 *3-41 (/,+%-+%)43.1 #".-%+21 2,.41 "21 !03"01./00'/+3&/'1(/,+%-+%)43.1#".-%+11 ."(1)+%$%-/1-4/1+/!/(/+"-3%(1%#1'%)"$3(/+!3.1 (/,+%(21 "('1 +/2-%+/1 $%-%+1 #,(.-3%(1 3(1 (%( 4,$"(1)+3$"-/1$%'/021%#1"+3(2%(21'32/"2/1 54/2/1 22-/$21 4"&/1 24%*(1 !+/"-1 )%-/(-3"01 3(1 +/!/(/+"-3(!1 '%)"$3(/+!3.1 )"-4*"21 "('1 %&/+.%$3(!1 .4"00/(!/21 +/0"-/'1 -%1 )+%-/3(1 '/03&/+1-%12)/.3#3.1+"3(1+/!3%(21%##/+3(!1(/*1 -4/+")/,-3.1 "&/(,/21 #%+1 -4/1 -+/"-$/(-1 %#1 (/,+%'/!/(/+"-3&/1'32%+'/+2 5%!/-4/+1 -4/2/1 /"$)0/21 ,('/+2.%+/1 -4/1 -+"(2#%+$"-3&/1 )%-/(-3"01 %#1 3((%&"-3&/1 '+,!1 '/03&/+122-/$21-%1"''+/221.%$)0/1.4"00/(!/21 3(1%(.%0%!1"('1+/!/(/+"-3&/1$/'3.3(/1-4+%,!41 $,0-3'32.3)03("+1"))+%".4/2 29 )(' )&%$#"%! #"%$ ) %##()#"%&"& #)%&$($% & )&%$#"%))(&%$" $#" #&"#$#" # Breitkreutz $#"! $$ !$"$ !$"$"$"! $!! $"! !! $$$ !! $" 1. 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FDA/CDER, Patient-focused drug B6C=D5>4=D@6C>=<: ;463: @>44D<47: D9: @2DC;4<9: >9;: approval-process-drugs/cder-patient-focused-drug-development. 72?1:9?:D90<4D?4D=/:=?:>:7/465:<.<9:65:=?:-,:0?4: EDCBA@?>=<;: BD9D=>8C<=7: @>9: 8<: 67<;: >7: development, ww.fda.gov/drugs/development-@2DC;4<9:?0:C<77:=2>9:-:/:>9;:65:=?:+**:0?4:):/: 2. FDA/CDER, Patient-focused drug ?C;: ?9<7: ('&%: $2<: ;DB<97D?97: @>9: 8<: <.<9: important to patients development: methods to identify what is , Guidance, 2022. <9C>43<;: =?: -%,: BB#: 7=DCC: =2<: @?B5?7D=<: <9;A 3. V. Klingmann, C.E. Pohly, T. Meissner et al., 5?D9=:B<=2?;:;=<:76D=>8DCD=/:8<=1<<9:+: Acceptability of an orodispersible film B:>9;:):/%:: compared to syrup in neonates and infants: A randomized clinical trial, Eur J Pharm Biopharm 2020, 151: 239-245. "4?;D75<47D8C<: BD9D=>8C<=7: 2>.<: 8<<9: 0D47=: 4. S. Stegemann, V. Klingmann, S. Reidemeister, 54?5?7<;:D9:-*++:(++&%:!9:=2<: :069;<;:54?<@=: J. Breitkreutz, Patient-centric drug product :12D@2:1>7:@?B5C<=@@?B5>9D<;:8/: development acceptability across patient ;=<7: ?0: =2<: 2DC;4<97: <>4=: E<;<4>=D?9: populations - Science and evidence. Eur J >9;: +%**: B3: <9>C>54DC>=: B>C<>=<: 1<4<: 5. %:04D>: 7@>C<4>#:%>49:<=: al., 76@@<7706CC/: ;<.=: ?64: 69D.<47D=/: >9;: =1?: =/5<7: ?0: ?4?;D75<47D8C<: BD9D=>8C<=7: *%-,: Pharm Biopharm, 2023. 188: 1-5. =4>970<44<;: =?: =2<: @?9=4>@=D93: B>960>@=64<4: for paediatric oral dosage form. endpoint method development on acceptability <=52>4B>%: "$7: 1<4<: 54?;6@<;: 69;<4: Involvement and Engagement, 2023, 9: 108. Research : @?9;D=D?97: >9;: 67<;: 0?4: >: @?B5>4>=D.<: 6. %: 9@2#: !%: <77C<4#: %%: ?77<: <=: >C%#: 8D?>.>DC>8DCD=/: 7=6;/: 67D93: 2<>C=2/: >;6C=: Evaluating the acceptability, swallowability, .?C69=<<47%: $2<: BD9D=>8C<=7: 72?1<;: and palatability of film-coated mini-tablet 52>4B>@?D9<=D@: 54?0DC<7: 7DBDC>4: >7: <9D=<@ formulatio in young children: results from an : 04?B: : >9;: 9?: =4>97B6@?7>C: >87?45=D?9: open-label, single-dose, cross-over study, @CD9D@>C:7=6;D<7:D9:5><;D>=4D@:5>=D<9=7:1D=2:2<>4=: 7. V. Klingmann, M. Hinder, T.H. Langenickel et al., Acceptability of multiple coated mini-tablets 0>DC64<7:D9:.>4D?67: 64?5<>9:?69=4D<7:D9:?4;<4: (+-&%:>7<;:?9:=2<7<:4<76C=7#:1<:D9D=D>=<;:=24<<: Pharmaceutics, 2023. 15: 1729. =?: >55C/: 0?4: >: ><;D>=4D@: 7<: >4<=D93: a randomised controlled study, in comparison to syrup in infants and toddlers: Arch Dis Child 6=2?4D7>=D?9: : >=: =2<: %: : : 2023, 108: 730-735. 4<6<7=<;: 064=2<4: D9A.D=4?: 7=6;D<7: ?9: 0??;: 8. N. Spomer, V. Klingmann, I. Stoltenberg et al., D9=<4>@=D?97: >9;: 67<: ?0: 9>7?3>7=4>C: =68<7%: <: Acceptance of uncoated mini-tablets in young 0?69;: =2<: 9>C>54DC: "$7: =?: 8<: 54<@D7children Results from a prospective ;?7<;#: 7=>8C<#: >9;: 06CC/: >@@<5=<;: 8/: >CC: exploratory cross-over study, Arch Dis Child @>4<=><47: 0?69;: =28C<: =2>9: 9. V. Klingmann, N. Spomer, C. Lerch et al., Favourable acceptance of mini-tablets 0?4B<4: 5><;D>=4D@: 7685?56C>=D?97%: 2DC;4<9: 2012, 97: 283-286. : <=9 B<;D@D9<7%: ?=2: 54?;6@=7: 1<4<: =4>970<44<;: =?: compared with syrup: a randomised controlled 4?.<@>:=;%#:!49;#:12?:4<@<9=C/:4<@Pediatr. 2013, 163:1728-1732. trial in infants and preschool children, J. B>4<=D93:>6=2?4D7>=D?9:0?4:8?=2:;?7<:7=4<93=2: 10. V. Klingmann, A. Seitz, T. Meissner et al., 0?4: 64?5<: >9;: D9=4?;6@<;: =2<: 9<1: : Acceptability of uncoated minitablets in 54?;6@=: D9: 7?B<: : B=<7: >C4<>;/%: neonates a randomized controlled trial, J. "=2<4:@?69=4D<7:1DCC:0?CC?1:7??9% Pediatr. 2015, 167: 893-896. 11. I. Stoltenberg, J. Breitkreutz, Orally disintegrating mini-tablets (ODMTs) A novel solid dosage form for paediatric use, Eur J 463A04<<: BD9D=>8C<=7: 2>.<: 8<<9: 72?19: D9: Pharm Biopharm 2011, 78: 462-469. 5>=D<9=A@<9=4D@D=/:7=6;D<7:=?:8<:76D=>8C<:;?7>3<: 12. %: <4#: %: >14;=: <=: >C%#: 0?4B7: D9: =2<: 5><;D>=4D@: 5?56C>=D?9%: E4?B: =2<: Enalapril and enalaprilat pharmacokinetics in <9=4/:?0:>:54?;6@=:1>7:4<>CD <;:+:/<>47:C>=<4:8/: cardiomyopathy and congestive heart failure after administration of an orodispersible 0?CC?1D93:>:@?97D7=<9=:5>=D<9=A0?@67<;:5>=21>/%: 8>7D@: D;<>: ?0: >9: "$: D9: -*++#: =2<: B>4<=: children with heart failure due to dilated enalapril minitablet (LENA-studies), Pharmaceutics 2022, 14: 1163. 31 /.- ,++*)(',)&%$&'#*"!%(+"%!* '(&%'**.!% *%,*/&",',)& '&!',+%* %*(.%$&",,+& %%$%!% %/% %"%$% ! %$% %"%" 1. 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Subramanian, P. Mucoadhesive Delivery System: A SmartWay to Improve Bioavailability of Nutraceuticals. Foods. 2021; 10: 1362. 3. ;07-'4 %4 7:9096'4 %4 !:78:/0394 324 Monolayer and Bilayer Buccal Films Containing Metoclopramide. Pharmaceutics. 2024; 16: 354. 4. 0.04 $'4 370.:74 '4 093!:4 '4 :8.<:;/96;4'47:90964%445/;:(423;4 paediatrics: How to administer highly dosed, bitter tasting paracetamol granules. Int J Pharm. 2021; 602: 120615. 44 765 43271707/.-,4+7/.-*).71)/-4(-'),4670&1%$-7/-3%27)/2+-#72"- 7/(6%11%24*!-$4#)6-,7+)%+) -"44+7/.-2")-*7."2- 3"%*1%47/)27-14,)6-(4*-*4&27/)-2")*%3)&27-,*&.- 14/724*7/. -' ! 1. INTRODUCTION =10?,-?,9/=1"7 2=:=>9,7 ?,7 @=2/=10/7 0?+9,=9/A27 =17 +9,.=1"7 (9.2%7 )A7 :?@A>27 (A,A7 ;/=>= BA@?>=<;:9876B5437=2797:?1?0>?19>791/=8?@.7 $?1/A7 9,>?7 2=:;>9/=?127 /?7 "A1A,9/A7 !7 9--,?+A@7 *?,7 /)A7 /,A9/:A1/7 ?*7 =1*>9::9/?,.7 -,?*=>A27 892A@7 ?17 0?+9,=9/A7 0?:8=19/=?127 8?(A>7 @=2A92A7 6'&53%7 $?@A>#=1*?,:A@7 2-A0=*=A@7 (=/)=17 /)A7 :?@A>27 91@7 /)A7 2/91@9,@7 -,A0=2=?17 @?2=1"7 6$'!537 =27 97 -?(A,*;>7 @?2=1"7,A"=:A1%7'179@@=/=?17/)A2A7:?@A>27(A,A7 9--,?90)7 /?7 =1@=+=@;9>= A/A,19>>.7A+9>;9/A@7;2=1"7 5$7:A92;,A:A1/27 ?-/=:= 91@7 -?-;>9/=?17 -,A@=0/=?127 892A@7 ?17 /)A,9-A;/=07 @,;"7 :?1=/?,=1"7 6 5$37 @9/97 91@7 0)9,90/A,=2/=027?*7?;,70?)?,/7?*7-9/=A1/2%791"A7 9@+910A7 5$#892A@7 @?2=1"7 =1@=+=@;9>=<9/=?1%7 ?*7 -,A@=0/=?17 A,,?,7 :A/,=027 (A,A7 A:->?.A@7 /?7 @?2A7 =1@=+=@;9>=<9/=?17 =27 ";=@A@7 8.7 @A/A,:=1A7 8=927 91@7 -,A0=2=?1%7 @@=/=?19>>.7 -)9,:90?:A/,=07 :?@A>#@A,=+A@7 0?+9,=9/A27 2=:;>9/=?1#892A@7 @=9"1?2/=027 (A,A7 ;/=>= ()A,A927 =1@=+=@;9>=<9/=?17 A+9>;9/A7:?@A>7-A,*?,:910A%7 =10?,-?,9/A278?/)7:?@A>70?+9,=9/A2791@7?1A7?,7 :?,A7?82A,+A@70?10A1/,9/=?127/?7,A*=1A7@?2=1"7 -,--',0-33-1 -,A@=0/=?12%77 0,=/=09>7 0?:-?1A1/7 ?*7 $'!57 =27 B547 0?10A1/,9/=?127 (A,A7 919>. /)A72A>A0/=?17?*7/)A7:?2/79--,?-,=9/A7-?-;>9/=?17 1?1>=1A9,7 :=A@#A**A0/27 :?@A>=1"7 9--,?90)7 -)9,:90?=1A/=076!?-!37:?@A>7*?,7-,A@=0/=1"7 =:->A:A1/A@7 =17 $$7 2?*/(9,A7 6'0?17 ?-/=:9>7 @?2=1"7 ,A"=:A17 892A@7 ?17 =1@=+=@;9>7 5A+A>?-:A1/7 ?>;/=?127 +A,2=?17 %37 ;/=>=<=1"7 @A:?",9-)=0791@70>=1=09>7-9/=A1/70)9,90/A,=2/=027 !?-!7:?@A>7@A+A>?-A@78.7?29,=?7A/79>%77 (=/)7)=")A2/7-,?898=>=/.7?*7,A90)=1"7/9,"A/7@,;"7 (=/)=17 /)A7 !'7 8>?0%7 )A7 922?0=9/=?17 0?10A1/,9/=?12%7 )A71;:8A,7?*7-;8>=2)A@7B547 8A/(AA17 B547 0>A9,910A7 637 91@7 !?-!7:?@A>27,A:9=127>=:=/A@7#791@7/)A=,7 @A:?",9-)=07 /,A9/:A1/7 91@7 0>=1=09>7 0>=1=09>7 9-->=098=>=/.7 )927 1?/7 .A/7 8AA17 0)9,90/A,=2/=027 (927 =1+A2/="9/A@7 ;2=1"7 917 2.2/A:9/=09>>.7 A+9>;9/A@%7 ?7 @A/A,:=1A7 /)A7 9;/?:9/A@7 2/A-(=2A7 0?+9,=9/A7 :?@A>=1"7 :?2/7 2;=/98>A7 :?@A>27 *?,7 B547 @?29"A7 9--,?90)% =1@=+=@;9>=<9/=?17 (A7 0?1@;0/A@7 917 A/A,19>7 A+9>;9/=?17 ?*7 9+9=>98>A7 :?@A>2791@7 @A+A>?-A@7 -*)+&62+-%/,-,7+&++74/ ?;,7:?@A>7;2=1"7,?;/=1A7 5$729:->A2%77 -+- -3 -3- -1%2)*7%6+-%/,-1)2"4,+ --',0-33- -1 ()=>A7 ="%7 7 -,A2A1/27 2=:;>9/=?127 892A@7 ?17 A7)9+A7A+9>;9/A@79>>7A=2/=1"7!?-!7:?@A>27 7 0?)?,/7 ?*7 7 '&57 -9/=A1/27 /,A9/A@7 9/7 /)A7 ?29,=?7A/79>%7:?@A>7%7 1=+A,2=/.7 $A@=09>7 A1/A,7 4+A<@9,97 =17 /)A7 A-;8>=07 ?*7 A,8=97 -,?+=@A@7 ?17 9+A,9"A7 #7 B547 /,?;")7 0?10A1/,9/=?12%7 5A:?",9-)=07 @=2A92A7>98?,9/?,.791@7-,=?,0;,,A1/7/,A9/:A1/7 0)9,90/A,=2/=027(A,A79+9=>98>A7927(A>>7% - 1 - - 1- - +- - 3 - 3- - )- 33-1 7 0?:-,A)A12=+A7 ,A+=A(7 =@A1/=*=A@7 2A+A,9>7 -;8>=2)A@7 !?-!7 :?@A>27 ?*7B547 #7 A90)7 45 .-, +*)('&%$#% ONMLKJIHGFEDFCBA@NKHF?J>HGFA=FMAGHKF? #%+! ;A>JBNAF HIF JK:F 987F @AKKA6N=5F 436HH2F GA>N=5F GLBN=5F MJN=IH=J=1HF N=F 0BA/=.>F GN>HJ>HF CACLKJINA=F 1. Rosario, M., et al., Population 1/JBJ1IHBN-HGF ?<,F +KH@I*F =AF CBH)NAL>F ?NAKA5N1F pharmacokinetics-pharmacodynamics of IBHJIMH=I(F ?AG vedolizumab in patients with ulcerative colitis J=IN?AGNH>F GHIH1IHG(F JK?LMN=F "&F 5! F J=GF +BN5/I*F . Alimentary CBH)NAL>F ?NAKA5N1F IBHJIMH=I(F ?AG Pharmacology & Therapeutics, 2015. 42: 188-GHIH1IHGFJ=IN3%$#FJ=IN?AGNH>(FJK?LMN=F&F5! :FKLHF 202. GAIIHGFKN=HFBHCBH>H=I>FKH)HKFA@F8F5!M : 2. Okamoto, H., et al., Population pharmacokinetics of vedolizumab in Asian and J?>AKLIHFCBHGN1INA=FHBBABF+E*F@ABF . Intestinal Research, 2021., 19: F 95-105. CBHGN1INA=>F ?J>HGF A=F I/HF CJINH=I>.F 1A)JBNJIH>F HGNJ=F CBHGN1INA=F HBBABF +$E*F J=GF MHGNJ=F non-Asian patients with ulcerative colitis and J=GF;A>JBNAFHIFJK:FMAGHKF987FJ=GF1JK1LKJIHGF?pharmacodynamic model of vedolizumab for 3. Hanzel, J., et al., Pharmacokinetic- 1AMCJBN=5F CBHGN1INA=>F 6NI/F MHJ>LBHMH=I>F targeting endoscopic remission in patients with BH>LKIHGF N=F )JKLH>F A@F :'F J=GF "':(F Crohn disease: Posthoc analysis of the LOVE-BH>CH1IN)HKCD study. Inflammatory Bowel Diseases, 2022. 28: 689-699. "#"#% ! '&'**% % !**% 4. A)J=A)N(F :(F HIF JK:(F Review and External &% Models for Vedolizumab in Patients with Inflammatory Bowel Disease: Assessing F /N5/F GH5BHHF A@FN=IHBN=GN)NGLJKF)JBNJ?NKNI.**% % &&&% % % Evaluation of Population Pharmacokinetic Predictive Performance and Clinical %$#F 1A=1H=IBJINA=>F 6J>F A?>HB)HG:F F Applicability. Biomedicines, 2024. 13(1): 43. ><>IHMJIN1FH)JKLJINA=FA@F1A)JBNJIH>FBH)HJKHGFJF 5. JB2A)N(FO:(FHIFJK:(FVedolizumab Clearance as >N5=N@N1J=IF GN@@HBH=1HF N=F %$#F 0 F N=F CJINH=I>F a Surrogate Marker for Remission in 6NI/F F HCA>LBHF IAF N=@KNNMJ?F ABF Inflammatory Bowel Disease Patients: Insights JGJKNMLMJ?F I/HBJC<(F 6NI/F 0 F ?HN=5(F A=F from Real-World Pharmacokinetics. J)HBJ5H(F :"F /N5/HBF N=F CBH)NAL>KPharmaceutics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ea Knez1, 2 1Faculty of Pharmacy, University of Ljubljana, Slovenia 2Pharmacy, University Clinic Golnik, Slovenia 1. INTRODUCTION 7#76725(#76,727#7;9,2:%2&65(722:62 25//:6(0;&2 9:2 9872 *:==:;2 :0/0932 *60976052 %:62(#76,72 >==<;:98765432 10982 0==<;72 /87/.4:0;92 #7;9,2 17672 67/:6(7(2 0;2 2 5;(2 $2 5;(2 0;80-09:6,2 +>*>)2 85,2 (65=590/5''32 /85;&7(2 9872 96759=7;92 (0,/:;90;<590:;2 0;2 2 5;(2 2 :%2 96759=7;92:%2;<=76:<,2/5;/76,$25/807#0;&2':;&" 45907;9,2 0;2 9872 =:;:">2 5;(2 /87=:">2 &6:<4$2 976=2 67,4:;,7,2 5''<(0;&2 9:2 9872 4:,,0-0'0932 :%2 67,47/90#7'32 !:2 96759=7;9"67'597(2 (75982 15,2 /<6725',:20;2965(090:;5''324::6246:&;:,0,2/5;/76,2 67/:6(7(2872:-,76#7(2,5%793246:%0'72/:=4567,2 5,2=795,9590/2=7'5;:=52:625(#5;/7(2;:;",=5''2 %5#:65-'32 10982 (5952 %6:=2*,$2 4:,,0-'32 5',:2 /7''2'<;&2/5;/762+5! **)2,2>*>285#72-77;2 (<729:2756'076296759=7;92(0,/:;90;<590:;2:%2>*>2 7;9760;&2987296759=7;92567;52:%2#560:<,29347,2:%2 5;(2 <;(76674:690;&2 :%2 5(#76,72 7#7;9,2 10980;2 /5;/76,$2#560:<,2(0,75,72,7990;&,25;(2=7(0/0;72 6:<90;72/'0;0/5'2465/90/72 /:=-0;590:;,$2 092 0,2 0=476590#72 9:2 5,,7,,2 5;(2 ,9675='0;72 (6<&2 98765432 9:2 0=46:#72 96759=7;92 87,72 (5952 5672 675,,<60;&2 5;(2 7,47/05''32 :<9/:=7,20;27#763(532/'0;0/5'2465/90/72 :=72:%2 0=4:695;925,2%06,9,25=:;&298:,72:60&0;597(2%6:=2 98727%%:69,2<;(7695.7;2-3267,756/876,2%6:=29872 9872 *7;965'2 5,976;2 <6:475;2 5675$2 ;0#76,0932 *'0;0/2 :';0.2 5;(2 5/<'932 :%2 /8565/97607(2-3252'5/.2:%2875'98/567267,:<6/7,2 856=5/3$2 ;0#76,0932 :%2 <-'5;52 10''2 -72 67,<'90;&20;252&5420;2/5;/762/:;96:'2/:=4567(29:2 467,7;97(210980;2980,2'7/9<67222 7,976;2<6:472 6759=7;92 10982 >*>2 6:<&8'32 (:<-'7(2 9872 65;,'590;&298727;/:<65&0;&267,<'9,25/807#7(20;2 ,<6#0#5'2:%245907;9,21098252! **2 90''$2:;'3252 987280&8'32(7%0;7(2%65=72:%265;(:=0,7(2/'0;0/5'2 9806(2 :%2 45907;9,2 67,4:;(2 9:2 96759=7;9$2 98<,2 9605',2+*)20;9:29872#5605-'725;(2<;467(0/95-'72 67,756/82 7%%:69,2 5672 (067/97(2 9:156(,2 ,09<590:;,2 0;2 6:<90;72 /'0;0/5'2 465/90/72 =532 -72 <;(76,95;(0;&29872=7/85;0,=,2:%20;960;,0/25;(2 /85''7;&0;&2 872 7%%0/5/3"7%%7/90#7;7,,2 &542 7960;,0/2 67,0,95;/72 9:2 >*>2 98765432 9:2 0(7;90%32 180/8256720=476590#720;29872/5,72:%20;96:(98592 5'9762 &<92 =0/6:-0:952 =532 0;976%7672 >*>2 7%%7/90#7;7,,2>;24:,9"8:/25;5'3,7,2:%2*,25;(2 :%2 ;712 96759=7;92 =:(5'0907,2 10982 52 #7632 =532 -72 0;%:6=7(2 :;'32 -32 675'"1:6'(2 ,9<(07,$2 153,29:2:#76/:=72092=:;&2:9876,$2=7(0/0;7,2 :-,76#590:;5'2,9<(07,274:,<6729:246:9:;24<=42 47/<'0562=:(72:%25/90:;25;(2<;0<729:0/0907,25,2 8<,2172/:;(0;2 45907;9,2 967597(2 10982 >*>2,2 =7(0/0;72 <,72 2/:;,7/<90#7245907;9,2109825! **$2967597(2 =532 (0%%762 0;2 #560:<,2 875'98/5672 ,3,97=,2 5;(2 10982 %06,9"'0;72 >*>$2 7098762 5,2 =:;:98765432 &7:&65480/267&0:;,$21725,,7,,7(298720=45/92:%2 >*>20==<;:9876543 =7(0/0;7,2176725,,:/0597(2109821:6,72,<6#0#5'2 0;80-09:6,2 +>)$2 5;90-0:90/,2 +59-)2 5;(2 :98762 +=:;:">2245907;9,)2:620;2/:=-0;590:;210982 ,7'7/97(2 /:;/:=095;92 =7(0/0;7,2 :;2 >*>2 /87=:98765432 +/87=:">2 2 45907;9,)$2 592 52 :<9/:=7,20;252/:8:692:%2;756'32 2/:;,7/<90#72 ,0;&'725/5(7=0/20;,909<90:;2 22872:-,76#7(2 45907;9,2 10982 5!**$2 967597(2 10982>*>2 0;2 5;32 =7(05;2 :#765''2 ,<6#0#5'2 += )2 15,2 2 ,7990;&2:62/:=-0;590:;2 =:;98,2 5;(2 2 =:;98,2 0;2 9872 =:;:">2 5;(2 /87=:">$2 67,47/90#7'32 87,72 67,<'9,2 '56&7'32 0980;2 980,2 50=2 172 5,,7,,7(2 9872 <,72 :%2 59-2 /:=456721098298:,72%6:=2987240#:95'2*,2:%2 10980;2 9872 37562 460:62 5;(2 <;90'2 9872 7;(2 :%2 >*>2 >*>,$2(7,40972:<62/:8:6920;/'<(7,245907;9,210982 96759=7;92 2 :,92 45907;9,2 +)2 17672 4::6762 476%:6=5;/72 5;(2 =:672 /:=:6-0(0907,2 467,/60-7(25;259-$2=:,92:%97;25247;0/0''0;2+)$2 985;298:,720;/'<(7(20;2*, 67/70#0;&252=7(05;2:%2274:,<67,25;(22(50'32 47 DCBA@CD? D>=C=? <;;;:? 9A87A@? 87C? >6=C5438A>@? 8>? 34>AD? +CDA/A@C=? 8738? +3.? 73+2C5? -,-? 2C5A>D1?05C=/5A28A>@?>B?386?9A87A@?>@C?.C35?25A>5? CBBC/8A4C@C==? =7>D? 6C? 25A>5A8A=CD? >4C5? 8>?-,-?93=?3==>/A38CD?9A87?=7>58C5?+*)?<386?4=? 5C3/8A4C?3225>3/7C=?3DDA@?+>5C?+CDA/A@C=?8>? @>@(386'?&%1$?4=?#"1!?+>@87= ?&1#$ ?"? >4C5/>+C?87C?735+?A+2>=C?6.?+CDA/A@C=?>B8C@? ,-'? &1$$(&1%:? A@? @A(? 68? @>8? +8A435A38C? 25C=+CD?3=?735+C==?3=?00-=1?? 3@3.=A=1?=C?>B?386?A=?/>++>@?A@?238AC@8=?9A87? 3),,?3@D?+3.?73+2C5?-,-?85C38+C@8?87=? =8538CAC=? 8>? +A@A+AC? @@C/C==35.? 386? =C? & 0CA/>@?1?*)('&1?%*'&$#"!& (")"*( =7>D?6C?C2>5CD?A@?B85C?5C=C35/71? "( ")*!'( #)( "!( #)")( 587C5?>@?9C?3==C==CD?87C?A+23/8?>B?87C?8A+A@? *")*!'( ( !)$&*( )!*')*)( "( 3@D? A@8C@=A8.? >B? 00-=? 85C38+C@8? >@? =54A43? *@/>? ' '* ("$'&&(*&&(&('*!1?5>@8? #$#? &'? D>A'? >8/>+C=? 1? 2C/8CD.? @C35.? %"? >B? &$1B>@/1#$#1&&#%!1 238AC@8=?73D?3//C==?8>?00-=?9A87A@?&?.C35?25A>5? $?D3.=?>B?-,-?=8358?<00-?4=?@>?00-'?&"1!?4=?#&1? 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'20 * * * * * * * * * * * * * * * * * * 0'6.%$ -++12006.% '#2 * * * * * * * * * * * * * * * * * ** * * * * * * * * 6"3-3-!' / 3/3&5-'6/. +602-02 (-* * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * (-16-)6 656' /. /1-5 +1&% * * * * * * * * * * * * * * * * * * * * * * * * * * ** * * * * * * * * 321 /1"-.!2 0 ! * * * * * * * * * 1. INTRODUCTION !(.)1 0( !2* + + + + + + + + + + + + + + + + + +-&( 1.##/*'*,-1 &,,! -"* 1#*%2$22% + + + + + + + + + , + + + + + + + + + & + + + + + + + + + + + + + + + + + + $+ + + + + + + + + #.(#01* 0$ -" + + + + + + + + + + + + + + + + + + "* 1-.! + + + + + + + + ++ + + + + + + + + 32110/.-20, -*1-2,) (*'&2,1 & %0(,,*(1-0,* 0$ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + ( + + + + + + + + + + + + + + + + + + #"&(' ( '&%*.-2%&/+ + + + + + + + + !* */0#'*,-,+ + + + + + + + + *,1.(2,)+ + + + + + + + + 0'#*,!2&/ / !2110/.-20, -*1-1 $&&2/ -0 %&#-.(* + + + + + + + + + + + + + + + + + + + + + + + + + + + $ + + + + + + + + + + + + + + + + + ++ + + + + + + + + %0,121-*,- !(.) (*/*&1* #(0$22/*1 &,,! #(*!2%-2,) + + + + + + + + + + + + + + + + + + + + + + + + + + + $ + + + + + + + + + & + + + + + + + + ++ + + + + + + + + !2 *(1* * + + + + + + + + + + + + + + + + + + *, 2(0,'*,- 0$ 1#*%2$22% #0#./&--20, + + + + + + + + + + + + + + + + + + $ + + + + + + + + + &+ + + + + + + + + 2, 2 0 !(.) #*($00(' + + + + + + + + + + + + + + + + + + + + + + + + + + + $ ( '&,%* *1#*%2&// $00( 10/2! + + + + + + + + + + + + + + + + + + $ + + + + + + + + ++ + + + + + + + + 0( !21*&1* )( + + + + + + + + + + + + + + + + + + (0.#1 (21 2,) .,!*(*1-2'&-20, 0$ & + + + + + + + + + + + + + + + + + + & + + + + + + + + + + + + + + + + + + + + + + + + + + + 0(&/ !01&)* $00(' + + + + + + + + + + + + + + + + + + $ ( '1 "2/* -(&!2-20,&/ '*-"0!1 + + + + + + + + + + + + + + + + + + + + + + + + + + ++ + + + + + + + + !(.) 2, 2 0 + + + + + + + + + + + + + + + + + + 0 #*($00(' + + + + + + + + + $ ( '&,,%* "*(*$00(* -"* * & + + + + + + + + + $ + + + + + + + + + + + + + + + + + ++ + + + + + + + + (*/ 0, 1-&,!&(!2*! %0,!2-20,1 !*$$22,*! + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + ++ + + + + + + + + %"&//*,)*1+ + + + + + + + + 2,+ + + + + + + + + !*12),2,)+ + + + + + + + + #*(10,&/2*!+ + + + + + + + + #"&(' ( '&%0#*2&+ + + + + + + + + & &,,!+ + + + + + + + + (*)./&--0( & (+ + + + + + + + + ).2!*/2,*1+ + + + + + + + + 20(*/* ,,-&+ + + + + + + + + !2110/.-20, -*1- 2,%/.! + + + + + + + + + + + + + + + + + +* 2!*,-2$2,) + + + + + + + + + $+ + + + + + + + + *'*())2,) (*1*&(%" "2)"/2)"-1 -"* ,**! $00( ( + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + $+ + + + + + + + + &,! 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(+ + + + + + + + + * * * * * * * * * +206%.6.%* * * * * * * * * -* * * * * * * * * 3210/.-56,2+* * * * * * * * * !2110/.-20, &(* &/-*(&-20,1 2, /.'2,&/ # . + + + + + + + + + + + + + + + + + + & + + + + + + + + + + + + + + + + + + + + + + + + + + ++ + + + + + + + +&,! + + + + + + + + + + + + + + + + + + )6/1252(-.' +600/5&'6/. 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" 987654321 0/1 .-1 57,763+3,21 *)/0,6*)(1 ") * 53,20)74*'3&1&*2204%+*0)1+32+1&32*()$ 7+*3)+253*/*1 .-1 0)&*+*0)21 7)1 5,0/0%)&41 Population/ Key considerations 7//3+1 &,%(1 &*2204%+*0)1 7)&1 0)23%3)+41 Disease/Condition (GI parameters) +"3,753%+*10%+0632$1"*21"*("4*("+21+"31)33&1 Paediatric patients Reduced stomach /0,153,20)74*'3&1&*2204%+*0)1+32+*)(1+"7+174*()21 volume; Altered gastric *)1 *+,01 60&3421 *+"1 57+*3)+253*/*1 .-1 pH, gastric emptying, 57+"05"2*040($1 -)1 57,744341 ,3(%47+0,1 GI motility pattern and /,7630,216%2+130431+0170660&7+31+"3231 luminal viscosity; *))07+*0)21 3)2%,*)(1 3%*+7431 &,%(1 3//*71 Reduced bile salts 7,0221&*3,231505%47+*0)2&*237231(,0%52$ concentration Geriatric patients " Reduced saliva excretion; Altered 1. Masada, T., et al., Bioequivalence of oral drug gastric pH; Decreased products in the healthy and special populations: GI motility; Reduced Assessment and prediction using a newly bile salts concentration Hypochlorhydria/A Reduced gastric acidity Pharmaceutics, 2021. 13(8): 1136. Diabetes Delayed relevant dissolution specifications (CRDSs) for gastric oral drug products: Virtual webinar series. emptying; Altered chlorhydria 2. McAllister, M., et al., Developing clinically (increased stomach pH) intestinal pH 3. Pharmaceutics, 2022. 14(5): 1010. #,0")!21&*23723 Reduced gastric acidity dissolution media in dissolution tests as a de Lemos, H., et al., Use of biorelevant (increased stomach pH); predictive method of oral bioavailability. Reduced gastric and Brazilian Journal of Pharmaceutical Sciences, increased small 2022. 58(3): e19759. intestine transit time; 4. Mire-Sluis, A., et al., Patient-Centric Quality Reduced bile salts Standards. Journal of Pharmaceutical Sciences, concentration 2024. 113(4): 837-855. 7,*)20)!21&*23723 Delayed gastric emptying; Increased )' Bariatric patients intestinal viscosity; /,7630,1 0/1+01 .,7)+1(,3363)+21 3+33)1 +"31 )*3,2*+1 0/1 34(,7&37%4+1 0/1 Altered motility pattern "7,671 7)&1 *)*2+,1 0/1 *3)31 Reduced fluid volume; Increased "*21 5,323)+7+*0)1 721 5,357,3&1 *+"*)1 +"31 gastric pH; Reduced GI gastric pH; Decreased 35%4*1 0/1 3,*71 (,7)+21 01 1 1 7)&1 01 gastric transit time; 1$ Altered gastric volume; Increased 3")040(*741 3340563)+1 7)&1 -))07+*0)1 stomach contractility " " '&" &##" $"# #0)+3650,7,1 ,3237,"1 *)*+*7+*321 "731 67&31 2*()*/*7)+1 5,0(,3221 *)1 &33405*)(1 *0,3437)+1 &*2204%+*0)1 63+"0&21 +7*40,3&1 +01 6*6*1 505%47+*0)1 0,1 &*23723253*/*1 .-1 0)&*+*0)2$1 "*21 5,323)+7+*0)1 *441 7&&,3221 20631 0/1 +"31 387654321&0%63)+3&1*)14*+3,7+%,3$1"*431+"3231 2+%&*321 &360)2+,7+31 5,00/1 0/1 0)35+1 /%,+"3,1 2+7)&7,&*'7+*0)1 7)&1 74*&7+*0)1 7,31 )33&3&1 +01 /%441*)+3(,7+3153,20)74*'3&1&*2204%+*0)1+32+*)(1 *)+01,0%+*)315,7+*317)&1,3(%47+0,1374%7+*0)$ 50 8765 43218780/.21.4-,+2*)00)(8028'&.80.%$'8*80)7.*)00)(8&.47)0+. %)+$38)7.#8).&278'"4-)&$.%8*32$!+3)*+820.)4432)*- Anna Roszkowska1,2 1 3,4 5 .. .#.+ , Joanna Bogusiewicz, Krzysztof Urbanowicz 6 3 5 6 2 , Patrik Schreiber , Barbara Bojko .3.+.& , Janusz Pawliszyn, +.(1 ,+*)('&%*$&#"!# ('%(*&(#*%&'#*(#$*'&#"!#(#(# "($ +*)('&%*$&#"!#*%&'#$*'&#"!#(&*'""#(&*'""#($(( ($$(#(&# $&*'$(&"$(#$(#**('#*$&'*#*#*) +*)('&%*$&#"!#(&'(#+'#(('#$*'&#*#*) +*)('&%*$&#"!# ('%("$(%#($#"# ('%(""#"(#")*'$#$*'&#"# "($ 6+*)('&%*$&#"!#"*%&'#*(#$*'&#"!#(#(# "($ 1. INTRODUCTION 4+81 9,51 :9#%987/5.;04,;.4651 /;%6:281 9,51 #%48.%6;:21 ,9027;!1 !:95,1 04331 362/,.;02,.1 <;:98765432109/.;2-,.4/,9;+1*<)('&193141+;+7 *777(<&1 2-54%3,9$21 3406:21 6.264.4,9;+1 ,2/5+9#%21 ,54,1 2+4":231,5212-,.4/,9;+1;!1:; 10;:2/%:4.1 295,1 .*20*7&820 /;06;%+831 !.;01 /;06:2-1 "9;:;9/4:1 04,.9/231 71+1,59313,%81<)('1 431;6,9092814+81 5216.;6;3281<)('7"4328102,5;8;:;9231/4+1 466:9281 "21 %3281 431 4+1 4889,9;+4:1 #%4:9,1 ,;;:1 !;.1 !;.1 89.2/,1 93;:4,9;+1 ;!1 65,;/4++4"9+;9831 *)3&1 !.;01 .; 9+1 0;+9,;.9+1,521/;+,2+,1;!164.,9/%:4.1)319+1,521 ($$(# )) 0289/9+4:1 /4++4"931 6:4+,1 /%:,9$4,9;+1 !4/9:9,9231 1 6:4+,31 * $# " 1 /;+89,9;+3&1 4+81 4:3;1!.;018.92816:4+,104,2.94:1 4+81 8%.9+1 /4++4"931 6:4+1 04,2.94:1 6.;8%/,9;+1 1 .%)+$38)7&.)0'.%$+-2'& .3$1$3$0*$& $# "1 <)('1 %,9:99+1 "9;/;064,9":21 1 /;%6:281 9,51 #%48.%6;:21 ,9027;!1 !:95,1 04331 use of SPME for drug analysis in clinical, toxicological, and forensic medicine studies . 362/,.;02,.1 *777(<&1 !4/9:9,4,281 ,521 6.;"231 4:;+1 9,51 :9#%987/5.;04,;.4651 1. 23/34112,14:1 Recent advances in the 2-,.4/,9;+14+814+4:3931;!14/989/1!;.031;!1)31 2. Talanta, 2024. 270: 125613. 9+1.; 9+10289/9+4:1/4++4"9316:4+,31 preparation technique for the monitoring of ;+9/4112,14:1SPME as a green sample- .3$&7+&.)0'.'8&*&&820 phytocannabinoids and endocannabinoids in complex matrices . Journal of Pharmaceutical 9+!:;.23/2+/231;!1, ;1$4.92,9231;!1 3. Roszkowska, A., et al., Monitoring of age- and ($$(#)) 1 gender-related alterations of endocannabinoid 6:4+,31 * <)('16.;"231 2.213,4,9/4::19+32.,2819+,;1,521 Analysis, 2023. 13: 1117. *1 78;09+4+,1 4+81 7 7 8;09+4+,&1 /%:,9$4,281 %+82.1 /;+,.;::281 SPME probes levels in selected brain regions with the use of /;+89,9;+31 521 .23%:,31 /;+!9.0281 ,54,1 ,521 4. . Metabolomics, 2023. 19(40): 1. ;+9/41 1 2,1 4:1 In vivo profiling of 82$2:;628102,5;81931416.2/93214+812!!9/92+,1,;;:1 phytocannabinoids in Cannabis spp. varieties ,54,1 2+4":231 89.2/,1 4+81 .46981 93;:4,9;+1 4+81 via SPME-LC-MS analysis. Analytica Chimica 4+4:3931 ;!1 )31 %+82.1 $# "1 /;+89,9;+31 Acta, 2024. 1306: 342621. 9,5;%,1 ,521 +2281 !;.1 /;::2/,9;+1 ;!1 6:4+,1 04,2.94:1+14889,9;+1<)('16.;"231 2.21%3281 !;.10;+9,;.9+1;!1,521/;+,2+,14+81/;06;39,9;+1 ;!1 )31 *4/989/1 4+81 +2%,.4:1 !;.03&1 4,1 89!!2.2+,1 3,4231;!1/4++4"9316:4+,104,2.94:154+8:9+1!.;01 54.$23,1 ;!1 6:4+,31 ,;1 !9+4:1 82/4.";-:4,281 6.;8%/,1'-,.4/,31;",49+281"1<)('1,2/5+9#%21 2.21 4+4:3281 4//;.89+1 ,;1 ,521 .2/;002+84,9;+31 ;!1 <1 )54.04/;62941 %,9:99+1 5951 62.!;.04+/21 :9#%981 /5.;04,;.4651 * )&1 9,51 1 82,2/,9;+1 51 /.- .,+*)(')&%$#*$.,)&'"!) )'$/$+)/ )*$+',)/&$ /, "')/&$)&%$,+/ /%)(" $*"'"$"&*$"()&+$ +",&)&%$ /*+ Teodora Tasevska1 1 1 2 , Nikola Geskovski , Katerina Goracinova , Riste Popeski Dimovski, Maja Simonoska Crcarevska1 "! ! 1. INTRODUCTION KEDJ 99J CPHIRPOPNAJ 2KFJ KFFQFFQD5J 9MOKCQEGFJ 2QHQJ LOKFFM@MQDJ KFJ (J :DHPI8;J /J :L?H3QD8;J PHJ J SRQPOPNMLKOJ IHPIQHGMQFJ KHQJ QFFQEGMKOJ MEJ :FGHKMNRGJ !J CC8;J 2MGRJ FGHKMNRGJ FGHMEN,OM>QJ DQGQHCMEMENJ GRQJ IHMEGKBMOMGAJ P@J IRKHCKLQ?GMLKOJ @MOKCQEGFJLPEFMDQHQDJIHMEGKBOQJ0=.5J9PHJ76JG2PJ ME>FJ MEJ = IQHIQEDML?OKHJOMEQFJ@HPCJQKLRJ@PHC?OKGMPEJ2QHQJ @PHCKGMPEJ :998J KEDJ OKAQHJ FGKL>MENJ :7685J 4J Q1GH?DQDJ GRHP?NRJ KJ %J EPOQJ PEGPJ KJ BKOKELQJP@JQOKFGML;J3MFLP?F;JKEDJLKIMOOKHAJ@PHLQFJ CMLHPFLPIQJFOMDQ;J@PHCMENJKJLHPFF;JKEDJKEKOAQDJ LPEGHPOJ @MOKCQEGJ @PHCKGMPE;J 2RMOQJ HKIMDJ ?EDQHJ FGQHQPCMLHPFLPIQJ :+66J 6GQCMJ =(!85J FGH?LG?HKOJHQLP3QHAJK@GQHJQ1GH?FMPEJMFJLH?LMKOJGPJ &KFQDJ PEJ OKAQHJ DQ@MEMGMPEJ KEDJ FGH?LG?HKOJ IHQ3QEGJOKAQHJDQ@PHCKGMPEJ0/.5J MEGQNHMGA;J @PHC?OKGMPEFJ 2QHQJ FLPHQDJ KFJ (J -RQJ KMCJ P@J GRQJ FG?DAJ 2KFJ GPJ KFFQFFJ GRQJ = :LPCIOQGQJ @?FMPE8;J /J :?ELOQKHJ BP?EDKHA8;J PHJ J IHMEGKBMOMGAJ IPGQEGMKOJ P@J FQCM,FPOMDJ Q1GH?FMPEJ :DMFGMELGJOKAQHFJ2MGRJLOQKHJFQIKHKGMPE8J0=.5J @PHC?OKGMPEFJ BAJ KIIOAMENJ CKLRMEQJ OQKHEMENJ CPDQOF;J ?FMENJ HRQPOPNMLKOJ DKGKJ KFJ MEI?GJ #$ $ $ $ $ $ 3KHMKBOQFJ KEDJ 99J KEDJ 76J HQF?OGFJ KFJ P?GI?GJ 3KHMKBOQF5 9QKG?HQFJ2QHQJQ1GHKLGQDJKEDJFGKEDKHDMQDJ?FMENJ 2. MATERIALS AND METHODS 6GKEDKHD6LKOQH5J 9PHJ / -$.$$ $$#*$ EQG2PH>J :8J CPDQOF;J LKGQNPHMLKOJ GKHNQGFJ -2QEGA,@P?HJ RADHPNQOJ @PHC?OKGMPEFJ 2QHQJ 2QHQJ PEQ,RPGJ QELPDQDJ 2MGRJ QHKFJ IHQIKHQDJ ?FMENJ DM@@QHQEGJ IPOACQHF5J +MNRGQQEJ GPLKGQNPHMLKOJ @?ELGMPE;J KEDJ MEI?GFJ 2QHQJ *)('J @PHC?OKGMPEFJ :&469;J %QHCKEA8J :/$5(#" HQFRKIQDJMEGPJ= )!5#/ JC8J2QHQJCKDQJBAJGRQJLPODJCQGRPD;J FGHKMEJFGQIF;JP?GI?GJ3KHMKBOQF85J9PHJGRQJSKEDPCJ KEDJ KDDMGMPEKOJ @PHC?OKGMPEFJ 2MGRJ GHKNKLKEGRJ 9PHQFGJ OKFFM@MQHJ :S98J CPDQO;J GKHNQGFJ 2QHQJ :' J KEDJ /( 8J KEDJ LRMGPFKEJ :/( 8J :6MNCK, QELPDQDJ ?FMENJ FLM>MG,OQKHEFJ HDMEKO+ELPDQH;J 4ODHMLR;J648JBAJDMHQLGJDMFIQHFMPEJMEJDMFGMOOQDJ KEDJGRQJDKGKJ2KFJHQGKMEQDJMEJMGFJPHMNMEKOJGKB?OKHJ 2KGQH5J 6M1GQQEJ *)('J @PHC?OKGMPEFJ MELO?DQDJ @PHCKG5J4J BKFMLJ J CPDQOJ :/ KLGM3QJF?BFGKELQFJLQ@M1MCQJ:GQE8JKEDJIHPBMPGML;J =J@MOGQHF;J>QHEQOJFMQJ;JCK1,IPPOMEN;JDHPIP?G;J KOPMDJ 4<;J 5JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ FP@GCK1J P?GI?G8;J KJ DQQIQHJ J CPDQOJ :G2PJ KLQDPEMK85 LPE3PO?GMPEKOJ OKAQHFJ 2MGRJ $)J KEDJ /#J @MOGQHF8;J KCIOMG?DQJ F2QQIJ CQGRPDJ 0.;J 2KFJ ?FQDJ GPJ GPJIHQDMLGJ99JKEDJ76JFLPHQF5J-RQJDKGKJ2QHQJFIOMGJ CQKF?HQJFRQKHJFGHQFF;JFGPHKNQJCPD?O?FJ:%8;JOPFFJ #((J @PHJ GHKMEMENJ KEDJ GQFGMEN5J PDQOJ CPD?O?FJ :%8;J OPFFJ @KLGPHJ :GKEJ 8;J GPH?QJ KEDJ IQH@PHCKELQJ 2KFJ Q3KO?KGQDJ ?FMENJ LPE@?FMPEJ LPCIOQ1J3MFLPFMGAJ:8J:*!(,/JLPEQJNQPCQGHA;J CKGHMLQF;J KEDJ IQH,LOKFFJ KLL?HKLA5J4DDMGMPEKOOA;J QHPJ NKIJ (5/()J CC;J !(5/;J KEN?OKHJ @PHJF;JOPFFJKEDJKLL?HKLAJL?H3QFJ2QHQJ?FQD5J @HQ?QELAJ/(JHKDJF;JFRQKHJFGHKMEJ(5/"/(( ;J=/J 9QKG?HQJ MCIPHGKELQJ 2KFJ LKOL?OKGQDJ @PHJ GRQJ S9J DKGKJIPMEGFJP3QHJ!((JFQLPEDF85J CPDQOJ GPJ KFFQFFJ GRQJ LPEGHMB?GMPEJ P@J QKLRJ ADHPNQOFJ 2QHQJ OPKDQDJ MEGPJ =J C7J IEQ?CKGMLJ HRQPOPNMLKOJ IKHKCQGQH5J -RQJ KEKOAFMFJ 2KFJ LKHGHMDNQFJ KEDJ IOKLQDJ PEJ GRQJ =DHPIP?G;J GHKMEQDJ @PHJ /!(J QIPLRFJ 2MGRJ QKHOAJ LPCIKLGJ HRQPCQGQHJ :SJ ;J 4EGPEJ *KKH;J FGPIIMEN8J 2QHQJ DQ3QOPIQD5J S9J CPDQOJ :/((J 4?FGHMK8J2MGRJKJ*QOGMQHJGQCIQHKG?HQJLPEGHPO5J-RQJ GHQQF8J2KFJKOFPJGHKMEQDJPEJFGKEDKHDMQDJ@QKG?HQFJ $, $$$ $$ BOPL>FJ2MGRJ&KGLRJPHCKOMKGMPE;J7QK>ASQ7;J KEDJ KEJ KD3KELQDJ J CPDQOJ :LPE3PO?GMPEKOJ :&MP ;J QOOME>;J 62QDQE85J-RQJ RADHPNQOFJ 2QHQJ Q1GH?DQDJ GRHP?NRJ KJ %J EPOQJ@PHJ =J FQLPEDF;J 53 CBA@?>= <;:9:876543C>= 2534<= 105C= ..= ;/A@435*= 8/= 8(5= *55@53= >= :8= A48@6/86:+>=4C*=<54+/3C) *5A/C<83485*=<83/C"=@53$/3A4C;5=:C=)=049:C"= 0(5= <8B*?= 4C46?'5*= &%= @/6?A53= $/3AB648:/C<= 4C46?<:<>=:8=;4C=+5=;/C<:*535*=4<=A/35== 8/= @35*:;8= != @/85C8:46=/$=5A:73. RESULTS AND DISCUSSION . = ;4@4+:6:8?= $/3= $548B35= :A@/384C;5= 466= 35 @3:C84+:6:8?=455*=8(3/B"(=..=4C*==;/35= 1,)=0(5=*48458=;/A@3:5*=%%%=:C@B8=*484= @/:C8)= = -535= ;(/5C= $/3= 8(5:3= 4+:6:8?= 8/= 5. REFERENCES -4= 565;85*= $/3= AB68:7;64= ;64:$:;48:/C= 4C*= ! ! )=56=!)==)= $548B35=:A@/384C;5=:C:"(8) ;4@8B35=85A@/346=*5@5C*5C;:5=-(:65=8(5=.= ) 433B64= ))= 58= 46)= "! 0(5=+4:;==A/*56=4;;B34856?=@35*:;85*=6/-= ) B884= ))= 58= 46)= @3:C84+:6:8?=164==..=)==)=@537;64= ! ! )= = 4;;B34;?,=+B8=83B""65*=-:8(=(:"(7B46:8?=@3:C8= *4C;5= )= 1!!,=!%!%)= 164= ,= @438:;B6436?= :C= ..= 1)!,= -(:65= !) /CC566=)=58=46)=! @53$/3A:C"=+58853=:C==1),)=8=46 ! )= A/*53485= @53$/3A4C;5= $/3= :C853A5*:485= @3:C8= 58(/*=:C=/65;B643=:/6/"?=)= % 7 164= ,=-:8(=+58853=4;;B34;?=$/3=..=1)%,=+B8= !!)= @//3= @53$/3A4C;5= :C= = 1),)= 0(5= *55@53= 4;;B34;?=:C==*5;3545*=1)%,)=8=460(5=4B8(/3=4@@35;:485=8(5=@3/5;8=A@3/:C"= 8(5= 85;(C:;46= ;/A@585C;5= /$= 8(5= 5C853= $/3= 5#835A5=+58853=164==..=)==),=+B8= @(43A4;5B8:;46=C4C/85;(C/6/"?=+?=:C83/*B;:C"= (/-5*=6/-53=4;;B34;?=$/3=64= =1..=),= B46:8?=@3:C8= 164= ,=:C=..= 1) ,= 468(/B"(= ACKNOWLEDGMENT =A/*56=:A@3/5*=8(5=@35*:;8:/C=$/3=(:"(7 8(/B"(==:A@3/5*=1),)=0(5=4*4C;5*== 84C*43*=4C46?8:;46=@3/;5*B35=:C=3(5/A583?=4C*= !=+:/@3:C8:C"=1 7!,)=**:8:/C466?=8(5= A/*56=$B38(53=5C(4C;5*=64==@35*:;8:/C=1..= 4B8(/3=-/B6*=6:95=8/=5#@35=8(5:3="348:8B*5=8/= )==)%,=+B8=$4;5*=*:$$:;B68:5=-:8(=64= 6946/:*== 9/@5= )= 4;5*/C:4= $/3= 8(5:3= =1..=)!==)!,)=./3=64= =:8=@53$/3A5*= "5C53/B=*/C48:/C=/$=;5$:#:A5) 6:"(86?= +58853=8(4C= 8(5= *55@53= = 1..= )= =)!,)=0(5=.=5#;5665*=48=@35*:;8:C"=(:"(7 B46:8?=@3:C8=164=,=:C==1),=+B8=(/-5*= 6/-53= @53$/3A4C;5= $/3= ..= 1)%,= ;/A@435*= 8/= 8(5= *55@53= )= 8= 4;(:55*= 83/C"= @53$/3A4C;5= $/3= 64= = 1..= )%,= +B8= -4= 6:"(86?= 6/-53= :C= = 1)!,= ;/A@435*= 8/= 8(5= *55@53=4C*=4*4C;5*==A/*56)=./3=64== 8(5= .= A/*56= -4= ;/C:85C8= 1..= )= = ),= *5A/C8348:C"= 8(5= (:"(58= /53466= 356:4+:6:8?= 4C*= @3/:*:C"= 46B4+65= $548B35= :A@/384C;5=:C:"(8)=./3=..=@35*:;8:/C=8(5=8/@= $548B35= :C$6B5C;:C"= @3:C84+:6:8?= :C;6B*5= = 1)= 46B5= /$= $548B35= :A@/384C;5,= *:;848:C"= $6/-=*B3:C"=5#83B:/C==1),=5C 3585C8:/C=4C*=8/3B5=1) ,=35$65;8:C"=8(5=$/3;5= 35B:35*= $/3= A4853:46= @3/;5:C")= ./3= = 8(5= A/8=:"C:$:;4C8=$548B35=-535==1) ,=1;3B;:46= $/3=A4853:46= $6/-= 4C*= 64?53= $/3A48:/C,=8/3B5= 1) ,=1:846=$/3=5#83B:/C,=4C*==1) ,=1"//*= 4*(5:/C=4C*=83B;8B346=:C85"3:8?=/$=64?53,) 4. CONCLUSION 0(5=*55@53==@53$/3A5*=+58=$/3=@35*:;8:C"= (:"(7B46:8?=@3:C8=164=,=:C=..)=68(/B"(=8(5= .=(/-5*=-54953=@53$/3A4C;5=$/3=64==:C= 54 :98 76:549:321640:4/4916.:-/,+*1240/),10(4:90+5+*+-'42+6104 9:321641%06$.+:-4#2496+-0+-'47:640/),1045/-$7/0$6+-' 9"! "4"1,2 1 1 1 3 , Simone Eder , Andreas Kottlan 4+"46"4!45" , Amrit Paudel, Patrizia Ghiotti3 3 3 1,2 , Thomas Quinten , Laura Martinez Marcos , Martin Spoerk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Aguilar-de-F;=2E82EFDE =A82 3D Printing Direct C=@ED6EBFEIHG?F=JF@E,.+EDI:FJEG6:=?F@ED6E***E Delivery Systems: State of the Art and Future )('8EHIDI=AEF=AF@E=EJI/HI4IG=HDE Perspectives., Pharmaceutucs, 2024. volume: 16.E @FG?F=JFEIHED=BAFDE:=JJE%*I/8E$7#E3IDCEDCFED=BAFDE 8$"E G6:=?F@E D6E DCFE D=BAFDE powder be used as feedstock to produce highly drug- loaded ethylene-vinyl acetate 3D printed tablets IH@IG=DIH/E IHG6HJIJDFHDE 63@F?E 4FF@IH/8E 6E 2. Samaro, A., et al., Can filaments, pellets and I:FE 4A63=BIAID;2E F?6JIA E %E 3#E 3=JE International Journal of Pharmaceutics, 2021, using extrusion-based additive manufacturing?; =@@F@8E 63FF?2E :IHI:=AE I:F:FHDE 3=JE volume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rbanowicz Magdalena1%%$#"! !%##1, Ewelina Baran2, Kulinowski Piotr2 1Falsified Medicines and Medical Devices Department, National Medicines Institute, Warsaw, Poland 2Institute of Technology, Pedagogical University of Cracow, Cracow, Poland 1. 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Norman, J., et al., A new chapter in 876 .124"0/.6 /-.6 4".(/4/6 7!6 /-.6 73*72("6 7)).5*7("4('6/76/-.6(.$16.3.)'."6*.060/66 products. Advanced Drug Delivery Reviews, 2017: 34(2/.564(6/-.6+6-)730/7')03%6/-.66 108, 3950. 0(015456 $056 *.)!7)3."6 /04(."6 5*./)06 2. Kulinowski P., et al. Development of Composite, ).#.01."6 06 "45/4(/6 *)7/7(6 54'(016 0/6 6 **3%6 Reinforced, Highly Drug-Loaded Pharmaceutical $-4-6 $056 05.(/6 4(6 /-.6 5*./)06 7!6 2(/).0/."6 Printlets Manufactured by Selective Laser 3./)7(4"071.6 4'6 SinteringIn Search of Relevant Excipients for 05."6 7(6 /-.6 -.34016 5-4!/6 0("6 54'(016 (6), 2142. Pharmaceutical 3D Printing. Materials 2022: 15 -0)0/.)45/45%6 /-.6 75.)#."6 5*./)06 $.).6 7(545/.(/6 $4/-6 06 (7$(6 ".')0"0/47(6 *)7"2/%6 3. U. Holzgrabe, et al., Quantitative NMR spectroscopyApplications in drug analysis, J. 68-45673*72("6456055740/."6$4/-6 /-.)3016".73*754/47(6*0/-$0567!6./60("6456 Pharm. Biomed. Anal, 2005: 38 (5): 806-81. (7$(6 /76 !7)36 2(".)6 -4'-/.3*.)0/2).6 # 7("4/47(56 8-456 !4("4('6 7(!4)356 /-0/6 517$.)6 -.0/4('6 )0/.56 *)737/.6 /-.6 !7)30/47(6 7!6 /-456 .5.0)-6 !4(0(."6 6 ')0(/6 43*2)4/ 6 !)736 /-.6 0/47(016 ,4.(.6.(/).%6710(" 58 432 10/.-,+0-*)(',4,%0$1.%/,4-,0(%)#"4!(%.-.)%)(',0( ,)%/, .."%,4(,"40%)(',)(%.'-)% Hadi Shammout1 1 1 2 2 , Krisztina Ludasi , 4,-Laczkovich ,$, ,%,/, %,/ 1 1 Institute of Pharmaceutical Technology and Regulatory Affairs, University of Szeged, Szeged, Hungary 2 Department of Optics and Quantum Electronics, University of Szeged, Szeged, Hungary 1. 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Kowsari K, Lu L, Persak SC, Hu G, Forrest W, Berger R, et al. Injectability of high concentrated suspensions using model microparticles. J Pharm Sci. 2024;113:3525 37. doi:10.1016/j.xphs.2024.09.026. 2. Marschall C, Graf G, Witt M, Hauptmeier B, Friess W. Preparation of high concentration protein powder suspensions by milling of 5432102/.102/.-,3+,4*).0*(.'34(.%*2/*2.-$). lyophilizates. Eur J Pharm Biopharm. #/1.24,/.("14*!.2 /.#41.55.%*%/*21024*.'2/ 2021;166:7586. *.2 /.%*24*""'.,(/.*.'4!*44%0*2.4*%1/0'/. 3. Agrawal P, Wilkstein K, Guinn E, Mason M, doi:10.1016/j.ejpb.2021.04.023. 4*..'"10''4*!..01.0'.'/1#/(.#/1. Serrano Martinez CI, Saylae J. A Review of 24,/. 0. %*'20*2. 432102/. 102/. 0'. ,04*204*/(. Tangential Flow Filtration: Process 42 4*..,4*"2/'.0.1/'4("03.0124%3/.!/*/1024*. Development and Applications in the 04(.10%24*...$.#+#.0'.1/0% /(.42 4*. Pharmaceutical Industry. Org. Process Res. . ,4*"2/'. %,3/2/. 04(. /% 0*!/. 0'. Dev. 2023;27:57191. 0% 4/#/(. *. 2 /. ,4/(. ,(/. 3"%2"024*'. 4*. doi:10.1021/acs.oprd.2c00291. . /1/. *2/(. ("/. 2. 2 /. 1/%/(4*!. %*%/*21024*. '2/. /#/1. 2 /. . */#/1. /%//(/(. . 01. . 1/'4("03. 04(. 10%24*. . $. #+#. 0'. /0,4*/(. 42 4*. 0. 2203. "144%024*.24,/...,4*"2/' /.#/ 4%3/.%*'",24*.0'..,3.1.2 /. ,4/(. ,(/. 0*(. . ,3. 1. 2 /. %*24*""'. ,/2 (."/.2.2 /.(43"24*.'2/.4*.2 /.,4/(. ,(/. 2 /. */%/''01. 1%/''. 24,/. 2. 1/0% . 0. 10%24*..04(."*(/1.$.#+#.0'.1/("%/(.. $.*.2 4'.,(/. /#/1.3"%2"024*'.4*.2 /. 432102/.102/.%%"11/(.,1/.1/"/*23 /.#41.55.'2102/!.1/1/'/*2'.0.1,4'4*!. 010% . 1. 2 /. %*24*""'. 0*(. '%0303/. /% 0*!/. 1,. 0124%3/. !/*/1024*. 04(. 2. '"'/*'4*. #/ 4%3/. 0*(. "%*%/*21024*. . (4'/1'4*'. 204*/(. 1,. 2 /. 0124%3/. !/*/1024*. 1%/''/'. 42 . 0. %3/01. "*(/1'20*(4*!. . %1424%03. 1%/''. 010,/2/1'. 0*(. 1,"3024*. % 010%2/14'24%'. 2 4'. ,/2 (. /*03/'. 24,/. 0*(. %'2/4%4/*2. (*'21/0,. 1%/''4*!. . '203/. '34(. ,4%10124%3/. '"'/*'4*'. 5"2"1/. 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";998878,5 +77+;#5 5 78:8#5 #;698,5 '87;5 ,';5 7+$#5 ;"%;6,5 $6#5 ,';5 (''##$# 78:8#589%+98,5'$#59!$77;"5$6#5$ "+8!$,;75 $9;#5 +65 ,';5 9,$,89,8%$75 $6$79895 +5 ,';5 ";9:7,95 9$!;598;#5867:;6%;5+65,';599,;!5;'$8+")5 %";$,;#5+"5;$%'5$,%')5038)5/*5(';5$%%; ,$6%;5 '+!"#+ $6#5:9865,;"6$"5#8$"$!95#;98659 $%;95;";5 %"8,;"8$5 +5 ,';5 #;9865 9 $%;5 ;";5 #"+ 7;,5 98;5 ('89599,;!$,8%586;9,8$,8+65+65,'895866+$,8;5 9!$77;"5,'$651--56!559!$77;"5,'$65-) 5$6#5 $6#5 %+! 7;5 #":5 #;78;"5 99,;!5 %$65 ;,$5 +,;6,8$75 ;7+5 /-5 !)5(':95 !$865 8,5 %+6,"8:,;5,+5;,,;"5:6#;"9,$6#865,';5"+7;5$6#5 +9987;5 ,+5 "; $";5 + ,8!8;#5 5 ;'$8+:"5+5,';586#88#:$75%+! +6;6,95865,';5 +"!:7$,8+69)5 +"!:7$,8+65 ;6$7;5 ,+5 ";#8%,5 ,';8"5 ;;%,5 +65 %+! ";998878,5'8%'58959:9,$6,8$75+"5:",';"5 #;;7+ !;6,5 86,+5 ,$7;,95 $6#5 +;"%+!;5 ,';5 %'$77;6;5+5+"$758+$$87$878,5+5 ; ,8#;95$6#5 "+,;869) '%$$%$$# 1. Drucker J. D., Advances in oral peptide therapeutics. Nature Reviews Drug Discovery, 2020. 19: 277-289 . 2. Verma S., et al., Challenges of peptide and protein drug delivery by oral route: Current strategies to improve the bioavailability. Drug Development ';98659 $%;5+5,';55 "; $";#58,'5 3. Anselmo C. A., et al., Research, 2021. 82(7): 927-944. ,';5%+!86$,8+65+5(;;651-5$6#5 $65-) strategies for biologics. Non-invasive delivery Nature Reviews Drug (' '#$# Discovery, 2019. 18: 19-40. 4. Mahmood A., et al., SEDDS: A game changing %%+"#865 ,+5 ,';5 %+6#:%,;#5 ,;9,95 ,';5 ;9,5 approach for the oral adminsitration of hydrophilic %+! ";998878,5$7:;95;";5+9;";#58,'5,';5 macromolecular drugs. Advanced Drug Delivery /./5.;:987865"$,8+5$6#5(;;651-5'$#5,';5 Reviews, 2019. 142: 91-101. !+9,5 $#$6,$;+:95 867:;6%;5 +65 5. Hassan, A.A.A.; et al. Hydrophobic Ion-Pairing %+! ";998878,)55!+#;77865$95 ;"+"!;#5 Complex of Lysozyme with Sodium Dodecyl Sulphate +"5 $5 #:$75 :" +9;.5 ,+5 ;5 $7;5 ,+5 ";#8%,5 ,';5 to Enhance Stability in Lipid-Based Carriers. %+! ";998878,5 $7:;95 +5 6;5 %+! +98,8+695 Pharmaceutics 2024, 16: 589. 0($7;5/*5$6#5,+5;9,8!$,;5,';5;;%,5+5$"8+:95 +!$$ $%,+"95 +65 ,';5 %+! ";998878,5 +5 ,';5 7+$#;#5 !;9+ +"+:95%$""8;"9)5 "+;%,5 6+5 (1-1/15 '$95 ;;65 8! 7;!;6,;#58,'5,';59: +",5 "+8#;#55,';5 8689,"5+5:7,:";5$6#566+$,8+65+5:6$"5 "+!5,';5$,8+6$75;9;$"%'5;;7+ !;6,5$6#5 66+$,8+653:6#586$6%;#5:6#;"5,';5(1-1/ 5:6#8659%';!;) 68 765 4342710/.-,+*6723)0-('*7-)%2$.,7+*$+1*,+#0(.,"$.,7+*7*+0!* -+$*102,#0-3*(3(.0)( # *1 1 2 1 1 *, * , Ferenc Fenyvesi , *- 1. INTRODUCTION ,+61(8+,%+'61%1+*,1$63,18!13'09/1(84(,,31 +*,')1$,6)1'%+,)%6'6+'8%17631'% ,3+'#6+,51-41 9876564321 09/.-63,51 +*,)6(',31 (641 6%1 $8%!8$61"'$)83$8(416!+,)11"'%2116%511*1 '%$),63'%#41'"(8)+6%+1)8,1'%1+*,1+),6+",%+18!1 +),6+",%+1 7'+*1 (84(,,31 !8)"6+,51 7'+*1 6)'831 5'3,63,31 3'09/1 '31 "6'%41 3,51 '%1 +*,1 !8),3$,%+41 6-,,51 /1 3'09/1 *,1 "6'%+,%6%$,18!15'!!,),%+1#,%,+'$15'3,63,3217*',1 '%+)6$,6)1 ,!!,$+1 8!1 +*,3,1 5,' ,)41 343+,"31 +*,1 "09/1 6381 3,51 '%1 +*,1 5, ,8(",%+1 8!1 7631'% ,3+'#6+,51 '61+*,1,(),33'8%18!1/1 6$$'%,31 ,1 +81 +*,1 6)#,1 "8,$6)1 3',1 6%51 ()8+,'%1 6!+,)1 1 *1 +),6+",%+1 -41 !8),3$,%$,1 (846%'8%'$1 %6+),1 8!1 09/21 +*,1 '%+)6$,6)1 "'$)83$8(41 %1 +*,1 $63,1 8!1 +*,1 "09/1 5,' ,)418!1+*,3,1"8,$,316),15'!!'$+1181 (84(,,31 +*,1 ,6"'%6+'8%1 8!1 +*,1 $,6)1 38 ,1+*'31()8-,"13, ,)615,' ,)41343+,"316),1 '%+,)%6'6+'8%1 7631 %81 ),, 6%+21 381 +*,1 6 6'6-,1 8)1 %5,)1 5, ,8(",%+21 7*'$*1 6),1 '%+)6$,6)1,!!,$+18!1+*,3,17631'% ,3+'#6+,51 '61 3641 -63,51 8%1 '(838",31 6%51 $6+'8%'$1 +*,1 ,(),33'8%1 8!1 #),,%1 !8),3$,%+1 ()8+,'%1 (84",)3213$*1',1$4$85,+)'%1(84",)311 1 6!+,)1 1 *1 +),6+",%+1 -41 !8),3$,%$,1 4$85,+)'%316),17'5,413,51,$'(',%+31'%1+*,1 "'$)83$8(416%51!871$4+8",+)41 (*6)"6$,+'$61 6%51 $83",+'$1 '%53+)',31 +81 '%$),63,1 +*,1 38-''+41 6%51 -'86 6'6-''+41 8!1 *-0(%2.(*$+1*1,(4%((,7+ '(8(*''$1 "8,$,321 -+1 +*,),1 (84",)1 5,)' 6+' ,316),16-,1+81$6))41"6$)8"8,$,31 *,1 1 (84",)1 7631 "8),1 $4+8+8'$1 +*6%1 +*,1 $6+'8%'$1$4$85,+)'%1(84",)1/$$8)5'%#1+81+*,1 %1+*,1(),3,%+13+5417,16'",51+81'% ,3+'#6+,16%51 1 6%51 ,+61 (8+,%+'61 ",63),",%+31 -8+*1 +81 $8"(6),1 +*,1 3'09/1 6%51 "09/1 $6))4'%#1 (84",)1 7,),1 6-,1 +81 !8)"1 (84(,,31 7'+*1 $6(6$'+418!161$6+'8%'$1$4$85,+)'%1(84",)11 3'09/1 6%51 "09/21 -+1 -63,51 8%1 +*,1 #,1 (84",)1 6%51 +*,1 (84,+*4,%'"'%,1 1 ,,$+)8(*8),3'321 +*,1 $4$85,+)'%1 (84",)1 '31 (84",)217*'$*1'313,51'%1+*,1()6$+'$, 6-,1 +81 !8)"1 61 3+)8%#,)1 (84(,1 7'+*1 "09/1 "8,$,1 84(,,31 !8)"6+,51 7'+*1 1 *)$.0-,$2(*$+1*)0.71( (84",)17,),16-,1+81,%+,)1+*,1$,3217*',1+*,3,1 *) !8)"6+,51 7'+*11 (84",)1 7,),1-8%51 +81 +*,1 *,138-,1$6+'8%'$1$4$85,+)'%1(84",)16%51 $,1 ","-)6%,1 84(,,31 7,),1 '%!,%$,51 '+31 !8),3$,'%.6-,,51 5,)' 6+' ,1 7,),1 +*,1 -8+*1+*,1/16%51#),,%1!8),3$,%+1()8+,'%1 ()85$+31 8!1 4$86-1 +51 56(,3+21 1 ,(),33'8%21 -+1 +*,1 '%$),63'%#1 %#6)41 *,1 (),5,3'#%,51 *"6%1 /1 $8%$,%+)6+'8%18!11(84",)17631$4+8+8'$ 3'09/21 +*,1 -)6%$*,51 (84,+*4,%'"'%,1 7,),1 *47+42%(,7+ %#6)421 7*',1 +*,1 ,6%6(1 1 "09/1 %1 3""6)421 7,1 *6 ,1 3$$,33!41 !8)"6+,51 5,)' ,51 !)8"1 '#"6./5)'$*1 +51 56(,3+21 $4$85,+)'%1 (84",).-63,51 09/1 5,' ,)41 7631 +*,1 ()85$+1 8!1 ,--'81 ,1 ,))64.,%. ,'%,321)6%$, (84",)31 343+,"31 84(,,31 !8)"6+,51 7'+*1 5'!!,),%+1 3*87,51 5'!!,),%+1 $,6)1 *,1$4+8+8'$'+41,!!,$+18!1+*,16((',51(84",)31 6-,1 +81 '%!,%$,1 +*,1 ,(),33'8%1 8!1 5'!!,),%+1 ()8+,'%316%51+*,1$6+'8%'$1$4$85,+)'%1(84",)1 7631 ,6"'%,51 3'%#1 +*,1 1 ",+*851 *,1 *) '%+,)%6'6+'8%1*,1 !8)"6+,51 (84(,,31 6),1 3'09/16%51"09/1(84(,,317,),1!8)"6+,51 '31"8),13'+6-,1!8)1#,%,18)1()8+,'%1,(),33'8%1 +*6%1!8)1#,%,13',%$'%#1 '%15'!!,),%+1%'+)8#,%(*83(*6+,191)6+'831!)8"1 1 +81 1 *,1 !8)"6+,51 (84(,,31 7,),1 $*6)6$+,)',51 -63,51 8%1 +*,')1 3',1 5'3+)'-+'8%1 3'%#154%6"'$1'#*+13$6++,)'%#116%51+*,')1 69 1.Kanasty, R.; Dorkin, J.R.; Vegas, A.; Anderson, D. Delivery materials for siRNA therapeutics. Nat. Mater. 2013, 12, 96777, doi:10.1038/nmat3765. 2.Chaturvedi, K.; Ganguly, K.; Kulkarni, A.R.; Kulkarni, V.H.; Nadagouda, M.N.; Rudzinski, W.E.; Aminabhavi, T.M. Cyclodextrin-based siRNA delivery nanocarriers: A state-of-the-art review. Expert Opin. Drug Deliv. 2011, 8, 14551468, doi:10.1517/17425247.2011.610790. ,+*)('% $*#% "!, % % (($% (($(%%(%*+%+*(%%(% $+%*% +(%$%$$**$%*%$+% +*%(%*$%((+'%((*($%$% $$**$%$%$$'(%$(+%(%"!, %$$%'((# 70 +*)( *'+*'+%$*$#"! $% #"+*$%"+%$+"+"%+ ' "*+ $%"+""$+' $" %$ Luca Casula1 1 2 1 2 , Simona Demuro , Rita Argenziano , Francesca Pintus , Alessandra Napolitano, Carla Caddeo1 1. INTRODUCTION " ! " " " =;:9:87;6=75:43:7247=1=627;70642=/29.=7;/294:7 247 2-=7 859:7 2-;:587 247 ,+*.987 ;)8460294:7 ;:(7 47064("/=7'/=64*90484!=8 79049(77 6=;/29.=741'=:780=/9=878/;.=:9:70640=629=8%7 !!7 ;:(7 '/=647 7 !!7 3=6=7 $10492;294:7 (980=68=(7 9:7 7 9:7 06=8=:/=7 467 ;)8=:/=7 4#7 4#7 8"/-7 0640=629=87 #467 (=6!4/48!=29/7 0"6048=87 )'7 "8=7 4#7 =;89'7 =7 !=;:9:7 7 !! 7 ;:(7 84:9/;2=(7 ;//=889)=7 8':2-=29/7 !=;:9:87 06=0;6=(7 #64!7 392-7 ;:7 "26;84":(7 (989:2=6;246%7 -=7 !=;:7 )948':2-=29/7 06=/"68468 7 0;629/";6'7 !=2-'7 (9;!=2=6 7 04'(980=6892'7 9:(=1 7 ;:(7 =2;7 =82=674#7 *(9-'(641'9:(4=**/;6)41'9/7;/9(7 042=:29;7 4#7 2-=7 .=89/=87 3=6=7 !=;8"6=(7 .9;7 = 7-;87847#;67)==:7-;!0=6=(7)'72-=7 (':;!9/==/2640-46=29/7 9-27 8/;22=69:%7 -=7 !;65=(79:84")992'74#72-=7!=;:9:8% =7!=;:9:79:7'/=64*90484!=873;87 ";:29#9=(7.9;7,+*.98780=/2640-424!=26'7;277 :7 2-987 82"(' 7 ;7 !=;:9:7 4)2;9:=(7 #64!7 :!7 ;:(7 2-=7 =:26;0!=:27 =##9/9=:/'7 3;87 =7.9;70=6419(;8=7419(;294:73;87 /; 9:/46046;2=(79:2470-480-4909(7.=89/=87392-72-=7 90484!=87 3;87 82"(9=(7 .9;7 /6'4=:9/* ;9!74#79:/6=;89:7928784")992'7;:(7064("/9:7;:7 6;:8!98894:7 $=/264:7 9/648/40'7 /6'4* ;"=4"8*);8=(7#46!";294:72-;27/4"(7)=78;#='7 $%7 -=7 ;:29419(;:27 ;/29.92'7 4#7 =7 ;009=(7 4:247 2-=7 859:%7 -=7 :;:4#46!";294:7 !=;:9:7 3;87 2=82=(7 .9;7 2-=7 7 ;:(7 7 /4:2;9:=(7'/=64 73-9/-73;875='7247064("/9:7 ;88;'8%7 =7 !=;:9:7 9:7 7 467 9:7 82;)= 7 8!; 7 -4!4=:=4"87 .=89/=8%7 -=7 '/=64*90484!=87 467 2-=7 =!02'7 '/=64* ;:29419(;:27;/29.92'74#72-=7=7!=;:9:7 90484!=873=6=7;((=(7247776=;=:27;:(7 '/=64*90484!=873;8782"(9=(7.9;77;:(7 ;:;'8=(780=/2640-424!=269/;'7;277:! 7 7 7;88;'8 7;:(72-=7)948;#=2'73;87;88=88=(7 7 6=;=:27 ;:(7 ;:;'8=(7 79:79!!462;9=(75=6;29:4/'2=8% 80=/2640-424!=269/;'7 ;27 7 :!%7 -=7 " $%"#" '+# 9:7 -"!;:7 5=6;29:4/'2=87 ;;7 =1048=(7 247 /'242419/92'74#7=7!=;:9:73;87;88;'=(7 )" =7 !=;:9:7 9:7 7 467 9:7 '/=64* 9049(7 84')=;:7 0-480-4909(87 3;87 90484!=8 7467247=!02'7'/=64*90484!=87#467 0"6/-;8=(7#64!79049(7!)7"(398-;#=: 7 7-%7=7.9;)992'73;87(=2=6!9:=(7)'72-=77 =6!;:' 7'/=6473;870"6/-;8=(7#64!7;=:47 ;88;'7;:(7=106=88=(7;87;70=6/=:2;=74#7/4:2647 ;6!9:;:4 72;' 70-480-;2=7)"##=6=(78;9:=7 ":26=;2=(7/=87% 7 07 %7 3;87 0"6/-;8=(7 #64!7 =6/57 " %"#"#$$+ 9;: 72;'%7=7!=;:9:73;874)2;9:=(7 .9;7 =:'!;29/;'7 064!42=(7 419(;294:7 4#7 92-72-=7;9!7247=1049272-=7#;.46;)=70640=629=87 =%7 #2=67 7 - 7 2-=7 !;19!"!7 4#7 =7 !=;:9:7 ;:(7 6=84.=7 2-=7 ;)8460294:7 4#7 2-=7 82;629:7 =7 8-9#2=(7 9:84")992'7 988"= 7 ;7 :;:42=/-:44'*);8=(7 );2-4/-64!9/;'7 )=/4!9:7 )64;(7 ;:(7 ;0064;/-7 3;87 =1046=(%7 '/=64*90484!=8 7 #=;2"6==88%7=.=68=*0-;8=77/4:#96!=(72-=7 3-9/-7 /4!)9:=7 2-=7 ;)992'7 4#7 90484!=87 247 /4!0=2=7/4:8"!0294:74#7=%7,04:7#"7 =:-;:/=7 84")992'7 ;:(7 )94;.;9;)992'7 4#7 2-=967 !4:4!=67/4:.=6894: 72-=76=;/294:7!912"6=73;87 /;647392-72-=70=:=26;294:7=:-;:/9:7;)992'74#7 ;/9(9#9=(72479:("/=706=/9092;294:74#72-=7!=;:9: 7 '/=64 73=6=7(=.=40=(%7-=79:/46046;294:74#7 3-9/-7 3;87 6=/4.=6=(7 )'7 /=:269#";294:7 ;:(7 =7 !=;:9:7 9:247 '/=64*90484!=87 '40-99;294:% 89:9#9/;:2'7 ;##=/2=(7 2-=967 !=;:7 (9;!=2=67 7 :! 7 3-9=7 04'(980=6892'7 ;:(7 8"6#;/=7 /-;6=7 71 ('% ONMN KJIHGNMNF EDIICHN BA@ D?N H=DPPH FRAP H98;7;6N7 F978HI=NF ?9? NJGMI886NJG N555999NJ= AA TE FE L L L L L L L L L L L L L L L L L L L L L L L L L L L I L L L L L L L L L 5 5 L L L L L L L L L Forrmulattion o a (%) ( g g//ml) ( g g//ml) E4432 DIICCHN BA@ L L L L L L L L L D I L L L L L L L L L MeDHICA mel lannin soluttion 87 a u 1.5 323 11 1744 136 MeDHICA mel lannin gly-lip a 844.0 31614 1749146 $##"! $ Empty gly-lip 472.4 1729.6 60054 L L L L L L L L L 1NIJJ F9 6 I L L L L L L L L L I 6NGNM E10A/ 8;H=F978NM79G== L L L L L L L L L L L L L L L L L L G L L L L L L L L L 9JFN. E-,A/ +NGI 8;GNJG9IH E*-A/ IJJF NJGMI886NJG L L L L L L L L L L L L L L L L L L L L L L L L L L L J L L L L L L L L L L L L L L L L L L I L L L L L L L L L J I JL L L L L L L L L N555999NJ= E)A ;5 H='@B@ D?N !# ' 1N0, 6 L L L L L L L L L 6NHIJ9J 7;HKG9;J OI7 J;G =G;G;.9 L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L L L ' E 3 IGG G?N ? L L L L L L L L L L L L L L L L L L I L L L L L L L L L ?9?NM ;J@ $A/ L L L L L L L L L J I L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L )68G= H=:H98%$#"@ @$"#@ I7 ONHH I7 L L L L L L L L L L L L L L L L L LL L L L L L L L L G?N 1N0, 6NH JJ9J H=@ $A@ L L L L L L L L L L L L L L L L L L 1N0,L L L L L L L L L @$"#@ 6NH JJ9J H=: I L L L L L L L L L( #$@ ! #@" 44#@% B H98 M==;: MD)1 69MI88?7 E9@ BA 7?;ONF G?IG L L L L L L L L L I L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L IL L L L L L L L L H= 8MNF;69JIJJGH= KJ9HI6 I L L L L L L L L L I 6NHHIM// CNH;O B J6/ IJF M L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L L L 7H9?GH= HIMNM O?NJ H;IFNF O9G?? 1N0, L L L L L L L L L M L L L L L L L L L L L L L L L L L L L L L L L L L L L G L L L L L L L L LL L L L L L L L L 6NHIJJ9J/ IL L L L L L L L L L L L L L L L L L 9J IH9J6NJG O9G? 79+N FIGGI EDIICHN BA@ L L L L L L L L L L L L L L L L L L L L L L L L L L L I L L L L L L L L L D L L L L L L L L L 99I IC9H9G== ;5 IID G L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L NF 5;;M $ L L L L L L L L L 5 L L L L L L L L L L L L L L L L L L ? O9G?? 1N0 L L L L L L L L L G L L L L L L L L L , 6NHIJ9J 9J 01 ;M 9J L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L H= H H98;7;6N7 E L L L L L L L L L E@BB 6 L L L L L L L L L 6HA@ )68G== L L L L L L L L L G L L L L L L L L L H= H98;7;6N7 ON L L L L L L L L L NMN GN7GNF I7 I ;JGMM;H@ ( L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L G L L L L L L L L L'@"L L L L L L L L L L L L L L L L L L D@ ( ( D?N MN7KHG7 7 L L L L L L L L L L L L L L L L L L 7?;ONF G??N 7K M==;:D)1 69MI8?7 ;5 1N M L L L L L L L L L L L L L L L L L L L L L L L L L L L N0,L L L L L L L L L IJG9;.9F JJG IL L L L L L L L L 6NHIJ9J H== L L L L L L L L L HH98;7;6N7 EA IJF N68G== L L L L L L L L L L L L L L L L L L L L L L L L L L L G L L L L L L L L L H== H NJ;KMI9J I 5KKMGG?NM 9JN7G9IG9;J G; IH9FIGGN L L L L L L L L L 5 M L L L L L L L L L I L L L L L L L L L L L L L L L L L L IL L L L L L L L L H98;7;6N7 EA IG 4/ 6I JJ9599 L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L 5@ G?N9M N555NN D?N IJJG9;.9FIJG I ( $ OI7 ?9? E%43 : L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L $ D)6H2 DIICCHN L L L L L L L L L L L L L L L L L L L L L L L L L L L D I N $A IJF L L L L L L L L L L L L L L L L L LL L L L L L L L L MN6I9JNF KJNF O?NJ JIJJ;5;;M6 D?97 7GKF= L L L L L L L L LL L L L L L L L L OI7 5KKJFNF C= )KM;8NIJJ J9;J : L L L L L L L L L 5 L L L L L L L L L L L L L L L L L L I L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L I L L L L L L L L L L L L L L L L L L I 5 M 6 6KHIGGNF@ I L L L L L L L L L 969HIMH=/ G??N 7I6 N GG NJNMI . 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INTRODUCTION *74-.*6413-*2/ *64-$-4#/ 0'1/ 45/ -4+/ *)-75/ ,35'!+/ *70/ 6*4-57-6/ +43'64'31 / 7/ *00-4-57"/ -4/ -70'61+/ 876574352210/.2110-7,/-+/*/)*(53/6*'+1/5/01*4%/ 4#!1/ / 6522*,17/ !350'64-57/ -7/ 4%1/ 5'70/ *70/ +1$131/ -7('3#"/ !*34-6'2*32#/ -7/ 65).*4/ 17$-357)174/*70/+'!!534+/4%1/!352-13*4-57/*70/ +617*3-5+ / 7/ +'6%/ 63-4-6*2/ +-4'*4-57+"/ 4%1/ )-,3*4-57/ 5/ -.35.2*+4+/ %-6%/ +'!!534/ 4%1/ *$*-2*.-2-4#/ 5/ *+4*64-7,/ %1)5+4*4-6/ *,174+/ -+/ 01,311/ 5/ %1*2-7, / 7/ *00-4-57"/ 53)-7,/ */ 1++174-*2/ -7/ )50137/ -3+4/ *-0/ !3*64-61+ / 65)!21/ +43'64'31/ -4%/ %1!*3-7+/ 4%*4/ !31$174/ 57$174-57*2/)14%50+/2-1/.*70*,1+/*70/,*'1/ .2550/ 62544-7,/ -7/ 5'70/ *!!2-6*4-57+/ *70/ 5417/*22/+%534"/1+!16-*22#/-7/65)!21/5'70+ / *661213*4-7,/4%1/%1*2-7,/!3561++/*31/*)57,/4%1/ +/ */ 31+'24"/4%131/-+/*7/ -7631*+-7,/01)*70/53/ +'!13-53/1*4'31+/5/4%-+/!52#)13/ *0$*7610/ %1)5+4*4-6/ +52'4-57+/ 4%*4/ *31/ '+13 3-1702#"/ 2-,%41-,%4"/ 65+41164-$1"/ +*1"/ *70/ 7/ 4%-+/ +4'0#"/ / *+/ .217010/ -4%/ / 45/ *00/ 6*!*.21/5/3*!-0/.2550/62544-7,"/%14%13/*!!2-10/ %1)5+4*4-6/ *64-$-4#/ .1+-01+/ -4+/ 5'70/ %1*2-7,/ .#/4%1/-7('310/-70-$-0'*2/53/*/-3+4/31+!57013/ / !35!134-1+ /*75-.13+/65)!5+10/5/"/"/*70/ 216435+!'7/ 7*75-.13+/ *31/ !35)-+-7,/ 03',/ !52#14%#2171/5-01/ /131/1216435+!'7/*70/ 6*33-13/ +#+41)+ / %1-3/ +43'64'31/ 625+12#/ 4%17/ 25*010/ -4%/ $*765)#6-7/ / 53/ -7/ 31+1).21+/ 4%1/ 143*6122'2*3/ )*43-/ "/ 65).-7*4-57/ -4%/ *74-.*6413-*2/ *,174/ 5/ -76/ %-6%/ -+/ 63-4-6*2/ 53/ +'!!534-7,/ 6122'2*3/ 7*75!*34-621+/ / 53/ 4%1/ -3+4/ 4-)1/ -7/ 4%1/ !3561++1+/ +'6%/ *+/ *44*6%)174"/ ,354%"/ 2-413*4'31 /%17"/4%1/!31!*310/53)'2*4-57+/131/ )5$1)174"/*70/0-13174-*4-57/ /%-+/)*1+/ 6%*3*6413-10/-7/413)+/5/)53!%525,#/*70/-.13/ 1216435+!'7/ 7*75-.13+/ *7/ 16122174/ 6*70-0*41/ +-1/ '+-7,/ / *70/ 03',/ 25*0-7,/ 1-6-176# // 53/714,1713*4-57/5'70/6*31/*!!2-6*4-57+ %17"/ 56. 4502-/ 6#4545-6-4#/ *+/ 01413)-710/ '+-7,/"/*70/4%1/%1)5+4*4-6/*64-$-4#/5/4%1/ -2/-.35-7//-+/*/!3541-7/-.13/!350'610/.#/ 7*75-.13+/*+/1$*2'*410/56.454-/'+-7,/!3*,'1 4%1/ +-253)/ 5).#/ )53- / / -+/ */ 7*4'3*2/ *21#/3*4+ .-5!52#)13/ -7/ !3541-7/ +43'64'31/ '+10/ -7/ 431*4)174/ -4%/ !%#+-6*2/ !35!134-1+/ +'6%/ *+/ "%!#(!"!$)"%(#-) 16122174/ .-565)!*4-.-2-4#"/ .-501,3*0*.-2-4#"/ *413/ $*!53/ !13)1*.-2-4#"/ *70/ )16%*7-6*2/ +"% " +4317,4% //+'!!534+/6522*,17/+#74%1+-+/*70/31 -253)/ 656557+/ 131/ 35)/ 5*/ -32-/ 1!-4%12-*2-*4-57 / %-45+*7/ / 1%-.-4+/ '3+*"/ 3-#1 / 5/ )5216'2*3/ 1-,%4/ 77 0/.- QPONMLKJI HGFEI DCBA@CI ?K=I 98KQ8N76KN895=I4M67H8NP768J8IM3O985IH210=IGFEI A=CCCI?K5=I/.2IH-ACCIJ,I4K+NOQ68ILO*85 -)+ ++ 0 ,+ G))=I H$D:IK#%8M%L5=IKJ9I"2G!BA CI,89O%,I8+8I "(%$#"! 4%+QPKL89I (+M,I O,KB69+OQPI HNI M%OL=I +%I 6MK9OJI 8((OQO8JQO8LI M(I NP8I (M+,%6KNOMJLI (M+,OQI KQO9I H'@;:5=I 6%NK+K698P798I LM6%NOMJI +-).)+ %(%$'(#%$#"!+ " + %!" #'(+ 5I.IKLIO(N89I7I1GI6KIH)+?O785I 8+8I 8N88JI @DB@;:I )P8I JKJM(O8+LI 8+8I 66IQP8,OQK6LI%L89I8+8IM(IKJK67NOQK6I+K98IKJ9I PM,M8JM%LI KJ9I JMJB8K989I 6LM=I NP8I (O8+I %L89IKLI+8Q8O89 LO*8LI M(I NP8I (M+,%6KNOMJLI +KJ89I 8N88JI $I *)*)+/('%(%$#"!+" +%!" #'(+"(%$#"! KJ9IDC IJ, =IKJ9I210I8+8I,O389IKJ9I$C:IH5I.I 66I M(I NP8I 4+84K+89I JKJM(O8+LI JMNI MJ67I KLI K9989I NMI NP8I 4M67,8+I 68J9I /.2I KLI L%44M+N89I Q866I L%+OK6I %NI K6LMI 4+M,MN89I Q866I )P8I(M+,OQIKQO9ILM6%NOMJLIM(INP8I4M67,8+LIM(I=I -)*)+ +$"$"##$+$ OJN8+KN89IOJNMINP8I4M67,8+I68J9IKNI$:IH5I 4+M6O(8+KNOMJ )P8I,O3N%+8LI8+8ILNO++89IKNICCI+4,IM8+JOPN=I KNI +MM,I N8,48+KN%+8=I NMI 8NI KI PM,M8J8M%LI -)-)+ +'"$%$#+$##$+$ LM6%NOMJI)P8J=IKJI868QN+ML4OJJOJI98OQ8IH.1B MNPINP8I6889OJINO,8IKJ9INP8I6889OJIK,M%JNI ACC=I !JM8JLM=I )+?O785I KLI %L89I NMI MNKOJI M(I NP8I (M+,%6KNOMJLI K446O89I MJI NP8I P8,MLNKLOLI 868QN+ML4%JI JKJM(O8+I (M+,%6KNOMJLI )P8I ,M986IQ+8KN89IMJINP8INKO6I8+8IQM,4K+89IONPI 9OLNKJQ8IKLI$IQ,I8N88JINP8IJ88968INO4IKJ9I NP8I QM,,8+QOK6I 4+M9%QNI 8,MJI,MJI NP8I NP8IQM668QNOMJI46KN8=INP8IM6NK8IKLI$I?=IKJ9I (M+,%6KNOMJL=I NP8I 46KOJI JKJM(O8+I (M+,%6KNOMJI NP8I (6MI +KN8I KLI AI ,6PI KLI 868QN+ML4OJJOJI +89%Q89I 6889OJI NO,8I NP8I ,MLN=I PO68I NP8I QMJ9ONOMJL JKJM(O8+I (M+,%6KNOMJI QMJNKOJOJI NP8I QM,OJKNOMJI M(I.I KJ9I /.2I KLI NP8I +M%4I M66MOJI 868QN+ML4OJJOJ=I NP8I MNKOJ89I NPKNI+89%Q89I6889OJIK,M%JNINP8I,MLN JKJM(O8+I ,KNLI 8+8I Q+MLL6OJ?89I ONPI 6%NK+K698P798I)P8IINMII+KNOMIKLIAEAI )+0 0 7I8OPNIOJINP8I(OJK6IJKJM(O8+L LIKI+8L%6N=IONIQKJI8ILNKN89INPKNINP8I9886M489I *)-)+ 46KOJ=I .I K6MJ8=I M+I QM,OJ89I ONPI /.2I %(%$'(#%$#"!+ " + %!" #'(+ 6MK989I JKJM(O8+I (M+,%6KNOMJLI K+8I OJJMKNO8I "(%$#"! 9+%IQK++O8+IL7LN8,LINPKNIQKJI8I%L89IOJI6889OJI )P8I L%+(KQ8I ,M+4PM6M7I KJ9I K8+K8I (O8+I QMJN+M6IKLIP8,MLNKNOQI(M+,%6KNOMJL 9OK,8N8+LIM(INP8IJKJM(O8+LI8+8I98N8+,OJ89I7I 1GIH11GI)+ 9+%I6MK9OJI8((OQO8JQ7IKLIQMJ9%QN89I AI%KJ=IF=I8NIK6=I *))+ +$"$"##$+$ I )MI 8K6%KN8I NP8I Q7NMNM3OQON7I M(I NP8I JKJM(O8+I 9KJQ89I 8K6NPQK+8I GKN8+OK6L=I $CA I D=I $;A ,KNL=I KJI OJ9O+8QNI G))I KLLK7I KLI QMJ9%QN89I $;$$I %LOJI NP8I P%,KJI 98+,K6I (O+M6KLNI Q866I 6OJ8I $/PKJ=I =I 8NI K6=I HBAA ?5I I M%+JK6I M(I *))+ 446O89I2M67,8+IQO8JQ8=I$C$IAAI +'"$%$#+$##$+$ )P8I +KNI NKO6I K,4%NKNOMJI ,M986I KLI %L89I NMI I M,OJI=I 8NI K6=I 98N8+,OJ8INP8IP8,MLNKNOQIKQNOON7IM(IJKJM(O8+I (M+,%6KNOMJLI 4+K%8BK687I +KNLI 8+8I %L89I I GKN8+OK6LI QO8JQ8I KJ9I (M+I NP8I 8348+O,8JNK6I +M%4LI 1NPOQK6I K44+MK6I 1JOJ88+OJ=I$C$CIAA (M+INP8ILN%97IKLIMNKOJ89I(+M,INP8I8*,OK68,I K?(I JO8+LON7I JO,K6I 1348+O,8JNLI MQK6I 1NPOQLIM,,ONN88=I!LNKJ%6=I)+?O78 0 )POLILN%97IKLIL%44M+N89I7I8K6NPI!JLNON%N8LIM(I )+?O78IH)1=I2+M8QNI.MEI$;$;5 78 ?= ;:?98765;473271;09/.-,4.;:?9;+-71+;1925;1+47*+9/);:9?8; 5+4,-7?6;1/6,870.;,61;.2.-,76+1;9+4+,.+;-?;(7?0?8,-7(747-/; ,..+..8+6- Mercedes Vitek 1 2 1 1 ';9%$#";9!# , Alenka Zvonar Pobirk ';8#;5$;8" 1 Department of Pharmaceutical Technology, University of Ljubljana, Faculty of Pharmacy, Slovenia 2 Department of Biopharmaceutics and Pharmacokinetic, University of Ljubljana, Faculty of Pharmacy, Slovenia 1. INTRODUCTION =;8"$#; BA@?=@9=897A?=6=4*:398296/9./9725A@?/=68/=6?6@;?@:9 45089/5 7/=//5A@ @03 %/1 B8/5 " /9 3:A97A82@?8A5 8A91 3A9@?825A950 ?= @: 45089/57/=/5?=/5A@%51 ##"%'/@: 7A=A47=@ /. 8:9/=?8 ;?6A661 -:9 .9/7./66$$1$A?=@(B9?6@19A=8"1=@ ;/6?=4 94?7= ?6 9,2?9;+ *:?6 ?6 8/7'?=A@?/= -?@: @:A=/5 %B:A97A8:71 638?A550 95)A=@ ./9 33@?;('A6; $5/)=?A" A=; 58?@:?= %?3/?; $( 1 ?3/?; @:9A32@?861-:?8:5?7?@;'09=@3//9 7'197A=0"6=@; + 6@A'?5?@0 A=; 6:/9@ 35A67A:A5.(5?)619,2?9?=4 .9,2=@3A9=@9A5Table 1. Composition of the studied precursor 62'82@A=/26%$#";3/@;924;5?)90606@76 formulations (m/m, %). /..9 A 39/7?6?=4 6@9A@40 @/ A;;966 @:6 ?6626 '0 9;28?=4 ;/6?=4 .9,2=80 A=; 5; 5 +" 4$" =A'5?=43A@?=@65.(A;7?=?6@9A@?/=+45 ( 7/=47=@65; +45 ( ./97?=450/@9/3?85?,2?;8906@A56%"1 +45 ( -:?8: A9 ./97; '0 @: 63/=@A=/26 65.( +45 ( A667'50 /. 398296/9 ./9725A@?/=6 ?=@/ +45 ( =A=/6@928@29; 76/3:A66 23/= 8/=@A8@ -?@: +45 ( A,2/26 .52?;6+ *:6 # 606@76 @03?8A550 +45 ( 8/=6?6@ /. /9;9; :A4/=A5 /9 82'?8 +45 ( 76/3:A66@:A@=A'5626@A?=;;92495A6 @:9/24:/26=A=/8:A==5617A?=4@:7 A@@9A8@?)35A@./97./9@:39/5/=4;;5?)90;"$#;"$;$" 5A'?5'?/3:A97A82@?8A56628:3@?;6A@923@A-A6)A52A@;49A)?7@9?8A550 /)5 6@ @?7 3/?=@6+ + 4 /. A8: 398296/9 ./97?=4-9;)5/3; )A52A@; '0 /29 96A98: 49/23 ./9 @: $# ./9725A@?/= -A6 ?=8@; ?=@/ A 7 )?A5 ;5?)908/=@A?=?=47:A@; A8?; ?772=/7/;25A@/90 33@?; -?@: A 6:/9@ 35A67A:A5.(5?.9,2?9?=4@-?8(-50;/6?=4+ ;;#$$$;"$" *:606@76;7/=6@9A@;:?4:3/@=@?A5./9$# B98296/9./9725A@?/=6% +744;'0@:45A@?/= ?=8@; ?=@/ 7 /. 95A6 7;?27 ?=@?86;'0-A@923@A 39:A@;@/#+77'9A=5667/;5-A6 95A6 /. * 1 @:6 9625@6 A9 =/- .29@:9 26;+ 2349A;;'0'?/8/73A@?'?5?@0@6@?=426?=4@: *$ A66A0 A=; 7/93:/5/4?8A5 A66667=@ '0 ;("%";$$" ?=)9@; 3:A6(8/=@9A6@ 7?89/68/30+ *: ?/8/73A@?'?5?@0 -A6 A6666; /= .?'9/'5A6@ /'@A?=; .?=;?=46 /..9 )A52A'5 ?=6?4:@6 ?=@/ % (@A"9A@?=/80@%#*#"8556'0 @: ;)5/37=@ /. 3A@?=@(.9?=;50 33@?; : 3/629 @/ 6A73561 ./55/-; '0 *$ ;924;5?)90606@76@:A@=:A=8@:9A32@?8 A66A0 A=; 7/93:/5/4?8A5 )A52A@?/= 26?=4 A= /2@8/76+ ?=)9@; 3:A6(8/=@9A6@ 7?89/68/3 %507326 # 1*/0/1 ;8,-+97,4.;,61;8+-?1. ;9+.24-.;,61;17.02..7?6 79 ,+*) ('*'%$#"! #$""$ $# ($@:8.6?1?8;901?=5?3871??8.=?38 /:3?1=#86;6?183;7745;:98! * ('('$##$""# Figure 3. Representative morphology of fibroblasts (top panels) and keratinocytes (bottom panels). Figure 1. Results of water uptake evaluation of the Untreated control cells (left) are compared with cells studied precursor formulations. following 24-hour treatment with the tested samples (right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igure 2. In vitro release profiles of the studied precursor formulations. 8 8 148?;+?1#8*8 (%$8*8 8 ?58 56:.?38 545=;9?38%8 1??=5?8 (';*8 $8 .;68 ($@:8 =938 ($@8 ,% =06;?+;98 6?8 6;6?58 ?+?58 34?8 :8 6?8 0:?";5=/?=1826=5?58=9386;68.=6;58 .:1,8 .=58 5422:1:+?9;=98 42=,?*86?81??=5?8.=585:.?187:1858.;68 ?5?=1068 =938 99:+=;:98 ?90#8 493?18 21?3:/;9=986?"=:9=8/?5:26=5?58?"0?287:18 ?5?=10680:1?8'493;98:*8%%8=938@1=98 :*8% %* 80 5432 NANOLIPID GEL-BASED LOCAL THERAPY FOR PERIODONTAL DISEASES: A DUAL-ACTION ANTI-INFLAMMATORY AND ANTIBACTERIAL STRATEGY Rabia Ashfaq1, 10-,+*0)(1'+%$),+#"! 1'+$!$,+-01'+ "-+#,)"%,1, Rita Ambrus1'+*, + +4" -2, 3 3 3 1 '+ -$,+%$ '+#,),$%+-$ % '+ -$,+,$-* ,+*)('((%$#"$! %('$% *##$*$%(#$""')$($#"$! $*'%)'($#"$%%$$ * #%$''($ $'##'$%)% $%*(%$* %#%('$%''*%$#(#$ ##$*'%)'($#"$%%$* %(%*($#"$%'$'#'##$*'%)'($#"$%%$* 1. 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SRQP ONMLKLJQIP HGFEP QDRLCLMQIP BA@O?ACQP P :MLCAPAAMLO:E. NR=ELP N?ONQ?MLQEP ;LMRP A:P A@Q?A9QP NA?MLQPELJQPOBP876P:K5PO;PNO=ILENQ?ELM=P4321P Zone of Inhibition for 48 hrs Formulations 0P /.-,+5P A:IP LKN?O@QIP JQMAP NOMQ:MLAP A actinomyce- S. mutans temcomitans 4*-).(K'+.P S%P NLC-ZnHA gel 6/.(PP/., N.A :A:ONA?MLQEP;LMRP$:LBO?KPILEM?LC$MLO:P4#L9P8+5P NLC-NaHAgel -).6PP8.- N.A. Q@Q:PABMQ?P6PKO:MREPOBPEMO?A9QP4#L9P-+. P NLC-HPMC gel 6/./PP/./ N.A ZnHA blank gel -).(PP8.- 8).-PP/.6 NaHA blank gel N.A. N.A. HPMC blank gel N.A. N.A. NLC-Melox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n vitro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ara Gigli , Nicola Demitri , Chiara Sabena , Carlo Nervi, Michele R. Chierotti3 4 5 6,7 6,7 1 , Serena Bertoni +.+ , Cecile Haberli , Jennifer Keiser , Dritan Hasa, Dario Voinovich1 1 , Beatrice Perissutti. 43210/.-2,.+*)+('2-%0$+0,#+"'0/-0%2!.%0$+ %2,%2+,2/.+*)+/2.2+.0$+ $2../0 ,%/*./*,2+/2.2+0*0/2.2+.0$ 3210/.-2,.+*)+('2-./+0,#+ +(2,./2+,2/.+*)+*/,*+.0$ *$**+,..!.2+*)+ %2,%2+0,#+2%',*$*+*%*+!0 3210/.-2,.+*)+"'0/-0%+0,#+*.2%',*$*+,2/.+*)+*$*,0+.0$ 3210/.-2,.+*)+/0,%+('2-./+0%!$.+*)+('2-%0$+,,22/,+0,#+2%',*$*+,2/.+*)+0/2+ (/*0.0 3210/.-2,.+*)+2#%0$+"0/0.*$*+ +/*1%0$+0,#+"!$%+20$.'+,..!.2+ .2/$0,# ,2/.+*)+02$+ .2/$0,# 1. INTRODUCTION strategies were explored using both individual coformers and preformed binary solids as ;:9876543321452:0/ :./ 5-:/ :8/ ,:8+/ 4952*+/ starting materials: )(48,49+'52943/208+%2+056/$#"!6 /(46/8+9+0537/ 420+%/455+052:0/46/40/200:*452*+/65845+7/.:8/5(+/ 1. From individual coformers; %+*+3:),+05/ :./ ,'352%8'/ 6:32%6/ #/ ,4:8/ 2. From preformed PZQ-NCM binary 4%*4054+/ :./ 5(26/ 4))8:49(/ 26/ 5(+/ 9: cocrystal, raw PZQ and AA; 4%,202658452:0/:./,'352)3+/#"!6/-25(20/4/6203+/ 3. From preformed PZQ-AA %:64+/ .:8,/ -(29(/ 940/ 62,)32.7/ 58+45,+05/ monosolvate, raw NCM and AA; 8+2,+06/ ,25245+/ 62%+/ +..+956/ 40%/ %8'/ 4. From preformed PZQ-NCM 8+6265409+/ 40%/ '352,45+37/ 2,)8:*+/ )452+05/ coamorphous and AA; 9:,)32409+// 5. From individual coformers with !0/4/)8+*2:'6/65'%7/-+/8+):85+%/5(+/6'99+66.'3/ preformed ternary cocrystal seeds. .:8,452:0/:./4/20487/4052)48462529/9:9876543/:./ )8412'405+3/ $" / 40%/ 0293:64,2%+/ $; / Slurry bridging routes %+,:06584520/ +0(409+%/ 405(+3,20529/ 4952*257/ /'23%20/'):0/5(+6+/.20%206/-+/42,+%/5:/ +'+ %+*+3:)/ 4/ 5+80487/ 9:9876543/ 6:3*45+/ 7/ 1. PXRD and Synchrotron X-ray diffraction; 209:8):84520/ 49+529/ 492%/ $## / 4/ 9:,):'0%/ 2. Thermal analyses (DSC and TGA); 8+9+0537/2%+052.2+%/.:8/256/405(+3,20529/):5+05243/ 3. Spectroscopic analyses (SS-NMR, 1H- NMR and FT-IR); 4. DFT calculations; +14*2(.4/3+4-%+12*)8%3 5. SEM analysis; 6+1 + 7. 6. Physical stability test; In vitro and in vivo activity. "/;/40%/##/-+8+/)8:*2%+%/.8:,/458:/ )4/ $:3:04/ !5437 / 2,4#3%829(/ $5/ +(23/*3+4-%+%.3'33.8-:'26/ # / 40%/ ;483:/ 84/ $ :%40:2340/ !5437 /8+6)+952*+37/#33/5(8++/9:,):0+056/-+8+/ (+/ 0+-/ )(46+/ -46/ :037/ 499+6623+/ *24/ '6+%/-25(:'5/.'85(+8/)'82.29452:0 ,+9(40:9(+,26587/46/63'887/+)+82,+056/.423+%/ + + 5+80487/ 6:32%/ -46/ 6705(+521+%/ 5(8:'(/ 433/ .2*+/ 8:'5+6/4/)'8+/)(46+/-46/:5420+%/7/,23320/4/ + 3+ + + + + 5:/ 72+3%/ 5(+/ %+628+%/ )8:%'95/ #35(:'(/ 5(+/ Mechanochemical routes: )8+.:8,+%/ "##/ ,:0:6:3*45+/ -25(/ 84-/ Mechanochemical grinding (25 Hz, 400 mg ;/20/4//,:348/8452:/.:8//,20'5+6/20/5(+/ total load) was carried out using a Retsch )8+6+09+/:.///:./##/-(29(/6+8*+%/:5(/46/ MM400 mill. Each strategy was tested at 30, 60, 4/ 6:3*45+.:8,20/ 4+05/ 40%/ 4/ 32'2%466265+%/ 40%//,20'5+6/-25(///:./##/4%%+%/5:/ 820%20/4%%252*+/(+/)'8257/:./5(+/0+-/)(46+/ each experiment. Five different synthetic -46/ 9:0.28,+%/ 7/ / 6)+958',/ -(29(/ 83 GFEEDGCDBA C@DA ?DGD=C@6=A C$;A ;8?;=D=CGA 9=C;A 6A G9=E7DA ,GC6779=DA 8;7D :9=B9=EA :FC@DA ;;-;6CDBA -,A +*)1/(A G?D;G;?,'A CFDA D7F9B6C9;=A %96A ,=@;C;=A ?;$BDA #)6,A ?6CCD=A 6=BA "! A ;?C9896C9;=A D%D67DBA 6A C979=91. Port A. et al., Cryst Growth Des 2019, 19, )+A G?6 @,B;ED=A-;=B9=EA!9E'A+2A6GAGF??;CDBA-,A! ) 2. Thipparaboina R. et al., Drug Discov Today 2016 , 21(3), 481-490. (AG?D;G;?,'A E;F?2A $9C@A C@DA C@DDA ;8?;=D=CGA 79=DBA -,A 3172 3182. 3. ".--F=;A 'A DCA 67'2A Int J Pharm 2023, 644, 123315. 4. Tan X. et al., Aquac. 2023, 577, 739993. 5. Sanphui P. et al., Cryst Growth Des 2012, 12, 4588 4599. .FC@;GA 6=;$7DBEDA 56;7;A 5D=E;A :;A ! )(A 6=67,G9GA GF??;CA 6=BA :F=B9=EA :;8A 0*'A /(A "9?6C98D=C9A B9A Figure 1. Capped-stick representation of the 2A 05A "+0+2A !55(A crystalline structure of PZQ-NCM-AA viewed along 5(12A 05A ""+2A 6=BA a axis. H-bonds showed with dashed blue lines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na Krese1 1 1 ! 2 ( ---------------------! -------------- 1SSaandooz, Lek d..d.., Dev oppment center Ljub d d d eloljana S , Sllovenia 1. INTRODUCTION *12,-------,2 3/# #02'2207/-------------- = :9876543 72 14 91307 085 (------- 2 2---------------------,+*) ) ( 53=7:. 2770 76=3=/851' -8. 5.3 ; ; ; ; ; ; ; ; ; ; ; ; ; ; 2 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; +4131/8 9/8 ; ; ; ; ; ; ; 8/33174. 72 9/017. '7564:1/8 / ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; '74.1:0 3= 41% 6=$.1787-1'/8 '74:13174. 72 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; /66/0/33.. /; ; ; ; ; ; ; 0977 .158/33 60'7044 ; ; ; ; ; ; ; / ; ; ; ; ; ; ; 0; ; ; ; ; ; ; '74:13174. +*) ; ; ; ; ; ; ; ) 53=7:. 5.3 5687$ 87 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 3.2. Optimiz zation of Test Parameters f 9785 :1..783174 5:1/ .=7033 3.314-; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 0 ; ; ; ; ; ; ;; ; ; ; ; ; ; :0/33174. /4: :$4/5 $ 6/0/5 ; ; ; ; ; ; ; 30. .'= /. :1..783174 5:15 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; / ; ; ; ; ; ; ; ; ; ; ; ; ; ; $ / 51' 2887 '74:13174.# ; ; ; ; ; ; ; 2 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ".613 3== 8/' 72 .3/44:/0:1: +*) '7567.13174 4 .02//'3/443 3$$6 /4: '74'430/33174 ; ; ; ; ; ; ; 2 / ; ; ; ; ; ; ; 3 ; ; ; ; ; ; ; ; ; ; ; ; ; ; /; ; ; ; ; ; ; ; ; ; ; ; ; ; 3 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; / ; ; ; ; ; ; ; ) 60737'78. ; ; ; ; ; ; ; ; ; ; ; ; ; ; 22770 76=3=/851' 607:'3. /: 66314- .51.781: 22887 0/33 ; ; ; ; ; ; ; /; ; ; ; ; ; ; //44: ./568 67.1317414- 0 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; / ; ; ; ; ; ; ; ; ; ; ; ; ; ; 6027705 .$.35/331'/8 / 88$ 763151:# 6=7.6=/33 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; /!2220: 2 2; ; ; ; ; ; ; 2 0 5/44' 3.3. ,#-# ( 72220. / / ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 2 2 ; ; ; ; ; ; ; ; ; ; ; ; ; ; 91/!!8 6/33= 37/0: :9 ./814 5:1 ; ; ; ; ; ; ; 5 13= 474 ; ; ; ; ; ; ; ; ; ; ; ; ; ;1741' .02//'3/443 /. ; ; ; ; ; ; ; 2 / ; ; ; ; ; ; ;; ; ; ; ; ; ; / ; ; ; ; ; ; ; / ; ; ; ; ; ; ; ; ; ; ; ; ; ; 87614- !17089/43 ; ; ; ; ; ; ; 4 ; ; ; ; ; ; ; 53=7:. 3=/33 0288'3 7'8/0 6=$.1787-$# .8'3: 37 ! ; ; ; ; ; ; ; ; ; ; ; ; ; ; !/8/4' .78!1813$ /4: 6=$.1787-1'/8 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; ; ; ; ; / ; ; ; ; ; ; ; 2 ; ; ; ; ; ; ; ; ; ; ; ; ; ; 089/44'# / ; ; ; ; ; ; ; ; ; ; ; ; ; ; 2887 0/33 72 55 2 ; ; ; ; ; ; ; / ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 514 /4: / 87 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ./568 :7. ; ; ; ; ; ; ; 68/': 74 / -8/.. ! (3,1*032 3/# (1,+7#2--------------------- 13=14 5 ; ; ; ; ; ; ; ; ; ; ; ; ; ;143.# 5 ( -------------- # ------- ' ------- .3 27705 ; ; ; ; ; ; ; 2 0 58/3174 /44: 0204' 81.3: :0- 0 / ; ; ; ; ; ; ; / ; ; ; ; ; ; ; 2 ; ; ; ; ; ; ; ; ; ; ; ; ; ; 0 ; ; ; ; ; ; ;; ; ; ; ; ; ; = 53=7: ; ; ; ; ; ; ;; ; ; ; ; ; ;:574.30/33: .4.131913$ 37 '0131'/8 / ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; .: 2770 '756/0/3319 9/8/33174# *//017. ; ; ; ; ; ; ; 2 ; ; ; ; ; ; ; / ; ; ; ; ; ; ; / ; ; ; ; ; ; ; *; ; ; ; ; ; ; %/813$ /333301! ; ; ; ; ; ; ; / 3 !3. ,.( 6/031'8/08$ 91.'7.13$ ; ; ; ; ; ; ; ; ; ; ; ; ; ; 0 ; ; ; ; ; ; ;; ; ; ; ; ; ; :1..783174 5:1/ /4: .02//'3/43. 0 /....: ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 2 4 ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ,1-#( /44: ; ; ; ; ; ; ; /; ; ; ; ; ; ; 6/0331'8 .1 :1.301!3174 ,1-#(# 0 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; 37 763151 :0- .78!1813$ /44: 08 /33'=. 13 / ; ; ; ; ; ; ;= /830: 678$50 '74343 70 :0-; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 0; ; ; ; ; ; ; '74:13174.# 4/8$31'/8-0/: 0 .!.3 44' / ; ; ; ; ; ; ; " .=7: :1.314'3 08 /4/8$31'/8 53=7:. 0 5687$: 3=07 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;-=73 ; ; ; ; ; ; ; '7421105 2 0 514- 3 ; ; ; ; ; ; ; 3= 5 ; ; ; ; ; ; ;3=7:. :1.'01514/3370$$ 670# ; ; ; ; ; ; ; / 0 ; ; ; ; ; ; ;; ; ; ; ; ; ; 3= .3:$ ; ; ; ; ; ; ; 790 0 ; ; ; ; ; ; ; " /4: 91.'7.13$ 5 ( ------- =1!13: 9 ; ; ; ; ; ; ; 94 -0= 53=7: :9876543 277'.: 74 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 2 ; ; ; ; ; ; ; ; ; ; ; ; ; ; 1:4312$$14-2; ; ; ; ; ; ; .13/!8; ; ; ; ; ; ; :1..783174 ; ; ; ; ; ; ; 5:1/; ; ; ; ; ; ; .02//'3/443. 2887 0/33. 44: ./568 67.1317414-2 / ; ; ; ; ; ; ; 2 ; ; ; ; ; ; ; / ; ; ; ; ; ; ; / ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; .30/33-1.# = 2114/8 53=7: /. .8'3: !/.: / ; ; ; ; ; ; ; ; ; ; ; ; ; ; 2 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; 74 13. /!1813$$ 37 60791: 0607:'1!8 ; ; ; ; ; ; ; ; ; ; ; ; ; ; 3 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; !17089 443 /4: :1.'01514/3370$$ :0- 08 85 *) ))(''%)$% "( ! $" ") ( # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # %'%% # # # # # # # ## # # # # # # # "( '% ! 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iri Zeman, Jakub Vyslouzil, Jan Elbl, Sylvie Pavlokova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"" $ # $#"! ! 80) 3 271750/ "3 ' 980123 '0210-0/,1)+3 ,+)757*3 %' # # !"! ! *0''9+)091359-0+7'3,) 02 753:36,+7'3%$02# .!43 ' " "' 9)71)0,++3 ,--7/)0123 ) 75,3 8 713 '7*3 #' /91/5571)+380) 3,1),/0*'3953013/91*0)091'3) ,)3 $ # $' 01/57,'732,')50/3:#3 ' ' $ # " % ! $' $$ $# $" $ $% $! $ $ " 0''9+)091359-0+7'39-3),+7)'3,)3:3#380) 3 ,+7)38702 )'3%133!43*0(71'091'43 ,5*17''3 '0(0+,50)376,+,)0913%133!# %133"!43,1*3/,+/+,)7*3)71'0+73')5712) # Sample Original 0'3 ')*3 /9(,57*3 ) 73 950201,+3 -95(+,)0913 Generic Generic Weight 80) 3)893271750/'43-9/'01239132,')5957)71)0913 727.90 845.10 714.90 ,1*3 *0''9+)0913 / ,5,/)750')0/'#3 3 59191/7*3 1 2 (mg) 10.40 9.00 8.40 *0--7571/73 8,'3 9'7567*3 01 ) 7 2,')5957)71)0673 Length 12.09 16.50 15.05 (7/ ,10'(3 ) 73 950201,+3 -95(+,)0913 ,1*3 (mm) 0.01 0.03 0.02 271750/ "3 )0+07*3 (,)503 '87++0123 ,1*3 Width 12.09 8.23 8.58 (/9,* 7'0913-95357)71)091438 757,'3271750/ .3 (mm) 0.01 0.02 0.02 +,/7*3 ) 7'73 5975)07'3 ,1*3 ,+'93 *0*3 19)3 -+9,)#3 Height 6.95 6.63 6.96 ,'7*3913) 7'73-01*012'43271750/3.3(,3)5,1'0)3 (mm) 0.03 0.02 0.02 ) 592 3 ) 73 ')9(,/ 3 (9573 5,0*+43 9)71)0,++3 F (N) 151.97 299.81 247.07 57*/0123 (7)-95(013 ,'95)0913 013 ) 73 753 6.69 15.47 8.48 2,')5901)7')01,+3 )5,/)#3 72,5*0123 *0''9+)09143 Ts 271750/3 "3 7 00)7*3 ,3 -,')753 57+7,'73 ,)3 :3 #43 1.15 2.45 n/a 2 8 0/ 3/9+*301/57,'73) 7350'39-3,*675'737--7/)'3 (N/mm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ndrej Grobin1/%/%1, Jurij Trontelj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'")($ A /-'* '*A $HDEGCH=5A /+//5A IED:@GHA@?A?EHD9GE=A4:E8AIEG?@GFEGA@8AIGC8DEDA ,A/,/ CFFEG=E6A C8A HA 6C==@9DC@8A FE6C9FA I@G@9=A $ $#AFHIAH@4=A=EIHGHDC@8A@?A(HGC@9=AIG@D@8A BCE8BEA E8DGEA 3@H86A .GH8D=A 89F'EGA $7 /+//""""++,A H86A $ I@@=-=C=5A HGEH=A /+,","""+/ 4CD:A6C??EGE8DA4HDEGAF@'CCD-A4EGEA6C=DC8.9C=:E6A '9''E=0:EA 4@GA 4H=A =9II@GDE6A '-A D:EA HDC@8HA @8E5A4HDEGAIE8EDGHDC@8A@8E=5AFHBG@ABGHB=5AHCGA 94 3221 0/.-,+/*)0/..-,(')%$),/$)0/.-,+/*)3/,.+#*-)3,%3-,.+-() +"!* -"#-)%3./*0+#)( (3-"(+%")#/ 1 1 1 1 1 eljko Agi , Nina Popovi , ) )+)3 , Andrea Roje 1Jadran-Galenski Laboratorij d.d., Rijeka, Croatia 1. INTRODUCTION 0++C@03=,DCA=@A1-=BAE@1/D=BACB E$0?-= FEDCBAC@?E= CD;E? 212BC?20E?= 02= = DEB0/= /EAD0/E2DCAE0@= 2E1D0E?= B32C@= )7= 12C@=DE=DAC=E/1++57= 7= = )77= =$2==20-?=E,=BEEA=212BC?20E?=2D;00D3= *)(6='?=@@0D0E?=F9=2%E$2=BAE+020?-=AC21D2=0?= E+CA D%C=DAC+0/= 212BC?20E?2=3=12C@=,EA=,EA+1),-( *.()/")+(# ((+%" BEEA3= 2E1;C= = 0?-AC@0C?D2=: '28=C?%0!CA3=;3= 1),) ) ) CDC?@0?-= E/1=3202=AC!CC@=@0,,CAC?/C2=0?=BC= '=BC2=AC+0?=0?=D%C=,EA?0=!C= 20C=E-3=E,=2DCA0C=C= 2E$3= 0+BAE!0?-= ;0E;00D3= *(6= '2= F96=%C=?E?.2DCA0C= '=/E?D0?2==20-?0,0/3= BC= 20C= 02= = C3= ,%0-%CA= BEAD0E?= E,= BC2= 2+CA= D% B%320/= 2D;00D3= 57)6= +8=$%0C=D%C=2DCA0C= '= 212BC?20E?26= %CAC,EAC= BC= 20C= %2==BC2=:57#6=+86= @02DA0;1D0E?=:8=02==/A0D0/=10D3= : 8=E,=21/%=@A1-=BAE@1/D26= A) %C=-E=E,=D%02=$EA=$2=DE=C!1 /A0D0/= += C= 2DCA0C= F9= 21/%= 2= BC= 20C= 2% 21A,1C?/C=D%C==0?=D%C= @A1-= BAE@1/D= $0D%= D%C= 0+= E,= EBD0+00?-= D%C= B%320/= 2D;00D3= E,= F9= 7654= C3C= @AEB2= 212BC?20E?6 )0/.-,+/*()/")0-.%( C!CA= ;C= :?0!CA2= B0?8=C=:'?@12DA0C=%0+0/=A= 'D38= F9= $CAC= 12C@= ,EA= BACB ;EAC.1B= ; BAE@1/D6= %C= 20C= E-3= E,= F9= BC2= $CAC= 32C@= 120?-= ;ED%= 2CA= @0,,A 3@AE==1?0D=!CA?=3D0/=FD@6=8= E-3=@0AC/DC@=+ := EAB%EE-0= .'= 232DC+= !CA?= 3D0/= FD@6= 86= %C= .3= E$@CA= !)1)"BD0/=+0/AE-AC=: 8= 0,,A2DCA0C=:8=F96 120?-= = %C= ?E?.2DCA0C= += /E?D0?2= +EAC= AE1?@= 7577= !CA?= 3D0/= FD@6= 86= BC2=:%0-%CA=/0A/1C= =0?=D%C=@A1-=BAE@1/D=$2=+C21AC@=;3=D%C= F9=BC2=E?-2CA= @0,,A=3202=2%E$C@=?E=@0,,CAC?/C=0?=/A32D= 3@AE==1?0D=!CA?=3D0/=FD@6=86= 2DA1/D1AC=;CD$CC?=2DCA0C=C=F96=E= E= 2/ACC?= ,EA= BEDC?D0= /% D0+C= +C21AC+C?D2= $CAC= BCA,EA+C@= 96 "!! ?=;:989?7654=395472?427:91=590/5:7.:-9-278;9 4:9,=2+=?4278;9*70?:5590+9525*:5805)9 (:.:7=39 3=/07=40769 /=4?:59 ?04=88;9 54:783:9 =-9 0'54:783:9 %9 1:7:9 *7:*=7:-$9 =-9 4:879 #("9 ==365:-)9 !339 /=4?:59 ?04=88;9 54:783:9 %$9 =-9 ;7:=4:79 #("9 4=9 /=4?:59 ?04=88;9 0'54:783:9%)9%.:9402;9" $9"$9=-9"9 1:7:950,:01958,83=7$94:9-8++:7: *7002:9"9=-9" 9#("9*=7=,:4:759 8;)9 $9 52;;:548;9 4:9 *7:5: =;;30,:7=4:59 89 4:9 /=4?:59 ?04=88;9 54:783:9 9#("9*=7=,:4:759" $9"9=-9"9+0795?=3:' !#)9 2*9 /=4?:59 ?04=88;9 54:783:9 (9 =-9 0'54:783:9 (9%)9#("9,:=527:-9=4947::948,:9*0845$95407:-9 =499=-997:3=48.:92,8-8469) %.:9 402;9 4:9 54:7838=4809 *70?:559 -8-9 049 7:52349 89 5472?427=39 ?=;:5$9 849 -0:59 :.0:9 ,07*030;8?=39 -8++:7: 0'54:783:9 7=19 ,=4:78=3)9 #=748?3:9 58:9 =-9 ,07*030;690+97=19,=4:78=398+32::9-72;9 *70-2?49#("9'90:90+94:9,=89 !590+9=9-72;9 *70-2?49 89 4:9 +07,9 0+9 525*:580)9 "72;9 *70-2?459+07,23=4:-9184954:783:9!#9:8/84:-9 ;::7=3369 3=7;:79 *=748?3:59 ?0,*=7:-9 409 405:9 9 #("9 *=7=,:4:759 "9 =-9 " 9 +079 ?04=88;90'54:783:9!#$91849=94:-: 3=/07=40769/=4?:59?04=88;954:783:9(9=-90' 54:783:9(9%) 8 485910791:9=.:950194=49#("9*=7=,:4:759 "9=-9" $9?=9/:9*7:-8?48.:90+94:9525*:5809 1095?=3:'2*9/=4?:5$90:9184954:783:9=-94:9 54=/83846)9 :5:9 *=7=,:4:759 =7:9 0+4:9 049 04:79 1849 0'54:783:9 %9 1:7:9 ,=2+=?427:-)9 8:9 -72;9 *70-2?49 079 7=19 ,=4:78=39 #("9 1=59 ,:=527:-9 +079 /049 /=4?:59 5*:?8+8?=4809 =-9 =7:9 049 89 4:9 5?0*:9 0+9 8,,:-8=4:369 2*09 /239 *7:*=7=4809 $9=+4:79 7:;23=40769;28-=:69?=9/:9=9 0:9 ,049 $9 =-9 =+4:79 589 ,0459 9 =49 :3*+239 40039 +079 +07,23=4809 5?7::8;9 89 4:9 9 =-9 9 7:3=48.:9 2,8-8469 )9 #("9 :=736954=;:90+9-:.:30*,:4) 7:523459=499501:-90958;8+8?=49-8++:7: 89" $9" $9=-9" 9/:41::9/=4?:59?04=88;9 54:783:9 =-9 0'54:783:9 %)9 :9 -8++:7: )9 !,0$9 )$9 :49 =3)$9 ! #("91=590369048?:=/3:989*=7=,:4:759"9=-9 " $9?0+87,8;94:97:5234590+93=/07=40769*=5:)9 )9 4:7=480=39 9==3658597:.:=3:-9=98*4=,030;6$9 )99' ) 0+9/=4?:59?04=88;954:783:9%$9183:94:95=,:9 )9:?,=$9)$9:49=3)$9! *:0,:09 1=59 049 0/5:7.:-9 +079 /=4?:59 )9 027=39 0+9 ?23=79 ?04=88;90'54:783:9!#)98598#=7,=?030;69=-9:7=*:248?5$9)99' 1=59 ,07:9 *7002 ) 024'0+'5*:?8+8?=4809 7:523459 +079 "9 899 8;)9 )90$9!)$9:49=3)$99 )994:904:79=-$94:9/=4?9?04=88;90' 54:783:9%9501:-9098:9#("9 )9 4:7=480=39 027=39 0+9 *=7=,:4:75) #=7,=?:248?5$9)99 97 /..- ,+*)('+,//%),$#+")+! !)!+)%,"!)*+)%+)%*)!$, Alaa Gamiel 1 1 1 , Csilla Balla-Bartos , Rita Ambrus 1 Institute of Pharmaceutical Technology and Regulatory Affairs, University of Szeged, Szeged, Hungary 1. INTRODUCTION EDCBDA@?= @; :D98B9 BDB;98CD@76 B9@5 @B46==9DC3 7C21 0/.-,+* )86DB1; 9D 9(8 DA@?= 1CD2:' ,9 @; ?D==DB63 2;87 4DC :@B ?DB7@9@DB; =2;?26D;86896 7@;DC78C; B7 :D;9D:8C9@%8:@B',9;=$DC:CD)68=;C89(8 4DC=269@DB ?(668B18; 728 9D :DDC #98C ;D62)@6@93B79(8:D98B9@61;9C@?;@788448?9; B7 26?8CD18B8;@;' "DB%8B9@DB6 6@!2@;D6@7 ;3;98=;B@BBD%9@%8B7:CD=@;@B1 ++C8:C9@DB=:684DC=269@DB' ::CD?( 4DC ?C89@B1 ;D6@7 7D;18 4DC=; 9(9 -+ + @B?DC:DC98 ;@1B@4@?B9 =D2B9 D4 6@!2@7 (3;@?D5?(8=@?6 :CD:8C9@8; D4 :2C8 E ;:8?@4@?663 )3 7@;:8C;@B1 9(8 7C21 #@9(@B B :(3;@?6 =@A92C8 %?22=57C@87 ;=:68 B7 ::CD:C@98(37CD:(@6@?B79@686@!2@7 4C88857C@87;=:68#8C8@B%8;9@1987'.2C4?8 =87@2=6@!2@77C21'(@;98?(BD6D13)D;9; =DC:(D6D13 #; B6387 %@ ;?BB@B1 ;8%8C6 )8B84@9; =D;9 BD9)63 8;8 D4 2;8 868?9CDB =@?CD;?D:3 0.*' (8C=6 44DC7)@6@93 ;2@9)@6@93 4DC 6C185;?68 )8(%@D2C #; ??8;;87 )3 +@448C8B9@6 =B24?92C@B1 B7 8?D54C@8B76@B8;;' ,B D2C ;?BB@B1 ?6DC@=89C3 0+."* ;9273 #8@=879D8=:6D39(8%@=89C@? -B63;@; 0-*' 5C3 9(@;98?(B@!28#@9(=D7@4@?9@DB;:8C9@B@B19D :D#78C 7@44C?9@DB 0+* B63;@; #; 9(82;189@B1=98C@6' 8=:6D387 9D 8A=@B8 9(8 :(3;@?6 ;9C2?92C8' -+,"'%!,+,*+'"#) .2C4?8 4C88 8B8C13 B7 :D6C@93 D4 #8C8 ?6?269874DC8%629@DB)@6@93' -++ (8 =D786 7C21 #; 6DCBDA@?= 0E* 05 -+!++ "(6DCD55(37CDA355=89(365/50:3C@7@B5536*5 ,B@9CD7@;;D629@DB98;9@B1#;:8C4DC=879: 59(@8BD 58 9(@@B855?C)DA=@78 ' B7C9@%8B63;@;#; 57@DA@78*0D3D"(8=@?6,B72;9C3 :B*' )89#88B D2C ;=:68 B7 9(8 ?D==8C?@6 D63%@B36:3CCD6@7DB80*0.@1=5 :CD72?9 2;@B1 9(8 :@C#@;8 ;@=@6C@93 4?9DC -67C@?( "(@B* #88B =D7860 0.@1=5-67C@?(*' "(8=@8=) +C=;979 8C=B3* ;D7@2= +%'"+,*+!$!)* )@?C)DB98B7-%@?86#8C8; 8A?@:@8B9;);87B7 ++ :2C@4@87#98C#;2;879D:C8:C89(8!28D2; E8A(@)@9876@=@987;D62)@6@939:''(8 ;D629@DB;' @B981C9@DBB7#88B#; --++ @B?DC:DC9@DB D4 ;D7@2= )@?C)DB98 !28D2; .D62)@6@93 D4 E #; ;927@87 @B 7@448C8B9 8B(B?8 ;D62)@6@93 @=:CD%8; ;D62)@6@93 #(@68 ;D629@DB 3@86787 9(8 D:9@=6 ;D62)@6@93 D29?D=8 ;D6%8B97@;9@6687#98C0:'* )2448C9: ' #88B #% !28D2; /" -+ + ;D629@DB#@9(#88B B7 #%!28D2; E ,=18; D)9@B87 %@ 0.* C8%8687 /";D629@DB' 18D=89C@? ?C3;96 ;9C2?92C8; 78=DB;9C9@B1 #@78 ;@8 7@;9C@)29@DB' "DB%8C;863 9(8 -+ + 4DC=269@DB; 8A(@)@987 :DCD2; =9C@A @12C8 C8:C8;8B9; 9(8 ;98:; D4 ;=:68 ;9C2?92C8 @B7@?9@B19(8:D98B9@6=DC:(@9@DB :C8:C9@DB' D4 9(8 ?9@%8 @B1C87@8B9' (8B ?D=:C@B1 %?22=7C3@B10+*#@9(4C8887C3@B10+* 9(8 + ;=:68; 8A(@)@987 =DC8 :DCD2; ;2C4?8' (8 E +." ?2C%8 8A(@)@987 98 )((' KJIHJGFH DCBHAD@? E E E E E E E E E @ ?JF D= =HH ;;:98 E E E E E E E E E = E E E E E E E E E = E E E E E E E E E 87 654 E E E E E E E E EE E E E E E E E E 150 IJ3GJ211JG3 2E E E E E E E E E JHIE E E E E E E E E KDFB?BIJHJBG4E E E E E E E E E 0AJFAE E E E E E E E E JIE E E E E E E E E ) FB@@B/B@=HHDK /1 .AD@? = E E E E E E E E E E E E E E E E E E @ ?B3@=--J?DH@JF , = E E E E E E E E E ,G=+1IJI E E E E E E E E E(% 100 *.),(9 .AJI D= 0=I =//IDGH JG HAAD E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E = E E E E E E E E E E E E E E E E E E H E E E E E E E E E I ? = ?+D E E E E E E E E E ved B0KD@I4 I'33DIHJG3 HAD 2BB@? E E E E E E E E E E E E E E E E E E E E E E E E E E E 2 @ ?=HHJBG = E E E E E E E E E B2 =GG E E E E E E E E E = E E E E E E E E E ol 50 =?B@AB'I IH=HHD9 .AD F@1IH=++JGJH1 @ E E E E E E E E E = E E E E E E E E E E E E E E E E E E E E E E E E E E E JGKJFDI E E E E E E E E E ss Di F=+F'+=HHDK 22@@B? = E E E E E E E E E E E E E E E E E E I=? = ?+D KJ222@@=FHB3@=?I @ E E E E E E E E E 2 2 E E E E E E E E E @D-D=+DK E E E E E E E E E @DK'FHJBGE E E E E E E E E F@1IH=++JGJH1 B2 HAD K@''3I *J3 E E E E E E E E E E E E E E E E E E E E E E E E E E E @ E E E E E E E E E 3'@D ;(9 E E E E E E E E EE E E E E E E E E 0 %DI'+HI B2 0 20 40 40 60 80 E E E E E E E E E E E E E E E E E E F=+F'+=HHDK B+=@JH1 =GK HAD = E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E I'@2==FD 2 E E E E E E E E E Tim e (min i ) LX Ph y si cal m i x t u r e 22@@DD DGD@31 I'33DIHDK E E E E E E E E E @ E E E E E E E E E E E E E E E E E E HHAA=HH HAD 2BB@? = E E E E E E E E E E E E E E E E E E 2 ?'+=HHDK = E E E E E E E E E v acu u m - dr i e d f r e e ze - d r r i e d I=?+DI E E E E E E E E E DCAJ/JH E E E E E E E E E 3@D=HHD@ = E E E E E E E E E 0DHH //J+JH H = H1 E E E E E E E E E = =GGK E E E E E E E E E A1K@BAJ+JF @BD@HJDI FB?=@DK HB HA E E E E E E E E E @ E E E E E E E E E E E E E E E E E E E E E E E E E E E AD '@D E E E E E E E E EE E E E E E E E E %$#"! E E E E E E E E E"JIIB+'HJBG @=HD @B2JJ+DI =H 9;9 E E E E E E E E E E E E E E E E E E 2 E E E E E E E E E E E E E E E E E E E E E E E E E E E K@'39 .ADE E E E E E E E E ?BKJ22JJDKE E E E E E E E E +J'JIB+JKE E E E E E E E E HDFAGJ' 'DE E E E E E E E E A=IE E E E E E E E E KD?BGIH@=HHD =K D222JJF=F1 JG DGA=GFJG E E E E E E E E E 2 2 E E E E E E E E E E E E E E E E E E G3 HAD E E E E E E E E EE E E E E E E E E IB+'/J+JH1 B2 E E E E E E E E E E E E E E E E E E 2 +B@GGBCJF=? @ = ? JG =FJKJF DG-J@BG E E E E E E E E E E E E E E E E E E E E E E E E E E E G?DGHI9E E E E E E E E E H@'FH'@=+ =GG E E E E E E E E E = G=+1IJI JGKJF=HHDI = @DK'FHJBG E E E E E E E E E = E E E E E E E E E E E E E E E E E E G JG HAD E E E E E E E E E E E E E E E E E EE E E E E E E E E F@11IH=++JGJH11 @ HE E E E E E E E E E E E E E E E E E B2 +B@GGBCJF=? @ = ? =+BG3IJKD JG E E E E E E E E E E E E E E E E E EF@D=IDKE E E E E E E E E A1K@BAJ+JFJ JH19E E E E E E E E E @DDD @ K@11JG3E E E E E E E E E IJ3GJ22JJF=GGH+1 =E E E E E E E E E J?=FHI HAD E E E E E E E E EE E E E E E E E E IH@''FH'@=+ JGHD3@JH1 =GK KJII @ E E E E E E E E E E E E E E E E E E E E E E E E E E E IB+'HJBGE E E E E E E E E @=HHD =E E E E E E E E E B2E E E E E E E E E +B@GBCJF=? = ?4E E E E E E E E E 1JD+KJG3E E E E E E E E E @B B?JIJG3E E E E E E E E E B'HFB?DI 2BB E E E E E E E E E 2 B@ HAD 2BB@?'+=HHJBG B2 GG =K E E E E E E E E E E E E E E E E E E 2 = E E E E E E E E E E E E E E E E E E = E E E E E E E E E K-=GGFDK =E E E E E E E E E %$#"! '9E E E E E E E E E $ $% %#" KJ222@@=FHB3@=?I B2 ,#!4 E E E E E E E E E 2 2 E E E E E E E E E E E E E E E E E EE E E E E E E E E A1IJF=+ AE E E E E E E E E B@=+ KBI=3D E E E E E E E E E D 2BB@? E E E E E E E E E 2 @ ? 0JHA @ JK KJIIB E E E E E E E E E E E E E E E E E E = E E E E E E E E E B+'HJBG4E E E E E E E E E ?JCH''@@D *# (4 -=F''? H ' E E E E E E E E E E E E E E E E E E ' ?K@JDK B0KD@ * E E E E E E E E E E E E E E E E E E "(4 =GK E E E E E E E E EE E E E E E E E E 'HJ+JJG3 = E E E E E E E E E E E E E E E E E E ?JGJ?=+ '=GGHJH1 B2 D E E E E E E E E E = E E E E E E E E E E E E E E E E E E2222DDFHJ-D 2E E E E E E E E E 22@@DDDK@JDK B0KD@ *"(9 E E E E E E E E E E E E E E E E E E DCFJJDGHI9 GE E E E E E E E E '@ 2''H'@D DGKD --B'@I E E E E E E E E E 2 E E E E E E E E E =E E E E E E E E E JGF+'KDE E E E E E E E E FBGK'FHJG3E E E E E E E E E IH //J+JH1 IH'KJDI =GGK = E E E E E E E E E E E E E E E E E E =E E E E E E E E E 22''@HHAD@ @E E E E E E E E E %" %#% JG-DIHJ3=HHJBG = GI HB DCHHDGK HAD =+JF=HJBG E E E E E E E E E E E E E E E E E E C E E E E E E E E E E E E E E E E E E = G B2 HAAJI E E E E E E E E E E E E E E E E E E HE E E E E E E E E .AD KJIIB+'HJBG @B2JJ+DI B2 HAD ,#! GGK E E E E E E E E E E E E E E E E E E 2 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E =E E E E E E E E E-=@JB'IE E E E E E E E E HDFAGJ'D HB E E E E E E E E E E E E E E E E E E B -=@JB'I K@''3 ?BKD+I9 E E E E E E E E E @ E E E E E E E E E 22BB@? @ ?'+=HHJBGI =@D KDJFHDK JG J3'@DI = E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E EE E E E E E E E E E E E E E E E E E 9 '@E E E E E E E E E 22BB@? @ ?'+=HHJBGI4 0AJFA 0D@D I'/DFHDK HB = E E E E E E E E E E E E E E E E E E E E E E E E E E E / E E E E E E E E E E E E E E E E E E 22@@DDD K@1JG3 E E E E E E E E E =GK E E E E E E E E E-=F''? E E E E E E E E E K@1JG3 E E E E E E E E E HDFA AGJ'DI4 E E E E E E E E E KD?BGIH@=HHDK FB?+DHD KJIIB+'HJBG 0J = E E E E E E E E E E E E E E E E E E E E E E E E E E EHAJG : E E E E E E E E EE E E E E E E E E KJIIB+'HJBG B/ID@-DK 0JHA HAD '@D K@'' =K=0J4 ,9, E E E E E E E E E ,999 I?=G4 ,99 =/J/ E E E E E E E E E E E E E E E E E E E E E E E E E E E /4 "9,9 E E E E E E E E EE E E E E E E E E ' E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E @ '3 E E E E E E E E E *=2HHD@ 2 E E E E E E E E E *)(''%$#"! ( + + + + + + + + + $$ ) ! # ##$%%)) !%--))%( $ + + + + + + + + + + + + + + + + + + $ + + + + + + + + + + + + + + + + + + $ % + + + + + + + + + % - (! )-- + + + + + + + + + + + + + + + + + + ) ?JG'HDI( ?JG'HDI4 JG FBGH@=IH HB HAAD +DII HA= 9E E E E E E E E E +DHB'A114 AE E E E E E E E E "9,9E E E E E E E E E )=?JD+4E E E E E E E E E ,9,9%9 ,E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E H E E E E E E E E E E E E E E E E E E =G E E E E E E E E E E E E E E E E E E E E E E E E E E E ' 9 .AJI DGA=GFD?DGH JG IB E E E E E E E E E E E E E E E E E E G E E E E E E E E E E E E E E E E E E B+'/J+JH1E E E E E E E E E ?=11 /D =HHHH@J/'HDK HB HAD JG2++'DGFD B2 DC ## !!% )) $ + + + + + + + + + !! *)"!$#) )% + + + + + + + + + + + + + + + + + + + + + + + + + + + )#(+ + + + + + + + + = E E E E E E E E E E E E E E E E E E = H E E E E E E E E E E E E E E E E E E E E E E E E E E E 2 E E E E E E E E E E E E E E E E E E CFJJDGHI E E E E E E E E E FABIDG9 '@HHAD@? E E E E E E E E E @ @ ?B@D4 HAD@D JI = IJ3 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E 3GJ2JJF=GH 2 GE E E E E E E E E $$##$ ( ((!##$!! $+ + + + + + + + + !!!! + + + + + + + + + ##$ !!$ $+ + + + + + + + + !!%$!+ + + + + + + + + $ + + + + + + + + + $$ $$ + + + + + + + + + + + + + + + + + + $## -$ --(($)% %%+ + + + + + + + + + + + + + + + + + BIJHJ-D FB@@D+=HJBG /DH0DDG HAJI 2JJGKJG3 E E E E E E E E E = E E E E E E E E E E E E E E E E E E E E E E E E E E E 2 3 =GK HAAD E E E E E E E E E E E E E E E E E E HE E E E E E E E E !%)) !!# %+ + + + + + + + + ----!!$ %(!$# !"!--$ E E E E E E E E E + + + + + + + + + !)''(!$ )+ + + + + + + + + 22BB@? @ ?'+=HHJBG HDFAGJ'DI D?+B1DK9 BHA = E E E E E E E E E E E E E E E E E E E E E E E E E E E A K@11JG3 E E E E E E E E E @E E E E E E E E E + + + + + + + + + !!)) E E E E E E E E E '(4E E E E E E E E E ,,4E E E E E E E E E ?DHABKI BIJHJ-D+1 =222DDFH KJIIB+'HJBG AB E E E E E E E E E E E E E E E E E E 2 2 E E E E E E E E E E E E E E E E E E B0D-D@4E E E E E E E E E KBJ97 7 8:9;9;;8;9 HAD 2@@DDD @ E E E E E E E E E 2 K@11JG3 ?DHABK 1JD+KI E E E E E E E E E E E E E E E E E EE E E E E E E E E GBH=//+1 =E E E E E E E E E ;9E E E E E E E E E BB B@D4 99 +=GGD@4 99 =HAAD E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E H D?=HJF=+E E E E E E E E E I'D@JB@ @DI'+HI * 5B?=@JIBG B E E E E E E E E E E E E E E E E E E B2 "JIIB+'HJBG #@B2JJ+DI9 E E E E E E E E E 2E E E E E E E E E%(+ + + + + + + + +$$!!)E E E E E E E E E E E E E E E E E E E E E E E E E E E * - -- ,, + + + + + + + + + 8;(9 .AJI =+J3GI 0 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E 0JHA HAD E E E E E E E E E E E E E E E E E E 22JJGKJG3I B2 .==+K=D- DH =+9 =I @DB@HHDK 4E E E E E E E E E,4 8 89 E E E E E E E E E E E E E E E E E E E E E E E E E E E . E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E @ E E E E E E E E E E E E E E E E E E JG HADJ@ E E E E E E E E E JGF@D=IDK I'@2==FD =@D= B2 HAD 2@@DD AD-+= E E E E E E E E E E E E E E E E E E J E E E E E E E E EE E E E E E E E E ))##--## #-$#) )--+ + + + + + + + + + + + + + + + + + ) E E E E E E E E E 2 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E 2 DK@JDK E E E E E E E E E $! + + + + + + + + + #)!(# + + + + + + + + + %))''!%$#! % ) + + + + + + + + + ) )-- + + + + + + + + + $#"! + + + + + + + + + B0KD@44 HAD =?B@AB'I IH@''FH'@D B2 $ @D-JD0 9E E E E E E E E E .==+K=D-4 ,9 #=G E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E . B-4 %9#9 E E E E E E E E EE E E E E E E E E E E E E E E E E E 4 0AJFA ?=11 /D DC+=JGDK E E E E E E E E E E E E E E E E E E = E E E E E E E E E E E E E E E E E E E E E E E E E E E /1 HAAD E E E E E E E E E H E E E E E E E E E @ E E E E E E E E E E E E E E E E E E E E E E E E E E E @ E E E E E E E E E E E E E E E E E E $ 4 =GGK E E E E E E E E E =E E E E E E E E E HADE E E E E E E E E AJ3AE E E E E E E E E B@BIJH1E E E E E E E E E B2E E E E E E E E E HADE E E E E E E E E +1BA AJ+J=HHD9 =E E E E E E E E E %(!$# %(!$# %%!!##!$ "# )''# + + + + + + + + + % $ + + + + + + + + + + + + + + + + + + ) ##$#) $9E E E E E E E E E ,KKJHJBG=++14 B'@ 2@@DDDK@JDK @BK' E E E E E E E E E E E E E E E E E E 2 E E E E E E E E E 'FH 0=I E E E E E E E E EE E E E E E E E E KBJ9A=@?=FD'HHJFI:; E E E E E E E E E '('4E E E E E E E E E 4E E E E E E E E E ; 4E E E E E E E E E ' 9 D-=+'=HHDK JG FB?=@JIBG HB /BHA IJ? = E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E +D =GGK E E E E E E E E E =E E E E E E E E E @ JK FB??D@FJ=+ H=/+DHI FBGH=JGJG3 $ = E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E $ 9 .AD E E E E E E E E EE E E E E E E E E KJIIB+'HJBG FA=@=FHD@JIHJFI B2 HAAD 2@@DD ' E E E E E E E E E E E E E E E E E E E E E E E E E E E H E E E E E E E E E 2 DK@JDKE E E E E E E E E @BK'FH 0D@D =GG=+B3B'I HB HABID B2 HAD @BK'FH 0D@D =GG=+B3B'I HB HABID B2 HAD @=JK E E E E E E E E E E E E E E E E E E = = E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E D @=JK E E E E E E E E E = = .AJI 0B@ 0=I I'B@HHDK /1 HAD ! .AJI 0B@ 0=I I'B@HHDK /1 HAD ! E E E E E E E E E @ @ E E E E E E E E E E E E E E E E E E @ @ E E E E E E E E E E E E E E E E E E E E E E E E E E EE E E E E E E E E ., E E E E E E E E E =FHJG3 FB??D@FJ=+ H=/+DH * E E E E E E E E E E E E E E E E E E E E E E E E E E E *---,,E E E E E E E E E (9 E E E E E E E E E 887 @BDFH9 E E E E E E E E E B 99 :998 765543210/./-5.3,5.376+401+*/)4(3/4'5*/7+*/)43)23*%53 /421054$53)23#+*563$)4*54*3+4.3:)'/.)45376+.5 /"! !3+1, *!371 1. INTRODUCTION CAE88AED +B8D 1/"D 6BAD B:9D =FED @ CAE88AED+B8D/D6BA/D GFEDCFBA@B?E=8=A7DFB8DAE?E:=;7D6EE:D 9E:=5=4A8D =4D 83 :!3 3 ! !3 3 ! 3 ?;@B=ED ?FB:3ED 210/D .:D AE?E:=D 7EBA8-D !3 CFBA@B?E=GFEDCBA==4:D+B8DE,B;B=E9D67D FBA9D=4DAE9?ED=FE=D 88:3DBD8E=D45D8=B:9BA9D8 =FED?FB:3E8DBAED8=;;D:E3;36;ED:DAE;B=4:D=4D=FED =FEDAB:3ED45DD@/DGFED5;4+DCA4CEA= 8ED 45D ?BA64:):=E:8,ED =AB9=4:B;D =B6;E=:3D@=AE8DCAECBAE9D+EAEDE,B;B=E9D67D @B:5B?=A:3D =E?F:4;43=FED C4+9EAD 5;4+D AB=ED A+E*BD 5;4+@E=EAD =7CED =FB=D +E=D 3AB:;B=4:-D +F(G-D A+E*BD (@6-D (EA@B:7#D B:9D 67D ?4@@4:;7D 8E9D @E=F49D :D =B6;E=D CA49?=4:-D ?B;?;B=:3D =FED BAAD :9ED 6B8E9D 4:D =FED ?4:=A6=E8D 83:58ED 3B8D 9E=EA@:E9D 6;*D B:9D =BCCE9D 9E:8=7D G D E@884:8/D (AEE:D 5;9'E9D 6E9D 3AB:;B=4:D "-D /D :3E;8@B::D(-D(EA@B:7#/DGFED=AED (%$(#D FB8D AE?E:=;7D 6EE:D :=A49?E9D B8D B:D 9E:8=7D +B8D 9E=EA@:E9D 8:3D BD FE;@D E:,C7?:4@E=EAD ??!7?D 1-D 3AB:;B=4:-D ?FBAB?=EA'E9D67D =FED B68E:?ED 45DBD #/ 9A7:3DCFB8ED2"0/DGFEDB@D45D=FEDCAE8E:=D8=97D +B8D =4D :,E8=3B=ED =FED :5;E:?ED 45D =FED +B=EAD 3 :!! 3 3 !3 ! 3 ! 3 ?4:=E:=DB:9D=FED=7CED45DC4,94:ED! !#D8E9DB8D 3"!! BD 6:9EAD 4:D =FED 5;4+B6;=7D B:9D =B6;E=:3D GFED ?4@CB?=8D +EAED CAECBAE9D 4:D B:D CA4CEA=;B=E8DCAECBAE9D67D(%$(/ :8=A@E:=E9D8:3;E)C:?FD=B6;E=DCAE88D(G!DD 8EA 5;B=D 5B?E9D C:?FE8/D GFED ?4@CB?=8D 1D @3#D +EAED ?4@CAE88E9D :9EAD ?4@CAE884:D ;4B98D :D 3+*56/+13+4.35*%). =FEDAB:3ED45DD=4DD*3DB:9DBD?4@CAE884:D 3!!3 8CEE9D45DD@@@:/DGFED9E=B?F@E:=D8=AE88DB:9D B?=48ED =FED EE?=4:D 8=AE88D +EAED ?B;?;B=E9D 5A4@D =FED @4:4F79AB=ED 3@B);9A (EA@B:7#D+B8D8E9DB8D5;;EADB:9DC4,94:ED1"D 54A?E)98C;B?E@E:=D ?A,E8D 46=B:E9/D GFED 4AD"D!;B894:EG ?4@CB?=D =E:8;ED 8=AE:3=FD +B8D ?B;?;B=E9D 5A4@D -D8F;B:9-D +='EA;B:9#DB8D 6:9EA/D ;48ED ,BCAD =FED=B6;E=D ?A8F:3D 54A?ED 9E=EA@:E9D 8:3D=FED 1"-D D !FBA@B-D #D B:9D ?4;;49B;D 8;A+E*BD G$D 1"D =B6;E=D FBA9:E88D =E8=EAD 949ED EA48;D "-D ,4:*D .:98=AA+E*BD (@6-D (EA@B:7#/D ;B8= (EA@B:7#D +EAED 8E9D B8D @48=AED B684A6E:=8-D "DFDB5=EAD?4@CAE884:#D+B8DB;84D9E=EA@:E9/D +F;ED?A48C4,94:ED!4;7C;B894:ED)1-D(%D GFED ?4@CAE886;=7-D ?4@CB?=6;=7D B:9D FE@8E9D B8D BD =B6;E=B6;=7DCA45;E8D+EAED?4:8=A?=E9-DB8D+E;;/ 98:=E3AB:=DB:9D@B3:E8@D8=EBAB=ED A/D!B;D 3. ! 3 4F@B:-D(EA@B:7#DB8DBD;6A3:!! 33! 3 CAECBAE9D +B8D 9E=EA@:E9D 8:3D =FED (AB:;B=4:D +B8D ?BAA=D :D =FED 7?A4;B6D 98:=E3AB=4:D=E8=EADA+E*BDG"-D(EA@B:7#/D 5;9D 6E9D CA4?E884AD GD ==;:-D (EA@B:7#DB??4A9:3D=4D=FED@E=F49D9E8?A6E9D67D D836501*3+4.3./$0/)4 GB*B8B* @ F-DB:9D=FED:;E=DBAED+B8D"D /D 83:!33 3! 3!3 GFED :,E8=3B=E9D 8B@C;E8D 8F4+E9D BD 8@;BAD GFED B@4:=D 45D +B=EAD 8E9D +B8D D 4AD /D GFED CBA==4:-D+=FD=FED@B:DCBA= +B=EAD 5EE9D AB=ED +B8D D 3@:-D =FED 8CAB7D B 8'ED5AB?=4:D6E:3D1"D@/DGFED:5;E:?ED45D =FEDB@4:=D45D+B=EAD4ADC4,94:ED,8?48=7D3AB9ED 100 ('' FE CBA @?CC=;A :=9A 8=:C=76C=FE 5?: EFC D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D D 5B=A D D D D D D:@FE8: CF CBA 0?;6A: AFEF6ED D D D D D =E CBA ;=CA?CC D D D D D D D D D D D D ?6A #) D D D D D D 11?00A8 =E CBA : / %% # # # # # # ! # # # # # # @F@FCC=FE F1 @?CC=;A: =E ;?3A @?CC=;A :=9A D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D D 5=CB .-. (BA =E0A:C D D D D D D C=3?CCA8 :?/ ? D D D D D D ? /@;A: :BF5A8 ?@=8 D D D D D D D D D D D DD D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D ?/@;A: @A@?A8 D D D D D DD D D D D D 11??: FE CBA 1;;F5 @F@AC=A: F1 CBA CA:CA8 8=:=ECA3?CC=F ? FE) (BA 8=:=ECA3?C=FE C=/A 0?=A8 D D D D D D D D D D D D ? D D D D D D D D D D D DD D D D D D D D D D D D D D D D D D,+*) (BA =E1;;6AE :?/@;A: 5?: ?;:F EFC @FEF6E7AC5AAE + D D D D D D ?E8 :4 8A@AE8=E3 FE CBA D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D D D D D D D D D D D D D D D D D D D D 1 D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D /=CC6A: CA:CA8 :BF5A8 ?11FF/ /6;?CC=FE ?D D D D D D 11??:D D D D D D ? ?EE8D D D D D D CBAD D D D D D ?@@@;=A8D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D ? D D D D D D @F@ACC=A: 5=CB %? =E8A 0?;6A: 7AC5AAE *$ A::=FE E @A::6A) E 3AEA?;4 1??: CB?CC D D D D D D D D D D D D D D D D D D D D D D D D 1 D D D D D D ?D D D D D D D D D D D D D D D D D D D D D D D D ? D D D D D D 1 D D D D D D ?E8 *#") ;A8 CF ? B=3 D D D D D D D D D D D D D D D D D D 3BA CAE:=;A :CAE3CB ?;:F ;A8 CF ? D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D ;FE3A 8=:=EC D D D D D D CA3?CC=FE C=/A) ? D D D D D D %$% " # # # # # # "!! # # # # # # !FF8 /AF@AC=A: F1 CBA D D D D D D D D D D D D D D D D D DD D D D D D @A@?A8 72 ! ! 5AA F7:A00A84 5=CB B=3BA D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D D % # # # # # # A::=FE @A::6A ;A?8=E3 CF B=3BA CAE:=;A = D D D D D D D D D D D D ; 8= D D D D D D D D D D D D B= B D D D D D D =;D D D D D D :CAE3CB =3) +) D D D D D D D D D D D D D D D D D D D D D D D D C 00A8 CB?CC CBA 6:A ? D D D D D D D D D D D D D D D D D D ? D D D D D D D D D D D DD D D D D D (BA A:6;C: @ D D D D D D D D D D D D @A:AECA8 :BF5 CB?CC ? EF0A;4 3AAE D D D D D D D D D D D D ? D D D D D D D D D D D D D D D D D DD D D D D D F1 .-. ,+* ;A?8: CF ? :F/A5B?C B=3BA CAE:=;A D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D ? D D D D D D D D D D D DD D D D D D 11;;6=8=9A8 7A D D D D D D8 3?EE6;?CC=FE A AAE3CB CB?EE .-. ,*4 5B=?=9A8 D D D D D D ? D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D D C?7;AC=E3 @F D D D D D D F@ACC=A: ?@=8 C?77;AC D D D D D D D D D D D D D D D D D D D D D D D D ?D D D D D D 72 =C: B=3BA 0=:?CC=F ? FE) C 5?: 1FF6E8 CB?CC CBA C2@A F1 D D D D D D D D D D D D D D D D D D 1 D D D D D D ? D D D D D D D D D D D D D D D D D D D D D D D D =: ?CC6A 8?CC? :6@@FCC=E3 CBA D D D D D D D D D D D D D D D D D D ? D D D D D D ? D D D D D D D D D D D DD D D D D D @F0=8FEA ??EE D D D D D D E8 CBA 5?CCA ?CA;2 D D D D D D D D D D D D ? D D D D D D D D D D D D ?D D D D D D 6:A F1 .-. ,+* 1FF CB=: ;F5 5?CCA F@ACC=A: F1 CBBA =E0A:C=3?CCA8 D D D D D D D D D D D D D D D D D D C D D D D D D ?D D D D D D 3 EE6;?CC=FE /ACBBF8 *) F CBA :?/@;A: ? ? D D D D D D C D D D D D D D D D D D D D D D D D D D D D D D DD D D D D D C?7;AC=E3 /= D D D D D D C6A:) @A@?A8 5=CB CBA :?/A C2@A F1 .-.44 ? B=3BA D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D . D D D D D D D D D D D DD D D D D D 5?CCA CAE:=;A ? D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D D :CAE3CB) % # # # # # # a) 3,0 +) A;B= 4 D D D D D D D D D D D D D D D D D D 4 ;/A;=3= ') D D D D D D D D D D D D %$#"! !!!# ! ! MP( 2,5 !"$ $# $$ ## $ #$$ $ # # !! $!# $ $# $) F6EE?; F1 %;A?EA .F86 D D D D D D D D D D D D D D D D D D D D D D D DD D D D D D en 1,5 trs 5% water, PVP K12 *) ???:?=4 D D D D D D ( D D D D D D )4 AC ?;)4 D D D D D D D D D D D DD D D D D D !! $ $ #! 3 thg 2,0 *$+) *+ + D D D D D D D D D D D D +)D D D D D D ile s 1 % wat a ter, PVP +* , 0 ## ##$ $$! ! # " # 5 % wa ter, P V P K2 5 en 3 % w at a ter, , P V P * ##$ $$! D D D D D D ECAE?C=FE?; F6E?; F1 D D D D D D D D D D D D D D D D D D 0 , 5 T .B .B?/? C= D D D D D D *$+ + * D D D D D D D D D D D D D D D D D D 1 0 0 1 2 0 1 4 0 1 6 0 1 8 0 2 0 0 Compression pressure (MPa) #) A?EA4 )4 AC ?;)4 D D D D D D D D D D D D D D D D D D D D D D D DD D D D D D #!!$ !!# $ ## #! ! % $$ $$# %$$$! % % !!# !#$ # # # # # # # # # # # # ( (??7;AC?7=;=C22 C D D D D D D @F1==;A: F1 CBBA =E00A 1 D D D D D D D D D D D D C D D D D D D E A:C=3?CA8 D D D D D D :?/@;A:) ! !!$ !!# D D D D D D .B?/? D D D D D DD D D D D D ?E8 (AA EEF;F324 *$+) *$+ +**+) D D D D D D ( B 3 D D D D D D D D D D D D D D D D D DD D D D D D (BA 0?;6A: 8ACA/ D D D D D D D D D D D D /=EA8 1FF CBA 8AC?A:: 5AA ?CCBA ;F5 +) " D D D D D D D D D D D D D D D D D D ? D D D D D D D D D D D D D D D D D D .? D D D D D D D D D D D D A3?8;A:: F1 CBA ?@@@;=A8 A::=FE @ D D D D D D D D D D D D D D D D D D ? D D D D D D D D D D D DA::6A4D D D D D D (B=: A:A?22 F1 D D D D D D D D D D D D D D D D D D 1 D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D =E8==F@ACC=A: ? D D D D D D D D D D D D ? D D D D D D D D D D D D F1 CBA D D D D D D D D D D D D =AEA2 F1 D D D D D D D D D D D D EEF0?CC=FE4 A@67;= F1 A77=? CBF63B C5F ? D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D CBA =E0A:C=3?CCA8 :?/@;A: 5?: 7AC5AAE D D D D D D ? D D D D D D D D D D D D D D D D D D D D D D D D + ?EE8 D D D D D D ? D D D D D D ! EEC '3AA/AEC: 5=CB E=0A:=C2 F1 A;3?8A ? D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D *" ?EE8 =EA?:A8 ?CC B=3BA A::=FE D D D D D D ?6;C2 F1 D D D D D D D D D D D D .B?/ /?2 F +$#+#*$*$# D D D D D D D D D D D D D D D D D D @A::6A:) *$$++ ??EE8 F +$#+#*$*$# *$$++) D D D D D D D D D D D D D D D D D D D D D D D D (BA A::=FE ?=9?C=FE D D D D D D D D D D D D ?D D D D D D F1 CBA D D D D D DD D D D D D C?77;AC=E3 /=CC6A @A@?A8 5=CB .-. , ? D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D ,+* EE8 D D D D D D ?D D D D D D #" 5?CCA A0A?;A8 1??0F 77;A A: D D D D D D ? D D D D D D D D D D D D 1 ? D D D D D D ::=7=;=C2D D D D D D ?E8 F1==;A:) E CBA A::=FED D D D D D @A::6A ?EE3A CA:CA84 CBA CAE:=;A :CAE3CB D D D D D D ? D D D D D D D D D D D D D D D D D D D D D D D D B 0?;6A: D D D D D DD D D D D D 5AA 7AC5 5AA 7AC5 D D D D D D C C 5AAE +) ?E8 *) .? ?E8 CB 5AAE +) ?E8 *) .? ?E8 CBA :F;=8 D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D BA :F;=8 D D D D D DD D D D D D 11?A =E CBA ?E3A F1 $)$)$4 D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D D 101 :998 765543210/./-5.3,5.376+401+*/)4(3/4'5*/7+*/)43)23*%53 /421054$53)23#+*563$)4*54*3+4.3:)'/.)45376+.5 /"! !3+1 1 , *!37 1. INTRODUCTION CAE88AED +B8D 1/"D 6BAD B:9D =FED @ CAE88AED+B8D/D6BA/D GFEDCFBA@B?E=8=A7DFB8DAE?E:=;7D6EE:D 9E:=5=4A8D =4D 3 :!3 3 ! !3 3 ! 3 ?;@B=ED ?FB:3ED 210/D .:D AE?E:=D 7EBA8-D !3 CFBA@B?E=GFEDCBA==4:D+B8DE,B;B=E9D67D FBA9D=4DAE9?ED=FE=D 88:3DBD8E=D45D8=B:9BA9D8 =FED?FB:3E8DBAED8=;;D:E3;36;ED:DAE;B=4:D=4D=FED =FEDAB:3ED45DD@/DGFED5;4+DCA4CEA= 8ED 45D ?BA64:):=E:8,ED =AB9=4:B;D =B6;E=:3D@=AE8DCAECBAE9D+EAEDE,B;B=E9D67D @B:5B?=A:3D =E?F:4;43=FED C4+9EAD 5;4+D AB=ED A+E*BD 5;4+@E=EAD =7CED =FB=D +E=D 3AB:;B=4:-D +F(G-D A+E*BD (@6-D (EA@B:7#D B:9D 67D ?4@@4:;7D 8E9D @E=F49D :D =B6;E=D CA49?=4:-D ?B;?;B=:3D =FED BAAD :9ED 6B8E9D 4:D =FED ?4:=A6=E8D 83:58ED 3B8D 9E=EA@:E9D 6;*D B:9D =BCCE9D 9E:8=7D G D E@884:8/D (AEE:D 5;9'E9D 6E9D 3AB:;B=4:D "-D /D :3E;8@B::D(-D(EA@B:7#/DGFED=AED (%$(#D FB8D AE?E:=;7D 6EE:D :=A49?E9D B8D B:D 9E:8=7D +B8D 9E=EA@:E9D 8:3D BD FE;@D E:,C7?:4@E=EAD ??!7?D 1-D 3AB:;B=4:-D ?FBAB?=EA'E9D67D =FED B68E:?ED 45DBD #/ 9A7:3DCFB8ED2"0/DGFEDB@D45D=FEDCAE8E:=D8=97D +B8D =4D :,E8=3B=ED =FED :5;E:?ED 45D =FED +B=EAD 83 :!! 3 3 !3 ! 3 ! 3 ?4:=E:=DB:9D=FED=7CED45DC4,94:ED! !#D8E9DB8D 3"!! BD 6:9EAD 4:D =FED 5;4+B6;=7D B:9D =B6;E=:3D GFED ?4@CB?=8D +EAED CAECBAE9D 4:D B:D CA4CEA=;B=E8DCAECBAE9D67D(%$(/ :8=A@E:=E9D8:3;E)C:?FD=B6;E=DCAE88D(G!DD 8EA 5;B=D 5B?E9D C:?FE8/D GFED ?4@CB?=8D 1D @3#D +EAED ?4@CAE88E9D :9EAD ?4@CAE884:D ;4B98D :D 3+*56/+13+4.35*%). =FEDAB:3ED45DD=4DD*3DB:9DBD?4@CAE884:D 3!!3 8CEE9D45DD@@@:/DGFED9E=B?F@E:=D8=AE88DB:9D B?=48ED =FED EE?=4:D 8=AE88D +EAED ?B;?;B=E9D 5A4@D =FED @4:4F79AB=ED 3@B);9A (EA@B:7#D+B8D8E9DB8D5;;EADB:9DC4,94:ED1"D 54A?E)98C;B?E@E:=D ?A,E8D 46=B:E9/D GFED 4AD"D!;B894:EG ?4@CB?=D =E:8;ED 8=AE:3=FD +B8D ?B;?;B=E9D 5A4@D -D8F;B:9-D +='EA;B:9#DB8D 6:9EA/D ;48ED ,BCAD =FED=B6;E=D ?A8F:3D 54A?ED 9E=EA@:E9D 8:3D=FED 1"-D D !FBA@B-D #D B:9D ?4;;49B;D 8;A+E*BD G$D 1"D =B6;E=D FBA9:E88D =E8=EAD 949ED EA48;D "-D ,4:*D .:98=AA+E*BD (@6-D (EA@B:7#/D ;B8= (EA@B:7#D +EAED 8E9D B8D @48=AED B684A6E:=8-D "DFDB5=EAD?4@CAE884:#D+B8DB;84D9E=EA@:E9/D +F;ED?A48C4,94:ED!4;7C;B894:ED)1-D(%D GFED ?4@CAE886;=7-D ?4@CB?=6;=7D B:9D FE@8E9D B8D BD =B6;E=B6;=7DCA45;E8D+EAED?4:8=A?=E9-DB8D+E;;/ 98:=E3AB:=DB:9D@B3:E8@D8=EBAB=ED A/D!B;D 83. ! 3 4F@B:-D(EA@B:7#DB8DBD;6A3:!! 33! 3 CAECBAE9D +B8D 9E=EA@:E9D 8:3D =FED (AB:;B=4:D +B8D ?BAA=D :D =FED 7?A4;B6D 98:=E3AB=4:D=E8=EADA+E*BDG"-D(EA@B:7#/D 5;9D 6E9D CA4?E884AD GD ==;:-D (EA@B:7#DB??4A9:3D=4D=FED@E=F49D9E8?A6E9D67D D36501*3+4.3./$0/)4 GB*B8B* @ F-DB:9D=FED:;E=DBAED+B8D"D /D 3:!33 3! 3!3 GFED :,E8=3B=E9D 8B@C;E8D 8F4+E9D BD 8@;BAD GFED B@4:=D 45D +B=EAD 8E9D +B8D D 4AD /D GFED CBA==4:-D+=FD=FED@B:DCBA= +B=EAD 5EE9D AB=ED +B8D D 3@:-D =FED 8CAB7D B 8'ED5AB?=4:D6E:3D1"D@/DGFED:5;E:?ED45D =FEDB@4:=D45D+B=EAD4ADC4,94:ED,8?48=7D3AB9ED 102 ('' FE CBA @?CC=;A :=9A 8=:C=76C=FE 5?: EFC D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D @A::6A ?EE3 D D D D D ? 3A CA:CA84 CBA CAE:=;A :CAE3CB 0?;6A: D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D @FEF6ED D D D D 5AA 7AC5AA D D D D D AE +) ?E8 *) .? ?E8 CBA :F;=8 D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D @F@FCC=FE F1 @?CC=;A: =E ;?3A @?CC=;A :=9A D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D 11?A =E CBA ?EE3A F1 $)$)$4 D D D D D D D D D D D D D D D D D D D D ? D D D D D D D D D DD D D D D 11?00A8 =E CBA : / D D D D D D D D D D D D D D D D D D D D D D D D D ? /@;A: @A@?A8 D D D D DD D D D D 5B=A D D D D D:@FE8: CF CBA 0?;6A: A?CC D D D D D D D D D D ?6A #) D D D D D :?/@;A: 5?: ?;:F EFC @FEF6E%$%$ " " " " " " " " " " 11??: FE CBA 1;;F5 @F@AC=A: F1 CBA CA:CA8 D D D D D D D D D D D D D D D 1 D D D D D D D D D D D D D D D D D D D DD D D D D /=CC6A: CA:CA8 :BF5A8 ?(BA =E0A:C D D D D D C=3?CCA8 :?/ ? D D D D D ? /@;A: :BF5A8 ?@=8 D D D D D D D D D DD D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D ? D D D D D @F@ACC=A: 5=CB %? =E8A 0?;6A: 7AC5AAE *$ 8=:=ECA3?CC=F ? FE) (BA 8=:=ECA3?C=FE C=/A 0?=A8 D D D D D D D D D D ? D D D D D D D D D DD D D D D D D D D D D D D D D D D D D D ? D D D D D 1 D D D D D ?E8 *#") 7AC5AAE + D D D D D ?E8 :4 8A@AE8=E3 FE CBA D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D 11FF/ /6;?CC=FE ?D D D D D 11??:D D D D D ? ?EE8D D D D D CBAD D D D D ?@@@;=A8D D D D D %$#$ ! " " " " " ! " " " " " A::=FE E @A::6A) E 3AEA?;4 1??: CB?CC D D D D D D D D D D D D D D D D D D D D 1 D D D D D ?D D D D D !FF8 /AF@AC=A: F1 CBA D D D D D D D D D D D D D D DD D D D D ;A8 CF ? B=3 D D D D D D D D D D D D D D D 3BA CAE:=;A :CAE3CB ?;:F ;A8 CF ? D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D @A@?A8 72 ! ! 5AA F7:A00A84 5=CB B=3BA D D D D D D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D ;FE3A 8=:=EC D D D D D CA3?CC=FE C=/A) ? D D D D D A::=FE @A::6A ;A?8=E3 CF B=3BA CAE:=;A = D D D D D D D D D D ; 8= D D D D D D D D D D B= B D D D D D =;D D D D D :CAE3CB =3) +) D D D D D D D D D D D D D D D D D D D D C 00A8 CB?CC CBA 6:A ? D D D D D D D D D D D D D D D ? D D D D D D D D D DD D D D D F1 .-. ,+* ;A?8: CF ? :F/A5B?C B=3BA CAE:=;A D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D ? D D D D D D D D D DD D D D D $ " " " " " :CAE3CB CB?EE .-. ,*4 5B=?=9A8 D D D D D ? D D D D D D D D D D D D D D D D D D D D D D D D DD D D D D 72 =C: B=3BA 0=:A:6;C: @ D D D D D D D D D D @A:AECA8 :BF5 CB?CC ? EF0A;4 3AAE D D D D D D D D D D ? D D D D D D D D D D D D D D DD D D D D =: ?CC6A 8?CC? :6@@FCC=E3 CBA D D D D D D D D D D D D D D D ? D D D D D ? D D D D D D D D D DD D D D D 11;;6=8=9A8 7A D D D D D8 3?EE6;?CC=FE A A CB=: ;F5 5?CCA F D D D D D F@ACC=A: ?@=8 C?77;AC D D D D D D D D D D D D D D D D D D D D ?D D D D D 3 EE6;?CC=FE /ACBBF8 *) F CBA :?/@;A: ? ? D D D D D C D D D D D D D D D D D D D D D D D D D DD D D D D 8=:=ECA3?CC=F ? FE) C 5?: 1FF6E8 CB?CC CBA C2@A F1 D D D D D D D D D D D D D D D 1 D D D D D ? D D D D D D D D D D D D D D D D D D D D @A@?A8 5=CB CBA :?/A C2@A F1 .-.44 ? B=3BA D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D . D D D D D D D D D DD D D D D @F0=8FEA ??EE D D D D D E8 CBA 5?CCA ?CA;2 D D D D D D D D D D ? D D D D D D D D D D ?D D D D D 5?CCA CAE:=;A ? 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' !$0..8:) 0) #'=#0'8) :" 0 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 66 6 6 6 6 6 6 6 104 10/.-0/, +*)(-'-*, +*%$-#+, ")!#, .0-#$, (, ,)0)'+-+,*")*,*"#,+,")!#,#*)-0#$,*"#-, ).".%+, +*)*#, .!#, *-#, -*", 0., # +*)''-)*-.0, .(+#!#$, +., ), , .0*"+, -0, ,*##)*%#,)0$,,, ")(#+ , "#, .%')*-.0, ., 1 +, %+-0/, , )0$, ,).".%+,+.'-$,$-+#+-.0,*# "0-%#+, +-/0-- )0*', -.!#$, #'# .-(+, +.'%(-'-*, ")) *#-+*- +, "-+, #+#) ", %0$#+ .#+, *"#, .*#0*-)',.,1 +,*.,.*--#,*"#,)'- )*-.0+, ., , .%')*-.0+, '#)$-0/, *., -.!#$, (-.)!)-')(-'-*, )0$, *"#)#%*- , #- ) , ., )*-#0*+, %*"#, +*%$-#+, )#, # .#0$#$, *., -0!#+*-/)*#, '-0- )', -'- )*-.0+, )0$, #-0#, .%')*-.0,+*)*#/-#+ 1. Register, E.C.T. Clinical trials for Celecoxib. [cited 2025 02.05.]; Available from: https://www.clinicaltrialsregister.eu/ctr- search/search?query=Celecoxibstatus=o ngoing 2. Desai, N., et al., How Do Orodispersible Tablets Behave in an In Vitro Oral Cavity Model: A Pilot Study. Pharmaceutics, 2020. 12(7). 3. FDA, Guidance for Industry: Orally Disintegrating Tablets;, F.a.D.A.C.f.D.E.a. Research, Editor. 2008, Silver Spring. 105 :998 7675432410/.73-:03,35+.32.*)62(.,4'/.%62,53)240.%30-7.-4(/.)%. 4.$37,)7/#5+:/.,4'1)"+-/5!+0,/0060)7/.*35!.(/":425!/2)0 .11 1 .'.- %$#"! $ "!"$ "$" "!"""!$! ! $$# 1. INTRODUCTION 3I9E DIH92E 35C9E F91@G?4E 5DCE 3@AE )C9CE =B9C3HAA1E;CCACFEI33EBGFE5I9CFE@GE=AICFE;AB5@=E JIHGFEDCBA@G?E@EBE=I;AC:E;9I=CE9C8H@9@G?EBE 7B?E 5IE B)B@5E 5C5@G?2E 0DCE =I;I@5@IGE I3E 5DCE 7BABG=CFE CG6@9IG5H9C4E ;9C;B9CFE3@AE@EH ;9C6CG5E @G3C=5@IG4E BGFE H;;I95E 5@HCE 9C;B@92E JD@ACE BF6BG=CFE 15C4E H=DE BE 5. .,I;I@5@IGEI3E=B5@G?EF@;C9@IGE-""E?*E ;I5CG5@BA4E5DC@9E=A@G@=BAEHCE@EI35CGE9C59@=5CFE71E CMC Dexpanthenol Sample Plasticizer (g) (g) GBGI5C=DGIAI?@C4E I33C9E 59IG?E 5DC9B;CH5@=E D@?DE=I54E9C?HAB5I91E=DBAACG?C4EBGFE5C=DG@=BAE (g) =I;AC:@512E0D@EDBEACFE5IE@G=9CBCFE@G5C9C5E@GE V-G-1-D 1.736 glycerin 1.736 0.4 @;AC94EH5B@GB7ACEE5DB5EB9CECB@C9E5IE V-M-1-D V-G-2-D 1.736 glycerin 3.472 0.4 ;9IFH=CEBGFEH@5B7ACE3I9E;9B=5@=BAEB;;A@=B5@IG2E V-M-2-D 1.736 macrogol 1.736 0.4 1.736 macrogol 3.472 0.4 /@=IC.FC9@6CFE =B97I:1CE JDCGE =I7@GCFE )@5DE FC:;BG5DCGIA4E BE )CAA. $ #!#$! $ 0DCE H9CE =B99@CFEIH5EH@G?EBGEB95@3@=@BAE)IHGFE5B7A@DCFE B?CG5E 5DB5E H;;I95E (@GE @AB;ACE-2EE2E=*E)C9CE)C@?DCFEIGE 9C?CGC9B5@IG4E 5DCE 9CHA5@G?E 3@AE ;9CCG5E BE ;IA12E ./...-,+,*E @E BE 7@IFC?9BFB7ACE BGFE 7@I=I;B5@7ACE ;9B=5@=BA4EC33@=@CG54EBGFEC=I.=IG=@IHEB;;9IB=DE BGE BGBA15@=BAE =BACE -E " 4E I55A2E EE#IDGE74EBA@G?CG4E*E@GEF91E 5IE E 5HF1E B@E -JF*E BGFE ;AB=CFE @GE 5DCE )IHGFE ;9C;B9CEBGFEC6BAHB5CE3@AEE E-4E4E4E ED*4E )CAA.C5B7A@DCFE@GE)IHGFE5DC9B;14E)D@=DEI33C9E )IAACGE3@AE)C9CE)C@?DCFE-J*2E0DCEFC?9CCE ;9I@@G?E;I5CG5@BAE)DCGE=I7@GCF2E0DCE3@AE I3E )CAA@G?E -#)*E )BE =BA=HAB5CFE H@G?E 5DCE )C9CE BCCFE )@5DE BE 3I=HE IGE 5DC@9E ;9B=5@=BAE C8HB5@IGE #)E E -JE E JF*JF2E !"$ # E B;;A@=B7@A@51E5IE5DCE)IHGFE'%2 @A8HB9CE-2EE2*E= 2. MATERIALS AND METHODS 5DCEB95@3@=@BAE)IHGFEEHCFE5IE =I6C9E5DCEC59@EF@DE5IE;9C6CG5E5DCEC6B;I9B5@IGE .- I3E5DCEA@8H@F2E5EFC5C9@GCFE5@E-4E4E 0DCE;B95@BAA1E=IC.51;CE 4E ED*4EH93B=CE;E)BEH9CF2E$"" IF@HCE -B,+,*E @GE #!#$! $ $$ $ 2E0CG@ACE59CG?5DE)BE 5DCE3@79IHE3I9EI75B@GCFE39I H9CFE@GE7I5DEF91EBGFE)C5E5B5CEH@G?EBE,0E ;IA2E 292I2E -,*2E $C:;BG5DCGIA4E ?A1=C9@GE BGFE 0C:5H9CE GBA1C9E -9II(3@CAF4E +@FFAC7I9I4E CFE +4E#*EBGFE0C:5H9C9IE,0EI35)B9C2E@A 39I2E-AI 59@;E- "EE"*E)C9CE59C5=DCFEB5E"2E .%. HG5@AE79CB(2EI9=CEBGFECAIG?B5@IGEB5E79CB(E;I@G5E 0DCE 3@AE )C9CE ;9C;B9CFE H@G?E 5DCE IA6CG5E )C9CE9C=I9FCFEBGFE9C=BA=HAB5CFE3I9EBE5D@=(GCE =B5@G?E ;C9@IGE I3E B,+,E -=H5E I3E""2 6@=IC.51;CE5C:5@ACE3@7C9*E)BE3I9 3. RESULTS AND DISCUSSION ;H9@3@CFE )B5C9E BGFE ;AB5@=@C94E 5DI9IH?DA1E DI)CAAE3I9EEDIH92E0DCE 0DCE3@AE)@5DEFC:;BG5DCGIAE)C9CEH==C3HAA1E =IG=CG59B5@IGE I3E ,+,E )BE "2""?=2E0DCE ;9C;B9CFE 71E 5DCE IA6CG5E C6B;I9B5@IGE F@;C9@IGE )BE B=@F@3@CFE 5IE ;E H@G?E -@?H9CE *2EAAE 5DCE B;ACE )C9CE 3AC:@7ACE BGFE D1F9I=DAI9@=E B=@FE HGFC9E =IG5@GHIHE @:@G?2E =IDC@6CEC6CGEB35C9E)C55@G?4E)D@=DE@EGC=CB91E #H7C8HCG5A14EBE"EFC:;BG5DCGIAEIAH5@IGE)BE 3I9E5DCE@G5CGFCFE)IHGFEB;;A@=B5@IG2E5@A@B5@IGE BFFCFE 5IE B=D@C6CE BE 3@GBAE =IG=CG59B5@IGE I3E I3E6@=IC.51;CEB,+,E@GE5DCE3@79IHE3I9 DCFEF@;C9@IGE)BE;IH9CFE BFF@5@IGBAEB=@F@3@=B5@IGEI3E5DCEF@;C9@IGEACFE5IE IG5IE59B1EBGFEF9@CFE@GEBEDI5EB@9EI6CGEB5E!",E 5DCE 59CG?5DCG@G?E BGFE @;9I6C 106 @?= ;;:9 7657?644:?3 52 4==? 2::9@3 3 1 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 13 935 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 !?= ;;:9 7657?64:?3 52 4=? 2::9@3 6? ? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 833?;4:9 ;8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 5;2::6@ 2 6 @?0 :; 5/6 76?.:5/3 34/0:?3 -,+* 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ;54 5;9 :; 06 344? /4 935 244?6 5;4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 1:4= 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 15/;0 ?/044? (1?44:;* :9@3 @ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 @/34 ? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 5=?3:.? ;;0 0/69? /4 299?:9? 2556 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 6 ?444?6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 48 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 0 77444:5; 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 45 4=? 15/;0* :/6? , 3=5 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 513 4=? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 @?= ;;:9 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 7657?644:?3 6?3/943* :9@ 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 83 1:4= 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 9?659 ?= 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 =::4?0 6?4?6 @?=;:9 3 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 346?;4=8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 5@76?0 45 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 5 4=53? 1:4= @6559 =5 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 51?.?6"" 68 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 :;6?3:; 4= 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 =? 7 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8934::?6 5;?;4644:5; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 89?0 45 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 6?0/4:5; :; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ; 346?;4==* 75; =064:5 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 5;" 4=? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 @?= ;;:9 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 7657?64:?3 52 4==? 2::9@3 0? 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 86?3?08 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 @6?09" 1 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 1=:9? =:= 299?::9:4 13 6?4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 :;?0" 3 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 )=? 2::9@3 ('%$#" '! # :;0:44?0 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 :;6?3?0 0?2556@4:5; . 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 9/?3 :; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ' 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ' # 5@76:35; 7 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 1:4= 4=? 06 3444?* 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 1?99:; 76 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 657?64:?3 :;0:44? 4=? :9:4 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 52 4=? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 06?33:; 45 7 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 765.:0? @5:34 ?;.:65;@?; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ;;4 :; 4=? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 15/;0 ;;0 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 356 ?/04?* :9@3 ?== 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ::4?0 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 @:90 0?6?? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 52 31?99:; (:/6? * )=? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ? 0?6?? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 52 31?99:; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 52 99 2::9@3 :;6?3?0 60/ 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 /99 /7 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 45 =5/63 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8* @ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 @79?3 1 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8:4= =:=?6 @ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 @ @5/;4 52 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 7934::?6 3 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 3=51?0 39:=49 951?6 0? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ?6?? 52 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 31?99:;* !?=;;:9 7657?644:?3 52 4==? 2:: 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 29@3 4. CONCLU USION )=:3 34//0 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 :@?0 45 0?.?957 ?222??4:.? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 28 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 15/;08 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 06?33:;3 /3: 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 :; 3:@79? @?4=50 ;;0 3/34 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4:;9?8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 @44?6:93 4= 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 =44 6? 96?0 1?99?349:3 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 3=?0 :; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 15/;0 6? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8* 3?0 5; 76?9:@:;6 ?. 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 9/44:5; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 22556 764: 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 9 7779:44:5;" 4=? 2::9@3 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 28 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 777?68 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 1 765@:3:; 0 765@:3:; 0/? 45 4=?:6 7 @50 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 0/? 45 4=?:6 7@50 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 /944:; /944:;8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ?222??43 ?222??43 2 2 2 28 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 1?99:; 7657?644:?3 52 4==? 2::9@ 1?99:; 7657?644:?3 52 4==? 2::9@ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 6 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 2 @3 @3 ;0 2..56 9? 56;;59?74: ;0 @? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 =;:98 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 7 ?.9/44:5; 3=513 4=? 06?33:;3 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 :9:4 45 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 7657?644:?3" ?.?; /;0?6 @5:34 5; 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ;0:4:5;3*8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 @:;4:; ; :0: ?;.:65;@?;4 4=44 3 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 3/7756443 68 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 51?.?6"" 2//64==?6 4?34:; :3 6? /:6?0* 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 6 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 15/;0 =?9:;* )9? 0?@5;34644? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ) 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ?3 4=44" 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 94==5/= 4=? 7 :;6?3?0 1:4=:; 4= 5. REFERENCES 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 =? 4:@? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 :;4?6.93" 99 4=? 2::9@3 6?4:;?0 :0: 7 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 .9/?3 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 $* '??6@" #* ?4 9*" 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ' 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8* ('(%%$#%"! %# # % ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) % % ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ?.?; 244?6 =" =:?.:; 7 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8.9/?3 2665 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 2 5@ * 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 %(( '$!$%$# "$! ( ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ! ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) )$ ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) *$ ('% # 45 *, * ('! 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ' 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ' ! #"8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 '# (($# %!$% $$#( * %$! ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) % ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) $ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 6?37?4:.?9*8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 %%#' '' ( %%# ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) " (( " ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) * , 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8$$* 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 87 94?64:5;3 :; 4:@ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 : @? * '::;965." * ?4 9*" 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ' 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 $! '# (( ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) $# $ ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) !%!) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) %# ( $ ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) "%( ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) #) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) $# #(!''"! !) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) S %" ! !!''!( %$ ! ! ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) * 5/6;;9 52 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8779:?08 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 a m p le 1 h 3 h 8h 2 4 h :5@?0::;?" $* $,$,, * 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 V-G-1-D *, * 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 * * 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 * * 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 * * 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 V-G-2-D * *$ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 * *, 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 * * 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 *, *, 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ,* )??;565." * ?4 9* 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ) 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ' %% $$# %# %!'' ! ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ! '%$# ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) V-M-1-D * *$ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 *, *$ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 *, *$ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 * *, 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 !) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) $!) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) '#) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) (($#) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) %( ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) #) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) V-M-2-D ,* * 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 * * 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 *, * 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 *, * 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 !!% ! # $$$$"$!!% %" ! ! ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) !!'!( ! !) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ) ! # 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8=6@?/44:3" * $ * / 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 )=:3 :;0:4?3 4=? :9:4 52 4=? 2::9@3 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 28 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 45 ??7 78 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4=?:6 7 :; 4=? :0: 6;;?" 35 1? ;; 5;3:0?6 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4=?@ 3 7 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8@50/944:; 15/;0 06?33:;3* 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 107 ,++* ),('('%)$%)%#%"$! $')%('#"!( )! )($')#)'!)%%"% Elzbieta Maria Buczkowska, Zoltan Mark Horvath, Kirils Kukuls, Alina Jaroslava Frolova, Liga Petersone, Valentyn Mohylyuk 1 1. INTRODUCTION DCBACB@?= 8C?26=5:A1?=7=0497?86B48A1?86 :C3B0? .@? ;:42498@? 90? :84,B@C? @? 499C,84,Mannitol Isomalt :C52C998@? C/? 0=37? 0 99C,84,@84C9@? 3@497? 2CBACB@? @? :C52C998@? ==8? C/? 86 8C24:?C/?0=37?0Sorbitol Xylitol 86=5:384:B?4903@8=A'? ))654:B?@8=3:83=@?C/?2CBACB@' 9?864@?=@=:6?%BBA?49,@84780? 26A@4:C:654:B? 2=C2=84@? 90? 498=49@4:? ), ) 04@@CB384C9?2=C/4B@?C/?$?:C55=:4BBA?,4B.B49? .@C=.0? %8=? +.C390? 2CBACB@'? #645? C/? 86=:6? %@? 868=(? 8=220? .A? 8690? .3B? %8=? :6=:8=4-84C9? C/?26A@4:C:654:B? 2=C2=84@? +/=8=(? +#.B'!'('? BB? 2CBACB@? 9BA@0? C/? :C55=:4BBA? ,4B.B=?90?6A0=C264B4:?+C7D?(') 3@@'? )$"!$'%)$ )"(% "))D6A@4:B?90?:654:B?2=C2=84@?C/?2CBACB@' "'!'? 8=4B@? Polyol Sorbitol Mannitol Isomalt Xylitol )2=A? 0=40? 7=0@? C/? 59948CB? ? 99C75? 0.03 0.78 0.41 0.02 KF ? 2B? )*? +C3881? @8=51=9:(? 90? 0.80 1.01 0.58 0.19 LOD )C=.48CB? + D""$$1? =:? 1? Mw 182.172 344.313 182.17 152.15 *=5@8081? =59A(? ? @C5B8? ? (g/mol) D+!"!"$!1? )0-3:=?1? .=476451? HBD/HBA 6/6 9/11 6/6 5/5 =59A(? 90? AB48CB? +*(? AB4@C=.? ? +"!C3881?@8=51=9:('? -3.73 -5.50 -3.73 -3.10 LogP "'"'? 86C0@? 90? 98=49@4:? *4@@CB384C9? 8pKa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s o mal t S o rbito l e 1, 30. 3 68"; 986=:60; "213 ?86586@ A A A A A A A A A A A A A A A A A A A A A A A A AA A A A A A A A A A 2. 98@5 M ann ito l X yl ito l =0@@4 C. 8:; =0@ .8 =8;;6028. ' A A A A A A A A A A A A A A A A A A A A A A A A A 0 A A A A A '".A A A A A ancb 986=:60; "213 0.+ 2.;2.@2= +2@@86:2 A A A A A A A A A A A A A A A A A A A A 28. ;0 A A A A A 0A A A A A 1, 0 "0@ 788:.+4 A A A A A 7 AA AA AA AA AA ors Ab "#$# * '* * % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % * '% % % % % 0, 5 '() *' % % % % % % % % % %% % % % % 0, Physico-chemical Suitability fo f o or oral drug 0 Characterization delivery syste ems 350 1350 2350 3350 ( (L LogP H Hiigh polar tyy and it Ideal foor foor f f rmulating Wavenumber, cm-1 hy roophilicity dr drugs with l low solubility +*)(' $# % % % % % % % % % %EDCB @?=;:9 87 ?86586@4 A A A A A A A A A A A A A A A 6 The reelease of polyols c reelates Control lled release r orr ffoormula ations over time with concentration D3 3213 ?0// 2..@22@ 87 ?86586@ 2. -B, A A A A A A A A A A 0 A A A A A A A A A A A A A A A A A A A A A A A A AA A A A A @?=;0 2.+2=0@ 0 3213 +1; 87 =;5@0662.25 A A A A A 0 A A A A A A A A A A A A A A A A A A A A A A A A AA A A A A Polyol gradual incrreease Rapid-r release *E214)4(4 rreeaching 85% to 100% dissolution at about 30 to 45 Iso minutes m a lt M a n n it o l S Polyols ree osmotically ar Osmotic Matrix o rb it o l Xy li to l (D) active solutes DDS DDS +*)(' # % % % % % % % % % %;8?8@+ 8#;##2" 87 ?86586@ 2. 8 A A A A A ; A A A A A A A A A A 6 A A A A A A A A A A 8;06 +;:1 A A A A AA A A A A + 62 +62##;55 @5@ 2 ; A A A A A 9@ !# % % % % % 35@2=8=39 92=06 ?;8?;2@ =8:6+ ' :@ A A A A A ; A A A A A A A A A A A A A A A @+ 0@ 0 A A A A A A A A A A A A A A A 886 788; 3 A A A A A 7 A A A A A 722;@ =382= 87 ?86586@ 2 A A A A A 7 A A A A A A A A A A A A A A A A A A A A A A A A A 2. +;::1 ;A A A A A +62#;5 @5@ A A A A A 9@4 86586@ =8:6+ 36? 8 2 A A A A A A A A A A A A A A A A A A A A A A A A A.=;0@A A A A A 3 +2@@86: A A A A A 28. ;0 87 ?88;65 "0;% A A A A A 0 A A A A A A A A A A A A A A A 0 %@86:'6A A A A A +;:1@4 0 500 1000 1500 2000 2500 3000 2 (degrees) + +*)(' "# % % % % %A A A A AB@:6@ 87 -B, 0.065@2@ 87 ?86586@ A A A A A A A A A A B A A A A A 6 A A A A A A A A A A ..20 +8"@/0 0 4 4 06 * E' (4 A A A A A A A A A A A A A A A A A A A A A A A A A A A A A A A A A A AA A A A A C.;2.@2=A A A A A D3 8'02.+ 2.;2.@2= +2@@86:28. ?;87226@ 87 A A A A A A A A A A A A A A A : A A A A A 7 A A A A A A A A A A .2#;@2554 ,2@@86:28. B A A A A A B0 *C,B( 87 86586@4 B210 A A A A A A A A A A A A A A A A A A A AA A A A A ;0+2.@A A A A A A A A A A 4$E E ?86586@ 0; ;0??2+ =89?6 0..+ ?86586% A A A A A A A A A A 0 A A A A A A A A A A 0 A A A A A E;868#0 4 A A A A A A A A A A A A A A A A A A A A A A A A A 4 06 * 0. (4 8@@ 8. A A A A A A A A A A A A A A A A A A A A . ,;52.1 A A A A A ;A A A A A =8.;866+ *E214$4(4 A A A A AA A A A A *8,(A A A A A 87 7A A A A A 86586@4A A A A A B210A A A A A ;;0+2.@ A A A A A .2#;@2554 4$E-C) /min) 50 E;868#0 4 4 A A A A A A A A A A A A A A A A A A A A A A A A A 06 * 0; (4 0;6 E2@=3 A A A A A A A A A A A A A A A A A A A A 3; *E( A A A A AA A A A A M ann ito l D2;;028. 87 A A A A A A A A A A 86586@4 B210 ;0+2.@ . 6 A A A A A A A A A A A A A A A .2#;@2554 A A A A A 40 I s o mal t 4$E $BC B 2 8;#03 4 ; A A A A AA A A A A 4 06 E8:;2;% A A A A A A A A A A A A A A AD D;;0.@788;9 7A A A A A C.7;;0;+ 7A A A A A m 30 Sorbitol *EDCB( A A A A A .:0+ D8806 A A A A A DA A A A A B766=0.= 7A A A A A *D DB(A A A A A mg/c 20 Xylitol ?=;8@=8?5 5 ;87226@ 87 86586@4 B210 A A A A A 7 A A A A A A A A A A A A A A AA A A A A ;;0+2.@ A A A A A .2#;@2554 A A A A A 4$E EDE DR,(I Polyvinylopyrolidone 10 0 D3 ?;8= A A A A A 8A A A A AC.;.06 =8.@862+028. 87 B A A A A A 0 A A A A A A A A A A B 0.+ A A A A AA A A A A 0 20 40 60 ;..06 =8. ; A A A A A .@862+028. 87 B "23 0 A A A A A A A A A A A A A A AA A A A A A A A A A Ti me , mi n ; .. :20 0 A A A A A 00''2625 87 @:10; 06=8386@ *?86 A A A A A A A A A A A A A A A A A A A A 586@( 0@ A A A A A A A A A A '2.+;@ 2. A A A A AA A A A A "2.%@=;" 96 1;0.:602 A A A A A A A A A A 0 28. 788; A A A A A 7A A A A A +*)('% % % % % !#% % % % % C,BA A A A A ?;87226@ 7A A A A A 87A A A A A ?865586@ 6A A A A A 0.+A A A A A ?;?0;028. ?;?0;028. 0 0A A A A A 87 3213% ; A A A A A +;::1%680++ 299 A A A A A 9+20% 0 ?865#2.5568?5;862+8. . ;60@ 0' A A A A A '6@ "23 @:?;28; 9= A A A A A A A A A A A A A A A =30..2=06 0A A A A A ?;8?;2@ * ; A A A A A *84 B% A A A A A %$%$ $(4 109 100/ .-,+*)('-%$(%#$"!(,("' ,('-%$-.$%#-+ '!("% "-%'(%%$+",-"(1*) Paula Cermakova1 1 1 1,2 , Miroslava Spaglova , Patricia Jackuliakova , Juraj Piestansky 2Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava, Slovakia 1. 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E 5AD;8?E :B7;:=FE ;ED;=E?FD;:E%E/:=F9E:DE1 $+ *;A4GF?;4:E @@B;4GF?2E '2E #:#BE *;7?:% $%#$$ BD9;4GE #GF?;F2E '2E ;DA?F=G:4;E *A+FHGFE 4A=F=E A4:@58B:=FDE /;=G;E =GFE :B84G2E 2E DF;A;+FDE /:=F9E *;A5:2E 1E $#"5E/:5E8:=;;FDE=AEF589FE=GFE:4489:46E:DE 9=;;4;:BE 7:5=9;4E 9F@9AD84;3;B;=6E A4:@58B:=;AE @9A4F55E 9A? E #B2E (:#BE *#F=9:B4GF?E 5191A12E 2E 85;7E ;4A=;:?;DFE DF;A;+FDE /:=F9E *;A5:2E 1E (;4A=;:?;DFE #:B;39:=;AE489CF5EAE/:=F9E:DE)('E;E *E F;3;A4GF?;4:2E 'E /:5E 85FDE E/F9FE49F:=FD1E%)E5@F4=9A@GA=A?F=96E 4A=F=EDF=F9?;:=;AEA*E E E ?E /:5E 85FDE =AE DF=F9?;FE =GFE $ .$ $ " $ $ 4A4F=9:=;AEA $#$ !FE @9F@:9FDE :E :8FA85E 5AD;8?E :B7;:=FE ';55AB8=;AE =F5=5E /F9FE 4AD84=FDE =AE 5=8D6E 5AB8=;AE *-1E E /CE /G;4GE FG;3;=FDE 9FBF:5FE @9A;BFE AE 4G:9:4=F9;5=;45E A=B62E D;=E@E4AD;=;A5EE/:=F9E:DE1EHGFE )('E/:5ED;55ABCFDE;E=G;5E5AB8=;A1EF@:9:=FB62E =F5=E/:5E@F97E3:5 F=E?F=GADE:=EEE /FE @9F@:9FDE #:#BE 5AB8=;AE *E E /CE 36E 1#E :DE :E 9A=:=;AE 5@FFDE AE =/AE D;55ABC;7E#:#BE;EDF;A;+FDE/:=F91EHGFE)('% C:9;A85E D;55AB8=;AE ?FD;:E E 0E :DE E 4A=:;;7E :B7;:=FE 5AB8=;AE /:5E F=98DFDE /;=G;EEGA8951 =G9A87GE:EFFDBFE;=AE=GFE#:#BE5AB8=;A2E/G;4GE /:5E 4A=;8A85B6E 5=;99FDE 85;7E :E ?:7F=;4E $, *)'$(%#$#"*-% 5=;99F9E.,1EA9E=GFE$#"5EE/FE85FDE :E :8=A?:=FDE @9A4F55E /;=GE :E 485=A?%38;B=E $$$$ :@@:9:=851E E HF98?AE =F9;4:E FFDBFE *)1'1E E 1E??E/:5E85FDE1E4F55EB;8;DE '$$;FCFE::B65;5EA 110 .--, 5 45C2=C 853@C=49==58 345 %4@ B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B @@$58B B B B B B B B B B B B B B C6522A@ A@ =5 9$A8A$ 5@0A4@35@= C=94==A@ B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 4 @ .443 B B B B B B B B B B B B B B .B B B B B B B B B B B B B B =5 : B B B B B B B B B B B B B B B B B B B B B B B B B B B B C=4' "@ 69==54++ 6654' B B B B B B B B B B B B B B = B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 5 C5 B B B B B B B B B B B B B BB B B B B B B B B B B B B B . @$5@=49==A@ . E9E2 B B B B B B B B B B B B B B B B B B B B B B B B B B B B @ 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B7B B B B B B B B B B B B B BC2=A@ 2A521 B B B B B B B B B B B B B BB B B B B B B B B B B B B B FEDC A@ ?= ;: 9@@8 ;7 6545 3C=21 4 45C2=58 A@ 345 =A=21 $4CC2A@58 C= B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B =4$=45+ 4 =B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 45=9A@58 B B B B B B B B B B B B B B @ CA505C 6A= 35C CA/5 . :7-,+ *:,+ 9 =545?1 458$A@ =5 C6522A@ $ %%9$A=11' B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 = B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 99@@8 ):, B B B B B B B B B B B B B B B B B B B B B B B B B B B B (3' 9C58 @ =5 C%5$A.AA$9==A@ . =5 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B . 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B 5 @55825 B B B B B B B B B B B B B BB B B B B B B B B B B B B B +** $%%!$ %$ ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( #"'!' B B B B B B B B B B B B B B B B B B B B B B B B B B B B ,' 33 9 %94==A$25 CA/5 459==54 = B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 4 B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B @@ , 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B (3 69C 9@@=A$A%9=58' %%4A39==521 -B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 9 B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B-, . B B B B B B B B B B B B B BB B B B B B B B B B B B B B %48$58B B B B B B B B B B B B B B FEDC 5 +*)* .'%$#"! $% $##" % ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( !% ""! )* ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( (05400A56 . %11CA$92 $949$=54AC9=A 4 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B @B B B B B B B B B B B B B B 45C2=C .4 ;: 9@8 ;7 B B B B B B B B B B B B B B . B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B $ )* ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( (!ACC C2=A@ %4.AA25 .4 ;: 9@8 ;7 A@ B B B B B B B B B B B B B B . B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B . B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B9 5CB B B B B B B B B B B B B B 358A3 3 9@8 B B B B B B B B B B B B B B B B B B B B B B B B B B B B ; ; 5 8ACC2= B B B B B B B B B B B B B B =A@ %4.AA25C . "! .443 FE B B B B B B B B B B B B B B . B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B . B B B B B B B B B B B B B B EDC 945 B B B B B B B B B B B B B BB B B B B B B B B B B B B B %45C5@=58 A@ B B B B B B B B B B B B B B B B B B B B B B B B B B B B=5 ;A' 7 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B ' "= $9@@ ?5 ?C540058 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B 4B B B B B B B B B B B B B B=9== =5 9 B B B B B B B B B B B B B BB B B B B B B B B B B B B B @= . 84 69C 45259C58 .443 @ B B B B B B B B B B B B B B B B B B B B B B B B B B B B 4 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B . 3 ;: A@= B B B B B B B B B B B B B B B B B B B B B B B B B B B B @B B B B B B B B B B B B B B =5 =5B B B B B B B B B B B B B B 69==54 69==54 #% 9 9 B B B B B B B B B B B B B B #%B B B B B B B B B B B B B B B B B B B B B B B B B B B B ) )'B B B B B B B B B B B B B B 4 AC A4 AC B B B B B B B B B B B B B BB B B B B B B B B B B B B B 9809@@=95C+ $@CA854A@ =9== "! AC 9CC 9 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B = 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B C$A9==58 9B B B B B B B B B B B B B B 6A= @354C 98054C5 5...55$=C A@ B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B . . B B B B B B B B B B B B B BB B B B B B B B B B B B B B 9$A8A$B B B B B B B B B B B B B B 5@0A4@35@= . C=39$' @ B B B B B B B B B B B B B B B B B B B B B B B B B B B B * ( ( ( ( ( ( ( ( ( ( ( ( ( ( 6 6 "! B B B B B B B B B B B B B B29858 3A$4$9%%C25 C93 B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B 9 3%25 5C 6545 B B B B B B B B B B B B B BB B B B B B B B B B B B B B 99?25 7 %45C5@=C =5 C3394A/58 45C B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B C2=C . B B B B B B B B B B B B B BB B B B B B B B B B B B B B C$$5CC.221 .4329==58+ 8A...554A@ A@ . B B B B B B B B B B B B B B . 9 B B B B B B B B B B B B B B . . B B B B B B B B B B B B B B @ 84 B B B B B B B B B B B B B B 4B B B B B B B B B B B B B B %1CA$92 %4%54==A5C' 9@@8 ! A@ B B B B B B B B B B B B B B 4 B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B @8A$9=58 9B B B B B B B B B B B B B B $@=5@=+ 92@ 6A= %29$5? 3A$4$9%C B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B C25C = B B B B B B B B B B B B B B B B B B B B B B B B B B B B F5$ @@A$92 45CAC=9@$5 =5C=C C658 5 092A89==5 =5 35=8' 5 %1CA$92 %4%5 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 54==A5C . 4 B B B B B B B B B B B B B B B B B B B B B B B B B B B B 3A@A392B B B B B B B B B B B B B B 8A...5545@$5C . .B B B B B B B B B B B B B B ?5=655@B B B B B B B B B B B B B B C93 3%25C'B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B $5225@=B B B B B B B B B B B B B B ;: @@8 ;7 6545 C9=AC.99$=41+ $3%949? B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 . B B B B B B B B B B B B B B ?25 9@@8 B B B B B B B B B B B B B B 9B B B B B B B B B B B B B B ..226 ??A2A=11 45C2=C 6545 853@C=49==58 $@CAC=5@= 6A= 5%5$=9=A@C' 5 9CC91 B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B B B B B B B B B B B B B B B B B B . "!B B B B B B B B B B B B B B 84 ??A2A=1 .4 ?= ;: 9@8 ;7' 9 B B B B B B B B B B B B B B . B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B $5225@= @B B B B B B B B B B B B B B 9 = B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 8' ! B B B B B B B B B B B B B BB B B B B B B B B B B B B B $@=5@= 8A8 @= 853@C=49=5 9 =6.2 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B . 28 84 B B B B B B B B B B B B B B 4B B B B B B B B B B B B B B %4%4==A@ . 184%$@=5@= A@ ;7' ;: C93 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 3%25 C658 C%54A B B B B B B B B B B B B B B B B B B B B B B B B B B B B A4 84 B B B B B B B B B B B B B B 4B B B B B B B B B B B B B B 69C 3 B B B B B B B B B B B B B B45 CA@A.AA$9@= A@ ;: 85 = B B B B B B B B B B B B B B . @ B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B B B B 85=543 45259C5 $949$=54AC=A$C A@ @=5@=B B B B B B B B B B B B B B :' =A35C 345 "! =9@@ ;:+ 92= 69C A85@=A.AA58 9C =5 345 %4 B B B B B B B B B B B B B B . B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 3ACA@B B B B B B B B B B B B B B 43A@9==A@ C658 =9== ;7 A@$4%% 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B 4 49==58 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B 65 B B B B B B B B B B B B B BB B B B B B B B B B B B B B ..43 4 329==A@' 9 5%5$=58 9 =6.28 A@$459C5' B B B B B B B B B B B B B B B B B B B B B B B B B B B B . B B B B B B B B B B B B B B * ( ( ( ( ( ( ( ( ( ( ( ( ( ( +*+* $"$ %!!$ "$ ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( : F 5= 92 ;9?4A$9=A@ 9@8 9%%2A$ : + F' B B B B B B B B B B B B B B B B B B B B B B B B B B B B ' 5= 92' ;9?4A$9=A@ 9@8 9%%2A$ B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B $9=A@ . B B B B B B B B B B B B B B B B B B B B B B B B B B B B $3%25 3A$ B B B B B B B B B B B B B B $4$9%C25C 9 450A56' "@ B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 'B B B B B B B B B B B B B B 7,7,' 2' : B B B B B B B B B B B B B B + @' + %' -), -*,' B B B B B B B B B B B B B B B B B B B B B B B B B B B B 7 D B B B B B B B B B B B B B B D D D ! + F' 5= 92' F2=A%25 @A= B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B = ?C5 B B B B B B B B B B B B B B 85=5445@= !C B B B B B B B B B B B B B B C95 ;43 B B B B B B B B B B B B B B 4 3 9C58 @ 8A3 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 3 B B B B B B B B B B B B B B 2A@9=5'B B B B B B B B B B B B B B "@B B B B B B B B B B B B B B ' 7,7' B B B B B B B B B B B B B BB B B B B B B B B B B B B B 2' ),+B B B B B B B B B B B B B B @' C:+ %' )- B B B B B B B B B B B B B B )*' 5C594$ 6 B B B B B B B B B B B B B B 69C .@858 ?1 49@= . E B B B B B B B B B B B B B B . B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 35@ACB B B B B B B B B B B B B B $ * ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( (6522A@ 49=A .4 ;: 9@8 ;7 B B B B B B B B B B B B B B B B B B B B B B B B B B B B . B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B @A054CA=1B B B B B B B B B B B B B B :,7:7,7-B B B B B B B B B B B B B B 9@@8B B B B B B B B B B B B B B B B B B B B B B B B B B B B :,,77' 6522A@ 49==A # B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B BB B B B B B B B B B B B B B B B B B B B B B B B B B B B . 92A@9==5?9C58 FEDC 69C 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B B B B 50929==58 A@ =6 358A9 ; @@8 69==54 #;A' : 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B 9 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 111 :998 TOPICAL TADALAFIL HYDROGEL: A PROMISING APPROACH FOR ERECTILE DYSFUNCTION THERAPY Rafal Chiczewski 1 1 1 , Zaneta Sobol , 76564321045.,+*3-)'%4$*6'+*3 6543210/4.0-,+-*)21/2(4'0(2%-$4().,%,#"!-*,/412.2.- 4(2%-.410"-.-(4(.!-*,%2. 1. INTRODUCTION B98@@-4C= ?B@?8B21= C;;8;;58620= 3C622C278= DCBA@?@= ;= A@C:98;78= C6:= ?9CB5C48324C1= :..3;@6=48!, C6C1;;= #3;6-= 'D!0= B88C;8= ;23:8;= #BC6= 68B2= ?@158B0= /C;= ;88428:= .@B= 2;= .C7@BCA8= ?91;4@49854C= C6:= B98@@-4C= ?B@?8B28;,= 2%$25%35346 CA;@B?2@6= 3;6-= ?8682BC2@6= 869C648B;0= +98= .@B53C2@6;= C6:= 4@5?@;2@6= @.= 2C:CC.= +98= .@B53C2@6= C5;= 2@= 869C648= 432C68@3;= 86CA6-= 5?B@78:= :B3-= :878B1= /98= -8=CB8=;9@/6=6=+CA8=,=B8.10=DCBA@?@== ?B8;8B76-=;C.821,=+98=;23:1=87C3C28:=2C:CC.= /C;=:;?8B;8:=62@=:;28:=/C28B=36:8B=;2 B88C;8=.B@5=298=91:B@-8=5C2B*=C6:=C;;8;;8:= 2@= .@B5= 9@5@-868@3;= ;@32@6,= %3A;8386210= 2;=?91;4@49854C=49CBC428B;24;0=4@6.C3*10= .@B=)(=298BC?1, C=9@5@-868@3;=5*23B8=/C;=@A2C68:,=+C:CC.= /C;= C::8:= C;20= A8.@B8= 626-= 2;=;32CA21=C;=C=2BC6;:8B5C=:878B1=78948= 8C49=B83= #"!5%2 2 '1:B@-8;= /29= 2C:CC.= .@B= 2BC6;:8B5C= ?B@48;;= A1= C:3;26-= 298= ?',=$=2C:CC.= -8;= /8B8=;2@B8:=6=C=.B:-8=C2=D=?B@B=2@=3;8=0 , 23,$%2 2 D@5?@;2@6= @.= 298= +C:CC.= 91:B@-8;= #65!$346 66+462624%2 7 :99 9 :353,%+ 3 !,%5 "%$ " " " " " " " " " " 9 98 =D==+(== =) 0 0 0 0 0 ,;, 9 98 =D==+(== =) 0 0 0 0 0 ,;, 9 98 =D==+(== =) 0 0 0 0 0 ,;, = D= = +(= = = ,;, 998 ===+== = 0 0 0 0 0 ) 998 = D= = +(= = = ,;, ===+== = 0 0 0 0 0 ) :B3-=:878B1=:1=2@=298=?86;=-C6;, 998 = D= = +(= = = ,;, ===+== = 0 0 0 0 0 ) = D= = +(= = = ,;, 998 ===+=== 0 0 0 0 0 0 22727 )== =) 998 = D= = +(= = = ,;, ===+=== 0 0 0 0 0 0 )== =) 998 = D= = +(= = = ,;, 234%53$+ ===+=== 0 0 0 0 0 0 )== =) = D= = +(= = = ,;, +C:CC.= C6:= 2;= B8.8B8648= ;2C6:CB:0= DCBA@?@= 998 ===+== = 0 0 0 0 0 ) 998 = D= = +(= = = ,;, C6:= ===+== = 0 0 0 0 0 ) 998 = D= = +(= = = ,;, D14@:8*2B6= /C;= ?3B49C;8:= .B@5= %-5C ===+== = 0 0 0 0 0 ) = D= = +(= = = ,;, $:B49= #%2869850= "8B5C61!,= $= 998 ===+== = 0 0 0 0 0 ) 0120 - 998 = D= = +(= = = ,;, 585ABC68= /C;= ?3B49C;8:= .B@5= 8B4= ===+== = 0 0 0 0 0 ) 998 = D= = +(= = = ,;, ===+== = 0 0 0 0 0 4#4.--21,3,%!---,'/-%4204!-$5--$22%2+%!-*--*!-$--$44.!---) 998 = D= = +(= = = ,;, 0)2.,%- !-*2124.-- 40)"%*2124.*1,3"%*2124.-.-2-0,02%-(,.(4.0120,.-,+-- = = = += = 0 0 0 0 ="== =) 0 ;@:35= :91:B@-86= ?9@;?9C280= :;@:35= .@B53C2@6; 91:B@-86= ?9@;?9C28= C6:= C482@62B80= /8B8= ?B@43B8:= .B@5= $$= # @B-0= @C6:!,= '-9=?3B8=/C28B=/C;=?B8?CB8:=A1=3;6-=298='= 22727 :8568BC8B0-#"1,%2-2332120'-5 22%4%5346 - 3'1+(20,.- "04/!,= $= @298B= 49854C;= /8B8=@.=%=@B=B8C-862=-BC:8=C6:=3;8:=/29@32= +98= ?'= @.= 298= 91:B@-8= ;C5?8;= /8B8= .3B298B=?3B.4C2@6, :828B568:0=C6:=/8B8=5@62@B8:=3;6-=C=:-2C= ?'= 5828B,= $= @.= 298= -8;= 9C:= 7C38= @.= 2%46+ ?',,, )C49= .@B53C2@6= /C;= ?B8?CB8:= 6= :3?4C280= C6:= 298= B8;326-= 91:B@-8;= /8B8= ;2@B8:= 6= C= 25%33,$4 B8.B-8BC2@B=#+=D!,=91;4@49854C=C6C1;;= @.= 298= -8;= 643:8:= ?'= 58C;3B858620= 3,$%22?'=7C38;=@.=2C:CC.=91:B@-8; 112 2110 Formulation Firmness Shear Stickiness Adhesion phase system consisting of sodium dihydrogen 01A0923 350.4018.39 347.2515,20 -404.313.21 -77.647.72 02A0923 824.6914.94 775.333,15 -600.919.33 -190.182.61 JVKHJVQIUT 'JF"=8@A%T QPDT QRUIKPOILONUT 'A!AT 03A0923 817.5818.48 895.1043.40 -594.16 -178.986.76 14.06 %@T WU0JULQI3LUT 2QHT HUIT QIT 04A0923 1165.09 14.23 1008.0927.41 -619.009.92 -232.526.69 05A0923 872.5913.67 877.7036.06 -749.3713.86 -174.429.72 retention time was found to be 3.21 minute at 06A0923 1147.01 0.88 1256.0053.66 -811.987.47 -325.5311.49 /./.-,+*)()'%$(-#*$"! *#*" flow rate of 1.2 mL/min. The detection wavelength was 285 nm [3]. $(*-/.-WVUTSURVQPORQNTMLKJULIOUHTKGTFEDLKCUNH Formulatio Hardness Adhesiveness Cohesion Resilience Springiness .- n (g) '"*% (%) (%) 01A0923 BA@B?T=T -@;:T=T A@??T=T B9@A8T=T ?7@T=T WLQPHDUL0QNT DL3CT DUNO1ULET HEHIU0T KGT IQDQNQGONT 1.219 8.234 0.005 2.581 0.980 02A0923 :A@?9T=T -8@8AAT=T A@?BAT=T 7@7;T=T 7:@B9;T=T 2QHT HI3DOUDT IVLK3CVT IVUT O0JLK1U0UPIT KGT 1.637 0.392 0.052 0.945 2.149 03A0923 8@?B7T=T -8@7:;T=T A@?B8T=T :A@A9BT=T 7B@?B8T=T HKN3+ONOIETKGTIQDQNQGONT3HOPCTIVUT0O)UDTHKN3IOKPT 0.904 10.080 0.035 0.624 1.559 04A0923 B@:7:T=T -8;@;7T=T A@?: BB@;BT=T 7@?BBT=T KGT 0QPET UPVQPRULT NO.UT FM,T MT 9AAT KLT 1.360 15.066 0.016 1.973 1.306 05A0923 9@9AT=T -8:@9;;T=T A@?89T=T B?@;AT=T 7A@:T=T I2UUPT ?A@T QHUDT KPT IVUT RKPD3RIUDT HI3DOUHT 0.180 13.674 0.019 2.286 1.662 06A0923 @978T=T -@78BT=T A@;:T=T B9@:B?T=T 7@?98T=T GKL03NQIOKPHTRKPIQOPOPCTIQDQNQGONT2ULUTHUNURIUDT 3.824 2.062 0.112 0.158 1.248 /..- )- !'-!")-!-!"'- $- QHTJKIUPIOQNTQNIULPQIO1UHTIKTKLQNTQD0OPOHILQIOKP@T !")-%*((" WVUT 0KHIT JLK0OHOPCT LUNUQHUT LUH3NIHT 2ULUT K+HUL1UDT GKLT GKL03NQIOKPHT RKPIQOPOPCT 6LQP54IEJUT DOGG3HOKPT RUNNHT 2OIVT QPT UGGURIO1UT RERNKDU)ILOP@ DOGG3HOKPTQLUQTKGTB@?BTR0:T2OIVTIVUTDOQ0UIULTKGT B8T00TQPDTQTLURUJIKLT1KN30UTKGTBB@?T0/T2ULUT .-,, 3HUDT IKT QHHUHHT OPT 1OILKT DL3CT JUL0UQIOKP@T WVUT B@TQNQLT@T@TQLQKI0QLNT1UPT@TTHIPDQT LURUJIKLT RK0JQLI0UPIT 2QHT .UJIT QIT ;-,@T WVUT .3LT@T':A:%@TMLUJQLQIOKPTQPDTRVQLQRIULO5QIOKPTKGT LURUJIKLTGN3ODT2QHTHUNURIUDTQHTJFT;@9TJVKHJVQIUT ,QL+KJKNT+QHUDTV3DLKCUNHTRKPIQOPOPCTDU)JQPIVUPKN@T +3GGULT HKN3IOKPT RKPIQOPOPCT B*T I2UUPT ?AT QPDT K3LPQNT KGT 6QR3NIET KGT MVQL0QRET KGT P.QLQT HIOLLUDTRKPIOP3K3HNET2OIVT0QCPUIORTHIOLLULTQITAAT PO1ULHOIET9;'%T;;A4;?@ LJ0@TMUL0UQIOKPTU)JULO0UPIHT2ULUTRQLLOUDTK3IT :@T QU.T @T (@T T ,VKT ,@T @T ':AB%@T WLQPHDUL0QNT 3PIONT:9TVTQGIULTQJJNORQIOKP@T(Q0JNUHT2ULUTIQ.UPT DUNO1ULETKGTIQDQNQGONT3HOPCTQTPK1UNTGKL03NQIOKP@ GLK0T IVUT LURUO1ULT RK0JQLI0UPIT QIT HRVUD3NUDT '%TB;BB;;@ IO0UTOPIUL1QNHT'ATBT:T9T8TV%TQPDTO00UDOQIUNET LUJNQRUDT2OIVTIVUTHQ0UT1KN30UTKGTGLUHVTLURUJIKLT @T IOJQOLOPT @T ,V3PVQRVQORVQPQT ,@T Q.JVUPCT GN3OD@T WVUT Q0K3PIT KGT IQDQNQGONT OPT IVUT HQ0JNUHT W@T ,VQPCHQPT @T (LORVQPQT W@T (Q2QIDUUT (@T 2QHTDUIUL0OPUDT+ETFM/,T$:#@ U1UNKJ0UPITKGTQT(ONDUPQGONT,OILQIUTSORLKU03NHOKP4 /KQDUDT FEDLKCUNT QHT QT MKIUPIOQNT (EHIU0T GKLT L3CT $(*- . UNO1ULET IKT IVUT MUPOHT QPDT IHT ,UNN3NQLT SUIQ+KNORT T MUL0UQIOKPT JQLQ0UIULHT KGT IQDQNQGONT GLK0T DOGGULUPITIQDQNQGONTCUNH@ SURVQPOH0@ :A:A TBA@ Formulation 2 Flux (mg/cm Accumulative amount number 2 /h) (mg/cm ) 01A0923 0.125 0.75 T HOPRULUNET IVQP.T L@T KLKIQT IL+H.Q4 01B0923 0.085 0.51 2OUINO.K2H.QT GKLT VOHT OP1QN3Q+NUT C3ODQPRUT 01C0923 0.022 0.132 02A0923 0.154 0.924 IVLK3CVK3ITIVOHTLUHUQLRV@T 02B0923 0.101 0.606 02C0923 0.056 0.336 03A0923 0.335 2.01 03B0923 0.224 1.344 03C0923 0.124 0.744 04A0923 0.221 1.326 04B0923 0.152 0.912 04C0923 0.101 0.606 05A0923 0.169 1.014 05B0923 0.106 0.636 05C0923 0.069 0.457 06A0923 0,225 1.350 /..-2-$$(""- The column is a reversed-phase column (Kinetex C18 ).TThe mobile 113 8767 543214023/ -0, . . . . . . . 0 ,+1* 1 *))2321, -(*3/'+ 23 . . . . . . . . . . . . . .. . . . . . . *5408(4+ +4)2% . . . . . . .. . . . . . . %2+8'0+243 %0223/ *--')'0* . . . . . . . 0 . . . . . . . 0 * *1 1'% +1* . . . . . . . 1 * *$2)21, 1'+123/ . . . . . . . +#"! + 1,2 1 1 2 1,2 . . . . . . . . . . . . . . , $#" #" . . . . . . . . . . . . . . , 5## Pajk #" . . . . . . . . . . . . . . . . . . . . . , Klemen Korasa 2F Faaculty off Pharmacy, University off Ljubljana, 100 o o 00 Ljubljana, Sllovenia S 1. INTRODUCTION +# . . . . . . . ## " . . . . . . . "J:O? MBSPK L L L L L L L KHM AHQH PQHPBQHI =@ =KHGIJG: ?OH L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L TSRQPORNM MRKJI IJMPHQMJRGM FTEDMC BQH L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L MPQ @@1IQJHI T B L L L L L L L TED AJ?O B ;Q@M?BKKJGH T* FT*C L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L AJIHK@ NMHI ?R JSPQRH ?OH MRKN=JKJ?@ BGGI L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L BL L L L L L L BGI 9%9 SJ L L L L L L L L L L L L L L?NQH JG B '/ QB??JR5 ?R R=?BJG T* L L L L L L L L L L L L L L L L L L L L L L L L L L L L B L L L L L L L L L L L L L L L L L L L L LL L L L L L L IJMMRKN?JRG QB??H R PRRQK@ MRKN=KH B;?JH N L L L L L L L B L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L ;RG?HG?M R L L L L L L L L L L L L L L 5 / 5 5 ' 5 5 BGGI L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L B L L L L L L LL L L L L L L POBQS Q SB;HN?J;BK JG:QHIJHG?M FT98MC7 6RAHHQ5 L L L L L L L L L L L L L L L L L L L L LL L L L L L L QHKB??JH ?R ?O B L L L L L L L L L L L L L L OH ?R?BK T* ;RG?HG? JG HB;O MBS L L L L L L L L L L L L L L L L L L L L L G L L L L L L L L L L L L L L L L L L L L L B SPKHL L L L L L L PO@MJ;BK M?B==JKJ?@ QHSBJGM B 4H@ ;RG;HQG5 BM L L L L L L L B L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L F#J:NQH /C7 L L L L L L L QH;Q@@M?BKKJ3B??JRG INQJG: M?RQB:H ; GG ;RSPQRSJMH Q B L L L L L L L L L L L L L L B L L L L L L L B L L L L L L LL L L L L L L PHQBJKB=JKJ?@7 B L L L L L L L L L L L L L LL L L L L L L 2 S B SBGG MPH;?QRM;RP@ JM PBQ??J;NKBQK@ MHGMJ?JH ?R B L L L L L L L L L L L L L L L L L L L L L Q L L L L L L L L L L L L L L L L L L L L L IJBKN ==KH SH?ORI JRNM JL L L L L L L M?NIJHM ? IJL L L L L L L O OBH BL L L L L L L I IHSRGM?QB??HI ? ? I BL L L L L L L J? J?ML L L L L L L HHGHMM 2 S B SBGG MPH;?QRM;RP@5 ;RS=JGHI AJ?O PBQ?JBK B L L L L L L L L L L L L L L L L L L L L L L L L L L L L QL L L L L L L 6 . . . . . . . . . . . . . . RSPRMJ?JRG R TED MBSPKHM AJ? L L L L L L L L L L L L L L L L L L L L L L L L L L L L ??OO BIIHI L L L L L L LL L L L L L L KHBM? M-NBQHM F9,EC QH:QHMMJRG5 L L L L L L L L L L L L L L L L L L L L LL L L L L L LABM PPPKJHI BM L L L L L L L B L L L L L L L L L L L L L L SJ?NNQQH R T ? N L L L L L L L L L L L L L L T* BGI 9%9 JG B '/ QB?JR7 L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L PQR;HMM BGGBK@?J;BK ?H;OGRKR:@ F9T L L L L L L L B L L L L L L L L L L L L L L 9 T T+ +C ?RRK * .. .. .. .. .. .. .. #! # " . . . . . . . . . . . . . . SRGJ?RQJG: ;Q@M?BKKJGJ?@ JG TEDM INQJG: L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L EBS B SPKHM AH L L L L L L LQH MN=H;?HI ?R B;;HKHQB??HI M L L L L L L L = L L L L L L L L L L L L L L B L L L L L L L M?B==JKJ?@ BL L L L L L L M?B==JKJ?@@ ?HM?JG:7 B ? L L L L L L L ;RGIJ?JRGM B L L L L L L L L L L L L L L L L L L L L L BB?? BGGI QHKB??JH ON B L L L L L L L L L L L L L L B L L L L L L L NSJIJ?@7 @L L L L L L L TGBK@MHM AH L L L L L L L HQH PHQ 5*1'02*)+ *3% 5'1(4%+ . . . . . . . . . . . . . . . . . . . . . BGI B 6 5# # . . . . . . . +# . . . . . . . #"# . . . . . . . TSKRIJPJGH SBKHB??H FT*C ABM 4JGIK@ MNPPKJHI L L L L L L L B L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L 2 S B SBGG SHB B L L L L L L L BMNQHSHG?M AHQH ;RGIN;?HI NMJG: G L L L L L L L L L L L L L L L L L L L L LL L L L L L L =@ )Q44B5 I7 I7 F(RR *HM?R5 EKRHGJBC7 L L L L L L L Q L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L 9OT+ T + PQR=H L L L L L L L H F)BJMHQ P?J;BK E@M?HSM5 8G;75 L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L 9RK@JG@KP@QQRKJIRGH5L L L L L L L :QBIHL L L L L L L )1'L L L L L L L F9%955 9L L L L L L L ETC5L L L L L L L ;RG GGH;?HIL L L L L L L ?RL L L L L L L 2BS B SBGL L L L L L L 2(/+*L L L L L L L SBQ4H?HI BM )RKKJIRG 'C5 ABM MRNQ;HI 1/ ETC7 EPH; L L L L L L L ;?QBK QBGG:H ABM /1 ;S L L L L L L L B L L L L L L L L L L L L L L7 +OH L L L L L L LL L L L L L L ABM MNPPKJHI =@ )Q44B5 I7 I7 FEKRHGJBC7 L L L L L L L L L L L L L L L L L L L L L Q L L L L L L L L L L L L L L L L L L L L L 8G?H:QB??JG: : B :L L L L L L L EPOHQH ;OBGGGHK ABM NMHI7 +A L L L L L L L B L L L L L L L L L L L L L L L L L L L L L + ARL L L L L L L +# ! " . . . . . . . . . . . . . . B;-NJMJ?JRGML L L L L L L AHQH BHQB:HI ?R R=?BJG B MJG:KH L L L L L L L B B L L L L L L L L L L L L L L L L L L L L L L L L L L L LL L L L L L L T MRKN?JRG ;RG?BJGJG: '7' T* BGGI /7/ 9%9 L L L L L L L N L L L L L L L L L L L L L L L L L L L L L L L L L L L L B L L L L L L L L L L L L L LL L L L L L L MPH;?QNNS5 A Q L L L L L L L AJ?O JG?H:QB??JRG ?JSH R / M7 L L L L L L L B L L L L L L L L L L L L L L L L L L L L L L L L L L L L JG H?OOBGGRK F C ABM MPQB@@1IQJHI NMJG: L L L L L L L ? B L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L B L L L L L L LL L L L L L L +OH 1QB@ P L L L L L L L L L L L L L L PRAIHQ IJHQGG 9BGBK@?J;BK5 )C5 NMJG: Q L L L L L L L L L L L L L L L L L L L L LL L L L L L L BB?? GG JGKH? BJQ ?HSPHQB?NQH R 5 AJ?O BG BJQ L L L L L L L L L L L L L L B L L L L L L L L L L L L L L B L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L N)L L L L L L LQBIJB?? B ?JRG F /7 L L L L L L L L L L L L L L L L L L L L LL L L L L L LC5 4% BGGI L L L L L L L L L L L L L L L L L L L L L B L L L L L L L L L L L L L L 'O BGGI KJ-NJI HQ GG BG:NKBQ QBG:H L L L L L L L L L L L L L L B L L L L L L LL L L L L L L T*9%9 QB??JR R '/7 L L L L L L L B L L L L L L L L L L L L L L R ' JG L L L L L L L L L L L L L L L L L L L L L L L L L L L L5L L L L L L LNMJG: GG 1 L L L L L L L L L L L L L L BHKHQB??RQ IH?H;?RQ7 B L L L L L L L 114 .-,- +*)* '%$#"! #!$ !!%# ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( !!% ,* ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( (3@C97 7BA7;6 ;. #" B;6BC6A@@?A7;6 76 :?)D C: A EDCBA@? =C@C D@C CD;:C9 A; ?B BC C@?AC9 :A?(7 7A%% AC:A7650 A :;.AA=?@C -,@8+C@ 4DA7+ *)(' *C@)?6%$0 3#" . #$#!%( ( ( ( ( ( ( ( ( ( ( . # $# ?69 3!E );9C : =C@C (87 A =7A ":DC6 ' + 2E"$0 C 3!E );9C =?: B@C?AAC9 8:765 A 26:B@?) ? )( C@ 0 -":DC6 CB6; ;5% 6B0 :?)D C: =7A +66;=6 #" B;6BC6A@?AA7;6: ?669 ? + 6 ? ? =?: AC6 ?DDD 7C9 A; D@C97BA #" B;6AC6A 76 ? 6 6 :?)D C: CD;:C9 A; ?BBC C@?AAC9 :A?(7 7A%% ? A B;697A7;6:0 * ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ** . ( ( ( ( ( ( ( ( ( ( ( *,* . !!%# ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( (( ( ( ( ( ( ( ( ( ( ( 6 758@C AC : ) ? )D C =7A;8A ?99C9 #" ?AA A ? 3#" :B;@C D ;A 3#" :B;@C D D ;A ;. AC B; CBAC9 ?) ;. AC B; CBAC9 ?) ? ? ) 66 :DCBA@? ) 66 ? ? :DCBA@? A 97:D ?%%: ?66 A 97:D ?%%: ?66 ? ? ? ? 6 3 D?AAAC@66 B?@?BAC@7:A7B ;. ?66 ? A @ ? :;=C9 A? A ??AA A :?)D C: 97...CC@C9 .@@;) . . . ) A;:C "E0 ".AAC@ . =CC+: 869C@ ?BBC C@?AAC9 :A?((7 7A% ? ? CD;:C9 A; ; ?BBC C@?AAC9 :A?((7 7A% B;6 ? ? 697A7;6: B;697A7;6: AC 97...@@?BA;5@?) . . ? ) @CC? : AC -758@C $0 7: :855C:A: A?A A ? :DCBA@? ) )?%% 6;A ? C)C@55C6BC ; @ ;. B@%%:A? 76C @C. CBA7;6: 7697B?A765 @ . ? @CD@C:C6A A C :A@C::C9 :?) ? )D C: =C =7B @CB@%:A? 7? ??AA7;60 C:C @C. CBA7;6: ?@C C:: . B;8 9 @C98BC C AC @C 7?((7 7A% ;. D@C97BA7;6 ? 6: )?9C :?@DD ?69 @ C:: 9C.776C9 B;)D?@C9 A; AC D8@C . (% AC 3!E ) );9C 0 #" 97...@@? . . ?BA;5@?) ? ) :855C:A765 AC .;;@) . @ )?AA7;6 ? ;. C::;@9C@ @C9 B@%%:A? 76C 9;)?76:0 @ # + # * ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( 3 97...@@?BA;5@?): ;. #" ?69 AC . . ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( +* 3#" :B;@C D ;A ;. ?)?6 :DCB D D BA@@?0 A :?)D C =7A; 7 8A ?99C9 #" @CB;@9C9 ?A A ?69 ?.AAC@ 99 9 #" 9 9 A 9 . . *+* . % ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( =CC+:0 C 9CC ;DC9 3!E );9C 76B 89C: A=; .??BA;@:0 . * ( ( ( ( ( ( ( ( ( ( ( C .77@:A .??BA;@ CD ?76: 1 . . ;. AC 9?AA? ? ?@7 ((7 7A% ?69 AC :CB;69 ?99: ?6;AC@ ? C 9CC ;D DC9 3!E );9C B?66 :C@C ?: ? 8:C.88 ? . 57765 ? A;A? ;. 0 C );9C C7(7A: ?66 A;; . .;;@ 8? 7A?A7C ? 9CACBA7;6 ;. 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INTRODUCTION Olivem 5.0 5.0 5.0 0 0 0 Xanthan gum 0 0.5 0 1.0 0.5 0.5 BA@?=;:987:;7:6:547:3498A927@5=29?:1907: Water up to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Thakur R., Sharma A., Verma P., Asha D. A 683%8//+=: 52;862@8: 8(: 2/@: (;30: review on pharmaceutical emulsion. AJPRD 2023, 11 (3). 2. Madawi E.A., Manaa H.M., Alattrach D.G., et al. Dry emulsions as a promising adaptation in pharmaceutical dosage formulations: A review of recent developments and biopharmaceutical significance. J. Drug Deliv. Sci. Technol. 2024, 96, 105712. 3. )6=-2-,-!.@7=-2: K., Laskowska E. Intracellular protective functions and therapeutical potential of trehalose. Molecules 2024, 29, 2088. 4. Greco L., Ullo S., Rigano L., et al. Evaluation of the filming and protective properties of a new trehalose and ceramides based ingredient. Cosmetics 2019, 6, 62. 5. https://cosmetics.specialchem.com/_/media/pro duction/cosmetics/dksh/tds/trehalose- 100brochurepim000000309.pdf 117 9876 95432410/.5-.,5+.9*54312.)52)324*043.02(.9'9%12.51/.)0%3.0,. 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Natalija Dimovska1 1 1 , Maja Todeska , Dobrinka Dimova 1. INTRODUCTION :?*A@433458 B90?8 ;98 ?';=80;/=40(8 !8 8 8**8B98 5B;*404@8?98;8@?0;@680;/=408A@4338;94306(88 CBA@?=?;:B98 765@?:7=?@B548 B38 ;8 ;958 32/3412490=68 :?;0458 B98 ;8 :?;0B9-8*;:7B948 =2?@?12B9?=?948 ;90B/B?0B:8 .B54=68 23458 ?@8 );@;8 ;/:?;0(8C;9;5;, 0@4;0B9-8/;:04@B;=8B94:0B?93,8+038?@*2=;0B?98;38 B=*):?;04580;/=4038497;9:43830;/B=B06(8B*A@?'438 "" A;0B4908 :?*A=B;9:4(8 ;958 4932@438 ;AA@?A@B;048 B3B904-@;0B?98 043038 ?==?.458 7,2@,88 5@2-8@4=4;34,8 -2B54=B943,8 748 0B*48 @412B@458 ?@8 :?*A=4048 0;/=408/@4;5?.98.;38@4:?@545, ::?@5B9-8 0?8 3?*48 @434;@:78 :BA@?=?;:B98 765@?:7=?@B548 B38:=;33BB458 ;38 ;8 %C$8 C=;338+#8 ," " 5@2-(8 B95B:;0B9-8 =?.8 3?=2/B=B068 ;958 =?.8 B33?=20B?98 3025B438 ?8 0748 B=*):?;0458 0;/=4038 A4@*4;/B=B06,8 "! 8 7B38 :=;33BB:;0B?98 32--43038 .4@48A4@?@*45823B9-8$8;AA;@;0238++8A;55=48 07;0807B38;:0B'48A7;@*;:420B:;=8B9-@45B4908+8 *407?5,8$;*A=438.4@48:?==4:0458;088*B920438 47B/B038A??@8;124?2383?=2/B=B06(8.7B:78:;98=4;58 ;958;9;=645823B9-8#83A4:0@?A7?0?*40@6, 0?8 A??@8 5B33?=20B?98 ;958 749:48 A??@8 /B?;';B=;/B=B06,8 "/""( ;@0B:=48 3B48 5B30@B/20B?98 ?8 :BA@?=?;:B98 7B38 302568 ;B*38 0?8 :?*A;@48 5B4@4908 A;@0B:=48 765@?:7=?@B548 .;38 ;9;=6458 23B9-8 =;34@8 3B438 ?8 :BA@?=?;:B98 765@?:7=?@B548 ;958 5B@;:0B?98 *407?58 A@?'B5458 /68 *;92;:02@4@8 5B4@4908 *407?538 ?8 A@?52:0B?98 ;958 4';=2;048 ?8+8;958/683B4'48;9;=63B3, 074B@8B*A;:08?985B33?=20B?98;9585B3B904-@;0B?9,8 3. RESULTS AND DISCUSSION 2. MATERIALS AND METHODS B4@4908A@?52:0B?98*407?538;9585B4@4908+8 -") A;@0B:=483B485B5908344*80?8;4:085B3B904-@;0B?9(8 CBA@?=?;:B98 765@?:7=?@B548 .B078 ';@6B9-8 3B9:48 ;==8 B=*):?;0458 0;/=4038 5B3B904-@;0458 ?@8 A;@0B:=48 3B438 58 *8 ;958 5!8 *8 =433807;988*B92043, .;38 ?/0;B9458 @?*8 ;@0B8 @2-3,8 :BAB49038 B9:=25458 B:@?:@630;==B948 :4==2=?348 @?*8 $8 748 :?@@4=;0B?98 ?8 0748 *407?538 ?8 A@?52:0B?9(8 #B';A2@ 0748 A;@0B:=48 3B48 ?8 0748+8 ;958 5B33?=20B?98 ?8 8 !!(8 ;B48 30;@:78 @?*8 ?12400438 @4@43(8?'B5?948;958C?A?'B5?948@?*8%$8 B=*):?;04580;/=4038B38A@43490458B980748?==?.B9-8 ?==B5?9 0;/=48;/=48!,8 8 8 ;958 ?==B5?9 8 C )(8 ;::?@5B9-=6(8 $B=B:;(8 :?==?B5;=8 ;9765@?238 @?*8 "-"407?538?8A@?52:0B?9(8+8A;@0B:=483B48;958 C;/?08 C;/)?)3B= (8 ;-943B2*8 304;@;048 @?*8 5B33?=20B?98@432=03, ?334=*;98 ;958 76A@?*4==?34)/;3458 B=*8 :?;0B9-836304*8@?*8C?=?@:?98A;5@6, Production API particle Dissolution ")""/ WG d908* 89.73 .?8 5B4@4908 *407?538 ?8 A@?52:0B?98 .4@48 method size (%) ;AA=B458 5B@4:08 :?*A@433B?98 C8 ;958 .408 WG d901708* 79.06 -@;92=;0B?98 ,8 +98 0748 C8 *407?5(8 *B .;38 A4@?@*458 B98 ;8 /B98 /=4954@,8 8 .;38 WG d901000 ** 81.12 A4@?@*458 B98 ;8 7B-78 374;@8 -@;92=;0?@8 .B078 DC d901000 ** 89.50 :BA@?=?;:B98 765@?:7=?@B54(8 *;B48 30;@:7(8 *B:@?:@630;==B948 :4==2=?348 ;958 A?'B5?948 +8.B078A;@0B:=483B485!8*8*B==45807@?2-78 /B954@8B90@;-@;92=;@=6(8;958;==8?074@84:BAB49038 !,8**83B4'4823B9-8;8@?0?@8*B==, B9:=25458 40@;-@;92=;@=6,8 748 0;/=4038 .4@48 122 #"! ! # 1 //5*4 *-2/5412544)3'(4"4 625534 -654 62+651-432110/.-20,4)('%$#"4 56 122,+4 7654 32110/.-20,4 214 62+65*4 )('%$#"4 !0*4 - /5-14 -654 20*- ,54 0!4 2,*5 1534 1.*! 54 *5 4 !0*4 *03.534 .12,+4 5-4 +* ,./ -20,4 2-64 4 5,6 ,534 32110/.-20,4 0,5*15/4 / *+5*4 *-2/54 1254 3 '(4 4 0 *534 -04 / *+5*4 *-2/54 12514 )3'%4 "4 ,34 5-4 *-2/54 12514 )3 '%4 4 %'#"4 76214 +* ,./ -20,4 0!4 2//534 4 160534 *53.534 1.++51-14 -6 -4 1 //5*4 *-2/54 1254 4 !0*14 32110/.-20,4/2 5/43.54-0435*5 15340*012-4 ,34 1 //5*4 +* ,./514 2-64 / *+5*4 1.*! 54 *5 4 0 -4+* ,./54!0* -20,42*5-40*51120,4 2/ /54 !0*4 32110/.-20,4 /5 32,+4 -04 5--5*4 0!442//534-6*0.+64 44412554*03.534 5*!0* ,5 0 * /54 32110/.-20,4 )('#"4 -04 5-4 2//534 -6*0.+64 4 4 4 12554 1604 0,/4 1.2- /54!0*432*5-40*51120,462/541 //5*4214 1/2+6-/462+65*432110/.-20,4)(#"40 *534 1.2- /54 !0*4 5-4 +* ,./ -20,4 765154 !2,32,+14 -044 *-2/5412543 ' %44)%'#" .,35*10*54-654,5534-04 *5!.//415/5-40-64-654 7 /5-14 *03.534 2-64 5-4 +* ,./ -20,4 0!44 1.++51-2,+4-6 -4/ *+5*4 *-2/5412540!4-6544214 +* ,./ -20,4 2-64 1 //5*4 *-2/514 )('%$#"4 # *-2/541254 ,34*03.-20,45-6034-042*054 7 /5-14 *03.534 2-64 32*5-4 0*51120,4 0!4 -654 20 2/ 2/2-4 0!4 2*0!/0 2,4 4 2//534 -6*0.+64 4 4 4 12554 625534 63*06/0*235 32110/.-20,40!4('� * /54-04 5. REFERENCES 5-4+* ,./ - # )3 4/25* 4445-4 /4 ' (44('%$#"476214350,1-* -514-6 -4 32*5-4 0*51120,4 65,4 /2534 -044 2-64 *-2/54 1254 3 0.*, /4 0!4 ' 4 4 ,4 2,- 2,4 5!!5-25432110/.-20,45*!0* ,54122/ *4-04-654 6 * 5.-2 /4125,51 4)"4$$ 5-4+* ,./ -20,4*051142-6442-64 *-2/54 12543'(4 ACKNOWLEDGMENT 56 ,2 /4*05*-25140!4-654+* ,./514 5 *14 7654 .-60*145*5114-652*4+* -2-.354-045/5 4!0*4 5-4 +* ,./ -20,4 62/54 0!!5*2,+4 5,6 ,534 -04 6 54 *53.534 32110/.-20,4 !0*4 4 2//534 !.,32,+4 ,34 1.0*-4 14 5//4 14 -04 -652*4 0//5 +.514!0*4-652*40,-*2.-20, -6*0.+64 44412554)(#"40 *534-04 32*5-4 0*51120,4 )('#"4 76214 32!!5*5,54 2+6-4 54 --*2.-534 -04 -654 0 -4 , -.*54 0!4 +* ,./514!0*5343.*2,+45-4+* ,./ -20,46264 0./34 /22-4 2532 -54 501.*54 -04 -654 32110/.-20,4532. # ,4 -6214 1-.34 2//2,+4 0!4 -654 4 2-64 / *+5*4 *-2/541254 14 /25345!0*545-4+* ,./ -20,4 7654 *51./-14 1604 -6 -4 2//2,+4 6 34 0,/4 4 2,2 /4 5!!5-4 0,4 32110/.-20,4 )%' 14 (#"4 5-4 +* ,./ -20,4 2-15/!4 4 05**2354 -6542,!/.5,540!42//2,+4 14-6544.,35*+0514 ++/05* -20,4 /-5*2,+4 2-14 32110/.-20,4 56 20.*4,0-65*45/ , -20,4!0*4-6214 154214 -6 -4.12,+4 4*0-0*42//4214,0-4 41.2- /54 4-04 *53.544 *-2/54125 4. CONCLUSION 76541-.3462+6/2+6-14-654*2-2 /4*0/540!4 *-2/54 1254 ,34 *03.-20,4 5-60314 2,4 0-222,+4 32110/.-20,4 *0!2/514 !0*4 6 //5,+2,+4 14 7 /5-14 *03.534 .12,+4 5-4 +* ,./ -20,4 2-64 123 ;:98 76574326160/.162-,520+*15)0,3)47()-2'-)76261,30)2/-%,)41 $+60).2-% .#"! !1"#1,2 1 2 1 , Froelich Anna , Schneider 0## , )#!1 # %$#"! # #! $#!#"# $ # "! " "# " # # 2% "! " !!#" -54000 Nancy, France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an Gajdziok1 1 1 !1- 19 !1/19 1 Department of Pharmaceutical Technology, Faculty of Pharmacy, Masaryk University, Brno, Czech Republic 1. INTRODUCTION -.379;:7=C=05ACC?=@"2,@;46C; ")2,45%6A:%=CA?;"2)-A7 :;79:A?=:8 37;A7;72 1B; 59=:?AC6; =0 ?B7 7?::;3 C 37?@@;3 -A?;:2 1B; 7=@4?=C -A7 DCBA@A?=C ?B;:A98 7 6=55=C@8 47;3 ?= ?:;A? 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"= 3;9=7?=C2 A99:=A6B-A7;3 +;8 9:=6;77 9A:A5;?;:7 =C 56:=9A:?6@; 1#-25.*-%/1-)+1#52$0+/ 9:=9;:?;72 DC 9;:5;C? D, A 04@@ 0A6?=:A@ 71# 3;7/CC0@4;C6;=0), 5ACC?=@ " ;C?A 72 :2 =2, :A/4;, -A7 $, 47C/ 7A59@;7 )2 #: 0@=- :A?; 7?:=C/@8 A00;6?;3 A;:=38CA56 =4?6=5;7, 6B=7;C;:=056:=9A:?6@;72#33C/ 59:=.C/ # AC3 ! ! A? B/B;: @;.;@72 @;46C;"89:=?;C, 9;:5;C? DD %4@? =C ?B;7; 0C3C/7, ;CBAC6;: =0 A;:=7=@A?=C 9:=9;:?;7 AC3 C?;/:A?C/A5;?;:"255 A55=C45%6A:%=CA?;" ;C?A72:2=2, :A/4;, A7 9=:=/;C2 D7=9:=9AC=@ " ;C?A 72 :2 =2, AC3 5=308C/ ?B; 0A6?=: 7;? "), $, C 7A59@;7 :A/4;,-A747;379;:7=C5;345 ?-=@;.;@04@@0A6?=:A@3;7/C721B;C0@4;C6;=0 0=:@A7;:300:A6?=C5;A74:;5;C?7,0;3 C3.34A@0A6?=:7-A7;.A@4A?;347C/##, 75=?6, :A/4;,2 -B@;3A?A7-A76=C346?;3.A7=0?-A:; -A?;:-A747;3 2 191 1 11 6:=9A:?6@;7-;:;9:;9A:;347C/ '79:A83:8;:"433;:70;@3,2#) 126 10/. -, *)('%( $#" "!( '((!# + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + ED'33/;DI; ' B B B B B B B B B B B B B B J4 884 HG @ /DF;G A); A;:A;F B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B B B B IHDFEAEHD:7 -,/, 1 + + + + + + + + + + + + + + + + + + + + + + + + + + + + JIIHGFEDC AH @? E=% % /, + + + + + + + + + + + + + +B B B B B B B B B B B B B B / //AA00/AA 2;G A / B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B B B B FG1EDC A;=0;G0GH0;GAE;: F;0 B B B B B B B B B B B B B B 0;DFEDC HD A)); H? 0 B B B B B B B B B B B B B B B B B B B B B B B B B B B B A B B B B B B B B B B B B B B B B B B B B B B B B B B B B =HG; :0);GEI3 0X1 X2 X3 X4 Y1 Y3 (ml/h Sample (m/s) (mm) Y (%) E=0GH2EDC ;GHF1D= B B B B B B B B B B B B B B =EI ;)2EHG7 J 3H(;G B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 2 ) 1 120 900 3. 0 1. 0 8. 11 1. 37 6. 62 0/=0 :0;;F 9%$-- =#")5 0GH2;F H0AE=34 /A B B B B B B B B B B B B B B B B B B B B B B B B B B B B , , , , , , , , , , , , , , B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 2 160 900 3.0 1.0 7.52 1.34 8.23 3 120 1665 3. 0 1. 0 7. 46 1. 34 9. 28 )EC);G :0;;F 3;F AH EGG;C/34 160 1665 3.0 1.0 7.65 1.35 8.69 !DIG;:;F 5 120 900 4.3 1.0 7.10 1.37 13.00 6 160 900 4.3 1.0 6.46 1.34 19.00 FGH03;A G;7 120 1665 4. 3 1. 0 6. 74 1. 36 16. 90 08 160 1665 4.3 1.0 7.12 1.37 13.30 5a 120 900 4. 3 0. 5 5. 84 1. 34 31. 70 F;'HHG= ' G =6a 160 900 4.3 0.5 4.91 1.34 52.20 67- AH -7 ==5 C;D;G7a 120 1665 4.3 0.5 5.53 1.39 39.02 8a 8a 160 160 1665 1665 4.3 4.3 0.5 0.5 5.76 5.76 1.36 1.36 34.11 34.11 :H3EFE'EE;F E 3EFE'EE F E ' ' B B B B B B B B B B B B B B EDAH :=33;G44 F;D:;G44 6 B B B B B B B B B B B B B B B B B B B B B B B B B B B B G11EDC A;=0 G B B B B B B B B B B B B B B 0;G A B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B /=0 :0;;F B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B JEG '33H( GB B B B B B B B B B B B B B ' B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B /DE'HHG= ' G = 0 H3; FE= B B B B B B B B B B B B B B B B B B B B B B B B B B B B =;A; ; ;G B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 6 J B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B @ B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 88 ;GH:H3 0;G B B B B B B B B B B B B B B G'HHG= ' G = , # ' + + + + + + + + + + + + + + '(!# IH=EDEDCB B B B B B B B B B B B B B )EC) FE= =;A;G4 ;3; B B B B B B B B B B B B B B ;24 3:;G FE'''GG / - F . 0J 9 7$6 =5 )EC);G 88 B B B B B B B B B B B B B B 8 97- 5 (;G; HAJ B B , B B , B B , B B , B B , B B , B B , B B , B B , B B , B B , B B , B B , B B , = + + + + + + + + + + + + + + . F;;0 3/DC B B B B B B B B B B B B B BB B B B B B B B B B B B B B F;3E2;G1 A);G 00;/AEI ; / B B B B B B B B B B B B B B ;'''EEI , *))*) + + + + + + + + + + + + + + )# )( / 1. Athanazi io R. Airwayy diisease: similarities and a d + + + + + + + + + + + + + + + + + + + + + + + + + + + + @? ? E= B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B = ddiifferenc ces between asthma, COPD and ' D bronchie ectasis. Clinics, 2012. 67: 1335-1343. -,, 1 2. Borghard dt J. M., et al., In I nhaled therap ap py y in + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + r esp ir a to or d ry y di i s ea s e: th e co mp m p lex in terp la ay y of of f J=HDC :0G B B B B B B B B B B B B B B 11 FG11EDC 0pp u lmo n a ry r y ki n etic p r o ce s s es . C an ad ian ''33H( GR esp ir ato o ry r y J o u r n al , 2 0 1 8 . 1 : 1 - 1 1 . : EA: EDIG; B B B B B B B B B B B B B B B B B B B B B B B B B B B B G ;:; 3;F AH DHA3; F;IG B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B G;:; ED B B B B B B B B B B B B B B B B B B B B B B B B B B B B 3. J F B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 7 7 Develo op p men t of o dr f dr ry y p o w de d er in h a ler J B B B B B B B B B B B B B B F fo f r mu la ti io n s fo r dr d r u g de d elive ry r y sy s ystem s . J o u r n al ;D)o f R esear r ch in Ph ar m ac y , 2 0 1 9 . 2 3 ( 6 ) : 9 7 3 - 9 8 7 . FE:0;G:EHD7 @=33;G DH3; FE= B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B =;A;G: EDA;D:E'EE;F B B B B B B B B B B B B B B 'B B B B B B B B B B B B B B 4. Munir M M., et al., Dev oppment off a spprayy-ddrried elo o s a A)E: ;''';;IA 1 0GHF/IEDC 'EED;G FGH03;A: formulati ion off pep dee-o tidDN NA A nanopparticles into o ;D:/GEDC a drryy po d r owdeer for pulmonaryy deeliveryy using d r d r B B B B B B B B B B B B B B =HG;; B B B B B B B B B B B B B B G B B B B B B B B B B B B B B /DE'HHG= ' G = B B B B B B B B B B B B B B 0 FE:AGE/AEHD4 factorial dde esignn. Pharmaceutical / B B B B B B B B B B B B B B ;D) 0;G'HHG= Research h, 2022. 39(6): 12151232. ' G = 127 0/.- ,+*+)(0'+%$(#$"$'!)% '(,")$+)+"+$, )(#+"$(, !'$ %")+%$! $+' () ,$+%! ($,$0 % Anastas Dimoski, Teodora Tasevska, Lina Livrinska, Maja Simonoska Crcarevska, Dushko Shalabalija, Katerina Goracinova and Nikola Geskovski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va Erak1 2 1 2 2 , 4)('7C%($#") 7)%($73%)'7)%$ 7%%'70)))-5() , Anita Hafner * )('%$#"! %%#%! %%% '%%$ % %%%% %%$%%% 1. INTRODUCTION Sample Melatoni PLGA Drug: # n conc. conc. polymer HGFEDFCBA@?=BG=;: DH987;: 6C5: 43: (g/mL) (g/mL) ratio 2@?FE:15?:0G/:@3.BAC4F:FG3=CBA@3:0G/-1FC= (w/w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ample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oltan Mark Horvath, Kirils Kukuls, Alina Jaroslava Frolova a, Liga Petersone, Elzbieta Maria Buczkowska, Valentyn Mohyly yuk 1. INTRODUCTION AB477 H 4 C C C C C C C C C C ?@A@ 4GG8= 7G 1@ 9(+ C C C C C C C C C C C C C C C C C C C C 4 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C B5GA96GF@ 76@ A@D7 G4 9GFEGFD ?@A@ C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C 96BA5 :AEED7BFF8@ A C C C C C C C C C C + !3 C C C C C C C C C C HGFEGFD BA@ ?=@FE 9@87D 8 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C A5B:@7:BF B88= 87AB:@7:BF 4GGA5 C C C C C C C C C C B C C C C C C C C C C C C C C C C C C C C 4 A 5G8D3 BC C C C C C C C C C 2 2BB1F@7D :G59AD@ B 5B00GA 9BA77 G4 76@D@ C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C B C C C C C C C C C C A C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C 44GGA5 A 5G8D/ 76G8 B C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C 4 4 C C C C C C C C C C C C C C C C C C C C 4 BC C C C C C C C C C 8-A@=@87D G8 76@ 9AG9@A7@D G4 76@ 4GGA5 8-A@=@87D G8 76@ 9AG9@A7@D G4 76@ 4GGA5 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C A A C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C 4 4 A A 5G8 5G8 B BC C C C C C C C C C 9G8 7B11F@78- D ,@AE 59GA7B887 =8- 9A@+ B88= C C C C C C C C C C B C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C A B C C C C C C C C C C C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C 44GGA5 A 5G83 BC C C C C C C C C C C C C C C C C C C C *8 76@ 5BA))@7 G4 96BA5 C C C C C C C C C C A C C C C C C C C C C C C C C C C C C C C A 5B:@7:BF 9@87D. 9GFEGFD BA@ A@9A@D@87@= 1E B ,BA@7E 8 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C G4 :6@5:BF D1D7B88:@D B8= 76@A -AB=@D3 C C C C C C C C C C C C C C C C C C C C B C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C 26D A@D@BA:6 B5@= 7G :G59BA@ 76@ 7B1F@78-+ C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C A@F@,B887 9AG9@A7@D G4 B,,BFB1F@ 9GFEGFD3 B C C C C C C C C C C A C C C C C C C C C C C C C C C C C C C C B C C C C C C C C C C ()% $( (# %) + (@ 77A8 ' 7AGD@ %$#". %$!#. B8= %$! ; C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C G@77@ %AA@D. @86@5. 7 C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C %A 88:@/ 88 ' B C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C AB477 . H. H + 4 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C CAB477 33. 4 C C C C C C C C C CC C C C C C C C C C A@E@. @F-5/ DG5BF7 ' -BF@8* #. !. C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C ##. !#. "# B C C C C C C C C C C C C C C C C C C C C . 88= $+H% + C C C C C C C C C C C C C C C C C C C C C C C C C C C C C CHBFB7787 BC C C C C C C C C C 51.C C C C C C C C C C @8. @A5 4 4 C C C C C C C C C C A 5B8E/ 5B8887GF ' C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C $B88G-@5 9BF ( BB88= 1E H* C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C H6BA5 + 3 9 C C C C C C C C C C ( ( 9AG4F@D G4 9GFEGFD !3 C C C C C C C C C C 4 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C A 5B. C C C C C C C C C C A 88= B,,@8. $*. . B C C C C C C C C C C B C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C HBA77@:) A C C C C C C C C C C $ !## B88= $ ## C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C $@A:) B. (BA5D7B=7. A C C C C C C C C C C B . C C C C C C C C C C .C C C C C C C C C C (444@@A@8:@D (444@@A@8:@D G4 9BA7:F@ D6 99@ B88= D 4 4 4 4 C C C C C C C C C C G4 9BA7:F@ D6 99@ B88= D C C C C C C C C C C A A C C C C C C C C C C B B C C C C C C C C C C B B C C C C C C C C C C @ ?@A@ @ ?@A@ C C C C C C C C C CC C C C C C C C C C @A5 A 5 88E. H@BAF7GF ## ( B C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C G@777@ 7 C C C C C C C C C C %AA@D. C C C C C C C C C C G1D@A,,@= 1E G97:BF 5:AGD:G9E 3 A C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C ,@AEE A C C C C C C C C C C @86@5. C C C C C C C C C C %A 88:@/ G B C C C C C C C C C C F-G4AA:7GD@ ' AB 4 A C C C C C C C C C C C C C C C C C C C C B477 H 4 C C C C C C C C C C C C C C C C C C C C-AB=@ G4 B,,BFB11F@ 9GFEGFD =@5G8D7A C C C C C C C C C C C C C C C C C C C C B B C C C C C C C C C C C C C C C C C C C C AB77@= B B C C C C C C C C C C C C C C C C C C C C + AB477 33. A@E@. @F-5 4 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C 5/ H C C C C C C C C C C C C C C C C C C C C D9@:4: H( 9AG4F@ 4 C C C C C C C C C C C C C C C C C C C C 4 C C C C C C C C C C . ?6:6 C C C C C C C C C C C C C C C C C C C C :B88 1@ B C C C C C C C C C C C C C C C C C C C C ### ' GFFDGF, H ### C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C % . C C C C C C C C C C C C C C C C C C C C =@D:A1@= 8 7@A5 C C C C C C C C C C C C C C C C C C C C A 5D G4 ( C C C C C C C C C C C C C C C C C C C C !# . ( C C C C C C C C C C # . 88= ( C C C C C C C C C C B C C C C C C C C C C # C C C C C C C C C C 3 =?-D6B4@@8. 4 C C C C C C C C C C @A5B88E/ D A B C C C C C C C C C C GA117GF ' HB C C C C C C C C C C C C C C C C C C C C AA77@:) * C C C C C C C C C C C C C C C C C C C C ! #. * ##. B88= * ## $@A:) C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C B C C C C C C C C C C C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C B B .C C C C C C C C C C HGFEGFD 6 ,,@ =444@@A@87 7A@ =@8D7@ C C C C C C C C C C B C C C C C C C C C C 4 4 C C C C C C C C C C C C C C C C C C C C @D3 %GA C C C C C C C C C CC C C C C C C C C C (BA5 A 5D7B=7.C C C C C C C C C C@A5 A 5 88E/ ;EF7GF ' EFDGA1 B C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C 1 # BB88= C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C @; 5 B 59F@ =@;7A8. ;EF7GF. DGA117GF. 5 C C C C C C C C C C ; C C C C C C C C C C C C C C C C C C C C A C C C C C C C C C CB887GF.C C C C C C C C C C ## C C C C C C C C C CG@777@ %AA@D. @86@5. % 7 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C %AB8:@.C C C C C C C C C C DG5BF7. 88. B88= D:AGD@ 6B,,@ !3 C C C C C C C C C C C C C C C C C C C C B C C C C C C C C C C C C C C C C C C C C B C C C C C C C C C C . !3 . C C C C C C C C C CC C C C C C C C C C ;EF7GF C C C C C C C C C C -5B C C C C C C C C C C F=A:6$@A:) C C C C C C C C C C B B . C C C C C C C C C C !3 !. !3 !. ! #. !3 . B8= !3"# -:5 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C 53 2A@ C C C C C C C C C C AC C C C C C C C C C (BA5 A 5D7B=7.C C C C C C C C C C @A5 A 5B88E/ D:AGD@ B B C C C C C C C C C C C C C C C C C C C C $ ;;5 5B *873 C C C C C C C C C CC C C C C C C C C C =@8D7E. 7G-@76@A ?766 DGF= D7B77@. 9BA77:F@ C C C C C C C C C C C C C C C C C C C C 7 C C C C C C C C C C C C C C C C C C C C B C C C C C C C C C C A @ D6B9@. C C C C C C C C C CC C C C C C C C C C G8-. F8766B3 7 B 9BA77:F@ AC C C C C C C C C C D@.C C C C C C C C C C B8=C C C C C C C C C C H(C C C C C C C C C C 84FF@8:@ 4 @DC C C C C C C C C C 76@C C C C C C C C C C =@8D4:B77G8 9AG4F@D G4 9GFEGFD 3 2 4 B C C C C C C C C C C 4 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C 26@ FBD7 C C C C C C C C C CC C C C C C C C C C HG?=@A + G8@ : 88 1@ 97G8 G4 C C C C C C C C C C B C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C 4 C C C C C C C C C C 7 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C 9G?=@AC C C C C C C C C C C C C C C C C C C C A EE (444AAB:7G8 9( !. B C C C C C C C C C C 4 4 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C G97:BF C C C C C C C C C C 5:AGD:G9E . 9 A 9AG9@A77@D 9G8 5;8-. :G8,@E8-A C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C-. B8= C C C C C C C C C CC C C C C C C C C C C C C C C C C C C C C C C C C C C C C C BA77:F@ D@ =D7A17G8 C C C C C C C C C C C C C C C C C C C C 8 H( C C C C C C C C C C C C C C C C C C C C . = 7B1F@78- 9A@:G59A@DDG83 C C C C C C C C C C C C C C C C C C C C @8D4:B77G8 . FGDD G8 =AAEE8- G 4 B C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C G( . C C C C C C C C C C C C C C C C C C C C 5@ 88 E@F= 9A@DD8 AB77@ C C C C C C C C C C BC C C C C C C C C C D@8D7,7E "3 C C C C C C C C C C C C C C C C C C C C %) (# $$# (@ 77A8 7AGD@ %$#" ; C C C C C C C C C C C C C C C C C C C C %$!#. B8= %$! . C C C C C C C C C C C C C C C C C C C CC C C C C C C C C C 88 AB477 B8 C C C C C C C C C C 4 C C C C C C C C C C C C C C C C C C C C= H. B88= GF-G4AA:7GD@ C C C C C C C C C C C C C C C C C C C C B C C C C C C C C C C 4 AC C C C C C C C C C 132 :1(A ?:@?:9:1-:*A "-)A *==:?:1-A +):0+/;A +0@.1*9A /1*A *==:?:1-A +00:?+ /2/;/;:A(?/*:9,A/A9@:+=+A(?/*:A=A@;8;9A+/1A :A +)9:1A =?A -):A=?0.;/-1A =A 1.-?/+:.-+/;A /1*A @)/?0/+:.-+/;A @?*.+-9A /9:*A 1A -):A *:9;*A 9-/-:,A @/?-+;:A 9:,A DCB,A *:19=+/-1A/1*A=;"/;-8A@?@:19-8,A/9A":;;A /9A 0:/1A 8:;*A @?:99.?:A /1*A 9-?/1A ?/-:A 9:19-2-84 1. ?2-),A44,A:-A /;4,A Dataset: Powder X- ray Diffraction (pXRD) Profiles of Polyols. 2025.01: Riga Stradins University, Riga, Latvia; DOI: 10.48510/FK2/TDBOKG. 2. Kukuls, K., et al., Dataset: Optical microscopy of polyols. 2025.01: Riga DCBA@?=;:9A=A@;8;9A7654 Stradins University, Riga, Latvia; DOI: 32:1A 1A 90/;;A /0.1-9,A 09-.?:A +1-:1-A +/1A 10.48510/FK2/RBPONG. 3. Kukuls, K., et al., Dataset: Particle size *?/9-+/;;8A+)/1(:A-):A0:+)/1+/;A@?@:?-:9A=A distribution (PSD) profiles of polyols . :'+@:1-9A.@1A-/;:-1(A7%54A$).9,A-):A#BA=A 2025.01: Riga Stradins University, Riga, +00:?+;8;9A"/9A*:-:?01:*A Latvia; DOI: 10.48510/FK2/GDGHY6. :=?:A-):1A1A-:?09A=AD8A/1*A 4. Frolova, A.J., et al., Dataset: Densification C!C4A 2:?/(:AD8A2/;.:9A=A@;8;9A":?:A=.1*A Profiles of Polyols. 2025.01: Riga Stradins 1A-):A?/1(:A=A6 D/,A");:A-):AC!CA":?:A University, Riga, Latvia; DOI: :-"::1AA/1*A6 10.48510/FK2/SLQKI9. A A754A-A0:/19A-)/-A *:=?0/-1A :)/2.?A =A @;8;9A .@1A drying (LoD) of polyols. 2025.01: Riga Stradins University, Riga, Latvia; DOI: 1+?:/91(A-):A-/;:-1(A9@::*,A-):A*==:?:1+:9A1A 5. Frolova, A.J., et al., Dataset: Loss on -/;:-1(A ";;A 1+?:/9:A 1A /++?*/1+:A "-)A -):A C!CA1+?:/9:4 6. 10.48510/FK2/XILW3Z. Kukuls, K., et al., Dataset: Mean yield pressure (Py) and strain rate sensitivity (SRS) of polyols. 2025.01: Riga Stradins University, Riga, Latvia; DOI: 10.48510/FK2/KMWDO6. 7. Paulausks, A., T. Kolisnyk, and V. Mohylyuk, The Increase in the Plasticity of Loaded with a Plasticizer. Pharmaceutics, 2024. 16(7), DOI: 10.3390/pharmaceutics16070945. $):A@?:+-A1-:?1/;A+19;*/-1A=A!CA/1*A :'-:?1/;A+19;*/-1A=A!CA"-)A#CD A4A 44444 # A(?/1-AC.-/;-8A =A9.(/?A/;+);9A@;8;9A/9A1*:?9A1A-"1 9+?:"A0:;-A(?/1.;/-1A=?A@?:@/?/-1A=A)() *?.(;/*:*A 00:* 9.@:??A 0:+)/1+/;A @?@:?-:9A 4A !C D A 9A =1/1+:*A "-)1A -):A =?/0:"?A =A -):A 3.?@:/1A 11A !:+2:?8A D8A/-A9;"A-/;:-1(A9@::*A294AC!CA754 /1*A!:9;:1+:A@;/1A/1*A-):A9-/-:A.*(:-4 133 2101 /.-,+*)('.%)$%#")-)#(!-' )('.%./%*'$.#)'!*.)$!$% ,'#-.!,+*'.%!* Patricia Jackuliakova1 1 1 1,2 , Miroslava Spaglova 2%# , Juraj Piestansky , 1 Comenius University Bratislava, Faculty of Pharmacy, Department of Galenic Pharmacy, Slovak Republic 2 Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava, Slovak Republic 1. INTRODUCTION 9= 1B3/:599!39#?##.3'593+=?(?=4# BA@?=;:9A87659=43 2=:93 1B0/9.3 5?=3 54-58@=43 ,+A@5:3 4?;23 4=:A-=?*3 9*9,=93 ,)5,3 (%%3")=3@+9A,A83(3B0/9 @6A8=3 ,)=3 6=8=(A,93 (3 =;:9A897 8589@5:=3 4?+:=,93 'A,)3 )A2)3 4?;27:54A823 @5+5@A,*7'A,)3 ,)=3 9,?;@,;?5:3 9,56A:A,*3 (3 2=:93 %$# ")=9=3(?;:5,A893,*+A@5::*3@89A9,3(3A:!3 '5,=?!3 9;?(5@,58,9 3 @79;?(5@,58,!3 5843 53 2=::A823 52=8,3$#3")=*3=8)58@=39A83+=?=5,A83')A:=3 5A8,5A8A82354)=9A83,3,)=34=?5:39;?(5@=#3?3 :A4@5A8=31.!353'A4=:*3;9=43:@5:358=9,)=,A@!3 B0/93 +?-A4=3 9;9,5A8=43 ?=:=59=!3 ?=4;@=43 9*9,=-=43 +5,A=8,3 @+:A58@=# 0%(% %%,!% 0%,)(!-')*%)$%,!(".$ ?3 @+5?5,A-=3 =-5:;5,A83 (3 :A6=?5,A83 (?3 2=:93 5843 B0/9!3 ?=(=?=8@=3 2=:93 'A,)3 9A 0%,% @+9A,A83593B0/9!36;,3'A,);,3,)=3A:3+)59=3 :=A@35@A431=8,?5:@)=+?+*:3*?A9,5,=31A257:4?A@)!30.3593583 :=A@3+)59=!34A9,A::=43'5,=?35843339:;,A83(3 %-!+*(%)$%$'#+'. :5=?3 3 1A257:4?A@)!3 ,#3 ;A9!3 %2 % .3 593 53 '5,=?3 +)59=!3 :*9?65,=3 3 ")=3 ?)=:2A@5:3 =59;?==8,93 '=?=3 ?=5:A=43 1=8,?5:@)=+)?3 ;84=?3 3 5843 (?3 (?;:5,A83 B0/%3 5843 31!3;4'A29)5(=8!30.!3:5=?3 B0/3 A,3 9)'=43 ,)A,?+A@3 6=)5-A?#3 ?3 %3 1B=?@!3 5?9,54,!3 0.3 593 53 9;?(5@,58,!3 (?;:5,A83 B0/3 2?5+)A@5:3 ?=+?=9=8,5,A83 "?589@;,:3A257:4?A@)!3,#3;A9!3.335843 9)'=43 ?)=+=*3 1A2#3 %.#3 "=,;?=3 585:*9A93 =,)58:31=8,?5:@)=7 4=89,?5,=43 ,)5,3 544A,A83 (3 3 4=93 8,3 9;?(5,58,!3 5?6+:3 3 1")=?3 9@A=8,A(A@!3 @)582=3,)=39,?;@,;?=3(3B0/!3=@=+,3(?3B0/!3 /==:!3 0.3 593 53 2=::A823 9;69,58@=!3 :A4@5A8=3 ')=?=3 ,)=3 544A,A83 (3 3 9A28A(A@58,:*3 1 B!3B=:9;82=830.3593,)=35@,A-=3 A8@?=59=43,)=38=25,A-=35?=5356-=3,)=3@;?-=3A83 +)5?5@=;,A@5:3A82?=4A=8,# ,)=3 (A?9,3 =59;?==8,3 @*@:=3 @??=9+84A823 ,3 ,)=354)=9A-=8=993(3,)=395+:=31A2#3.# 00%2%% % BA@?=;:9A893 '=?=3 +?=+5?=43 6*3 53 +)59=3 ,A,?5,A8#33 '5,=?3 +)59=3 '593 ,A,?5,=43 A83 583 A:3 +)59=3 @8,5A8A823 5:93 9;?(5@,58,3 5843 @7 9;?(5@,58,3 19==3 A83 "56:=3 %.#3 83 ,)=3 +?@=993 (3 +?=+5?A823 =;:9A89!3 ,)=?=3 '593 8,A@=43 9=:(72=::A823 +?@=99!3 93 A83 ,)A93 @59=3 83 ,)=?3 2=::A82352=8,3)5-=3,36=3;9=4#3?3,)=3+)*9A@5:3 @)5?5@,=?A5,A83 (3 ,)=3 B0/9!3 ?)==,?*3 18,8355?!3)=:56.!35843,=,;?=3585:*9A93 1,56:=3 BA@?3 *9,=3"7"3 +:;9.3 '593 ;9=4#3 44A,A85::*!334?;23?=:=59=3,=9,3,)?;2)3 134 +)85)9 699539 (:5 (:9 0/. 18864 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 6 45(()8 5 ( 3:8 538 $9 8(9 (:5(( 8 9(9(5$$ 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5 $9 67 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 62 2 2 2 2 2 2 2 2 2 2 2 2 2 2 699539 *53 8$3969 16684 (:9 84565 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5()9 79 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 81 18864 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 6 45(()8 , 6)7 (:9 :86 3(% 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 296)82 2 2 2 2 2 2 2 2 2 2 2 2 2 2 (:96918869! )(3 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 3 5((5 569 8( )9 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 %%$ $ $ $ $ $ $ $ $ $ $ $ $ $ $ #"! ! 699539 3(% 16684 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ( 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 4 0/.3 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 # $ $ $ $ $ $ $ $ $ $ $ $ $ $ $7932 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ; ; ;::9876543 81 0/.3- 0/. 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ., +*)(:: 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 (2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 '8%386$$5(9 62 2 2 2 2 2 2 2 2 2 2 2 2 2 2 #"! 769% ! 0/. +*)(:: 69 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ( 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 948:862 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ; ; "! $9 ! 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 20/. +*)(:: '885496 ,##! 76 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ( 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 699 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 :9 3(% 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 6939(3 5 )85(()9 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 52 2 2 2 2 2 2 2 2 2 2 2 2 2 2 964 6 452 2 2 2 2 2 2 2 2 2 2 2 2 2 2 118864 118864 6 6 45(()8 45(()8 5 52 2 2 2 2 2 2 2 2 2 2 2 2 2 2 11886 54))3(65()8 81 11886 54))3(65()8 81 :9 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 :9 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 69569 1886 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 664 45(()83 3:8*9 (%)5 :%3)5 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 :565(96)3()3 1886 793! *:): ) 8( :579 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 99 51((96 ) 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 )86865(()8 81 )()8 81 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ) 8( 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ( 511199( (9(69 81 69569 0/.3 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 3)7)1))5(% 1 5 % :9 3(% 81))649 (:5(( 0/.3 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 45% )468 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 92 2 2 2 2 2 2 2 2 2 2 2 2 2 2 "% #"! $ $ $ $ $ $ $ $ $ $ $ $ $ $ $2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 67 699539 16684 (:9 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 83579 18864 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 6 4 4 ,2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 56%%5$! 62 2 2 2 2 2 2 2 2 2 2 2 2 2 2 0!2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 9(2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5!2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 !"% $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ %$ $ $ $ $ $ $ $ $ $ $ $ $ $ $ ! 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( 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 )199 $% (::9 5)()8 81 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ( 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 %% #"! $ $ $ $ $ $ $ $ $ $ $ $ $ $ $2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 67 699539 3()93 6 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 9483(65((9 52 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5 :)3 693956 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 6: *53 19 $% 5(% 81 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 )956 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 699539 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 681))9 896 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 :863 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 8456)38 81 (:9 (8(5 67 699539 51((96 ':564 6 45%!2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 849)3 )963)(% 65()3 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5 52 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 6 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 :863 +)7 ))5((93 (:5(( 5,"2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 552 2 2 2 2 2 2 2 2 2 2 2 2 2 2 85 594% 81 )993 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5 0/. 18864 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 6 45(()83 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 8(5))7 '8%386$$5((9 #" 86 6948:86 /. ,"" 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 " 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 6 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ;" 5:)99 7695((96 67 699539 (:5 (:9)6 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 6 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 866938)7 84565(()9 793 8(653(! 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 5 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 22 2 2 2 2 2 2 2 2 2 2 2 2 2 2 11886 88 4 5 496$5392 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 188645(()83! 52 2 2 2 2 2 2 2 2 2 2 2 2 2 2 (:92 2 2 2 2 2 2 2 2 2 2 2 2 2 2 84565(()9 52 2 2 2 2 2 2 2 2 2 2 2 2 2 2 792 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 155))(5((9 52 2 2 2 2 2 2 2 2 2 2 2 2 2 2 :)7:962 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 135 .-,+ *)(')(%$#"! ('*$)""%$)%#(' '$ '#').%'*#.#* $$%(' %'*##('% *'*#. $$%#(' *%.% ($')'*)$ '$'%' )"#%.%( ' $") Emilia Jakubowska1,2 1 , ' 1 Department of Chemical Engineering, University of Chemistry and Technology Prague, Czech Republic 2 Chair and Department of Pharmaceutical Technology, Poznan University of Medical Sciences, Poland 1. INTRODUCTION '+'16.6=8;;+.=707=,68;=7;0,+# ?= ;:9876;658 4:53 29;1 ;5 158;680507 Excipient Grades 3980491;0:68/ 68 .9:391=0;619- 68,07;:+ 97 (suppliers) 2==868;=876*=-+,671077=,68:=1=8;+=9:7)(6; MCC PH 101,102,301, 301 45107 58 158;680507 ,6:=1; 153.:=77658 (Asahi Kasei, JRS 302, 12; UF 702,711; '%$# "5(=*=:) 398+Pharma, Chemfield) KG802,1000 ;67 .:51=77 ,0= ;5 .55: .5(,=: 4-5( 98,5: Low-substituted L-HPC: LH-11, LH-153.:=77658 .:5.=:;6=7# ?=6: HPC (Shin-Etsu) 21, NBD-021 3980491;0:926-6;+ 198 2= 63.:5*=, 2+ 15 Pre-gelatinized Lycatab: C, CT .:51=7768/ '% $ (6; =16.6=8;7) :=70-;68/ 68 starch (Roquette ) =8/68==:=,153.576;=.9:;61-=7(612=9*=68 Lactose Lactopress 9 7636-9: (9+ ;5 20- =16.6=8; # (monohydrate spray Granulated; %:+7;9--679;658.:51=77=79:=9.:536768/9:=9;5 dried or SuperTab: 11SD, 2= =.-5:=, 45: ;67 .0:.57=# =-+68/ 58 ;= agglomerated; 22AN, 24AN 30GR; .=853=85854;=3.-9;=,=;=:5801-=9;65858 lactose anhydrous) Sheffield: Spray 975-6,70:491=);=+9:=,=76/8=,;5.:535;=;= ( DFE Pharma, dried Foremost 315, 1:+7;9--679;658 54 98Kerry) Spray dried FastFlo .9:;61-=7) 97 5..57=, ;5 4:== 20- 1:+7;9-7# 5 316, Anhydrous 49:) 15.:51=7768/ 7;0,6=7 =3.-5+68/ 155-68/ 60M, Anhydrous 1:+7;9--679;658(6;9-636;=,:98/=54=16.6=8;7 Impalpable 9*=2==8,=71:62=,'=#/#$# Mannitol Granutol: F, R, S (granulated, spray-?=96354;6771:==868/(5:(97;5;=7;;= Pearlitol: 100SD, 200 dried) 4=97626-6;+5498;675-*=8;1:+7;9--679;65845:15 SD (Freund, Roquette) .:51=7768/ 93-5,6.68= 2=7+-9;= 97 35,=-(SPI (6;9(6,=*9:6=;+54153358%=16.6=8;7 Advantose 100 Pharma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ario Jug1 1 2 2 #-6"! -; #-4-! #-. -) # "! !!!!! ! 2Chemistry Department, University of Zagreb Faculty of Science, Croatia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ample Grinding RDC time DE (%) (min) ? ;9,7%? 9;%7(? 95? -=7;:7)*)7(? CNZ - 100 - A)*%)60?#" ?? CNZ/HPCD 80???;70?-0? 120 9.21 10.91 GR 20 Hz CNZ/HP CD ? 40 0 22.13 -=7;:7)*)60?#"?"?? GR 30 Hz -;9(9%8$? 8;*%$*%8? 7? (7$? 5;%)*6? ,;9*$6? =*8=? %;8+? *%,? 9? =;7$? *%7%)*7(? 6,,9;? 5;9:? = 6+6:0? ,9%*7((+? ;*88;*%8? = $8;7$7*9%?.9;0?=*%? ==+? 95? 378;?95?=*7%?A)*%)*9%?7%$? *%? = 7;(+?7)%9($8$ $:9%6;7*%8?%9?$;8?$8;7$7*9%?*;;6,)* 95?=;6?:,(9+$? 139 '%$ #"! "!" '' "! " ' !" #"' '#! !' " "!'' Paula Kaufelde1, Rita Maria Soares2, Oxana Brante1, Konstantins Logviss1 10/.-,/+-,*)-()'%$#"!) -$/")'-,$)'/+*)-()#/,/*)/)+,/!)%",$+*)0/+/ '/+*)-()#/,/*)%",$!/!")!")0$.-/)''0)-,+/ 1. INTRODUCTION /.8)7= ;0= 7B8CD= (D= D-7D49= 2?6D,= '686D7:=82,=*?@?23=6.D*=*82)8CC4 EDCDBA@?= ?6A-9= ?7= 8= 2A276D-A?,8C= 826?1 Flowability: Tapped density, Hausner ratio ?2+C8**86A-4= ,-)3= ('%$:= BC877?+?D,= )2,D-= (HR), and Carr Index (CI) were determined 6.D= #?A".8-*8BD)6?B7= EC877?+?B86?A2= '476D*= using a 100 ml graduated cylinder with the #E':= EC877= %%= ! = $)D= 6A= ?67= "AA-= 8)DA)7= USP2 method at a 250 taps/min speed using 7AC)?C?649= 76-86D3?D7= 7)B.= 87= 8*A-".A)7= 7AC?,= tapped density tester Erweka SVM II (Langen, ,?7"D-7?A27='$7:=.85D=DD2=?,DC4=8,A"6D,= Germany). 8""-A@?*86DC4= = A+= *8-D6D,= "-A,)B67= Preparation of ASD: The Vacuum Compression ?2= 6.D= ".8-*8BD)6?B8C= ?2,)76-4= (A68C49= -D)?-?23=7AC)?C?64=D2.82BD*D26=D6DD2=0= Molding (VCM Essentials) type M-01-04-1 (MeltPrep GmbH, Graz, Austria) was used for 82,=00=)6?C?D,='$=6DB.2ACA34=!0= the preparation of the ASD samples. 0.75 g of *A23= 6.D= 58-?A)7= *D6.A,7= +A-= '$= the PMs previously prepared were placed into a "-A,)B6?A29= .A61*DC6= D@6-)7?A2= := ?7= 25 mm diameter sample holder. The samples "8-6?B)C8-C4=8,582683DA)7=,)D=6A=?67=7AC5D261+-DD= were heated 86= = E= +A-= 0= *?2)6D7= 86= 6.D= 286)-D9= 7B8C8?C?649= D++?B?D26= ,-)31"AC4*D-= vacuum, followed by cooling, obtaining *?@?239= 82,= 8?C?64= 6A= D2.82BD= ,-)3= 7AC)?C?64= transparent glasslike discs with well-defined 82,= 768?C?64= ?2= 8= BA26?2)A)7= 82,= -D"-A,)B?CD= geometry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ot melt extrusion (HME): Co-rotating twin- screw extrusion was carried out using a Thermo % Fisher Pharma 11 extruder (Thermo Electron .D= *86D-?8C7= )7D,= ?2= 6.?7= 76),4= ?2BC),D,= GmbH, Karlsruhe, Germany). The extruder was equipped with a volumetric feeder (feeding rate ""-. = 86B.= E9= #$= .8-*86DB.= 0.3 g/min) and with conveying screws and 6,9=DC8238289=%2,?8:=?6.=0=")-?64=82,= mixing elements, the rotation speed was set to ) = 86B.= =-"*=-ABD77?23=6D*"D-86)-D7=D-D= == 9= '= .8-*89= A7D2D-39= in all seven zones, except for the first zone at 60 D-*824: = .D= 6A-)D= ,?,= 2A6= D@BDD,= = A+= 6.D= %% *8@?*)*= 8CCAD,= = (*:= .D= +?-76= = 3= A+= extrudates were discarded. ,"/,/+-%)-()#*$/)+,"$)=7=A+= EE;= 82,= = D-D= "-D"8-D,= 4= ?2,?5?,)8CC4=D?3.?23=D8B.=BA*"A)2,=)7?23=82= $ #"!!#" 140 .-,+ HHG EDCBA@?=;: 9A8?88 A7 CCB;C6 = 2 8 (3003 F F F F F F F F F F F F 3F F F F F F F F F F F F "F F F F F F F F F F F F (2=8' F +*)* '%$#"!! ( ( ( ( ( ( ( ( ( ( ( ( .F F F F F F F F F F F F 9D::8=;84 35CDC=8 2120 F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F 85 EECBF F F F F F F F F F F F ' (H' 8 F F F F F F F F F F F F ! 8 F F F F F F F F F F F F F F F F F F F F F F F F D F F F F F F F F F F F F F F F F F F F F F F F F 1 1/ / E=D9;: DA,BCB @ADEE5A;A+:4 ,;?*C 5C /2) F F F F F F F F F F F F F F F F F F F F F F F F D F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F FF F F F F F F F F F F F ! ! (B== A 85A3' %?+5CD /2) 9A DC8,; == ! ! ! ? ?9DC=8CB %$ =B H) #=;,C8 ("==--;C !'4 F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F " = F F F F F F F F F F F FF F F F F F F F F F F F = F F F F F F F F F F F F CCB;C6;?*CF F F F F F F F F F F F E=D @ADE5A;A+:F F F F F F F F F F F F A7F F F F F F F F F F F F HHG /;; F F F F F F F F F F F FF F F F F F F F F F F F 5DCCF F F F F F F F F F F F 2 8F F F F F F F F F F F F +*+* % ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( !% ( ( ( ( ( ( ( ( ( ( ( ( BC@A8 "5CF F F F F F F F F F F F F F F F F F F F F F F F (HHG2121/ 1 /'F F F F F F F F F F F F 7 7AAD@,;= =F F F F F F F F F F F F ##" )* ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( (%$ =B H) A7 2 8 F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F F BC@A8 E=D @ F F F F F F F F F F F F F F F F F F F F F F F F . D F F F F F F F F F F F F = @C ! 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'"' " ! ! !! '%8 7,(5,!3-/1!, .#8$8% (03512.,#8 # 3/8 , ' $ %$#! %$%% $! % % ! (-48 3 13(#8 $8 8 1 73 5-.#8#83/, #8738(,10383938/3./8 -(3986'8*7/7, 56 6498$$8% 0,6".,+6.21#8#83/, #8 ! $ 143 ,+*) ('(%$#"#!, ( "%"$'( $'!"($(%"" ($#%" ( $$'",(( "#'!$'$'"!$("'!%!"% Gabriela Koutna1 1 2 3 3 , Katerina Kubova , Jan Kotoucek , Martina Urbanova , Larisa Janisova, Ivana Sedenkova3 1 1 1 1 3 , Jan Muselik , Jakub Vyslouzil , Miroslava Pavelkova , David Vetchy , Jiri Brus %$#"! $ "!"$ "$" "!"""!$! ! $$# %$#"! $ "!""$ $!"!$$"! $ $$# 3 Department of Structural analysis, Institute of Macromolecular Chemistry, Czech Academy of Sciences, CZ 1. 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(*).$)*- #- ) ! ("#$- )$) 2-"-$- ) !2 $ )$ #*$" (2. #--2*(- ) Agata Grzejdziak1 2 3 3 , Mateusz Kurek , Konstantinos Raftopoulos , Krzysztof Pielichowski, Aleksander Mendyk2 *)('(%$#"!' ((#"("'$#"$"$# "!''"$##($"%"'$#"(##"%$("(#$ $%"(" $%$''$#" ' (#("$"( $%$''"$'#"(" $%$'"$##($"%" '$#"(##"%$("(#$ )$%"(" %"$" ' (#("("(#%"$'#"(" '$#"%"$" ' (#("%$(" %"(" ' (#("%$("(#$ 1. 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(;FDD00D((*D5DC2@A5;D@9DD6D(C?D 23C4;D @9D ;FED =CD 256D 37@4E??D @5D ;FED 67@A58D5DCD7;?4FD/A1E72@E4AC7D ?;7A4;A7ED @9D ;FED 3@?C4@5C.@ED C58D 3C5E;C7:D=CD2DA?56D899E7E5;D;2E?D97@2DD 4@5972D;FEDC2@73F@A?D5C;A7ED@9D;FED37E3C7E8D 146 )(' ;:987678;543;8:2;1 0;6/.26-46;4426826,8+*346)16.996 2'382;8/6 78;;:9*'8:26 3.'4;6 #:36 5345.3476 5345.3476;(;'4;6%4346826.:35$:*;6#:31 #:3*9.'8:2;1 Intrinsic dissolution rate Formulation ( 2 Improvement +/cm /s) factor processed raw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INTRODUCTION 84)*9;;14/6456376*4'.9;6':;6.4/6-;1/(6:6 +:,49:3496#8 =;:9876 5496 84;325583106 :/.6 ;-;3:1/:,+6 *+:/32,:;.6 ):391:+;6 5496 *7:9):8-318:+6 4!#$22:,+36549)-+:314/684)*4;1314/61/6''6% :**+18:314/6 7:;6 (94'/6 *4*-+:96 4096 376 *:;36 :9;%6 $4/;3:/36 /.6 5496 1)*940)MCC MCCLAC SPC PuOC *7:9):8-318:+6 549)-+:314/6 :/.6 379:*-3186 D1 ad 100 - - - 4-384);6 7:;6 89:3.6 :6 /.6 5496 /40+6 D2 - ad 100 - - *7:9):8-318:+6 #81*1 76 :1)6 456 371;6 ;3-.6 ':;6 346 1/0;31(:36 376 ;;20#!# 22 $2$2 $ *4;;1,1+136 456 *948;;1/(6 ,14*4+)9;6 26 ;46 76 ,-+6 :/.6 3:**.6 . *9431/6 84/8)1#3-9;654963:,+31/(6'963;3.61/6:8849.:/86 .910:36 :/.6 *-)*1/6 41+6 8:6 -$ 6 376 '1376376!!-94*:/67:9):84*41:67%6-9%6 %$! 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DCBAA@?= :=A9==8 7A6=B @ @5D7BAAC8A4 96@= B ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; B 8 ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; DCBAA@?= A7 - "6@ @A00 71 +*) C83 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; A ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; 96=8 3=A=B5 ; ; ; ; ; ; ; ; ; B 5@8@82 C82= 71 B=D7=4 A=83=8?0 71 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ,33@A@78 71 +*) C883 *.() @8?B=C=3 C882= 71 *.() A7 : ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; := DB7?==3 C DCBAA 71 AC:=A ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; B ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; B ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; B ; ; ; ; ; ; ; ; ; B=D7= @8 177B5 1177B5 B 5.CAA@78 C; ; ; ; ; ; ; ; ; 1177B 3@B=?A ?75DB=@78 5=A673-; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;C ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;C ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 71 '))4 % 883 $ #"C:= ! - (:AC@8=3 AC: ; ; ; ; ; ; ; ; ; :=A 5=A A6= B=.@B=5=8A 71 A6= *6-; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;1 B5.CA@78 9@A6 6@26=B DB7D7BAA@78 C ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; B; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;C ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; " ; ; ; ; ; ; ; ; ; .B- 177B ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 1; ; ; ; ; ; ; ; ; .8@177B5 1 B 5@A00 71 5C 883 1BB@C:@@A0-A ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; C ; ; ; ; ; ; ; ; ; 1; ; ; ; ; ; ; ; ; ! ; ; ; ; ; ; ; ; ; *793=B 5@A..B= 179C:@@A00 =C.CA@78 ; ; ; ; ; ; ; ; ; A ; ; ; ; ; ; ; ; ; 1 A ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; .BAA6=B :C=3 78 A66= :./ C83 ACDDD=3 3=8@A@= ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; A ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; C ; ; ; ; ; ; ; ; ; D6CB5 B 5C?=.A@ @?C B7= 71 A6== :@7D705=B4 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; HR CI [%] Flowabi ilty 7DA@5C DB7D ; ; ; ; ; ; ; ; ; D7BAA@78 @8 A6= 177B5 B ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 1 B 5.CAA@784 C 9= C C ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; D1 1.40 28.43 poor A6=@B D7A=8A@ ; ; ; ; ; ; ; ; ; 8 @C @81.=8?= 78 A6= 3@@8A=2BCAA@784 ; ; ; ; ; ; ; ; ; 1 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 8 C; ; ; ; ; ; ; ; ; D2 1.65 39.23 verryy v ryy poor er 3@7.A@78 C . ; ; ; ; ; ; ; ; ; CC883 @A0 71 A6= D6CB5C?=.A@?C ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; D3 1.42 29.33 poor 37 D4 1.47 31.96 very poor D5 1.66 39.86 verryy v ryy er y poor D6 1.55 35.38 very p poor - CB?@C4 -; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;-4 =A C-4 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; " "!! ! ! ! :C=3 78 C82= 71 B=D7= ; ; ; ; ; ; ; ; ; *793=B 5@A..B= 179C:@@A00 = ; ; ; ; ; ; ; ; ; A ; ; ; ; ; ; ; ; ; 1 A ; ; ; ; ; ; ; ; ; C.CA@78; ; ; ; ; ; ; ; ; ! ; ; ; ; ; ; ; ; ;7.B8C 71 B.2 =@=B00 +?@=8?= C83 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; @ B ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ! ! ! ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; " "==?6877 04 2 ; ; ; ; ; ; ; ; ;- $! -; ; ; ; ; ; ; ; ; Flowabi ilty D1 ! *C83C4; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 4-= A C 4- ! ! ! ! ! ! ! !! 39.94 ffaair (aid not needed r d) D3 !!! ! !! !! ! 37.24 ffaair (aid not needed r d) ! C A = ; ; ; ; ; ; ; ; ; C 8 3 ; ; ; ; ; ; ; ; ; @ 7 5 C ; ; ; ; ; ; ; ; ; C C 7 B @ C A @ 7 8 4 ; ; ; ; ; ; ; ; ; D4 D2 ! 39.29 ffaair (aid not needed r d) 39.94 ffaair (aid not needed r d) -%%$ % %$ $! !- D5 44.90 pas abble (may hang saup) D6 A6; ; ; ; ; ; ; ; ;.B7D=C8; ; ; ; ; ; ; ; ;*6CB5 B 5C?7D7=@C4; ; ; ; ; ; ; ; ;)7.8?@; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 7 1 1.B7D=4; ; ; ; ; ; ; ; ; 52.76 poor (must agitatte, vib a brate) +ABC:7.B24; ; ; ; ; ; ; ; ;!-; ; ; ; ; ; ; ; ; =79C:@@A0 71 ; ; ; ; ; ; ; ; ; 1 1 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 1 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; D793=B; ; ; ; ; ; ; ; ; 5@AA.B=4 C AC::=A 177B5 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; C ; ; ; ; ; ; ; ; ; 1 B 5.CAA@78 5 C ; ; ; ; ; ; ; ; ; 5=A A6= ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; "6@ $.()7.'-%'0+*)+77*0/+'0.%)0%)$0!- # .1-)/.,0%+'0.%) $.(,"*+'0.%1 Agata Nowak1 2 1 1 , Monika Rojewska Janina Lulek , Marcin Skotnicki $#"! "M"!" " """ !" """! 2 Institute of Chemical Technology and Engineering, Poznan University of Technology, Poznan (Poland) 1. INTRODUCTION , IB48HJ??GKE M M M ,3M A7?9M )EBEM /GK=?4M 7.HJGKE=M6B7AM D3M*M7373M LKMJIHGFEMDCJBAJIE@HGIJ?MGKBE=GEKHM ;7?JK=93M GKM JM 87?G=M =78JEM 67BAM IJKM E5G8HM EGHCEBM GKM JM IB48HJ??GKEM 7BM JKM JA7BDC7@8M 8HJHE3M 2CEM 55),)#)1) JA7BDC7@8M67BAMG8MJMCGC1EKEB4M8HJHE0M=G66EBGKM LHHEADH8M )EBEM AJ=EM H7M 7.HJGKM I71JA7BDC7@8M 6B7AMGH8MIB48HJ??GKEMI7@KHEBDJBH8MGKMHCEM?JI/M76M L!::M JK=M :M I7A.GKJHG7K83M 27M ?7K1BJKEM A7?EI@?JBM 7B=EBM JK=M GKIBEJ8E=M 7.HJGKMJA7BDC7@8MJK=MI71JA7BDC7@8MAJHEBGJ?8M A7?EI@?JBM A7.G?GH43M LA7BDC7@8M =B@M GKMJM,,MA7?JBMBJHG70MJM!GKGMDBJ4M%B4EBM1(M 8@.8HJKIE8M JBEM GKIBEJ8GK?4M JDD?GE=M GKM HCEM =E8GKM76M87?G=MDCJBAJIE@HGIJ?MDB7=@IH8M=@EMH7M )J8M@8E=3MDBJ4M=B4GKM)J8MDEB67BAE=M@8GKMJKM HCEGBM 8@DEBG7BM 87?@.G?GH4M JK=M =G887?@HG7KM L;:MJK=ML;:1L;:M87?@HG7K8MGKMJM87?FEKHMAG5H@BEM ICJBJIHEBG8HGI8MI7ADJBE=MH7MIB48HJ??GKEMAJHEBGJ?8M 76MAEHCJK7?MJK=M=GIC?7B7AEHCJKEM,,0MFF9MJHMJM -,+3M2CEGBMAJGKM?GAGHJHG7KMG8MDC48GIJ?MGK8HJ.G?GH4M I7KIEKHBJHG7KM 76M ,M M JK=M JKM GKCEBEKHM HEK=EKI4M H7M BEIB48HJ??G*E3M 27M HEADEBJH@BEM76M $3M 8HJ.G?G*EM HCEM JA7BDC7@8M DCJ8E0M JM I71 JA7BDCG*JHG7KMDB7IE88MG8MEAD?74E=0M)CEBEGKMHCEM 5)')+)' + L;:M G8M I7A.GKE=M )GHCM EGHCEBM JKM E5IGDGEKHM 7BM 2LMI@BFE8M)EBEM7.HJGKE=M@8GKMJM2M(MM JK7HCEBM L;:3M :KHEBA7?EI@?JBM GKHEBJIHG7K8M 2JB8@8M M HCEM8HJ.G?G*JHG7KM76MHCEM848HEAM-(+3M2CBEEMA7=E?M D?JIE=MGKMJMIEBJAGIMIB@IG.?EMJK=MCEJHE=M6B7AMM , H7MM$MJHM, $AGK MBJHEM@K=EBMJMKGHB7EKM JIHGFEM DCJBAJIE@HGIJ?MGKBE=GEKH8M ME?.0MEBAJK493M, AM76MHCEM8JAD?EM)J8M HCEM JKHGD84IC7HGIM =B@M I?J88M 'M JAG8@?DBG=E0M 6?7)M76MMAAGK,3 8@?DGBG=E0MJK=MBG8DEBG=7KEM'M)EBEM8E?EIHE=M67BM 5) )1)/) HCEM8H@=4M=@EMH7MHCEGBM?7)M87?@.G?GH43M2CE8EML;:8M 1/ E5CG.GHM8GKG6GIJKHMD7HEKHGJ?M67BMHCEM=EFE?7DAEKHM %$M I@BFE8M )EBEM 7.HJGKE=M @8GKM JM %$M (,M 76M6G5E=1=78EMI7A.GKJHG7KM%$9M67BA@?JHG7K83M ;7?4AJM M 2CEM DBGAJB4M 7.#EIHGFEM )J8M H7M 7.HJGKM I71 0ME?.0MEBAJK493M( H7 MAM76M8JAD?EM)EBEM JA7BDC7@8M 848HEA8M @8GKM HCEM 8DBJ4M =B4GKM D?JIE=M GKM JM DGEBIE=M J?@AGKG@AM IB@IG.?EM JK=M HEICKG"@E3M 2CEM DB7#EIHM JGA8M H7M =EFE?7DM JM CEJHE=I77?E=M6B7AMMH7M, $MJHMJMBJHEM76M,M 8HBJHE4MH7MEK8@BEMHCEMDC48GI7ICEAGIJ?M8HJ.G?GH4M $AGK , 0M@K=EBMJMKGHB7EKM6?7)M76MMAAGK 1 76M HCEM BE8@?HGKM 8HB@IH@BE80M H7M GKFE8HGJHEM HCEGBM , 3 HCEBAJ?M.ECJFG7BMJK=MEFJ?@JHEM/GKEHGIMDB7IE88E8M JK=MH7MJ88E88M8@B6JIEM)EHHJ.G?GH43 5)7)!#) )7!( ;7)=EBM 1BJ4M =G66BJIH7BJA8M )EBEM 7.HJGKE=M @8GKM JM B@/EBM %(M ;LM 5),+'-(0+*1)+% ),-'3. 1 A.0MJB?8B@CE0MEBAJK493 5),) 5)) LAG8@?DBG=EM @B6JIEM DB7DEBHGE8M 76M HCEM A7=E?M AJHEBGJ?80M M 3M A7? , GKI?@=GKM )EHHJ.G?GH4M JK=M 8@B6JIEM 6BEEM EKEB40M 90M 8@?DGBG=EM M IB48HJ??GKE M M M ,3 , JBEM .EGKM GKFE8HGJHE=M 2CEHJM GH?EM ,,0M M A7? 9M JK=M BG8DEBG=7KEM :9 M 150 JIHGFEDICBGCA@?ED=E;I:E98=9:7FE6C H7G:8=7H7C:E4E;7E93A7=:EG=7E9BGCC7AE8:IC5E ;7E :7::IB7E A=9E H7;AE 21I5HGE 0/.FE -1,E JIHGFDICBGCA@? +I447=7C=IH7=*E 2+1)@E GCGB*:I:E 4E ;7E :9=G*(A=I7AE '%E GCAE 1$#E =7"7GB7AE;7E9=7:7C67E4E5BG::E=GC:ICEA8=IC5E ;7E4I=:=EGHI:8B9=IA7E2 '% 5EE!E /?EE/?/E)FE . 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iljana Makraduli1,2 1 1 1 , Maja Gavriloska , Sevda Sofronievska , Marina Bakalinova, Natasha Churevski1 1 1 1 , Katerina Zafirova Georgievska , Jasna Bosnjakovska , Angelina Zimbakova, Vesna Tegova3 *)('%$#"! $!#!$#('((%"(#)"!#$("$ $%# "($%(("((%((!# "!#$("$ $%# !$()(%($!#"()"!#$("$ 1. INTRODUCTION */('+-'*,!"*".'0''.)! CBA@ A?B?=@ B;@ =:9@ A:8?78695=684@ 3251=?91@ * 0* $$* $$* * * 832@=:9@?90548=B?@/B291@1@=:9@.584=-@18;9=-@ *$*%0,$#*$*$$$$* 832@ 9;;686@ B;@ 2?50@ A?B256=1-@ 866B7A41:92@ #1@811911793=@,81@5192@3@293=;30@6?=684@ ,=:@ +584=@ /@ *9103@ )+/*(@ 8AA?B86:@ 832@ A:8191@B;@=:9@7835;86=5?30@A?B6911@=:8=@:89@8@ '%@+$@)#"(-@'%@+!-@'%@+ @05294391@ -@ 103;683=@ 7A86=@ B3@ A?B256=@ 6?=684@ .584=@ @*?50@A?B256=@7835;86=5?30@1:B542@=89@=:9@ 8==?/5=91@)+1(@)C8/49@ (@ 6?=684@ A?B6911@ A:8191A8?879=9?1@ )1(@ 3=B@ 6B3129?8=B3@3@?908?21@=B@4994@B;@=:9?@7A86=@ -**?=684@A?B6911@A:8191@832@4994@B;@7A86=@ B3@ A?B256=@ 6?=684@ .584=@ 8==?/5=91@ )+1(@ B3@+1@B;@=:9@2?50@A?B256= 4932@:B7B0939=@832@6B3=93=@53;B?7=@8?9@ CQAs of Critical process phases B;@ A?78?@ 7AB?=8369-@ 91A96844@ ;B?@ 2?@ drug Film 730@832@2?96=@6B7A?911B3@"@ Mixing Tableting product coating 2*,-(/.,+'*,!"*(-)"' Dissolution low low medium 2*$* Assay low low low '@ 69==39@ 2:2?B6:4B?29@ )4B6:97@ '3251=?91-@'328(6A93=1@6?B1AB2B39@832@ Content 6BAB2B39@ )-@ 9?783(-@ 694454B19@ Dosage medium medium low 76?B6?1=84439@ 832@ : :8?78-@ 9?783(-@ 486=B19@ 7B3B:2?8=9@ uniformity )90049-@9?783(-@1468-@6B44B284@83:2?B51@ )8/B=-@ 94057(-@ =846@ 832@ ==8357@ 2B29@ 4=:B50:@ 69=?39@ :2?B6:4B?29@ /94B301@ =B@ )9?6-@ 9?783(-@ 786?B0B4@ @ )48?83=-@ @64811@ @):0:@1B45/4=@832@A9?798/4=(-@ 9?783(-@279=:6B39@)*B,@B?330-@(-@ =:9@6B3=93=@53;B?7=@832@53;B?7=@B;@=8/49=@ 78039157@1=98?8=9@)B1194783-@94057( 6B?91@78@/9@6?=684-@/968519@B;@=:9@4B,@2B19@ B;@'@832@=:9@2?@730@2?96=@6B7A?911B3@ 22*** ** C:9@0BB2@6B3=93=@53;B?7=@832@53;B?7=@B;@ *?@730@832@:B7B0938=B3-@=8/49=30@832@ =8/49=@ 6B?91@ 515844@ ?9154=@ 3@ 8@ 0BB2@ 6B3=93=@ ;47@6B8=30@:0: 1:98?@79?@)@"@ @48==-@ 53;B?7=@ B;@ =:9@ ;47@ 6B8=92@ =8/49=@ 5=-@ =:9@ 9?783(-@79? /49329?@)64BA@2=57/49?@ ;47@ 6B8=30@ A?B6911@ 78@ 3;459369@ =:9@ '@ @78-@'=84(-@=8/49=@A?911@)3A?911@*87B32@ 211B45=B3@;@=@1@3B=@A?BA9?4@A9?;B?792@ "@ @8391=-@?98=@?=83(-@;47@6B8=9?@) C@ @%-@?98=@?=83(@ 2*0*4$$*4$*%044$#* C:9@1@;B?@986:@6?=684@7835;86=5?30@A?B6911@ 2******* A:819@ ,9?9@ 293=;92@ 832@ 84591@ ;B?@ 986:@ *11B45=B3@ =91=-@ 118@ =91=-@ 3;B?7=@ B;@ A8?879=9?@,9?9@19=@)C8/49@"( 2B1809@53=1@#'-@8??@3@@ '@ A96=?BA:B=B79=9?@)8=@""@@"@37( -*2*9=@84591@;B?@986:@@ Process Critical Process Set Phase Parameters values 152 '%$ Dry mixing, Mixer speed/ rpm 150 start 5.5, middle 5.5, end 5.5 trituration Chopper speed/rpm 500 Duration/ minutes start 2.0, middle 2.0, end 2.0 10 Dry mixing Speed/ rpm 16 !)3,/)6,2$; 21; 6'8; (5,6,()3; %52(877; %')787; )7; before Duration/ minutes 10 /2$8;"9)*38;0;9)*38;;9)*38; ; lubrication Tableting Pre-compress./ mm min. 3.0 !"!9876;5874367;;;1,3-;(2)68/;6)*3;*)6('; Compression/ mm min. 1.0 Dissolution: min 80% (Q) /f. c. t. expressed Tool loading/ kN max. 18 as % of the declared content in 45 minutes Film coating Inlet air temp./;: C 53-58 start 95.71%, middle 97.33%, end 96.14% Outlet air temp./;: C 40-42 Flow rate/ ml/min. 150-300 Assay: 9.5 mg - 10.5 mg/f. c. t. or 95% - Pan speed/ rpm 4-7 1.8- 105% of the declared content Atomization p/ bar 2.2 start 97.51%, middle 97.90%, end 100.87% Spray pattern width 2.0-2.4 pressure/ bar Uniformity of dosage units: ((8%6)$(8;#)348;"! ;;;" #"#"!' !! start 4.5, middle 5.5, end 2.0 !#"!9876;5874367;21;0;/5.;-,+64587;*)6('87 "! Triturate 9'8; 7; (5,6,()3; %52(877; %')787; )$/; 7; Assay: 11.41% API (w/w) (10.84%-11.98%) 858;,/8$6,1,8/;,7;38#837;858;)778778/;9'8; top 10.95%, middle 11.04%, bottom 11.38% 6876;5874367;125;7;858;,6',$;6'8;7%8(,1,8/; top 10.97%, middle 11.02%, bottom 11.08% 3,-,67; 9'8; 74((877143; 65)/,6,2$)3; #)3,/)6,2$; "EudraLex, Vol. 4, GMP guideline) 21; (5,6,()3; top 11.51%, middle 11.69%, bottom 11.38% -)$41)(645,$;%52(877;%')787;21;86,5,,$;1,3-; (2)68/;6)*3867;;-; (2$1,5-7;6')6;7;)$/; Dry mixture before addition of lubricant 7;858;833;,/8$6,1,8/ Assay: 7.78% API (w/w) (7.397%-8.175 %) top 7.415%, middle 7.481%, bottom 7.653% "! top 7.464%, middle 7.592%, bottom 7.489% ; ')$; ; 86; )3+* )('%$#* "#* ! %* * * $ %( **'' ($%'*!($*! $; top 7.846%, middle 8.076%, bottom 7.906% $6833,8$6;')5-)(.;;;; %"! )5)/43,; ; 86; )3; $* * $ $* !"!9876;5874367;21;0;6)*386;(2587;*)6('87; (%%$#* %* ! * ! * " !* %$%'* Dissolution: min 80% (Q) /tabl. expressed as ('$%* '!* * '%'" )(8/2$,)$; % of the declared content in 45 minutes ')5-)(846,()3;43386,$;;;;; start 96.12%, middle 96.56%, end 97.28% #"!985;2576;;86;)3+* $'$%**)('%$#*"#* start 98.69%, middle 94.90%, end 96.30% ! %*)"*%% *%* ('$#*% ** !%%'* !($* %* $ * ( '* %* * start 93.23%, middle 94.96%, end 93.87% $ **'!*;9'85)%846,(;$$2#)6,2$;; 843)625.;(,8$(8;;"0 0 Assay: 9.5 mg - 10.5 mg/tabl. or 95% - 105% start 95.99%, middle 96.77%, end 99.77% of the declared content ;8+%5877;-.;7,$(858;5)6,64/8;62;-.;74%85,257; start 99.48%, middle 97.15%, end 97.08% )$/;(2338)487;125;6'8;74%%256;)$/;(22%85)6,2$ start 100.87%, middle 102.45%, end 102.28% Uniformity of dosage units: ((8%6)$(8;#)348;"! ;;;" start 4.5, middle 4.5, end 4.5 153 )('( %%$#"!# "))$"%!"%" $! "!#"! "%"!$% ) %!"%)$ $)!"!$%$$%(( !" ! Roza Markovski 1 1 2 3 , Anita Vasilevska , Malgorzata Strozyk , Chhanda Kapadia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ejan Mihajlovski1 1 1 2 , Grozdana Leshevska , Nena Smiljanovska , Irena Slaveska Spirevska, Kristina Grncharovska2 2Quality Control Department, Replek, North Macedonia 1. INTRODUCTION 6@??'6.-16?=1./=?%-+9?.5:.8?=:7"-77-(4+ -7-;0+?.@48-6*/# "9.:847?71?24 A@? =? :5 @43@2?55414?710 9/.4541-64:76?1@74,/?+-7=6@?:96484*-64:7:5 :9?.-6473 1:7=464:7+ 6: ?7@-71? 6@? 04?;=) A@?9.?9-.-64:7:56@?1./=??'6.-16%-+=:7? +?;?164460 -7= ,/-;460 :5 1-77-(4+ ?'6.-16+) -6 (0=4++:;4734647?6@-7:;47-.-64::5 6@?94;:6+1-;?-.?1./14-;+6?9+5:.+1-;473/96@? 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A#" +6.-47) A@?+-8?9.:1?=/.?%-+9?.5:.8?=%46@- 9.:=/1?=-66@??9;?9.:=/164:79;-76/+473 .-64: :5 A#" "./=? ?'6.-16 =4;/6?= 47 +/9?.1.4641-; " 6?1@7:;:30 :. 6@? ?6@-7:; A@? %476?.4*-64:7 -7= .:6-.0 9/.4541-64:7 -7= 1:71?76.-64:7 9.:1?++?+) ?-9:.-64:7 9.:1?++?+ %?.? 9?.5:.8?= 47 6@? -7-;0641-;23.-=? ?6@-7:; $ %-+ /64;4*?= +-8? 8-77?. -+ 6@? 9.?4:/+ 9.:(? 7 6@? A@? %476?.4*-64:7 9.:1?=/.? %-+ 1-..4?= :/6 54;6.-64:79.:1?++6@?:7;0=455?.?71?%-+/+473 /+473 - / 8:=?; .?5.43?.-6:. 6@.??;-0?.+:554;6.-64:79-9?.))-7=8 4;6.-64:7:56@? 6 6@? ?7= :5 6@? 9.:1?++) +-89;?+ %?.? ?'6.-16%-+9?.5:.8?=/+473- @43@2?55414?710 54;6.-64:7 8-1@47? 8:=?; 1:;;?16?=-7=6@?79.:9?.;0-7-;0*?= 9:;;:+64;; 8-7/5-16/.?= (0 :. 6@? 54;6.-64:79.:1?++%-+/+?==?=41-6?=54;6?.9-9?. 5:.9:;;:+64;;%46@-.04739:.?+4*?+$8) 8) -7= 8 ==464:7-;;0) 6@? +:; %-+ .?8: /+473 - 6?.:3;-++ .; .:6-.0 ?-9:.-6:. $8:=?; 6.4?A@? -7-;0+4+ :5 156 $#"! " '/30.(*612 +.12'34(6112."'346120+62/*.0#2452 765436210/30.-,20+625.*03/0.4-2/-)2(4-(6-03/0.4-2 (4"'4-)12*.62!/%61$2*.'.)12/-)25/0122 0+62 5.-/*2 '34)(02 #2 36"4.-,2 -!/-06)2 '34(6116124520+62(3)626%03/(0$2/-2/-/*#1.124520+62 .-.0./*2 1/"'*612 !/12 '63543"6)2 /-)2 0+62 -40+63236/14-254320+62+.,+632'3616-(624525/012.-2 '63(6-0/,62 452 $2 7$2 /-)2 72 !/12 0+62 1/"'*62 "/#2 62 0+62 ).*0.4-2 !.0+2 60+/-4*2 )6063".-6)2+62361*012/3621+4!-2.-20+620/*6 +621/"'*60414*6-023/0.42.12/26#2'/3/"60632 .-5*6-(.-,2 0+62 655.(.6-(#2 452 (4*)2 60+/-4*2 ! -.0./*2(/--/.-4.)2(4-06-02452(3)626%03/(0 5.*03/0.4-2.-2(/--/.12'34(611.-,22(4"'/3.14-2 Cannabis extract THC- Mataro Blue 60!66-2 2 /-)2 2 3/0.412 366/*6)2 0+/02 0+62 66.08% THC +.,+632 14*6-02 4*"62 2 .-(36/16)2 0+62 Obtained results (4-(6-03/0.4-2 452 2 )).0.4-/**#$2 0+62 2 2.84% CBD 3.20% CBN 3/0.42 361*06)2 .-2 ."'346)2 '3.5.(/0.4-$2 *.6*#2 506320+62340/3#26/'43/0.4-2'34(611$2 534"20+62 )62042"43626556(0.62'6-603/0.4-24520+6214*6-02 5.3102'346223/0.420+62/"4-02452260+/-4*2 .-042 0+62 6%03/(02 '/30.(*612 /-)2 600632 /506325.3102/-)216(4-)2+4324526/'43/0.4-2!/12 14*.*./0.4-2 /*14$2 5/(.*.0/0.-,2 0+62 36"4/*2 452 "6/136)2 +62361*0124520+62/-/*#6)21/"'*612 -)61.36)2(4-10.06-012 /362,.6-2.-20+620/*6 4106/'43/0.4-2(4-(6-03/0.4-12 +.12 "60+4)2 .12 6556(0.62 .-2 '3.5#.-,2 /-)2 Results 68.55 % (4-(6-03/0.-,2 (/--/.12 (3)62 6%03/(02 /(04312 72.32% THC After 1 h After 2 h 0.76% CBD THC 0+/02 1.,-.5.(/-0*#2 /556(02 0+62 655.(.6-(#2 452 0+62 0.96% CBD '3.5.(/0.4-2 /-)2 (4-(6-03/0.4-2 '34(6112 /362 0+62 +62 361*012 452 0+62 1/"'*62 (4**6(06)2 534"2 0+62 /"4-02 452 /*(4+4*2 16)2 042 ).*062 0+62 .-.0./*2 16(4-)2'346223/0.42/3620+6254**4!.-,2 6%03/(0$2 /-)2 0+62 0#'62 452 5.*06312 16)2 .-2 0+62 '34(6112 .016*52 +6162 5/(04312 -66)2 042 62 " 4106/'43/0.4-2(4-(6-03/0.4-12 (4-1.)636)$2 !+6-2 610/*.1+.-,2 0+62 .-)103./*2 After 1 h After 2 h '34)(0.4-2452'3.5.6)2(/--/.126%03/(01 70.12 % 76.01 % Results THC THC +625.-/*240/.-6)2361*02534"2'34622.12 2 0.66% CBD 0.72% CBD 1. Gonzalez, R. (2025, March 12). Cannabis Winterization Process: The complete guide. Root 2/-)2534"2'34622.12 2 Sciences.https://www.rootsciences.com/blog/winter ization-process-in-cannabis-extraction/ " 2. ErtelAlsop. (2019, September 12). Proper 50632 36(6..-,2 0+62 361*012 452 0+62 (+6".(/*2 https://ertelalsop.com/proper-cannabis-and-cbd-oil- cannabis filtration - ErtelAlsop ErtelAlsop. /-/*#1.124520+62(/--/.12(3)66%03/(02'3462$2.02 filtration-whitepaper/ !/12 -406)2 0+/02 /2 (630/.-2 /"4-02 452 *.'.)12 /-)2 3. Addo, P. W. et al. (2022). Cold ethanol extraction 40+632(4"'4-6-01236"/.-6)2.-20+626%03/(02 +62 of cannabinoids and terpenes. Molecules, 27(24), 36/14-254320+.12"/#2620+621624524-6 8780. https://doi.org/10.3390/molecules27248780 */#6325.*0632 '/'63$2!+.(+2/**4!1214"624520+625/012'3616-02.-2 0+626%03/(02042'/1120+34,+2 +62'34612.-).(/06)2 0+/020+621/,624520+3662*/#63124525.*03/0.4-2'/'632 +62/0+4312!4*)2*.620426%'361120+6.321.-(6362 $2 2 /-)2 2 "2 .12 "4362 6556(0.62 0+/-2 0+62 ,3/0.0)620420+6206/"2/026'*62'34)(0.4-2'*/-02 1/,624524-62*/#6324525.*03/0.4-2'/'6322"2 5432 '34.).-,2 0+62 (/--/.12 (3)62 6%03/(02 /-)2 +625.*03/0.4-2!.0+24-62*/#6325.*0632!.0+2'43612452 06(+-.(/*2 1''4302 )3.-,2 0+62 '3.5.(/0.4-2 "2 36"4612 4-*#2 */3,632 '/30.(*612 .-632 '34(6112 /-)2 042 0+62 /-/*#0.(/*2 06/"2 5432 5.*06312 "2 /-)2 2 "2 /362 16)2 5432 530+632 '63543".-,2 0+62 /-/*#1.12 /-)2 '34.).-,2 0+62 '3.5.(/0.4-2 452 0+62 6%03/(02 534"2 0+62 1"/**632 361*01 157 :987 865:4321065:/.-,+**)+4360('+,065*+:,.0,5%/45*/./2(,:0*3%3*5 64$560.3#04- "! 5 1 2 1 2 2 , + !5 , Jan Elbl , Roman Svoboda , Kevin Matzick, Jana !2 2 1 , !52 , +5%! 1Department of Pharmaceutical Technology, Masaryk University, Czech Republic 2Department of Physical Chemistry, University of Pardubice, Czech Republic 1. 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INTRODUCTION +"!) 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INTRODUCTION 3J= LKJIHGFEDCBAC@?AJCAHGCEAI=ECEDHAJCA HEA E2H=J?H##% "#% CG9JHA J=EA ?GDCE9IAHEA8GK7AJKJIHAJDKA6G=5ADGD4 1E=EA E3J9@JHEK:A EA 9(@ CIE?6?GD?EDH=JHEKA3 HJHA ?JDA ?J@CEA CH=ECC4=E9JHEKA C75IHG5CBA DKEA H @CGKC%A 3J?@@54K=7 +HJDKJ=K*EKAE2H=J?HCAG6A ;J8A G5IJDA G=EJ#A JDKA 6=EE*E4 ##%"#% ;:AJ3EA 8EEDACH@KA K=7 5J7A 8EA 5IG=HJDHA 6G=A IJH2+ 59KA HGA CE3E=EA JFE4=E9JHEKA DE@=GKEFEDE=JH3EA KEA A I=E3J9ED?EA G6A =GK K7CIJFEA FE=HJ89EHEKA@CHGIA?G5IJ?H E=A =G0A JA +;DEAJDGBA!EKE9IJ=5BA=JD?E#: DE@=GKEFEDE=JH3EA K1++++.!! 5IJ=5EDHBA HA 8EA 8EDE6 H=JKA KGCJFEA @JDH 6G=5CA HJHA KGA DGHA =E(@=EA C1J99G1G9EA HJ89EHCAG=A?JIC@9ECA/',: IE=6G=5EKA @CA $"BA E=5GA +? +.%/).-(+.'*+%0*( 1J3E9EDFHCA G6A -A D5A JDKA -- A D5BA +! =ECIE?H3E97:A =G5JHGF=JI J?ED79A CHJHJCEA ##%"#% =ED79A?G9@5DBA. AA:A55BA A;:AI9JDHA5JHE= $EI@89-:A 5BA .A A JHE=CBA +L#:A @J9 L99A =EJFEDHCA JDKA JDJ97HI7HG?E5EA CHJDKJ=K*EKA JII=GI=E2H=J?HA 1JCA ?J== "2?J=5J?E@HJD(@A 9E2A C7CHE5A ?G@I9EKA 1A =8 JCA ?G5I95EDHJ=7A CJ5I9ECA 6=G5A =EI@HJ89EA "2I9G=E=5GA E=A +? 5JD@6J?H@=E=CA G=A I=G?@=EKA H=G@FA CHJDKJ=KA GIE=JHEKA A IGC ?G55E=?JDDE9C: +4!+.!!++ ! +%++ +- 7C )=*G5EA G6A ##% #%A ;:A @C 1E=EA599EKBA@C ) $EHC?BA E=5JD7#A JDKA CA JDJ97H @C CBA E=5JD7#:A "$"LBAE=5JD7#: EA 6=J?H E2H=J?HJDG9A JHA 162 %$#" #! Talc 2.0 Magnesium stearate 0.5 #!! :987656865432 87810/5/2 .-2 6,32 15+95*2 3)6(8'6/2 5*3765-53*2 %$2 36,87.12 8/2 6,32 #./62 3--3'65432 #2,0/5'812',8(8'63(5/65'/2.-2/286',2 3)6(8'68762 -.(2 "5/#8'3(865.7!2 0531*57 2 6,32 Test Result ,5 ,3/62'.7'376(865.7/2 .-2 (./84572 $287*2 Weight (mg) 500 /815*(./5*32 %$2 .6,2 48'99#2 *(057 2 87*2 Extract (mg) 100 10.,515865.72(3/9163*2572/5#518(2'.7'376(865.7/2 Diameter (mm) 11 8"132 2 .343(!2 6,32 48'99#*(53*2 3)6(8'62 Hardness (N) 88.4 Disintegration time 1:59 8/2',./372-.(2/687*8(*5865.72*9326.282,5 ,3(2 .-2 6,3/32 8'65432 '.#.97*/2 572 6,32 *(02 3)6(8'6/2 Thickness (mm) 5 (min) .43(81123)6(8'620531*2,3248'99#*(53*23)6(8'6/2 3(32 ..13*2 87*2 /687*8(*53*2 6.2 $2 (./84572 '.76376287*29/3*25726,32-.(#91865.7/2.-2/ "! ! 2 ! 2 6,32 287810/5/2(3/916/2-.(2*(023)6(8'6 /687*8(*53*2 48'99#*(53*2 2 2 Extract Rosavin Salidroside 3)6(8'62 *3#.7/6(863*2 /9568"515602 -.(2 6,32 Vacuum-dried 4.70 1.95 ,5 ,2 0531*2 87*2 '.7/5/63762 8'65432 '.#.97*2 extract '.76376 (%) (%) *3431.#3762 .-2 2 -.(#91865.7/!2 .--3(57 2 Lyophilized 4.66 1.93 Standardized 3.02 1.70 -.(#91865.7/!2 2 *3#.7/6(863*2 /93(5.(2 68/63 vacuum-dried #8/57 2 87*2 3(32 /313'63*2 -.(2 68"1362 extract 2 2 #.7 2 %2 63/63*2 extract (38(865.72'.7685757 2%%2# 2.-2/687*8(*53*2 :98156865432 87810/5/2 "02 2 (343813*2 6,32 3)6(8'62 .(#91865.7/256,2/1.3(2*5/5763 (865.72 (3/37'32.-2#.(326,872%2'.#.97*/!257'19*57 2 65#3/2 3(32 8//.'5863*2 56,2 5#(.43*2 68/632 ,37.15'2 '.7/6569376/2 /9',2 8/2 -184.7.5*/!2 3('365.72 86',2 !2 872 8/8(68#3-(332-184.7.5*2 10'./5*3/!2'5778#5'28'5*2*3(5486543/!2 -.(#91865.7!2/,.3*26,32"3/628*,3(37'326.26,32 87*2876,.'0875*57/ ,0/5'812'(563(58 #! ! ! .(#91865.7/2 '.7685757 2 %%2 # 2 .-2 1. Tinsley, G.M., et al., Rhodiola rosea as an /687*8(*53*2 3)6(8'62 87*2 3)'55376/2 /9',2 8/2 adaptogen to enhance exercise 681'!2 87*2 #8 73/59#2 /638(8632 (.*9'3*2 68"136/2 J Nutr, 2024. 131(3): p. 461-473. 2. ("31-Biedrawa, D. and I. Podolak, Anti-56,2 5#(.43*2 81868"515602 .343(!2 *932 6.2 '(./'8(#311./3!2'56(5'28'5*!2*3)6(./3!28/8(68#3!2 performance: a review of the literature. Br Neuroinflammatory Effects of Adaptogens: ..(2 ,0/5'812 (.3(653/!2 6,32 8/8(68#3-(332 A Mini-Review. Molecules, 2024. 29(4). -.(#91865.72 86',2 2 8/2 /313'63*2 8/2 6,32 3. Chinwala, M., Recent Formulation .65#812-.(#91865.728"13/2287*2 Advances and Therapeutic Usefulness of Orally Disintegrating Tablets (ODTs). 22-.(#91865.7!286',2 Pharmacy, 2020. 8(4): p. 186. Component Content (%) Extract 20.0 Dextrose 74.5 Citric acid 2.0 Croscarmellose 1.0 163 210/ 2.-,+*)+('2%$#"-.,.!'$% ,+('* 2 *+)#!+(' (-( %$!+$+)#2 . "-)-. 2 1 1 1 1 , Artjoms Iljicevs , Konstantins Logviss , Liga Petersone 1. INTRODUCTION ;3-16/:789,753;185.-34843;8/1+18;316/:78 ?=;:98 7698 58 463218 0/.-41,63478 16/1.8 6.316/:78 -3+18 -4.1978 416348 3+*.1;31+698 .18 0.7:8 =61.8 97,)718 +;(8 .8 +/:;.3;184:47/1:784/7.3;1" 0991998 5968 .14.:96773'63+8 =/34/8 3+8 6,.+8 .1;,4198;.,*8)3237)3736:8%$#"8!8;;.19986/398 +91+8 97,)3736:8 0.-161.98 =1.18 196)739/1;8 0.)71-8 7698 58 973;8 -.0/,98 ;3901.93+98 ,93+*8956=.18 /218 )11+8 ;121701;8 % #"8 -.0/,98 ;.,*8 3828 """88=989,)1461;8 ;39972198)1661.8+;8.14.:9673'198761.8)14,918 68 9721+698 +;8 .19,7698 ;399723+*(9=1773+*8 58)91+41858+,47163+841+61."8!84/31218973;8 *.;1;898$8+81551468*.;1;8988=1.181+61.1;8 -.0/,98 ;3901.93+8 2.3,98 07:-1.;.,*8 3+68 956=.18 +;8 97,)3736:8 0.-161.98 =1.18 ;9*185.-98.18-;1"8?68+7:86/68),68798 474,761;8 ,93+*8 *1+16348 7*.36/-"8 .8 -;178 ;.,*8;17321.:821/347198+;83-07+68;1234198.18 ;.,*98 --68 -714,7.8 ).18 8 5.-,761;8 98;.,*8+;807:-1.84-)3+63+98 474,763+98 =1.18 ,91;"8 ;178 ;.,*98 =1.18 %#"816853+;3+*8+1=85.-,763+98-3*/68)1863-18 9171461;83+89,4/8=:868)18=36/80.-161.9847918 +;8.19,.4184+9,-3+*"8!/1.15.18=183-1;8 6885.85.-8368+;83+8)16=11+" 68 196)739/8 0.1;3463218 -;178 5.8 ;.,*8 7;3+*8 40436:83+807:-1.8684/31218-.0/,98;.,*8 0/91"818,91;8+91+897,)3736:80.-161.9868 0, !=8 *.-98 58 8 =98 ;399721;8 3+8 8 -78 4/3121873+1.80.1;3463218-;1785.87;3+*8$8 4/7.5.-8$-78586/39897,63+8=98;;1;8688 -;178;.,*983+6807:74634843;88980.58 0.1=13*/1;8-,+6858-;178;.,*8+;82.618 584+4106" -31;8 ,+6378 ;399721;"8 .8 ;.,*98 6/68 ;3;+68 ;39972184-071617:8$87858!8=98;;1;"8 " )-.+ $ (,"-)%, !/18 97,63+8 =98 0,.1;8 +8 *7998 973;18 +;8 77=1;868120.6183+85,-18/;8+;8;.:85.8 8 =98 )63+1;8 5.-8758 /1-396.:8 8/"8 " $"8 ;178 ;.,*98 58 0/.-41,63478 2$*2$ *.;18 160.51+8 .-7).8 )1+'43+18 8 /.-614/8 -)8 0.1;+397+18 -0718 =13*/68 58 8 -*8 =98 61+8 +;8 3*-8 ;399721;8 3+8 $8 -78 4/7.5.-8 6/1+8 8 -78 58 7;.34/8 -16.+3;'718 !8 +0.1+8 39:+6/8 3),0.51+8 9;3,-8 -16/+78=98;;1;8684.9/8,68"8-07198 ,01748 417143)8 8 /.-614/8 =1.18 9+3461;8 5.8 $8 -3+,6198 41+6.35,*1;"8 ;34751+48 9;3,-8 *.+8 94.)348 43;8 3,3;80/918=9853761.1;8,93+*8" 8-89:.3+*18 3*-8 7;.34/8 03.416-8 3*-8 7;.34/8 53761.8 +;8 +7:91;"8 /.-6*.0/348 +7:9398 0.416-78 3*-8 7;.34/8 73;43+18 =98;+18,93+*8.121.9180/9184/.-6*.0/:8 /:;.4/7.3;18 9:961-8 1,3001;8 =36/8 8 ;16146.8 -+/:;.618 39:+6/8 3+;-16/43+8 !"8 721+698 58 +7:63478 !/1.-394/1.8 +,39/8 1.-+:"8 !/.118 *.;18 4/7.5.-8 ;34/7.-16/+18 $$ 910.618-16/;98=1.18;121701;868,+635:8778 ;34/7.16/+18 $8;.,*8-714,719" +373+18 616.5,.+8 416+36.3718 4/7.)1+'1+18 67,1+18 416+18 ;3-16/:78 5.--3;18 16/:78 416618 -16/+78 164 +*)( '( $#" ! ! + % % % % % % % % % % % % % % % % % % % % % % % % IHG EDECBA@A ? A = 600 F F F F F F F F F F F F E =y = 1470,5x + 20,917 8;CC;76ED7; =?9; 5A@D4 3@D@2C;1 0//.- F F F F F F F F F F F F F F F F F F F F 500 ,H@4E++5 -?8400 EE66 !/# =@D* A6; A@; //!#* CE= F F F F F F F F F F F F F F F F F F F F E = ?C6E4; /+** F F F F E F F F F F F F F 300 CE= E = 7588;D6 ! =* !'# 9@;844;D7; AC@6* -5 F F F F F F F F F F F F F F F F F F F F 8 F F F F F F F F F F F F F F F F8E9@E66@?D :@6 -5 E F F F F F F F F F F F F1! @@C6;8 8 F F F F F F F F G8554 C?E9;9 F F F FF F F F 200 AE=66;8 =E+@D4 6;= F F F F F F F F E F F F F F F F F F F F F F F F F F F F F 100 ) # " " % % % % % % % % % % % % 0 0 0,1 0,2 0,3 0,4 1/D^2 ) ! % % % % % % % % % % % % EDA;D A?C5@C@6B ??8 4@;D F F F F F F F F F F F FF F F F % % % % % % % %G855 8 54 C?E9@D4 A A?C5@C@6BB F F F F F F F F F F F F 5 6 F F F F :;8; ;A6EC@A;9 5A@D4 '/ A?C;D6A @D 6?6EC* F F F F E F F F F F F F F F F F F F F F F F F F FF F F F 4@@D4F F F F 0 , F F F F ( :@6 F F F FF F F F % % % % A?C5@C@6B A;8; 8E9@5A H 00 EDD9 @@6 ? 6; F F F F F F F F F F F F F F F F F F F F F F F F E F F F F F F F F F F F F F F F F =?9;C !///* D? :8?D4 A?C;D6A @D ?8 ?56 ? 6; ?? 8;9@76 9 F F F F F F F F 9854. A?C5@C@6B A;8;* :@7 @D9@7E6;A :;CC. 6? ??8=5CE66; F F F F E ; E= F F F F E =?8 ;A6 C@A;9 A?C5@C@6B A?C5@C@6BF F F F 8;@?5ACBF F F F 6;A; 7EC75CE66@?DA 4E; E45; F F F F E F F F F F F F FF F F F )' $#" ! ! + % % % % % % % % % % % % % % % % % % % % % % % % @DA@46 :;6 @DA@46 :;6 F F F F 6;8 =@A7@@C@6B 7 DD ; E7@;;9* 56 F F F F F F F F E F F F F F F F F F F F FF F F F E= E =??8 F F F F F F F F F F F F 8 F F F F F F F F F F F F F F F F @A :?8++ F F F F 8F F F F 8;A;D6A :EBB ? ;A6EC@A@D4 D F F F F E F F F F F F F F EF F F F AE= A?:;9 ?DCB = F F F F F F F F E =?8 A?=; 7EA;A 78BBA6ECC@D; )) + ! ! ! 7?88;CE66@?D ? E F F F F F F F F ? H'F F F F F F F F /* 56 6? EC@9E66; 6@A F F F F F F F F F F F F E F F F FF F F F % % % % % % % % % % % % % % % % % % % % % % % % =?9;C* =?8 F F F F 8; ;1;8@=;D6A :@6 =?8; 98554A* F F F F D F F F F F F F F F F F F 8F F F F % % % % ?E9@D4 7E??8 98554A :;8; ;18;AA;9 EA F F F F 8 F F F F F F F F F F F F F F F F 9854 =?C;75C;A ?C5=; @D ! 48 = 9?D; F F F F F F F F F F F F F F F F F F F F E = ? * F F F F F F F F F F F F 4@@D4 5D@6A F F F FF F F F F F F F ? C 4 D;8A; A5E8; ? 6?6EC F F F F F F F F F F F F F F F FF F F F 9@;;8;D7; @D A?C5@C@6BB F F F F F F F F 6 F F F F E =;6;8A ;6:;;D F F F FF F F F ( ## % % % % C?E9 7 !F F F F @5BED H 5 F F F F HED* ;6 EC* E9ED7;A ? 7?=@DE6@; F F F F F F F F F F F F F F F F F F F FF F F F F F F F E F F F F F F F F F F F F F F F F F F F F F F F F E F F F F F F F F DED?78BBA6EC EDD9 =?9;C 9854 :EA ; 9E66E A?:A C@D;E8 7?88;CE66@?D ;6:;;D F F F F E F F F F F F F F F F F F E F F F FF F F F 6; @DA?C5C; 8 F F F F 8;??8 @=8?@D4 F F F F 4 F F F F F F F FF F F F F F F F ; 9854 A?C5@C@6BB ED9 @?EE@CE@C@6BB* F F F F F F F F 6 F F F F F F F F E 6F F F F A DD9 C?E9@D4 7E'F F F F EC5; ? F F F F F F F F 78BBA6ECA* '/'' 8 F F F F ' ?C5=; !' ,"( !'// F F F F F F F F F F F F F F F F / /* :@7 @D9@7E66;A 6E66 6@A =?9;C 8;9@76A F F F F @ @ 9@ 6 F F F F E F F F F 6 6 6@ E F F F F F F F F 9 C F F F F 9@ 6F F F F 'F F F F)E4E9;;A E5 * ;6 EC* A?C5@C@6BB E9ED6E4; ? F F F F F F F F F F F F F F F F F F F F 6 F F F F F F F F F F F F 9854 C?E9@D4 7E 98554A ??8 ? 98554A F F F F F F F F F F F F F F F F 8 F F F F F F F F F F F F F F F F 8F F F F E=?8 F F F F F F F F F F F F 8 =E7;56@7EC 7?78BBA6ECA* F F F F 8F F F F 5A;9 @D 6@A =?9;C ,2@4 !( F F F F F F F F F F F F F F F F F F F F $F F F F @;+; ;=@D* ;6 EC* AA;AA=;D6 ? 9@;;8;D6 F F F F F F F F F F F F F F F F F F F F F F F F F F F F ??8 55A;9 9; F F F F F F F F F F F F F F F F F F F F F F F FF F F F =?9;C@D4 ? 9854 C?E9;9 @= F F F F F F F F F F F F F F F F F F F F F F F FF F F F ? E8=E7;56@7A ED9 @? F F F F F F F F F F F F 8 =E7;56@7A* '/!* ?C F F F F F F F FF F F F !! ."$ F F F F F F F F 165 CBA@ C?=;:98786::9;:75?:74639267619290/:6/9?27 ?5762/998:?.967C-=/?/-;:6C;,/980 +*)('%7C%$'#)%"1 1 2 1 1 , Lucie C+ *)%" 74*74+*)%" , Jakub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ample Phytotherapeutic content [mg/g] DCA07A48DE(G 3AD?G 6911A4AE@DG 0H7'6HC58DEG -%G $@G 416.70 Alg_C H:EC@A2?DG497D9CEGH1G $#"! G4E776G386G6960E@FEFG FA79DAH@6GA@G 8G BA4CHDADECG 078DEG/.--,+*)3E77;G A@G Carvacrol Eugenol Thymol 1.48 A@G8 #?H60?8DEG"911ECEFG!87A@EG/#"!(;G8F 96DEFG 389.03 Alg_E DHG --G -%G 8@FG 19CD?ECG FA79DEFG A@G 8DAH@G 2.59 $F 96DEFG 77ECA@DH@G 5CHD?G BEFA9BG 407.06 Alg_T /$"(%GH79BEGH1G-G7G386GDC8@61ECCEFGDHG 3.59 E84?G3E77GH1GD?EGBA4CHDADECG078DEGDHGH5D8A@G1A@87G 214.51 210.90 Alg_CE 4H@4E@DC8DAH@G .-G )B7%G #78DE6G 3ECEG 1.51 1.43 6E87EFG96A@2GD?ECB87G8F?E6A:EG6E87A@2G1A7BG8@FG 203.72 205.22 Alg_ET 0.63 0.79 A@4958DEFG 1HCG .-G ?H9C6G8DG G %GI?EG= 3. RESULTS AND DISCUSSION HC0?H7H2A487G 08C8BEDEC6;G 6AEG 8@FG 60?ECA4AD'G !EDECBA@8DAH@GH1G= H1G08CDA47E6G8CEG6?H3@GA@GI857EG%GI?EGH56EC:EFG MIC against Phytoterapeutic @E8C7'G 0EC1E4DG 60?ECA4AD'G A6G A@FA48DA:EG H1G E. coli [mg/ml] 18:HC857EG 0CH0ECDAE6G 1HCG 6956E9E@DG 08CDA47EG Carvacrol 2.5 B8@A0978DAH@GDE4?@A9E6% Eugenol 3.9 !HC0?H7H2A487G4?8C84DECA6DA46 Thymol 3.9 Equivalent Sphericity Carvacrol/Eugenol 4.0 Sample diameter SD [m] Eugenol/Thymol 1.5 factor SD Alg_C %G% -%G-%- Carvacrol/Thymol 1.0 Alg_E %G.-%. -%G-%- Carvacrol/Eugenol/Thymol 0.4 Alg_T .-%-G% -%G-%- Alg_CE 4. CONCLUSION .-%.G%---%G-%- !8DA6184DHC'G68B07E6GH1G?'FCH2E7G08CDA47E6G3AD?G Alg_ET .%G.--% -%G-%- 0?'DHD?EC80E9DA46;G 3ECEG 6'@D?E6AEF%G 6A@2G Alg_CT %G.% -%G-%- 8 DCA07EG4HB5A@8DAH@GH1G84DA:EGA@2CEFAE@D6G6?H36G Alg_CTE 0CHBA6A@2GD?EC80E9DA4G0HDE@DA87% ..%G%-%G-%- I?EG E@4806978DAH@G BED?HFG EB07H'EFG 5. REFERENCES 6944E661977'G 84?AE:EFG ?A2?G 4H@DE@DG 7E:E76G /A@G .%G #8:E7H: G ACH678:8;G EDG 87%G " " I857EG(;GB8A@2GADG69AD857EG1HCG19CD?ECGCE6E8C4?%G ! !" !"" !" " " " " " " " " " $@DABA4CH5A87G " #G#?8CB84E9DA46;G-.;G$/(G E11A484'G 69C:E'6G H1G 0?'DHD?EC80E9DA46G 828A@6DG .% $#" ! G 6?H3EFG 6'@EC2A6DA4GA@DEC84DAH@6G8BH@2G@E8C7'G877G84DA:EG 866A@EG 7G $DA%;G EDG 87%G !" $!" " A@2CEFAE@DG 4HB5A@8DAH@6;G 3AD?G D?EG 6DCH@2E6DG " " " $" " " E11E4DGH56EC:EFG3?E@G877GD?CEEG4HB0H9@F6G3ECEG "! "$ " " #"H9C@87GH1G E0HCD6G A@G #?8CB84E9DA487G !4AE@4E6;G --;G /.(G .-.-% 167 ,+*) ('%$#"! (#%,"!("%$$(%#%('%,'#'!% ,"#,"(%#$%("#(%, %$#"%##,$% "%#$%'"#,'%#!(" Sylvie Pavlokova1 1 1 1 , Nicole Fulopova , Katerina Bruckner , Ales Franc 1 Department of Pharmaceutical Technology, Faculty of Pharmacy, Masaryk University, Czech Republic 1. INTRODUCTION *%% =;:9;876;5::743254;9610;9=;3:3/.,5,;78;7'235:9;9323;5,;*65-365/.;'3,9; *3625)1/36/.;876;2=;,-3//;3:9;/360;5:2,25:,+;5,; 7:; 3; )7-'5:3257:; 78; *65:)5*3/; )7-*7::2; *6,,5:0;2793.(;=5,;160:).;5,;1:96,)769;'.; 3:3/.,5,; %; 3:9; 3:3/.,5,; 78; 43653:); 2=; 9-3:9; 876; '72=; /7)3/; 3:9; ,.,2-5); ;712*12,(;,;3;-1/25436532;2)=:51+; 39-5:5,263257:; 876; )7:95257:,; /5; 1/)63254; %; 3-5:,; 5:269*:9:)5,; '2:; )7/525,+;%67=:$,;95,3,+;3:9;)7/7:;)3:)6;#"!( 876-1/3257:;*363-26,;;)7325:0;2.*+; )3*,1/; 2.*; 3:9; , =5,; 6,36)=; 35-,; 27; 9,50:; 3:9; 43/132; 3:; )=363)265,25),(;=;,78236;+;46,57:,;( ("; :265) )7329; =369; )3*,1/; 97,30; 876-; 876; 3:9;((+;3,;-*/7.9;876;9323;*67),,5:0( 236029;9 8)3/;-5)67'5723;263:,*/3:2;76;/54;-5)67',+;27; 2=; 95,23/; *362,; 78; 2=; (; =; )3*,1/,; 36; *%" (%% # 9,50:9; 27; '; 5:26:3//.; *672)29; 3035:,2; 9,261)257:; '.; =.967*=5/5); 85//5:0; 9165:0; =;85:95:0,;'3,9;7:;2=;,2325,25)3/;3:3/.,5,;78; *3,,30;2=6710=;2=;950,254;263)2( *=.,5)7)=-5)3/; 83216,; 36; *6,:29; =6(; 76; 9235/9; 5:876-3257:; 603695:0; 2=5,; %!("%%!('# 6,36)=;5,;5:)/199;5:;2=;*1'/5,=9;,219.;# !( %!% % *%,%% ;%;45,13/53257:;5,;*6,:29;5:;2=;,)76,; 369;0 */72;50(;";3:9;/7395:0,;*/72;50(; ( '3,9; 7:; %; 3:9; 0 :)3*,1/32; 3; =.967*=5/5); '79.; 2-*63216 /518.5:0; 0 5:,7/1'/; 567:; 759; -5216(; 5886:2; )7-'5:3257:,;78;*7/.-6,;876;2=;5:26:3/;; 3:9; 26:3/; ; )7325:0; 6; 2,29(; ; 2=./); ; 1+; 3:9; *7/.45:./; 3)232; (; ; 1963052; ; 1+;)6./ ;)+;3:9;) =; ; *672)2,; 3035:,2; 2=; 03,265); :4567:-:2+;=5/;2=;;*672)2,;3035:,2;2=; /5159;=.967*=5/5);85//5:0;9165:0;*3,,30( %,% 3-*/,; 6; 43/1329; 876; 2=56; *=.,5)7)=-5)3/; *67*625,; 2=5):,,; 78; ; )7325:0; 926-5:9; 1,5:0; ,)3::5:0; ; 2=5):,,+; $% % %; ,)76,; */72(; .-'7/,; 3:9; )7/76,; 50=2; 5:)6-:2; 78; ; )7325:0; 95886:2532; 2=; )3*,1/; 2.*+; 3:9; *75:2; /3' ;50=2;5:)(+;3-71:2;78;;695886:2532;2=;;)7325:0;2.*( 3826;" ;3:9;;-5:;78;95,,7/1257:;95," +; 95,+;3:9;95,5:2063257:;25-;95,5:2(;25-( 168 %$# 1; 0 =+0! :9698407!;498 49,0 )5 3# ,;98=":.04410:0-969 0:9//0!08-0*04+008 6;,5=06 +941 ; %. %$2 -;4987 98 *41 -;56.=04"5063?=6=+0!%$2;8:('2%$2 +09714 98-3 )5 3# +0!0 !0 0;=0: / %.%A? -,5;!0: 4 410 %4-6;,5=063 ?'?;=6-8/9!,0:4106;,00//0-4/('2 +0971498-3;8:('2%$2419-8066)5398 ;==-;606#3 $ ?=41.71 410 6;,5=06 +941 :9//0!084 4"506 %$2-;4987;!0!;8:,=":964!9*.40:9865;-0 )973 # 410 ?'? !0 0;=6 410 /==+987 4!08:6?- %.@0/%$2419-8066;8:%. @0?- / :9698407!;498 49,0 )5 3 / "! A@?=;:98765=43 *41#3 410! 98/=.08-06 ;!0 6,0+1;4 210 98/=.08-0 ,0;8 +09714 98-3 *41 ;,*97..6 ;6 9840!;-498 0//0-46 64!87=" 98/=.08-0410,3 -;49876 )('2%$2# ;8: -;56.=0 4"50 +;6 -8/9!,0: ;6 64;49649-;==" 69789/9-;84 / 410 $ !!! /==+987 ;=.06 ('2%$2 419-8066 %$2 +0971498-3:9698407!;49849,0;8::96)5 07;!:987 410 !0.9!0,084 / ; 1971 3/;==-;606#3:968="410('2 :9698407!;49849,0;8:;=+50!-084;70410 4"505=;"6;69789/9-;84!=0)53#+19=0 !0=0;60:;,.844106.*64;8-0;4;8: *41 -;4987 4"506 )('2%$2# 1; 0 ; ,98 ;8 ;!0; +941 6;,5=06 :069!;*=0 69789/9-;84 0//0-4 8 410 ('2 +09714 98-3 )5 5!50!490696=-;40:;55!9,;40="98410.550! 3#3410?'?!0 0;=0:;64!87 !9714 .;:!;84 )973 #3?4 410 6;,0 49,0 410 0//0-4 9840!;-4986 ;,87 410 / 1971 ('2%$2 419-8066 ;8: %$2 ;8: 5;!;,040!6/;=,64;==,0;6.!0: ;!9;*=063 ('2%$2 +09714 98-3 ;=6 - 4 4196 :9!0-4983 $ '0 0!410=06694-;8*064;40:41;480849-0;*=0 )973 #3 (8 410 .550! 5;!4 410 7!;51 69789/9-;84 540849;= / 4;!7040: :!.76 0650-9;==" 2 +;40! 6.6508698 4 410 (2 6;,5=06;!0=-;40:+19=006;,5=06-;8 5!0/0!;*=" 410 -=83 210 ;:,98964!;498 *0 /.8: 98 410 =+0! 5;!43108 9840!5!04987 :9//0!0849;4986;,5=06*;60:8-;56.=04"50 (88 ;49 0 ;8: 0;6" -;56.=0 -;49876 - 4!08: 96 5!06084 8;,0=" 410 410 -880-498 +941 4196 :9!0-498 98 410 ,041:96!*.64;8:84-869:0!;*=";//0-40: *"410.;8494"9840!8;=040!8;=-;49876 =;:9876 5=4 )973 # 94 -;8 *0 6;9: 41;4 8=" *" -,*98987 ('2 ;8: %$2 5=",0!63 6;,5=06 -,5;!0: 4 06;,5=06 1; 0 2196 ,041: -;8 *0 50!/ 98 !07.=;! 19710!('2%$2419-8066;8:=+0! ;=.06 =;*+941.4650-9;=90:98:9 9:.;=;8: :96 ;8: :963210 ;=.0 :96 96 50!68;=90:4!0;4,0840.95,084,;98794; 410 ,64 69789/9-;84 0//0-4 / :9//0!0849;4987 5!;-49-;=;8:/0;69*=0,041:/:!.7:0=9 0!"3 6;,5=06;--4410-;56.=04"503 $$ " 210:9//0!0849;4984106;,5=06*;60:8410 3 A19=95?3 04 ;=3 ('24"50966=9714="849-0;*=098410:9!0-498 3,;80:9-;=.!8;=3 410/9!64A@3A?6;,5=06;8:%.%6;,5=06 3 ;!0=-;40:8410=0/469:04107!;513?4410 6;,0 49,0 410 %4-6;,5=06 ;!0 :964!9*.40: .=5 ;'304;=3 5;!4 410 7!;51 )973 #3 %.%6;,5=06 ;8: 3 , ;!9;*=" 98 65;-0 *.4 !;410! 98 410 !9714 A?6;,5=06 -,5;!0: 4 410 %4-6;,5=06 3 (840!8;498;= .!8;= A1;!,;-0.49-6 3 169 =; :987657439979821064810/756072.3032180-,31-6+7657.83173+*7 43997103+95807*:0-+)75088,8(*0/-+)7-+7061821-'873)9 4%$#"%7! # 1 1 1 1,2 , Pegi Ahlin Grabnar , 4%"%7"!!#7#!% 7#"%%7%#, Klemen !%2 1. INTRODUCTION 743180-3973+*7481.6*9 DCBAB@?A==;:9 CB?9 B;89 7B669 ?AB;65A49 B32;:9 74%$!#% 1=?C9 ;60A=;:9 0;=52A79 ?CA7B39 C=6?2A/9 80A=;:9 /2.A2?@?9#B:69B;896?B=;36696?39?AB/691A9 .A=7BA/98A/=;:49=6966;?=B3952A9B@C=-=;:9C=:C, .A2-=889 #/9 '@3;9 (0?#A0;;49 B/A;49 +0B3=?/9 5A,8A=89 .A280@?6*9 )59 ?C9 .A280@?9 A7B;/*9 'C9 #B:69 @2;6=6?9 259 B9 ,52=349 ?7.AB?0A9('.9%@869@A=?=@B39?CA6C238649=?9 6B73663/9 2=;89 ?29 B;9 %.B;88,'9 @B;9 3B89 ?29 6?A0@?0AB39 85@?69 60@C9 B69 @233B.69 77#AB;9 3B/A49 60..2A?89 #/9 B9 6.0;,#2;889 B;89A603?9=;9%@66=-92A9=;@2;6=6?;?9A6=80B39 ;2;,12-;9 9 6?A0@?0A9 *9 339 60#3=7B?=2;9 72=6?0A9 3-369 =;9 ?C9 5=;B39 .A280@?*9 $0A=;:9 ?6?69 1A9 @2;80@?89 06=;:9 9 79 ?/.9 )9 6@B3,0.92A95A,8A/=;:9259?2%=@952A703B?=2;69 #2A26=3=@B?9 :3B669 ?0#=;:9 -=B369 .BA?=B33/9 6B389 =;9.A2?@?=-9#B:649=;-=?B#39B3?AB?=2;69=;9?C9 1=?C9=:32296?2..A6*93?AB,.0A91B?A91B690689 CB?9?AB;65A9@255=@=;?9("-9B;89?2?B39A6=6?B;@9 52A9B339%.A=7;?6* (! ?9 @B;9 6=:;=5=@B;?3/9 =7.B@?9 '.49 ;@66=?B?=;:9 @BA5039.A2@669@2;?A239B;892.?=7=75!! !(# 339%.A=7;?691A9@2;80@?8906=;:9/26?BA9 9 ( 9 @=;?=5=@49 BA7=;6?A49 *9 B@C9 A0;9 =;-23-89 9 -=B3649 B@C9 5=3389 1=?C9 9 79 259 1B?A49BAAB;:892;9?C97=88396C354960AA20;889 #/9 7.?/9 -=B369 ?29 ;60A9 @2;6=6?;?9 ?CA7B39 =@ 99+0=3=#A=079-B.2A9.A660A92-A9=@9 @C 99 @2;8=?=2;6*9 'C9 3=+0=89 1B69 5A2;9 ?29 ,D9 @CB7#A9.A660A9 6 99@A266,6@?=2;B39BAB9259 2-A;=:C?9 #52A9 .A=7BA/9 8A/=;:9 B?9 ,D9 B;89 ?C9-=B3 9#BA952A99C20A6*920A98=6?=;@?9%.A=7;?B39 - ? 49B;89! 1. 878?49" 6?0.691A9?6?89=;9520A9=?AB?=2;69?29B;B3/9 Vials placed directly on the shelf 699CB?960#3=7B?=2;9259=@9'6996C35960A5B@9 2. Vials placed on stainless-steel tray ?7.AB?0A9 ' .9 9 .A280@?9 ?7.AB?0A9 69 9 3. Vials placed on stainless-steel tray with @A266,6@?=2;B39BAB9259?C9-=B3 single layer of PP-foil attached to the bottom of the tray =;8=-=80B39 -=B39 1=:C?6=69 ?C9 726?9 .A@=69 in protective bags 7?C28952A98?A7=;=;:960#3=7B?=2;97B66953219 74%7!$!$ 'C9 :AB-=7?A=@9 B..A2B@C7B60A=;:9 4. Vials placed on stainless-steel trays sealed (878?*921-A49=?9=69B3629?=7,@2;607=;:9B;89 =;-B6=-*9 0A9 :2B39 =69 ?29 8-32.9 B9 ;2-349 3669 'C97B66925960#3=7B?891B?A91B698?A7=;89 3B#2A,=;?;6=-49 /?9 6055=@=;?3/9 B@@0AB?9 #/91=:C=;:9?C9;?=A9?AB/9#52A9B;89B5?A9?C9 ?@C;=+09 52A9 878?9 8?A7=;B?=2;9 1C=39 ?6?*9 'CA20:C20?9 ?C9 60#3=7B?=2;9 .A2@6649 ?C9 6/6?7B?=@B33/9 B6666=;:9 ?C9 =7.B@?9 259 -BA=2069 1B?A9 -B.2A9 @2;@;?AB?=2;9 =;6=89 ?C9 @CB7#A9 .A7B#39 #B:9 @27.2;;?69 2;9 7B669 B;89 CB?9 1B69 @2;?=;02063/9 72;=?2A89 06=;:9 B9 7B669 ?AB;65A980A=;:960#3=7B?=2;9 * 6.@?A27?A*9'C9BAB90;8A9?C9@0A-95A279?C9 6.@?A27?A98B?B91B69?C;9@2AA3B?891=?C9?C9 7B6698=55A;@49B3321=;:952A9?C9@B3@03B?=2;9259 878?90;8A96?B8/,6?B?9@2;8=?=2;6* 7)%#!$#7%% 170 10/. ?=;=:987658743935210.-98=08.:9 ,=";:07( 41-34 15219 687,4 :18= ' 4,/75+8:5-= :- 0:9+5= 38:9 7-44* 04"5: 8=:5;5"8:0 298:9 18: :98=4.9 5= )005:5-=877(' 41-9:9 4,/75+8:5-= :4:4 39 0, :- 8005:5-=87 18: 98058:5-= .9-+ :1 ;-=0,;:0 :- ,8=:5.( :1 +844 -. 4,/75+8:0 ;18+/9 8=0 ,""9 417.* 18+/9785= : 87* 38:9' /.-9 4:80(%4:8: ;-=05:5-=4 39 .-,=0 :18: 15219 68"-9 "944,9 5= /824 98;10'=8/75=2:1;87;,78:5-=-.4,/75+8:5-= =18=;4 18: :98=4.9 /( 5+"9-65=2 38:9 0+$0:5=4:80(%4:8:;-=05:5-=4* 68"-9;-=0,;:565:(-69=5:9-2=-9859*=+844 /-,+*)('),%$#,#")!())" $ 452=5.5;8=:05..9=;4'3157 0-4=-:05..9 4";:9-+:9(' 4:,"4 ' ' 8=0 41-3 /--,),,,, 452=5.5;8=:7(.9-+ 8=0 ':1-,21847521:7( 15219 94"-=4 5= ;-+"890 :- 54 -/4960* ?-:87 9454:8=; : ;-=454:4 -. 4:-""9 9454:8=; 4' 0950 "9-0,;: 9454:8=; "' 8=0;18+/99454:8=;;*"54=27525/7 35:1 38:9' 3157 4 8=0 ; 9+85= ;-=4:8=: 8;9-44 4:,"4* = :1 29865+:95; 8""9-8;1' : 05..94452=5.5;8=:7(8+-=2 ' '8=0 '35:1 ;-+"898/7 :- /,: 054:5=;: .9-+ * = +844 4";:9-+:9( ' ' 8=0 41-3 =- Figure 1 452=5.5;8=:05..9=;4' 31984 4:8=04 -,:* . Mean and standard deviation for dm/dt versus setup (1-4) and technique (G-gravimetric ?1152199454:8=;5=4:," 544-77(0, approach / MS-mass spectrometry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his work was supported by the Slovenian ,*#8=8=04:8=08900658:5-=.-968=0: Research Agency under grants P1-0189, and co- 694,4 4:," % 8=0 :;1=5, %29865+:95; 8""9-8;1$#%+8444";:9-+:9(* founded by Sandoz d.d. 5:19 6 =-9 : 05..9 452=5.5;8=:7( 8;9-44 4:,"4 /840 -= :1 0:9+5=8:5-= +:1-0* = :1 29865+:95; 8""9-8;1' 6 687,4 .-9 877 4:,"4 05..9 452=5.5;8=:7(* :," 171 .-,+ .*)()'%'$#%'"! )#'% !"%)#!'!*)#!*%!'#(%# %()#')#"! ()" %#1 1 2 3 4 , Veronica Rizzetto , Lara Gigli , Serena Bertoni , Michele R. Chierotti, Dritan Hasa 1 1 , Beatrice Perissutti* 543210/.3-/,+*,)(3.'1%,1-$,#(10.13"/'1%,!'3-3 ,-'30 '/,+*,0'3 /3,#%3,"0+21,5,5,0'3 /3,/1% %3//01!'-0+/0+-3,0'3 /3,!!,5,.,5,'-,031,!'3-3,#10,1 +'10'3 /3,/1% 43210/.3-/,+*,#(10.1,1-$,'+/3(-+%+,-'30 '/,+*,+%+-1, ,!,4+-1/+,5,5,+%+-1,/1% 43210/.3-/,+*,)(3.' /0,1-$,!,)3-/03,-'30 '/,+*,+0'-+, ,'"0'1,,55,+0'-+,/1% 230' "//'"-'/ '/ 1. INTRODUCTION :97;?3=;'32B;?BCA.AB;: E!7; 5;A); EDCBA@?D=; :E9876; A5D4D3?BB2; =1 /.;-,+*@;?@;?3;?3/.D3/DC;?43/6;.?@;5 ## =D15@; /.5?0(/DC; ?5@AB1?/@; ?@; 05)A5=; D3; ?/A52; 4?D3=;?//3/DA3;)A5;D/@;0A/3/D?B;50(50A@D34;D3; .; DC?B; @23/.@D@; A); E98; /53/@; %-$#"; !A 156; D/@; 0AA5; ?(/ A;$+;;@/?D3B@@;@/;; @AB('DBD/2; ?3=; /.; /3=3C2; /A; CA315/; D3/A; ?; 1D'5?/DA3?B;DBB;:?327;(D00=;'2; ' .D4.B2; D3@AB('B; A3A.2=5?/; )A5; .D3=5; D/@; .; DBBD34; )5(3C2; )A5; CBD3DC?B; =13/; ?3=; 'DA?1?DB?'DBD/2; %#"; ?D3/?D3=;?/;$+;!6;/.;1AD=;1AB(;D3;/.;?5; 3; 05AD@D34; @/5?/42; /A; A15CA; /.@; ?@;?B@A;0/;CA3@/?3/;:D""6;?;/A/?B;A);**;4;A); C.?BB34@;D@; /.; )A5?/DA3; A); @AB1?/@6; .DC.; 0A =5;05;$+;;?57;?3=;/.;DBBD34;/D; ?@; C?3;@/?'DBD;E98;D3;A5;@AB('B;)A5@;?3=; 0/;)D=;:D""6;*;D37" C.?33=D))53/; @AB13/@; (@D34; / A; =D))53/; E98 @AB13/;AB?5;5?/DA@6;D""6;--;?3=;-$"; ?@=;A3; @/5A34; .2=5A43; 'A3=D34; D/.; @AB13/; /.;5@(B/@;A'/?D3=; D/.;/.;?'A13/DA3=; ABAB@; E98; ?@; 45A(3=; D/.; ?'A1; 3/DA3=; -$; 0513/; A3A.2=5?/; CA315@DA3"; (; /A; D/@; A3A.2=5?/@; ?5; ?B5@/ADC.DAAB?5; 5?/DA@; :D""6; -*"+6;--"+6;-7; 5;(@=;D3;BDD/=;C?@@" @/(=2;?D=;/A;0?3=;/.;@ABD=)A5;B?3=@C?0; A5; CA0?5D@A3; 0(50A@@; @B(552; '5D=4D34; A); E98; '2; @C53D34; -$; @AB13/@; /.5A(4.; 05D3/@; :=(5?/DA3; A); ; =?2@7; 5; DC?B; ; 5; @(CC@@)(BB2; A'/?D3=6; 5;C.?5?C/5D=;(@D34;?;5?34;A);@ABD=@/?/; , ## ?3?B2/DC?B;/.; @AB1?/@; 5; C.?5?C/5D=; (@D34; ?; @/?'DBD/26; 05A1D=D34;1?B(?'B; D3@D4./@;D3/A; /.CA05.3@D1;@ABD=@/?/;?3?B2/DC?B;?005A?C.6; 0A/3/D?B;)A5;D05A1D34;E98;)A5(B?/DA3;?3=; D3CB(=D34; 0A =5; 5?2; =D))5?C/DA3; :76; /.5?0(/DC;05)A5?3C" =D))53/D?B; @C?33D34; C?BA5D #)*% "#'#)!" /.5A45?1D;D3)5?5=;@0 ## /A/?B; 5)B ; 5@A3?3C;:E87";==D/DA3?BB26;/.;@AB1?/@; :8!7; .0?C/DA3; ?3=; @/A5?4; ?/; :!E76; 'D3/;/05?/(5;)A5;@D;A3/.@;?3=;(3=5; 76; E98; D/.; ?; =-! E8; ?3=; @ABD=@/?/; 3(CB?43; D@A05A0?3AB; =D; .(D=; CA3=D/DA3@; /A; ?@@@@; /.# /.?3AB; :/!7 ; @/?'DBD/2; ?3=; 0A/3/D?B; CA315@DA3;/A; /.; E98; ?C;;?CA3A.2=5?/; )A5"; A5; /.; @AB1?/; A'/?D3=; 172 ! DCBA@ ?=;:@ 9876A54B547@ 34D215@ 05/8@ @ 2C..5/6BC47@2/B/@D151@64--16B12, +C*1@4)B@4.@BA1@BD1-*1@94-*17B9@519)-B12@C7@('@ 94-*/B19:@ C7@ DAC6A@ BA1@ 94-*17B@ D/9@ %4)72@ B4@ ('@ C7@ 2C..1517B@ D/89@ /72@ DCBA@ */58C7$@ 9B4C6AC4#1B58,@"AC9@519)-B12@C7@BA1@2C964*158@4.@ .C*1@71D@94-*/B19@DCBA@94-*17B9@64##47-8@)912@ =0;@ 3/BB157@ 4.@ 7479B4C6AC4#1B5C6@ C7@BA1@3A/5#/61)BC6/-@.C1-2,@!/912@47@BA1@/%4*1 94-*/B19@64#3/512@B4@74D7@('@94-C2@.45#9, #17BC4712@ 1 B179C*1@ 6A/5/6B15C/BC47@ 4.@ BA191@ 64#34)729:@ BA1@ 71D@ 989B1#9@ D151@ 6-/99C.C12@ #345B/7B-8:@/--@.C*1@('@94-*/B19@D151@.4)72@ C7B4@BD4@9)%$54)39@ B4@%1@9B/%-1@.45@/B@-1/9B@9C @#47BA9@/72@519C9B/7B@ B4@647*159C47@C7B4@BA1@)7219C5/%-1@#474A825/B1@ @ "A1@ .C59B@ 9)%$54)3@ C76-)219@ BD4@ 1)C#4-/5@ .45#@4.@('@/72@B4@64#35199C47, 9B4C6AC4#1B5C6@ 94-*/B19@ 7C6-49/#C212C#1BA8-@ 9)-.4 C21@ ('@ /72@ BA1@ 74*1-@ 7C6-49/#C21?38554-C2471@ ('?=;,@ "A54)$A@BAC9@1 315C#17B/-@9B)28:@D1@1 3/7212@ "A1@ -/BB15@ 51351917B9@ /@ 71D-8@ C217BC.C12@ 94-C2@ BA1@ 94-C2.45#@ -/7296/31@ 4.@ 7C6-49/#C21@ %8@ .45#:@ DA491@ 6589B/-@ 9B5)6B)51@ A/9@ %117@ 94-*12@ C217BC.8C7$@ %4BA@ 9B4C6AC4#1B5C6@ /72@ 747 A151@.45@BA1@.C59B@BC#1@)9C7$@9876A54B547@34D215@ 9B4C6AC4#1B5C6@ 94-*/B19:@ DAC6A@ 74B@ 47-8@ 4..15@ 05/8@2C..5/6BC47@2/B/,@/%=0;@3/BB1579@/51@ 91*15/-@ 34B17BC/-@ /2*/7B/$19@ .45@ C72)9B5C/-@ 51345B12@C7@+C$)51@, /33-C6/BC479@ C7@ 3A/5#/61)BC6/-@ .45#)-/BC47@ @ 21*1-43#17B@ %)B@ /-94@ 1..16BC*1-8@ 351*17B@ BA1@ .45#/BC47@4.@BA1@)7219C512@#474A825/B1, ,@ 'A17:@ ,@ 1B@ /-,@ '%$#"! # ! $ %@'1--@C$7/-@?:@:@ ,@ 0):@ @ 1B@ /-,@ #! "% !!$ ! %$#"! ! " !% #!$@ '@ =0;@ 3/BB157@ 4.@ BA1@ 9B4C6AC4#1B5C6@ 7.16B@C9@??:@:@ ,@ 94-*/B19@64#3/512@B4@74D7@('@94-C2@.45#9 , /7@ "47215:@ ,',@ 1B@ /-,@ =!!# ! ?@ "A1@ 916472@ 9)%$54)3@ 6479C9B9@ 4.@ 747 "%#% %!$ #" # %$#"! 9B4C6AC4#1B5C6@ 94-*/B19:@ 3519)#/%-8@ !!!# ##!",@7B@@=A/5#@?:@ C949B5)6B)5/-@ 6A/771-@ 94-*/B19:@ $C*17@ BA1@ ? :@?, 9C#C-/5CB8@C7@=0;@/72@+";@/7/-8919,@"A191@ /77:@,,@1B@/-,@"#$!##%"""%!$ C76-)21@ 7C6-49/#C21/61BC6@ /6C2@ (':@ "#$!"#%$#"! @4-@=A/5#@??:@?:@ 7C6-49/#C21/61B471@ (''":@ /72@ ? 7C6-49/#C21%178-@/-64A4-@('!(,@ 513542)6C%C-CB8@ C7@ BA1C5@ .45#/BC47@ /72@ */5C/%-1@ ## % "$!"" $ ! !!# # %$#"! ! ;96@ 2*@ ??:@ :@ 94-*17B@ 647B17B@ DCBAC7@ BA1@ 9B5)6B)51@ )347@ "A191@ 94-*/B19@ %4BA@ 21#479B5/B1@ -4D@ )/C:@ ,,,@ 1B@ /-,@ !# !%#" :@4C, 5/%, 9B45/$1,@"AC9@C79B/%C-CB8@D/9@647.C5#12@%8@':@ "@ /72@ (;@ /7/-8919:@ /-47$@ DCBA@ 4*/$4:@ ,@ 1B@ /-,@ ! "%!$ C949B5)6B)5/-CB8@ 1*C217612@ BA54)$A@ =0;@ $!"%!#%$#"! !#$ ## .C$)51@?@/72@+";, "#$!"! ##!!6B/@'589B/--4$5@ '@B5)6B@'A1#@?:@:@ 1@BA/7@/BB14@)9C@7C*159CB8@4.@C#15C6@ .45@)91.)-@2C96)99C479,@ 173 :989 765643:2610/3./-1/,1135-0,56/7+*/:3)76+/,1(-46+/.3+/'*'062,/ 764,56+*/3./-1/-10,:'*(30,/7+%$ #"!/,/:"1 1 1 , Rita Ambrus , Petra Party 1. 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"$%))"$ Maja Todeska, Martina Gjorgjevska, Daniel Velickovski, Dragana Petrusevska, Nikola Geskovski, Katerina Goracinova, Teodora Tasevska, Lina Livrinska, Maja Simonoska Crcarevska 1. INTRODUCTION 4=5 )5 .625 =17#7=6*5 $=9=5 293!%0=62=25 :5 =6*(9=5 (67;:985 27*97%(7:6 55 %762=95 *:0(7:65 =;:987654329:140:972=/5.5-,+5,0.**5)))529('5 :037630#399:072:6=5"5"/5=0.765"5 47'45*:0(%7073/50:$5#=98=.%7073"/57*5.5;79* .625 +.9145 5 "5 765 #(97;7=25 $.=95 $.*5 4=9.#35 ;:95 27.%==*5 8.6.'=8=6 5 =*#7=5 '9.2(.0035.22=25(62=951:676(:(*58776'5(6705 0787=25107671.0578#.152(=5:57*5#=98=.%7073! .51:4=*7=58.**5$.*5;:98=2 54=5$=58.**5$.*5 0787=25 .%*:9#7:65 14.9.1=97*71*/5 27**:0(7:65 #.**=2549:('45.55885*7==5:5;:985'9.6(0=*5 7876'58.35*7005%=59=0=.652(=5:58=;:9876*5 .625297=25.5 ,5;:95.##9:78.=0355 6.99:$5 .%*:9#7:65 $762:$5 :95 =.1=9%.7:65 :;5 876(=* 5;=95 29376'/5 9='9.6(0.7:65 $.*5 2:6=5 )5*72=5=;;=1* 5 :5=6*(9=5(67;:985#.9710=5*7= 4=5 :%=17=5 :;5 4=5 *(235 $.*5 :5 =.0(.=5 $4=4=95$=00!2:1(8=6=2576=9.17:6*5%=$==65 5 ,:8#:*77:65 :;5 9.#72!9=0=.*=5 8=;:98765 4=5 %762=9/5 19:*1.98=00:*=5 *:27(85 .*5 '9.6(0=5;:98(0.7:6*5 27*76='9.6"/5 .625 8=;:98765 4329:140:972=5 F1 F2 F3 .17=5 *(%*.61="5 *7'67;71.6035 .;;=15 '9.6(0=5 Component (g) (g) (g) 14.9.1=97*71*5 .5 4=5 *#=17;7=25 1:61=69.7:6*5 Metformin HCl 50 50 50 .625761:9#:9.7:658=4:2* 54=5*(235.78=25:5 27.2 27.2 27.2 Lactose 2==9876=5 4:$5 4=*=5 76=9.17:6*/5 #.971(0.9035 5 5 5 4329:'=65 %:6276'5 %=$==65 1:8#:6=6*/5 Microcrystalline 10 10 10 Cellulose 76;0(=61=5 19771.05 '9.6(0=5 #9:#=97=*5 *(145 .*5 #.9710=5 *7=5 27*97%(7:6/5 ;0:$5 14.9.1=97*71*/5 Croscarmellose 2.5 2.5 2.5 Sodium 1:8#9=**7%7073/5 .625 27*76='9.7:65 %=4.7:9 5 Sodium Starch 62=9*.6276'54=*=576=9.17:6*57*5=**=67.05;:95 5 5 5 Glycolate 97*5 .6.03*7*5 .625 8.6.'=8=65 .*5 $=005 .*5 ;:95 Polyvinylpyrrolidon 2.5 / / :#78776'5 ;:98(0.7:65 #.9.8==9*5 .625 e (PVP) =6*(976'51:6*7*=65#9:2(15#=9;:98.61=57654=5 Gelatin / 2.5 / ;76.052:*.'=5;:985/ Starch 1500 / / 2.5 Sucrose 2 2 2 Talc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ence Sipos1 1 1 1 1 , Petra Party '' , Rita Ambrus ','+ 1Institute of Pharmaceutical Technology and Regulatory Affairs, University of Szeged, Hungary 1. 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"$ %!$#"$$#"$$ $#" !!!$#$ ! "$ 1. INTRODUCTION 2. MATERIALS AND METHODS 43210/..2-, +*3/)(2+, '4%+$, #32, "0!2), ! 2, ("1!", ), ! /+, +!-*, 2, !/./2-, ".2.#3, -*)#/+, "/2, 0"3, 3/#3*, ##(/)(, 0"3, /"."(/#., '$,+/.#!/")+,!",2."32,! 2,/)!23#!/")+,"0, -3(+, -2, !", ! 2/3, ")2)/2)2, 0"3, +2.01 #3/"+, ")".")#., #)!/"-/2+, '+$, #!, ! 2, #-/)/+!3#!/"), 2+2/#..*, 2), /)"3"3#!2-, %,,.//-,/)!230#2,%,#+,"-2..2-,#+,#,+.#, /)!",#!"/)2!"3+,"223,/++2+,#),#3/+2,/0, "0,".*-/2! *.+/."#)2,'4%$,".2.2+,/), +/./")2, "/., '%$, +2-, !", .3/#!2, ! 2, +*3/)(2, ")!#!, /! , 3"!2/), +".!/"), -#)2-, #332., -2.2!2+, /)!", ! 2, +".!/"), .2#-/)(, !", +/.#!/"), !2 )/2+, + , #+, 32..#, +*3/)(2,#.0)!/"), 2,-2.2!/"),/+,#+2-,*, +#./)(, 232, !/./2-, !", +/.#!2, +, /)!23#!/")+, 2!22), %, #)-, ! 23#2!/, #-+"3!/"),!",! 2,%, 2+2,2! 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RESULTS AND DISCUSSION 32+")+2,/),#!/2)!+,,,2,!",.//!2-,!2+!/)(, 3".2+, "0!2), (", ))"!/2-, )!/., , .#!23, $/$$ 2), ! 2, 3/#3*, ##(/)(, -22."2)!, 2(/)+,!,! #!,"/)!,/!/(#!/"),"0,+ ,/++2+, 2"2+, #, #..2)(/)(, !#+, #+, #)*, #)(2, /), 0"3.#!/"), ".-, 32/32, +/()/0/#)!, 32-22."2)!, 200"3!+, 332)!.*, ! 232, /+, )", #3#!2,#)-,32./#.2,2! 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CONCLUSIONS )0 6 70 670 50 5750 %$(50 460 2,60 4,50 3)343%(0 8766.0 6.0 *6)*584'0 *6).37-3)!0 4,%40 -6(5* (%70 -6+5((3)!0 *%)0 $50 25+0 460 24 +0 0 +58(5436)0 + 50 460 3)457%*436)20 34,0 876453)20 3)0 873-%70 8%*%!3)!10 0 " 74,570 7525%7*,0 5..67420 3((0 $50 2,3.45+0 46%7+20 .3)+3)!0 86223$(50 *6775(%436)20 $5455)0 58573-5)4%(0 752 (420 %)+0 4,57-6+)%-3*0 87685743520 6$4%3)5+0 34,0 -6(5* (%70 23- (%436)20 %)+0 !5)57%43)!0 75(3%$(50 %)+0%** 7%450875+3*436)0-6+5(1 5. REFERENCES 10 6(('0 "11'0 540 %(1'0 "! 0 7685%)0 6 7)%(0 6.0 ,%7-%*5 43*20 %)+0 368,%7-%*5 43*2'0 100 1 10 % 7%$,'0 1'0 540 %(1'0 ! 183 -,+* )('%$#"! !!)-( -(%($#" %%" $%) )(( %(!(%) !-%(!%)-( "%! % Barbara Sterle Zorec, Rok Dreu 1. 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" " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " 53? 558* @?7; 75873244; 4?24 4? 2 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 2 = = = = = = = = = = = = = = = = = 2 = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = ;02//6= = = = = = = = = = = = = = = = = $## ! " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " " "" " " " " " " " " " " " " " " " " 8:(5 :6 470* ;707/26 ,2444655 7; :. ,264473/; 9::60 6 = = = = = = = = = = = = = = = = = 9 6 0 3/;46; 4?24 6282//1 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 2 = = = = = = = = = = = = = = = = = 1 .62++ = = = = = = = = = = = = = = = = = 2= = = = = = = = = = = = = = = = = @? ; :9 87999654 326676; 9::6 861 0 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 05 7; ;8 2; 2 326676' 6268/; = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 24 6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = .;68 6= = = = = = = = = = = = = = = = = ?2; 2 3:5;7862./ 759//53 :5 4? ;7- = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = :9 4? = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = ./25+ 75+ = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = ; :9 4? = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = ,264473/; 4?244 26 687;,6;8 75 8799965 6 = = = = = = = = = = = = = = = = = 2 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 9 54 ./255+ = = = = = = = = = = = = = = = = = 2= = = = = = = = = = = = = = = = = 99644?6 ;,,:6448 .1 2552/1;7; % = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 0870 ;8* @?7; :.;62 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 6 22447:5 7; = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 75+ 0872* )?5 /234:; (2; ;8' 4? , = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = ,264473/; 6= = = = = = = = = = = = = = = = = (?73? ;?:(; 4?244 .:4? /234:; 255 6 = = = = = = = = = = = = = = = = = 6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 2 = = = = = = = = = = = = = = = = = 7* "' = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = 562//1 027542758 ;707/26 087255 ;7- = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 2 = = = = = = = = = = = = = = = = = ; %8$#" = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = ,264473/; 757472//1 9::60 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 2 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 258 = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = !3,4 9 = = = = = = = = = = = = = = = = = 29446 .75 687;,6;8 75 2 # = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = %" = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = .62+ 2,2644 8675 6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 60 /266 26244 = = = = = = = = = = = = = = = = = 6 = = = = = = = = = = = = = = = = = 2 2 4; 4?244 = = = = = = = = = = = = = = = = = 2= = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = =;4:6 * 2 * 6 4? ,264473/ ;7- 7; /26 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 6 ;4 2558 = = = = = = = = = = = = = = = = = 2= = = = = = = = = = = = = = = = = 75362;; (74? 470 258 (74? 753 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 362;75= = = = = = = = = = = = = = = = = 40,62446 258 ?078741 8675 ;4:62 2 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 2 * # " " " " " " " " " " " " " " " " "= = = = = = = = = = = = = = = = = :6,,?:/:11 :9 ,26473/; 3:54275 6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 575 %2" = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = /234:;' %." = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = /234:;$$ %((" 258 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 8 %3" = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = % *# *#" = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 2; 8611 06* 6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 6= = = = = = = = = = = = = = = = = # " " " " " " " " " " " " " " " " " 5 4? ,6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = ,;54 ;481' 8611 06 = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = 8/:,8 = = = = = = = = = = = = = = = = =;75 4?6 87999654 ,26473/ 326676; = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 9 = = = = = = = = = = = = = = = = = 6 = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = 234:;' = = = = = = = = = = = = = = = = = 234:; 258 * @?; 326676; = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = !?7.748 5 = = = = = = = = = = = = = = = = = 5:42../ 87999653; 75 ,264473/ 2 = = = = = = = = = = = = = = = = = 9 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 6= = = = = = = = = = = = = = = = = 0:6,?:/:1 1 2; (// 2; 75 4?76 687;,6;75 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = .? 7:6 75 2 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 5 4?6 ./255+ 75+ 0872' 23? (74? = = = = = = = = = = = = = = = = = 2 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = 57 ;6922 = = = = = = = = = = = = = = = = = 9 23 45;7:5 2558 7;3:;741 ,6:,647;* = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 2 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 6 %# ! ! " " " " " " " " " " " " " " " " " * 46/ :6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 63' *' @ = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = ?260 6 02347 732/ 8/:,054 258 43?5:/:1 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = #' :/0 = = = = = = = = = = = = = = = = = 0 ' ,* * = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = # " " " " " " " " " " " " " " " " "= = = = = = = = = = = = = = = = = @? ? ;7- %8# %/:(6 .:!,,/:4 /7 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = ! = = = = = = = = = = = = = = = = =5"' 8$# = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = 44?? 687;,6; = %0788/ .:!, = = = = = = = = = = = = = = = = =/:4 /75" 258 8 # %,,,6 .:!,/:4 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 4 /75"" :9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 8 ,264473/; 87634/1 29446 ,6:83 = = = = = = = = = = = = = = = = = 6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = 347:5 258 = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = /234:;' ." = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = /234:; 258 3" 2; ,26473/ = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 3266676 = = = = = = = = = = = = = = = = = 6* 99775 55372/ ; 29446 :5 0:5 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 5544?? :9 ;4:62 75 87999654 0872 4 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 9 = = = = = = = = = = = = = = = = = 2' (74?? 2" = = = = = = = = = = = = = = = = = 4 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = @? 24?: = = = = = = = = = = = = = = = = = 26; 624498 4? = = = = = = = = = = = = = = = = = 2 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = 2 = = = = = = = = = = = = = = = = = ,,:644 ,6:788 .1 4? /:57255 6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 2= = = = = = = = = = = = = = = = = )?5 ;263? = = = = = = = = = = = = = = = = = 531 %6:620 = = = = = = = = = = = = = = = = = 2 0 # "* = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = /234:; 7; ;8' 4? 8 # ,2647 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = ' = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = , 6 73/ ;7- = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 458; 4: 8362; :6 2 :5 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = =0:54? ;4:62 = = = = = = = = = = = = = = = = = 2= = = = = = = = = = = = = = = = = ,67:8' (?7/ 4? 8$# 2558 8# 2/; 60275 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 2 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = 185 3210 /.-,+*3)-(3)'-%$,.#%"./",! -,# /. -./", !,-",' /)/, /.! -#.*,./,$.3.%(),! -#$%. /,))$ 3#)/ Elizabeta Atanaskova1,2, Johannes Poms2, Matthias Wolfgang2, Carolina Alva2, Nadia Mussner2, Nikola Geskovski1 3 2,3 , Andreas Zimmer , Sharareh Salar-Behzadi 1Ss. Cyril and Methodius University in Skopje, Faculty of Pharmacy, Institute of Pharmaceutical Technology, Skopje, Republic of North Macedonia 2Research Center Pharmaceutical Engineering GmbH, Graz, Austria 3University of Graz, Institute of Pharmaceutical Sciences, Department of Pharmaceutical, Technology and Biopharmacy, Graz, Austria 1. INTRODUCTION ,#%)-.%$(,%/,#)+ ( BAA@?=;A:?:A9879765?7:??4@76749?4399;21;?72? ,#, 03@/4;6749? .;-;9A0/;26,? +707.*8:;.? ;6@1954;@A9?0@679?;:6;@7)7;.?763?09/7674?2.? )A@/967A2:?A));@?)9;(7879765?72?:A98797'721?A)? :6;@74? 47.:? +70A9;2? B%? .7*%? 6@7*%? 35.@A03A874?467-;?03@/4;6749?721@;.7;26:? 6;@6@%? 2.? 3;(1954;@A9? B%? B%? B%? 2.? 76372? 63;? 9707.? /6@7(%? 9;.721? 6A? 724@;:;.? B? ;@;? 72.95? 0@A-7.;.? 85? ""? +$? .@1? :A9879765? 2.? 8:A@067A2? )@A/? /8,?;6@35.@A)@2%?.7/;6359:9)A(7.;%?* 1:6@A726;:6729? 6@46%? 967/6;95? ;2324721? 0@A02A9%? /A2A8:74? :A.76;%? 87A-79879765,? $2:@721? 27)A@/? /A9;49@? :A.76;? .78:74? 2.? :A.7@59? .7:6@7867A2? 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A::5 ABB'=AB +* C C C C C C C C A C C C C C C C C C C C C C C C C /$( " C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C 0= :: 6=;?5 A C C C C C C C C 5D5 =A@D5 A@'D9 ':5D=9B;=D ?9 6=DB?9?;:$ FED 5DD@;6D5 C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C CC C C C C C C C 4;;= 6=; C C C C C C C C C C C C C C C C C C C C C C C C 4 C C C C C C C C ;?5?:0C C C C C C C C 7;5D@C C C C C C C C D:A@D5 4;;=7 C C C C C C C C 4 = 7'@A 6A=A7 A 7D ?:B;=6;=A 7;@DB'@A= 5?9=?' 187 -,+* STABILITY EVALUATION OF KETOCONAZOLE SHAMPOO: INFLUENCE OF pH AND ANTIOXIDANT ADDITION Maja Todeska1 1 1 , Dobrinka Dimova , Natalija Dimovska 1. INTRODUCTION @3: 6933B/BA: ?@?BA/=A9@8: @3: 8@6940: +56/@96B': 669A9@=;;51:A7@: $: -BA@?@=@;B: CBA@?@=@;B:98:=:796B;5:48B6:=A9342=;:=2BA1: 8+=0*@@8: 7B/B: */B*=/B6@B: ?@A=992: 9A:98:=:9096=@;B:6B/9.=A9.B1:=:7B=-:,=8B:79A+: ,4A5;=AB6: +56/@5A@;4BB: : =8: =: A7@:*C=):.=;4B8:('%:=6:$'#"':!/@64?A:8A=,9;9A5: =A9@96=A1:=6:@B:79A+@4A:': 98:=:90*@/A=A:9884B:9:*+=/0=?B4A9?=;:9648A/5': : A+@/@42+: -@7;B62B: @3: A+B: ?+B09?=;: =6: ('')#$ #)# " *+589?=;:8A=,9;9A5:@3:6/428:=6:6@8=2B:3@/08:98: A=,9;9A5:8A469B8:@3:-BA@?@=@;B:8+=0*@@:7B/B: ?/9A9?=;: 9: A+B: 6B.B;@*0BA: =6: B.=;4=A9@: @3: ?@64?AB6:9:=??@/6=?B:79A+::2496B;9B8': *+=/0=?B4A9?=;8': CBA@?@=@;B: 98: ?+B09?=;;5: +B: 8+=0*@@: 8=0*;B8: 7B/B: 8A@/B6: 9: 8A=,9;9A5: 48A=,;B1: 79A+: @96=A9@: =6: +56/@;5898: ?+=0,B/8:=A: ::$::(::::: /B*/B8BA92:A+B:0=9:/@4AB8:@3:9A8:6B2/=6=A9@') 3@/: =: 64/=A9@: @3: %$: 0@A+8': =0*;B8: 7B/B: 4/A+B/:9.B8A92=A9@8:=/B:BB6B6:A@:90*/@.B:A+B: ?@;;B?AB6: =A: 9AB/.=;8: @3): 1: 1: (1: #1: =6: %$: 8A=,9;9A5: @3: -BA@?@=@;B: 9: =4B@48: 0@A+8:3@/:*+589?=;:=6:?+B09?=;:B.=;4=A9@'$ 3@/04;=A9@8:=6:A@:B*;=9:+@7:A+B:*:@3:A+B: 8+=0*@@1:A+B:=669A9@:@3:=A9@96=A81:=6:@A+B/: ('+)-!")"# *+589?@?+B09?=;:*=/=0BAB/8:=33B?A:9A8:8A=,9;9A5': +B: 8+=0*@@: 7=8: B=09B6: A@: *+589?=;: : ?@0,9=A9@: @3: 9@9?: =6: @9@9?: ?+=/=?AB/98A9?8:A+=A:93;4B?B:A+B:4=;9A5:@3:A+B: 84/3=?A=A8:+=8:,BB:8+@7:A@:*/@.96B:A+B:0@8A: 39=;: */@64?A1: 9?;4692: ?;=/9A51: .98?@89A51: 8A=,;B: A+9@A/@*9?: 858AB0: ,5: @*A90992: 493@/09A51: *: .=;4B1: ?@;@/: 9AB89A51: =6: 9AB/3=?9=;: 39;0: 3@/0=A9@: =6: 0@;B?4;=/: ?@898AB?5' 9AB/=?A9@8':% 9.B: A+B: 796B8*/B=6: 48B: @3: -BA@?@=@;B: ')) ) 8+=0*@@8: 9: A/B=A92: 6=6/433: =6: 8B,@//+B9?: '') - $) "$!$") $) $) 6B/0=A9A981: B84/92: A+B9/: 8A=,9;9A5: 98: B88BA9=;: "# 3@/:0=9A=992:A+B/=*B4A9?:B339?=?5:=6:*/@64?A: +B:8+=0*@@:7=8:+@0@2B@481:493@/01:?;B=/1: 8+B;3;93B' .98?@48:;9496:79A+:=:?+=/=?AB/98A9?:@6@/:=6:=: +B:=90:@3:A+98:8A465:98:A@:B=09B:A+B:8A=,9;9A5: 9AB8B: *9-@/=2B: ?@;@/': 4/92: 8A=,9;9A5: @3: -BA@?@=@;B: 9: /B;=A9@: A@: ?+=2B8: 9: A+B: 8A465: A+B/B: 7=8: @: 892939?=A: ?+=2B8: 9: 8+=0*@@8:*:=6:?@?BA/=A9@:@3:=A9@96=A: *+589?=;:B.=;4=A9@8' A@:6BAB/09B:7+BA+B/:A+B:=669A9@:@3:=:8A=,9;9'(') "") ) ) ) "#$") =33B?A8:9A8:8A=,9;9A5' #$ #)) +B: *8A=,9;9A5: @3: $: -BA@?@=@;B: 98: 2. MATERIALS AND METHODS */B8BAB6:9:=,;B:%': @:.989,;B:?+=2B8:7B/B: ('')%$#"! $) @,8B/.B6: 64/92: A+98: *B/9@6': +B: 0@8A: 8A=,;B: CBA@?@=@;B1: @6940: ;=4/5;: 84;*+=AB1: 3@/04;=A9@: 7=8 =A: * ': =6: 3@/04;=A9@: =A: 698@6940: ;=4/5;: 84;3@84??9=AB1: @?@4A: *: (': 7=8:A+B: ;B=8A: 8A=,;B1: 8+@792: =: "'$: 69BA+=@;=09B1: 90964/B=1: +56/@?+;@/9?: =?961: 6B?/B=8B: 9: -BA@?@=@;B: ?@ABA: ?@0*=/B6: A@: 8@6940:+56/@96B1:8@6940:?+;@/96B1: 4A5;=AB6: 9A8:99A9=;:?@?B/A=A9@': 56/@5A@;4BB': ;;: 0=AB/9=;8: 7B/B: *+=/0=?B4A9?=;:2/=6B' $")')33B?A:@3:*:@:: 88=5:@3:CBA@?@=@;B: 64/92:A=,9;9A5: ('()%"#))-!"$!$# @/:A+B:*4/*@8B:@3:A+B:8A4651:$:-BA@?@=@;B: 8+=0*@@8: 7B/B: */B*=/B6: 4892: 8=0B: */B*=/=A9@:0BA+@6':+B:3@/04;=A9@8:.=/9B6:9: *1:=648AB6:A@:('1:'1:=6:':,5:A+B:=669A9@: 188 10/. Form Initial Initial Final Visual '% , + + + + + + + + + + ;;;;==%4 :; ; $746544=, ,9:4:67=2= 7 5 ulatio Assay pH Assay appearanc 6="=6. :2 ..5 :; =4:%:25:6= ,7932# 5 n (%) (%) e 4 $$3634 5 F1 99.78 8.5 100.84 No visual 4. CONCLU USION changes F2 97.74 7.5 95.21 No ?= .45$$ 63 43 4 :; =4:%:25:6= 32 .58 8-:: visual ;: ; 9 : 98 87654 534:2. . -* 52, 4= changes -9=.=2%= : ;52 5 2 34:3,52 2 '4 = ;: ; 9 : 98 87654 534:2 54 5 4-* F3 101.37 6.5 96.99 No ' =3$34= =, 4= 3#=.4 .45$$3 364) /36= 4= visual ;: ; 9 : 98 87654 534:2 54 5 4-* ' .:/=, 4= #9=54 5 4=.4 changes ,=#95,54 534:2) ) / 43 5 '( ,=%9=5.= 32 =4:%:25:6 6= %:24=2 '4 ?= 5,,3 34:2 : ; '0 -,/, *)(''%)$% #( "! '!% # !#'%)% + + + + + ( + + + + + + + + + + + + + + ++ + + + + $74654 5 4=, ,9:4:67=2= +*? ,3, 2:4 #)+ + + + + !%+ + + + + !'! + + + + + #(+ + + + + %!#$#)#'%+ + + + + .3#23;3 ; 3%52 5 2 64 =252 5 2%= .45$ 5 $ 63 43) -594 9 43%76596 54 5 4 #'!#) 6:/=9 -*' ?=.= ;3 ; 32,32#. .7##=.4 4 4 5 54 ?= ;::98 ; 9 8765443:2 10 /5. -9=-59=, /34 +*?)) 5 ? :-438332# 4= -* :; ;: ; :987654 5 43:2 3 . 8:9= 52, 4= ;::98 ; 9 8765443:2 1( /5. -9=-59=, /34:74 5 %9 433%56 45 52 5 2 5,, 433:2 :; 52 5 2 34:3,52 24 ;: ; :9 +*?' ?= -.3%56 5---=59522%= :; $:4 5 5 8532453232# =4:%:25:6= .45$$3 364 3 258 5 8-:: ;;::98 9 8765443:2. 9=8532=, 72%52#=, :"=9 5 0(! 5 -9:,7%4.' %6=59)) 52, "3.%:7. 63 73, ;::98 8:24 -=93:,' +:4 9=4532=, 5 :8:#=2:7.) 9 ; 98 /34 5 %595%4=93.43% :,:9 522, 22 324=2.= -32! 5 5 5 :9 22#= 0' 3$5) ) ') 3$5! 3 223) ') 59%53.) *') 5 5 %:6:9' 32:9 9=,7%43:2. 32 -* 52, 9=65443"= 5 7%6:.) ') 9257,) ' ,=2.34 /=9= -9=.=24=, 32 ?55$$6= (' 2 ? 5 = 5..5 :; 5 ' =4:%:25:6= ,=%9=5.=, $ (' 32 24=9225443:25 9 56 :79256 :; -598 9 85%=743%.) ( 10 52, ' 32 1() 32,3%5432# 5 #95,756 6:.. :; 5 00 0' ' 5%43"= %:24=24 :"=9 438= 13#.'0' (,+ + + + + * 0 ( + + + + + ,,32# '0 :; $746544=, ,9:4:67=2= 5 ' 9:%==,32#. :; 4= 5243:3, 224 ,:=. 2:4 .==8 4: 38-9:"= 4= 5 324=925443:256 5 6 %:2;==9=2%= :2 598 ; 9 8:23.5443:2) 5 .45$$3634 :; =4:%:25:6= 32 4= 5 7=:7. 5 4 =2="5) ( ' ;;::98 9 8765443:2 -59443%76596 544 -* '' 5 9 5 98 = /:76, 6 63= 4: =4=2, :79 #95443477,= 4: 4= 5 4 ays 96 %:66=5#7=. ;9 ; ;;99:8 4= =.=59% 522, ="=6:-8=24 5 (% 32"5675$$6= . 5 .7--:944 52, %:2493$743:2. 4: 43. 9 92 .47,' As =-59448=24 ) 9 5522, 4= %:8-522 =-6= ;::9 4=39 5 ; 94 0 5 5 10 1 0 15 T im m e (M on th s s )) F1 F2 % , + + + + ++ + + + + (((%%$! #( " #) #( ( ( + + + + + + + + + + + + + + + + + + + + + + + + ++ + + + + %!#$#)#'% ) !'! , %!#$#)#'% ) !'! , + + + + + + + + + + Formulation F1 F2 s Parameters 0 12 0 12 month months month months pH 6.47 6.05 6.50 6.08 Relative 1.05 1.05 1.06 1.06 density % Assay of 96.3 94.36 96.8 93.24 Ketoconazol 2 3 e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amples M (mg) #" ! $# ! ' ' %$ ' %$ SD 68697)61 6 1/-- 87+297+ )/ G E E E E E E E E E E E E E E E E E E E E E E E E E E E EFC 6 ::+ % % % % % % % E E E E E E E 6 E E E E E E EE E E E E E E HPC 3L GF D C E E E E E E E E E E E E E E BA D @ E E E E E E E E E E E E E E FA D G E E E E E E E E E E E E E E )/E E E E E E E ?A@@ @=B!E E E E E E E 523375)4:3E E E E E E E )(6)) 6E E E E E E E +823E E E E E E E HPC 6L ?? D @ E E E E E E E E E E E E E E @@C D ?? E E E E E E E E E E E E E E ?GC D @ E E E E E E E E E E E E E E 4:9/8*/86))4/ 6: 16+7 )(7 .44-15 76#78 6:+ -755 E E E E E E E E E E E E E E E E E E E E E . E E E E E E E E E E E E E E E E E E E E EE E E E E E E HPMC 3L G= D C E E E E E E E E E E E E E E B D C E E E E E E E E E E E E E E F= D = E E E E E E E E E E E E E E 7-65)49E E E E E E E HPMC 6L ?CG D B E E E E E E E E E E E E E E CF? D B E E E E E E E E E E E E E E F=? D ?= E E E E E E E E E E E E E E HPC/HPMC ?? D E E E E E E E E E E E E E E C? D B E E E E E E E E E E E E E E @@A D ?@ E E E E E E E E E E E E E E # # 79( (6:496- *8/*78)475 /. C *84:)7+ 561*-75 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E 3+3L Placebo Mix 6L Placebo ??A D G E E E E E E E E E E E E E E @BG D ?@ E E E E E E E E E E E E E E ? E E E E E E E E E E E E E E Samples TS (MPa) ! ' YM (MPa) HPC/HPMC ?FC D = E E E E E E E E E E E E E E A?C D ? E E E E E E E E E E E E E E @@A D ? E E E E E E E E E E E E E E ' %$ SD ' %$ 3+3L+PAR HPC 3L C D ? E E E E E E E E E E E E E E ?C= D G E E E E E E E E E E E E E E ? D ? E E E E E E E E E E E E E E Mix 6L+PAR ?FF D C E E E E E E E E E E E E E E CB D ?@ E E E E E E E E E E E E E E ?@ D = EEE E EEE E EEE E EEE E EEE E EEE E EEE E HPC 6L n/a 700 n/a ;:987654:3 :21078 /. 6 E E E E E E E E E E E E E E E E E E E E E- ,,785E E E E E E E6:+ 4:9/8**/86))4:3 E E E E E E E 8 6E E E E E E E HPMC 6L HPMC 3L B D ? E E E E E E E E E E E E E E =B D @ E E E E E E E E E E E E E E C@B D @? E E E E E E E E E E E E E E *68697) 1 6 1/- -7+ )/ (43(78 .44-1 1655 'GF E E E E E E E E E E E E E E E E E E E E E E E E E E E E . E E E E E E E E E E E E E E HPC/HPM ?B D @ E E E E E E E E E E E E E E ? D @ E E E E E E E E E E E E E E =? D ?G E E E E E E E E E E E E E E HPC/HPMC B D @ E E E E E E E E E E E E E E ? D = E E E E E E E E E E E E E E @F D G E E E E E E E E E E E E E E ?FF 13$ ::+ )(49#:755 'BAA?C "1$! 4)( 6 % % % % % % % E E E E E E E 6 E E E E E E E E E E E E E E % % % % % % % E E E E E E E E E E E E E E E E E E E E E 3+3L Placebo 9/8875*/:+4:3 4:987657 4: +454:)7386))4/: )417 E E E E E E E E E E E E E E E E E E E E E : 6 E E E E E E EE E E E E E E Mix 6L Placebo = D C E E E E E E E E E E E E E E @= D E E E E E E E E E E E E E E ?G D E E E E E E E E E E E E E E 'FA@@A 5$ F % E % E % E % E % E % E % E E E E E E E E-6,,78 .44-15 5(/ 7+ )((7 6 E E E E E E E . E E E E E E E E E E E E E E )E E E E E E E HPC/HPMC G D ? E E E E E E E E E E E E E E ?A D A E E E E E E E E E E E E E E BC D ?G E E E E E E E E E E E E E E-/:375) +454:)7386))4/:! (4-7 C E E E E E E E : 6 E E E E E E E E E E E E E E-6,78 .44-15 E E E E E E E E E E E E E E .E E E E E E E 3+3L+PAR R +454:)7386))7+ )((7 .665)75) 6 E E E E E E E ) E E E E E E E .E E E E E E E 454:)7386))4/: )417 : 6 E E E E E E EE E E E E E E Mix 6L+PA AR F D ? E E E E E E E E E E E E E E @ D F E E E E E E E E E E E E E E ?A D ? E E E E E E E E E E E E E E 9/887-6))7+ 4)( .44-1 )(49#:755 6::+ 9/1*/54)4/:! 6 E E E E E E E E E E E E E E . E E E E E E E E E E E E E E 6 E E E E E E EE E E E E E E :6 :/) 6** E E E E E E E E E E E E E E E E E E E E E-4960-7 561*-7 7977+7+ )(7 176528717:) E E E E E E E E E E E E E E E E E E E E E E E E E E E EE E E E E E E 4:+496))4:3 )(6)) )(49#78! +8223 96*694), /. )(7 E E E E E E E E E E E E E EE E E E E E E+7497 6 E E E E E E E 6 E E E E E E E E E E E E E E 8-/6+7+ .44-15 E E E E E E E . E E E E E E E +454:)7386))7+ 1/87 5-/ -, */5540-, +27 )/ 6 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E EE E E E E E E ) % 87+297+ 6)78 *7:7)86))4/: 6::+ 4:987657+ 16))84 E E E E E E E 6 E E E E E E E 6 E E E E E E E 6 E E E E E E E E E E E E E E 6E E E E E E E +7:54),E E E E E E E 6:+ E E E E E E EE E E E E E E 0657+ C *84:)7+ .44-15 E E E E E E E E E E E E E E E E E E E E E .E E E E E E E 7(404)7+ + E E E E E E E45)4:9) 179(6:496-! +454:)7386))4/:! E E E E E E E E E E E E E E 6E E E E E E E 100 6:+ 87-7657 E E E E E E EE E E E E E E 07(64/85 4:.--27:97+ 0, */-,178 E E E E E E E . E E E E E E E E E E E E E EE E E E E E E 80 ),*7! -6,,78 E E E E E E E 6E E E E E E E 9/:.443286)4/:! 6::+ +8223 -/6+4:3 . 6 E E E E E E E 6 E E E E E E E 8 E E E E E E EE E E E E E E 60 7:( E E E E E E E (6:97+ ..44-1 E E E E E E EE E E E E E E 5)87:3)(! ((4-7 E E E E E E E E E E E E E EE E E E E E E 9/:)8402)7+ 2E E E E E E E E E E E E E E )/ .--7404-4), ;:987657+ )(49#:755 . 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E E E E E E E 87-7657+E E E E E E E ? @E E E E E E E8745! ! 7 6-! /1*686)47 5)2+, /: :/ E E E E E E E E E E E E E E E E E E E E E E E E E E E E 4 E E E E E E E E E E E E E E E E E E E E E /7- )75) E E E E E E EE E E E E E E /. )(7 +8223! (4-7 )(7 E E E E E E E E E E E E E E E E E E E E E 8 E E E E E E E E E E E E E E E E E E E E E-6,,787+ 6 E E E E E E E .44-15 E E E E E E E .E E E E E E E'CC -6,,785$ 87-7657+ E E E E E E E 6 E E E E E E E + B E E E E E E EE E E E E E E /8/+45*78540-7 .44-15 /28::6- /. (6816 5,5)715 )/ +7)7814:7 +454:)7386)4/: ) E E E E E E E E E E E E E E E E E E E E E E E E E E E E417 /. E E E E E E EE E E E E E E E E E E E E E . E E E E E E E 8 E E E E E E E E E E E E E E 69, 6:+ E E E E E E EE E E E E E E 4)(4: )(7 561 E E E E E E E E E E E E E E 6 17 )417 .8861 E E E E E E E E E E E E E E . 6 17! 4:+496))4:3 E E E E E E E 6E E E E E E E /786--E E E E E E E (68169/-/3,! @?A! FF! ??@?? 3 E E E E E E E E E E E E E E E E E E E E EEE EE EE EE EE EE EE 5414-68 87-7657 07(64/8 E E E E E E E E E E E E E EE E E E E E E (7 5-43() +4. E E E E E E E ( E E E E E E E ....7787:97 .E E E E E E E 16,, 523375) )(6))! $# 6 E E E E E E E E E E E E E E 6 E E E E E E E (4-7 0/)( .44-1 ),, E E E E E E E E E E E E E E . E E E E E E E ) ,*75 687 E E E E E E E E E E E E E E 524) 00-7 .//8 *8/-/:37+ +7-478,! * 6 E E E E E E E . E E E E E E E E E E E E E E E E E E E E E */-,178E E E E E E E (45 8757689( 65 .22:+7+ 0, )(7 4:4 E E E E E E E E E E E E E E E E E E E E E . E E E E E E E E E E E E E E E E E E E E E 45)8,, /. 8 E E E E E E E E E E E E E E +45)8402)4/: 6...779)5 87-7657 +,:61495! E E E E E E E . . E E E E E E E E E E E E E E E E E E E E E 4)( )(7 E E E E E E E E E E E E E E 947:97! 779(:/-/3496- 77-/*17: E E E E E E E E E E E E E E :) 6::+ E E E E E E E 6 E E E E E E E 1/87 (/1/37:7/25 16))84 4: )(7 14 E E E E E E E E E E E E E E 6 E E E E E E E E E E E E E E E E E E E E E 7+ .44-1 E E E E E E E . E E E E E E E ;::/6))4/:! 7*20-49 /. 780046 )(8/23 6 E E E E E E E E E E E E E E E E E E E E E 8 E E E E E E E 3( ) / E E E E E E E E E E E E E E-4#7-, .6694-4)6))4:3 768-478 +8223 E E E E E E E . 6 E E E E E E E E E E E E E E 8 E E E E E E E +4...2254/: . . E E E E E E E 6600-7 @ 6 E E E E E E E E E E E E E E 8 ::) 387717:)5 4)( :47854), /. 7 6 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E 7-386+7 5(/ 5 )((7 179(6::496- *8/*78))475 /. )(7 5 E E E E E E E ) E E E E E E E 6 E E E E E E E 8 E E E E E E E E E E E E E E E E E E E E E 661 1*-75 E E E E E E E 692-), /. (681 E E E E E E E E E E E E E E 8 169, / AG?C?CF@ E E E E E E E E E E E E E E @GC E E E E E E E F E E E E E E E-6,,78 .44-15 5(/ 7+ )(7 (43(75) 6 E E E E E E E . E E E E E E E E E E E E E E E E E E E E E ) )7:54-7 E E E E E E E E E E E E E E @?F? ::+ / AG?C?C=@@GC @ E E E E E E E 6 E E E E E E E E E E E E E E E E E E E E E ?F? 5)87:3)( E E E E E E E '?$ 47+ 6:+ -6,787+ E E E E E E E . E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E EE E E E E E E *-6970/ .44-15 (6+ 1/+786))7 5)87:3)( ' =A E E E E E E E . E E E E E E E E E E E E E E 6 E E E E E E E 3 E E E E E E E E E E E E E E 191 *)(' %%$#"!%"*! "$!$##!""! $ "#"!"#" $#!*"*!$ Luca Eva Uhljar1 1 1 , Adam Kajari , Rita Ambrus 1Institute of Pharmaceutical Technology and Regulatory Affairs, University of Szeged, Hungary 1. INTRODUCTION "# ""#! 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INTRODUCTION ::/= ! #$!/% #/ GFEDECBA@?ABC=DEF;B:GG9;8B7F6BD=6BAF?7F5B ,=6BAFED62D.6B2E7DC3B;E0@DECB)7;BAF6A7F64B5B 4F@3;B@;64BDEBDF67DB7241F607D64B4;67;6;BE/BD=6B 3F74@7005B 744C3B AF61)63=64B GB DEB D=6B @AA6FB 37;DFECD6;DC70B DF72DB 4@6B DEB D=6FB =3=05B A@F/64B)7D6FB)=06B2ECDC@E@;05B;DFFC3B@CD0B ;6062D.6B C=DEF5B 6//62DB ECB D=6B 7241/EF?C3B 2E?A06D605B 4;;E0.64B 7C4B =E?E36C+64-B"/D6FB /@C2DECBE/BD=6B;DE?72=-B,=6BAF?7F5B4F@3B/FE?B D=7D*BE0@DECB)7;B;AF7564BECDEBD=6B72D.6B2EF6;B D=6B GG9B 3FE@AB ;B E?6AF7+E06*B )=2=B )7;B CB /0@4+64B 64B 2E7D6F;B @;C3B D=6B ;7?6B ;5CD=6;+64BCB(''B7/D6FB4;2E.6FC3B7B@C%@6B A7F7?6D6F;B7;B@;64BCB;D6AB9*B"2D.6B2E7DC3- 6C+5?6*B$#1;D?@07D64B",G7;6*BCBA7F6D70B2600;B :/8 !#/% # 4@FC3B6!A6F?6CD;BECBD=6B37;DF2B?@2E;7B (-BB F;D*B 7B DD7C@?B 4E!46B ;@;A6C;ECB )7;B 9CB*BE?6AF7+E06B)7;B/E@C4BDEB6BD=6BD=F4B AF6A7F64B 5B 744C3B 7B )63=64B 7?E@CDB E/B ?E;DB2E??EC05BAF6;2F64B2=6?270B;@;D7C26B DD7C@?B 4E!46B DEB D=6B A@F/64B )7D6FB 7C4B CB C307C4*B )D=B D=6B 4F@3B 0;D64B ECB ?!C3B @CD0B 6C3B =E?E36C6E@;-B 9CB EF46FB DEB 7AAFE!?7D605BB?00ECBAF6;2FADEC;B -B,=6B ECB D=6B 4;A6F;ECB /EFB 6CD6F2B 2E7DC3*B 0E270+7DECBE/BA=7F?726@D270B4F@3BAFE4@2DECB @4F73DB1*BG07;"2F50*B7C4BD=6BAF6A7F64B ;BEC6BE/B@FEA6;BAF?7F5B;DF7D636;BDEB6C;@F6B DD7C@?B 4E!46B ;@;A6C;ECB )6F6B 74464B DEB D=7DBD=6BAEA@07DECB=7;B7226;;BDEB7//EF4706B7C4B )63=64BA@F/64B)7D6FB)D=B2ECDC@E@;B;DFFC3B 3EE41%@70D5B ?642C6;-B ,=;B ;D@45B 46;2F6;B @;C3B 7CB E.6F=674B ;DFF6FB /EFB 7DB 067;DB (B D=6BD62=CE0E3270B46.60EA?6CDBE/BE6%@.706CDB ?C@D6;- 46075641F6067;6BE?6AF7+E06- :/.# #/ " :/,098'103/05./,8964. ;;E0@DECBD6;D;B)6F6BA6F/EF?64B722EF4C3BDEB :/0 #$!/% # D=6B GB?ECE3F7A=B5B@;C3BD=6B;D6BB 4;;E0@DECBD6;D6FB: GBD5A6B99B7AA7F7D@;B)D=B7B 9CBD=;B;D6A*BE?6AF7+E06B;@;A6C;EC;B/FE?BD)EB A744068B7DB7B;DFF6FBFED71DECB;A664BE/B(BFA?B ;@;A6C;EC;*B9"B7C4B9*B)6F6B2E7D64BECBC72D.6B 7C4BD=6B4;;E0@DECB?64@?BD6?A6F7D@F6BE/B-B ?2FE2F5;D700C6B2600@0E;6B2EF6;-B@;A6C;ECB9"B B-B-B"CB724BF6;;D7C26BD6;DB)7;BA6F/EF?64B )7;B AF6A7F64B 5B ?!C3B D=6B /E00E)C3B CB 7B 4;;E0@DECB ?64@?B E/B B ?0B E/B -(B B C3F646CD;B?6AF7+E06*B072DE;6B?ECE=54F7D6*B =54FE2=0EF2B 724B :08B AB (-B /EFB B =E@F;-B ;E4@?B07@F50B;@0/7D6*B7C4B41;E4@?BA=E;A=7D6B "/D6FBD)EB=E@F;*B/EFBD=6B4;;E0@DECB@//6FB;D736*B 4E4627=54F7D6B CB A@F/64B )7D6F-B ,=6B ;62EC4*B B?BE/B-BB47;2B;E4@?BA=E;A=7D6B 9*B7B;E0@DECBE/BGB7C4BG*B)7;BAF6A7F64B )7;B74464BDEBD=6BB?BE/B-(BB=54FE2=0EF2B 5B4;A6F;C3BD=6B2E?AEC6CD;BCBA@F/64B)7D6FB 724B ?64@?B CB D=6B .6;;60B 7C4B 74@;D64*B /B =67D64B DEB B B /E00E)64B 5B ;0E)B 2EE0C3B 7DB C626;;7F5*B)D=BBB=54FE2=0EF2B724BEFBBB FEE?BD6?A6F7D@F6B/EFBB=E@F;-B ;E14@?B=54FE!46BDEBD=6BABE/B-BB-- G7FD;B9"B7C4B9B)6F6BD=6CB?!64B7C4B;AF7564B ECDEB C72D.6B 2EF6;B CB /0@4+64B 64B 2E7D6F;B :;/*#!#$%!!/ " 64AEF6*B +62=B 6A@02B B /EFB 07,=6B E62D.6B )7;B DEB 2E?A7F6B D=6B F7D6B 7C4B ;2706B 7D2=6;*B "6FE?7D216046FB "*B 6!D6CDB E/B 7;EFADECB /E00E)64B 5B 7B E1 )D+6F07C4*BBE;2=BA0ED076%@.706C26B 7;;6;;?6CDB E/B D=6B 36C6F2B 7C4B /EFBA0EDB;2706B7D2=6;- E;62B:EF3C7DEF8B4F@3;B7/D6FB7B;C306B4E;6BE/B 672=B4F@3B7DB7B4E;6BE/BB?3- 197 ('% $#"! "" "7851"168168561# $1(%#1681888%'81# $1 "58'1(6111515")%1%1$1- $##""" 987786543221 0322/.-21 ,+*1 )(51 '(%$8#1 6"1 98778654.221 0.22/-2221 ,+*1 !8'( 581 "1 681 5+(771581"1681877865178#16"156'16"$8681 $#%#"(" 81 (%661 "1 91 )(51 %'8(58#1 "+1 ..16"1.16"1('881 (1!86681"68'681 7(81 $#$#" " 81 (%661 "1 #($641 2/331 )(51 %'8(58#1 6"1 ('881 !86681 8%68'1 7(85181 %(71 (76(681(%#1 (%66(681'"+"56"%1"1 681#887"8#1"+ 7(6"%151858%68#1%1(!781 " #" 9"+(5"%1 "1 #55"7 6"%1 "7851 "1 681 - 68561# $1(%#1681888%'81# $1"58'1(611 " " #" $#$#" " 81 %(71 (76(681 (%#1 (%66(681 '"+"56"%1%1(1'(5 78 $1 1 5")51 681 (8($81 (+('"%86'1 "7851"1"+8("781(681681(#+%56(6"%1 Components Function Quantity "168168561(%#1888%'81# $51 in mg Omeprazole API 40,00 Lactose Filler 34,00 monohydrate Sodium lauryl Solubilizer 1,20 sulfate Sodium pyrophosphate Buffer agent 5,20 dodecahydrate Film-former/ 39,60 HPMC Binder Film-former/ HPC 2,00 Binder " #" 8($8#1 (+('"%86'1 "7851 "1 681 Cellets 700 68561# $1(%#1681888%'81# $ Neutral Cores 285,24 Titanium #" Colorant 2,00 dioxide 1!"8 (78%61#87(8#/878(581"+ 7(6"%1"1 Talc Lubricant 0,80 "+8("781(51!88%1#887"8#1"1681+(86 Eudragit L30- Enteric D55 (dry 39,96 coating agent PlasAcryl (dry #"! ! Anti-tacking 6,80 matter) matter) /Plasticizer -1('511 1(+('"68(1-.1-.0-*1 /. $##" " 1 )(518(7 (68#1(61(15%$781"%61%1- 21+% 6851 "+1 665)))56(656('"+56(656'5.385'6 81(+" %61"1"+8("781878(58#1(611- 1 1 (7(!781 8168561(%#1888%'81# $51878(58#1"+121 "%/'"56/(%(755/6"/6)8%6/'8+'(75/!/68+5/%/ 6"1-21(%#1-16"13.1"1681"+8("781 8%$7(%# ''8558#1 318!1 2 3 858'6871 1 '"+(5"%1 "1 #55"7 6"%1 198 4321 04/.-.,+/.0*)0()'+.+%)$#/'"!.0*) ./)!4$'0*.,+/.0*)"!.*) ++*$)$#4$'.-$*/+%)$!.*! +)1, 2, Klemen Korasa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able 4. Most desirable solution from numerical +0=@?-5'01+.1'0.14*)5'20=.'-++ optimization. @1/011+0.14?+.'0+.(+@.'05@?'0@+0= Factor Optimal value )5.-5'5@(.='525.-5'?.141+.1'0 MCC 49.7 mg 2@10/.-1.2575'@@+51+('! Lactose monohydrate 28.3 mg HPMC 2.0 mg Table 2. Significant factors in screening DoE. Water quantity 66.9 mg Response Significant factors Spheronization time 167 seconds Width of particle Water quantity (p = 0.0006) Spheronization speed 615 rpm size distribution Spheronization speed (p = 0.0123) Sphericity Water quantity (p = 0.0138) Spheronization speed (p 0.0001) !# Spheronization time (p = 0.0485) Particle size Spheronization speed (p 0.0001) -1@ @.2= =4'0@.5+.= .- @2@@2( 0.6371.0 mm +??(1+.1'0'+1.2575'@@+51+(' Process yield Water quantity (p =0.0829) +??5'+- .' @142(.+0'2@( '?.141 '542(+.1'0 '4?'@1.1'0 +0= ?5'@@ ?+5+4.5@ '5 .- =('?40. ' +5=1('( )!"!# $,$# ''%$# # # ?((.@! ?5'@@ +51+(@* )+.5 2+0.1.* $ @?-5'01+.1'0@?=*+0=.14*)51=0.11= ' @142(.+0'2@( '?.141 '.- .- .-5'2/- + (+,..7254+0 @5010/ =@1/0* '542(+.1'0 +0= ?5'@@ ?+5+4.5@* + 6 6 )-1('542(+.1'0+0=@@'?.141+.1'0)+@ 1.2575'@@ +51+( ' ; =@1/03 +-1= 2@10/ + '410= ?5140.+( )+@ 2@= ;('%$# )3! - =@1/0 10(2== $ =@1/0! - +??5'+- 0+(= 5'2@. ?5140.+(520@$:4'=(0.11+.1'0 ' 51.1+( +.'5@ +0= ?5'=2= 1.0.5?(1+.@! ?((.@ )1.- =@15+( 2+(1. +..512.@* 10(2=10/ 0+55') @1 =1@.512.1'0* -1/- @?-511.* +0= -1/- 1(=! -@ 10=10/@ Table 3. Factors in KCV design. @2??'5. .- +@11(1. ' 10./5+.= Mixture Process parameter '542(+.1'07?5'@@ =('?40. 10 (10 )1.- components @+0=-1/-(1/-..-0.1+('.-1@ MCC (40 7 50 mg) Water quantity (40 7 70 mg) @.5+./ '5 2.25 ?-+54+2.1+( ?5'=2. Lactose monohydrate Spheronization speed =('?40.! (24738 mg) (4007800 rpm) HPMC (276 mg) Spheronization time Total = 80 mg (607240 s) *!# +- 5@?'0@ )+@ 101.1+(( +0+(= 10=11=2+((! 2451+( '?.141+.1'0 )+@ .-0 :!'50=2@.5 5+4)'5, ?5'54= '5 00'+.1 -+54+2.1+( ('?40.* .' =.5410 .- '?.14+( '410+.1'0 ' '542(+.1'0 +0= ?5'@@ +02+.2510/*+0=2+(1.@@25+0*$88#! ?+5+4.5@! - '.1@ )5 .' 410141 .- )1=.- ' ?+5.1( @1 =1@.512.1'0 +0= 4+141.-@?-511.*@ ( 10 =@15= @1 5+0/ +0= ?5'@@ 1(=! - -1@5@+5--+@0@2??'5.=5,+*=!=!* 4'@.55=@'(2.1'0)1.-=@15+1(1.@'5 ''4@.'! ' 8!99 1@ ?5@0.=10 ('%$# ! - '?.14+( 200 7654 32120/7.2-,+/*+-/-)32(,'%,$12+.2,#/3+*/'+"((2(.2-,+/*+ %/-,2-,+-$*/'.$,!+/*+.2,/%0/7'".$32+$-+ %%"0+*$0.(+ "(23+ /-+-$'+(72%,'/(%/7! +1 1 , /+7 1. INTRODUCTION BH@JB4G 36G :DEA;AJ2G IB9DG ;E@?J?A=G EFG #G G ?EICAJE@G EFG $G B@=G 8B?G KJIHG FEDHCIBAJE@?G J@A@==G FEDG ;B?A=G E2DG A6G =DC7G ;E@ABJ@J@7G IB9DG BAG B::IJ;BAJE@G BDG @EAG 9AG BG 8J=I9G J@A7DBA=G A6J;/@??G EFG -###G H4G 36G DG FJIHG 8B?G =E?B7GFEDHG J@G :6BDHB;95GBIA6EC76G A69G 8DG =DJ=GBAG#G GFEDG!#GHJ@GB@=G;CAGJ@G#*# HHG J@ADE=C;=G=;B=?GB7E4G36DGBDG:69?JEIE7J;BIG ?J4GG B@=GA;6@EIE7J;BIGIJHJABAJE@?GA6BAG:D2@AGA6JDG 8J=DG B;;:AB@;4G 36J?G ?AC=9G FE;C?=G E@G A6G +, +-$'++ A;6@EIE7J;BIG B?:;A?G EFG G ('G ?:;ADE?;E:9G EFG DG FJIH?G 8DG :D:BDBAJE@4G1C;;BIGFJIH?GBDGC?CBII9G:DE=C;=G :DFEDH=G C?J@7G ('G $G %1DC/DG :AJ;?5G 9G A6G ?EI2@AG ;B?AJ@7G HA6E=5G J@G 86J;6G BG DHB@9GJ@GDFI;AB@;GHE=4G36GFJIH?G8DG 2J?;EC?GIJ0CJ=G;E@ABJ@J@7GBGFJIHG;EH:E?JAJE@GJ?G :IB;=G E2DG A6G HB?CDJ@7G 8J@=E8G EFG ;E@AJ@CEC?I9G ;B?A=G E@G A6G HE2J@7G IJ@D4G 36G ?:;ADEHADGB@=G;E2D=G8JA6GFIBAGBICHJ@JCHG A6J;/@??GEFGA6G;B?AJ@7G=FJ@?GA6G=E?GEFG=DC7G 8J76AG A6BAG DFI;A=G A6G IJ76AG G :DG?CDFB;GBDB5G86J;6GJ?GBG;DJAJ;BIG:DE=C;AJE@G =A;AED4G36G ('G ?:;ADBG 8DG E=G BFADG :BDBHADG .-,4G 36G J??CG EFG =E?G C@JFEDHJA9G --!G B;0CJ?JAJE@?G BAG D?EICAJE@G EFG -!G ;HG J@GA6G A8@G FJIH?G BI?EG =:@=?G E@G A6G =D9J@7G -DB@7GFDEHG-##GAEG!##G;H4G :DE;??4G+6@GA6G=DC7G;E@A@AG:DGFJIHGHB??GJ?G *:D??=5G A6G D?J=CBIG HEJ?ACDG J@G A6G FJIHG 4+3++ :IB9?G B@G JH:EDAB@AG DEI4G )CDJ@7G A6G =D9J@7G 36G =DC7G ;E@A@AG J@G A6G FJIH?G 8B?G 2BICBA=G :DE;??5GA6DGJ?GBGDJ?/GEFGC@2@G2B:EDBAJE@GEFG C?J@7G A6G ;IB??J;BIG B::DEB;6G EFG =J??EI2J@7G A6G HEJ?ACD5G 86J;6G IB=?G AEG C@2@G HB??G EFG A6G FJIH?G J@G 8BADG B@=G =ADHJ@J@7G A6G FJIH?4G36G BJHG EFG A6J?G ?AC=9G 8B?GAEG =2IE:G BG ;E@;@ADBAJE@G EFG $G J@G A6G ?EICAJE@4G 36G HA6E=GFEDG=ADHJ@BAJE@GEFG=DC7G;E@;@ADBAJE@G BG 8B?G HB?CD=G BAG #G @HG C?J@7G BG J@GA6G;ADE?;E:94 J?G?:;ADEHADG%BDD9G!#5G7JI@A5G4G +.",2'$"0(+"-3+.2,#/3( 36G ?:;ADBG EFG BIIG *;J:J@A?G 8DG 2BICBA=5G B@=GA6JDGBGDB@7G=J=G@EAGJ@ADFDG8JA6G +.+ A6GHB?CDH@AG8B2I@7A64 36G=G?E=JCHG BI7J@BAG :EI9HDG %$DEAB@BIG -#"!#5G K G 5+7+ +++ 1JE$EI9HD5G 5G *9IJAEIG %9IJ?ED36G;E@;@ADBAJE@?GEFG $GE=G9G2J?G E0CAA5G KDB@;G B?G :IB?AJ;J?DG B@=G A6G =DC7G ?:;ADE?;E:9G8DGC?=GB?GBGDFD@;G2BICGAEG HAE;IE:DBHJ=G 69=DE;6IEDJ=G HE@E69=DBAG G ;BIJGA6GHE=IGFEDG:D=J;AJ@7G=DC7G;E@A@AG $G %1JE?J@A65G IE2B/JB4G $DEA;AJ2G IB9DG FDEHG ('G ?:;ADB4G 36G :D=J;AJE@G HE=IG 8B?G ;E@?J?A=G EFG 69=DE*9:DE:9IG ;IICIE?G %IC;IG G C?G EFG CB@AG G ;6HEHADJ;G K5G?6IB@=5G4 HE=CIGEFG?EFA8BDG$G4-G%1DC/DG:AJ;?5G +7+ ++ DHB@94G 3EABIG EFG G DFD@;G ?:;ADBG 8B?G 36G?EI2@AG;B?AJ@7G?EICAJE@G;E@ABJ@=G?E=JCHG IJ@/=G 8JA6G A6G $G ;E@;@ADBAJE@G 2BICG E@G BI7J@BAG%G5G*9IJAEIG%-GGB@=G $G%#4 86J;6G A6G ;6HEHADJ;G HE=IG 8B?G G -4G 4G CIAJ:IG ;EH=J@A?G E:AJHBIG DB@7G EFG 8B2I@7A6?G 8B?G =ADHJ@=G 8DG CAJIJ=G AEG E9G ;DE??2BIJ=BAJE@G C?J@7G A6G IB2G E@G ECAG BHEC@A?GEFGA6G=DC74G36G?EICAJE@G8B?G?:DB=GE@G B::DEB;64G BG 7IB??G :IBAG C?J@7G B::IJ;BAEDG ###4G %6@A@D5G8JADIB@= G BAG-##GHGA6J;/@??4GG KJIH?G8DG=DJ=GJ@GBG;E@2;AJE@G=D9DG%$G5G 201 .-,+ *)('%$#"%(! (%$%% -17= 0?;6?276= :567= '?9= A976= 25= ?4?+7= 217= ( ;3A/7=.=-17=/79A29=67:5492/?27=21?2=217== BA@@?=;:9=8?976=54=956;A:=?3;4?27=1?07=217= @54@742/?2;54=@?4=0?/+=?2=217=76379=5=217=;:= 92/A@2A/7=5=?=37.=-17=,5+:7/=A9A?+=@542?;49= 8?2@1(= '1;@1= '?9= 7*,7@276.= )4= 3747/?(= 217= *))( ( ( ( @54@742/?2;54=:?,=;9=915'4=?9=?=@5425A/=,52=;4= 9,7@2/?= 5= 7?@1= 8A@@?= ;:= ,/56A@76.= -17= ?= @7/2?;4= ?:5A42= 5= :5;92A/7= 25= :?;42?;4= ;29= 8A@@?= ;:9= ?2= 217= 7637= 5= 217= 8?2@1= ?/7= 7*;8;;2+= ?46= 91?,7.= )4= 5A/= 5/:A?2;54(= 217= 6;9@?/676=?9='?927=?46=54+=217=5/:A?2;549=;4= '?27/=@542742=;4=217=;:9=0?/;76=872'774=.%$ 217=@7427/=5=217=8?2@1=?/7=:=54= 217=3?99=,?27.=-17=:5;92A/7=@542742=?7@29=217= :?99= ?46=21;@ 4799= 5= 217=;:= 8A2= 915A6= 452= ?7@2=217== 6597= ,7/=;:.=-17=A@2A?2;43= :5;92A/7= @542742= '?9= 217= :?;4= @54@7/4= 5/= @5//7@2= :7?9A/7:742= A9;43= )= 9,7@2/5:72/+.= -'5=?9,7@29='7/7=;4=217=5/73/5A46.=;/92+(=217= )= 9,7@2/?= ?/7= ?7@276= 8+= 217= ,/7974@7= 5= '?27/= ?46= 97@546+(= 217= ;:9= ';21= 1;317/= :5;92A/7= @542742= ?/7= 21;@ 7/(= 95= 217= ;312= ,?21= 21/5A31=217=9?:,7=;9=7*274676.=== *))( % ( ( ( ( ( ( ( 707/?=8?2@179=5=8A@@?=;:9='7/7=,/7,?/76= 54= ?= 3?99= ,?27.=-17= ;:9= '7/7= ,59;2;5476=;4= 217=91?,7=5=217=:?2/;*.=-17=)=9,7@2/?='7/7= 582?;476= = ;:9(= 8A2= 217= = @542742= '?9= 6727/:;476= 8+= $;9= 9,7@2/59@5,+= 5/= 54+=97074=;:9=,7/=8?2@1.=-5=977@2=217=:592= ;:=8?2@1=,/7974276=?9=?=@5425A/=:?,.== ( )( 54@742/?2;54= 5= = ;4= 8;?+7/= 8A@@?= /7,/79742?2;07= ;:9= 5/= @542742= :7?9A/7:742(= 217=9?:,79='7/7=,59;2;5476=54=217=6;?354?=5= +)( #$% '?9=@5:,749?276.=-17=:5;92A/7=3/?6;7429='7/7= /7@534;76=?9=?=A97=:72156=5/=6727/:;4;43= = @54@742/?2;54= ;4= 8A@@?= 8;?+7/= ;:9.= ,/79742=87@?A97=217=;:9='7/7=6/;76=;4=217=6/+7/= 79,;27=217=0?/;?8;;2+=;4=:5;92A/7=?46=7*@;,;742= ';21= 5/@76= @5407@2;54.= -17= 9;67= 5= 217= 3?99= 3/?6;742= ;4= 217= *$= 5/= +$6;/7@2;54= 5= 217= :?2/;*= -17= A97= 5= )= 9,7@2/59@5,+= 1?9= 8774= 217= :?2/;*.= )4= 21;9= '?+(= 217= ,599;87= :5;92A/7= ,?27= @597/= 25= 217= 17?2= 95A/@7= @542?;476= 799= @542742(=217=:567=915'76=1;31=/7;?8;;2+.=-17= :7?9A/76= ,/7@;9;54= '?9= .!(= '1;@1= ;9= :5;92A/7= 21?4= 217= 9;67= 7= 5/= ?= 454$6792/A@2;07= :72156.= -1;9= 6/+7/.=-5=6=97/07=?9=?=,/5@799=?4?+2;@?=255= @542742='?9=45/:?;76=25=217=:?99=5=217=;:.===== 5/= 54;47= A?;2+= @542/5= ;4= 217= ,/56A@2;54= 5= *)*)(((( ( 8A@@?=;:9.= 2/?;4;43= ';21= 2792= 6?2?= 8+= /?465:+= 9?:,;43= ,)('' %-17=@?;8/?2;54=5=217=:567='?9=,7/5/:76=8+= 547=5=217=9,7@2/?=?9=?=2792=587@2.=-17=5,2;:?= .= 1;,,(= . = ;A(= . = 7/?;$?/9?4;(= . = 9,7@2/?= /?437= '?9= 6727/:;476= 8+= 217= @/599$ 'A59?(= -..= BA@@?= ;:9= = 70;7'= 5= 0?;6?2;54= 7*,7/;:7429= ;4= '1;@1= 217= :567= -17/?,7A2;@= ,,5/2A4;2;79(= 5/:A?2;549= = 915'76=217=8792=;2=25=217=2792=6?2?.=-17=9,7@2/?= 770?42= 0?A?2;54= ,,/5?@179.= /?437= ;4?+= @15974= '?9= 872'774= "= ?46= =-(=(=""% =@:$.=-17=:567='?9=8A;2=54="=9,7@2/?= ! # " ,5;429= ?46= @5//7?276= ';21= 217= :7?9A/76= @54@742/?2;54=0?A7.=-17=5,2;:?==0?A7='?9= -17= ?A215/9= ?@ 45'7637= ;4?4@;?= 9A,,5/2= ".%%=';21=?4==5="."#.= 5074;?4= /797?/@1= ?46= ;4450?2;54= ?374@+=/797?/@1=@5/7= *)+)= ( ( ( .( ( ( 202 = ;:9;876:5432856106;/2.854/1;/6-,+58.96*5).10(*;/2.845).15 *2:.7;2865:610;*5:6*2961, Feria Hasanpour1 1 1 2 2 3 , 0'$#"5! "#-4 5;#"53' 5$54# *5/$# , Eszter Ducza Adrienn Seres-Bokor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va Rotar Pucko , Rok Dreu $#"! " " " " " "" 1. 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(.-% ,+-*(-)#(. %$/#* 274-1::*9647.6?48=,9?695?=1=9 7-9 =219 =1-15,9 ?69 =219 347;/1:9 =?9 :134137=19 (.( -;439=7./1=39"1419817:;41-9?497::1::1-$ !219 347;/1:9 5?::=1-9 ?69 9 6 8??2,-47=19 81:29 9 9 *9 9 .-14:9 (, *"%*(-)#(#+*0"**+/) 9854?54,:=7//1951//;/?:19+51/9#99 (4( (( @@*9 9 0?/,+,/0,44?/-?19 9 9 9 !219 =,019 ?69 04?51::9 7-9 7/:?9 52731:9 9 =219 7-9 9 ?69 =2198?-1/9 -4;39 =74=4719 9!$9 04?51::9074781=14:96/;1519=2198?402?/?3,9?69 18147/:1-9 "7=149 "7:9 ;:1-9 7:9 =219 =219347;/1:97-9=2149:19-:=4.;=$969=219 347;/7=39/(;-$916?419=7./1=3*979/;.457=9 =24119 347;/7=9 81=2?-:9 9 04?51::39 8731:;89:=1747=19939"7:97--1-9=?987 19 41:;/=1-9 9 =219 :87//1:=9 347;/1:9 "=29 =219 ;09$9?69=21967/9=7./1=398)=;41$9 744?"1:=9 *9 "239 41:;/=1-99=219/7431:=9347;/1:9"=29=219"-1:=9 *9 "=29 =":541"9 347;/1:9 67//39 9 .1="1199=148:9?69:197-9:19-:=4.;=$99 205 ,+*) 98765 32 6371103/3.- ,+* .7)(('/)993( 7 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 7 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 ) ) 4 4 4 4 4 4 4 4 4 4 785'/98%4 4 4 4 4 4 4 4 4 4 1)55 9008 10)76 4 4 4 4 4 4 4 4 4 4 9 4 4 4 4 4 4 4 4 4 4 7 6)!318)/ 277)/9- 9859$ ) 4 4 4 4 4 4 4 4 4 4 2 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 )5 53684 4 4 4 4 4 4 4 4 4 ( 908 6359 778.'/)7 50)118% .7 (('/85 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 ) 4 4 4 4 4 4 4 4 4 4 ) 5$ #0/8 4 4 4 4 4 4 4 4 4 44 4 4 4 4 4 4 4 4 4 32 908 9)/895 #878 738( %'7(. 9008 9859 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 ) 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 9 4 4 4 4 4 4 4 4 4 4 9 ))/89 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 0.0"508)7 .7)(('/)93( 785'/98% ( 4 4 4 4 4 4 4 4 4 4 ) ) 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 44 4 4 4 4 4 4 4 4 4 63784 4 4 4 4 4 4 4 4 4 6)55 )7 /9- #)5 .8(87)//- /3# 4 4 4 4 4 4 4 4 4 4 ) 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 2337 )// 4 4 4 4 4 4 4 4 4 4 2 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 !3(53/%)98% .7)('/85 #900 0.087 510 ) 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 9 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 087!9- 4 4 4 4 4 4 4 4 4 4 5)61/85 +390 '(23376 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 2 7 69- 32 !3(98( 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 (9 )((% 4 4 4 4 4 4 4 4 4 4 )4 4 4 4 4 4 4 4 4 4 # #("5!78# .7)(('/)993( )/53 785'/98% 4 4 4 4 4 4 4 4 4 4 ) ) 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 ( 63784 4 4 4 4 4 4 4 4 4 '(23376 2 7 69- 32 %35).8 '(95 689 10)76 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 7 6) )!318)/4 4 4 4 4 4 4 4 4 4 !3(53/%)98% .7)(('/85 '9 #90 ) ) 4 4 4 4 4 4 4 4 4 4 ) 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 .7 ) 8)9874 4 4 4 4 4 4 4 4 4 78'7868(95 08 98(%8(!- 32 .7 ((' 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 ) '/85 93 4 4 4 4 4 4 4 4 4 44 4 4 4 4 4 4 4 4 4 /8(.90#%900 7)993$ )5 503#(( ( ,.'78 9 4 4 4 4 4 4 4 4 4 4 ) 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 # 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 58.78.)998 %'7(. 9)/89(. #)5 )/53 /3# ) 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 ) 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 # 4 4 4 4 4 4 4 4 4 44 4 4 4 4 4 4 4 4 4 22337 )// 5)61/85 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 44 4 4 4 4 4 4 4 4 4 " " " " " " " " " " 08 !03!8 32 908 9-18 32 .7 (('/)993( 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 ) )4 4 4 4 4 4 4 4 4 4 173!8554 4 4 4 4 4 4 4 4 4 )(% 95 173!855 1)7)6 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 ) 689875 0)5 ) 5.( 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 (2!)((9 2 )4 4 4 4 4 4 4 4 4 4 (2//'8(!8 3( 908 637103/3.- 1)799! 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"# (' %$#"! # #"## ,4/ /BA= ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; =7+:8/B@5B7/ B45BB ?3?7@@ 3759 5 ; ; ; ; ; ; ; ; ; ; 5 ; ; ; ; ; ; ; ; ; ; @ ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; ('' 7 BB@52B :?B5 # ; ; ; ; ; ; ; ; ; ; 587= A/86 ?7B587A@75 5 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; 7#4?B/ /B ; ; ; ; ; ; ; ; ; ; B@:86 58B ; ; ; ; ; ; ; ; ; ; 8BB@:/85/7; ; ; ; ; ; ; ; ; ; 588= 588B ; ; ; ; ; ; ; ; ; ; 5 ,47 @7/A3B/ :9 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; 10 .1-,++ / / / / / / / / / / ,; ; ; ; ; ; ; ; ; ; 5588BB@:/85/7 5//5 ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; 453A@:8=5/ /7; ; ; ; ; ; ; ; ; ; 52BB7/1; ; ; ; ; ; ; ; ; ; 464364B86; ; ; ; ; ; ; ; ; ; B/; ; ; ; ; ; ; ; ; ; *6A@7 ( / ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;4:07= B445BB ?3?7@@ 3759 7#B@52B/ 45= ; ; ; ; ; ; ; ; ; ; B 5 ; ; ; ; ; ; ; ; ; ; @ ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; # ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; .:B78B53 5/ 8 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 5 /A/B585?371 588B ; ; ; ; ; ; ; ; ; ; 55686 86@7=78B ; ; ; ; ; ; ; ; ; ; 8; ; ; ; ; ; ; ; ; ; 4647@ 588B ; ; ; ; ; ; ; ; ; ; 5B@:/85/7 52BB B4588 9@@AAB ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 5 ; ; ; ; ; ; ; ; ; ; 9 @; ; ; ; ; ; ; ; ; ; 7 BB@52B/- 99::@ =7@+:2: ; ; ; ; ; ; ; ; ; ; :/+7B2 5..325B:8/-; ; ; ; ; ; ; ; ; ; 5 # ; ; ; ; ; ; ; ; ; ; )3B4:A64 B47@7 07@7 8: /B5BB/B2533 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 5 ; ; ; ; ; ; ; ; ; ; /6892588B =99977@7827/ 5+ 9 5 ; ; ; ; ; ; ; ; ; ; 9 9 ; ; ; ; ; ; ; ; ; ; 5 +:86 9@@AAB 7#BB@52B ; ; ; ; ; ; ; ; ; ; 9 @ ; ; ; ; ; ; ; ; ; ; #; ; ; ; ; ; ; ; ; ; 52BB7/1 B47 24:27 :9 /:378B /689258B3 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 8 ; ; ; ; ; ; ; ; ; ; 9 8; ; ; ; ; ; ; ; ; ; 5999772B7= 3757/ # 9 9 ; ; ; ; ; ; ; ; ; ; 5 ; ; ; ; ; ; ; ; ; ; 7#B@52B 52BB- ,47 4647/B ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 6; ; ; ; ; ; ; ; ; ; *' %$$%$ $$#- !@A !@A 1 )- - 7B 53-; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;**00 )11-,,+ (++'%$$ #++, "0 11! / / / / / / / / / / )-/ / / / / / / / / / ( ' % / / / / / / / / / / # / / / / / / / / / /- 52BB 05 ; ; ; ; ; ; ; ; ; ; 5/ :?/7@7= 8 B47 ?7B5 ; ; ; ; ; ; ; ; ; ; @ ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 87A@75; ; ; ; ; ; ; ; ; ; " 110 ####11 / / / / / / / / / / # # "$$$$0- " ,1-1% '- 10 / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / ' / / / / / / / / / / / / / / / / / / / / 5 375 ; ; ; ; ; ; ; ; ; ; 7/ # ; ; ; ; ; ; ; ; ; ; 7#B@52B-; ; ; ; ; ; ; ; ; ; ,++$--1 $, / / / / / / / / / / ,; ; ; ; ; ; ; ; ; ;!:/+7B2/1 - *($ ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; ''- - ,55=1 -- - 7B 53-; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; %' %0- / / / / / / / / / /-/ / / / / / / / / / / / / / / / / / / / +$$11 11 11$ %-/ / / / / / / / / / 11%'' 1%,, )101 ,-1%% $ % / / / / / / / / / / , / / / / / / / / / / ) / / / / / / / / / / % / / / / / / / / / // / / / / / / / / / ,1 / / / / / / / / / / 10/ / / / / / / / / / ++1% +/ / / / / / / / / / 110 / / / / / / / / / / ,,-1+0 --; ; ; ; ; ; ; ; ; ; )8B:#=58B/1 1 - *($ ' -; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; - + + +5058BB 5+5=:8 58B58 +5058BB1 - - -1 5+5=:81 -1 58B581 8 8 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; "- ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; AA8+ +1 )-; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; - ,% --1 ++%..0-$$ / / / / / / / / / / / / / / / / / / / /-/ / / / / / / / / / %-/ / / / / / / / / / " " / / / / / / / / / /-+1% -,,-1+0 ,##++,0 / / / / / / / / / / -/ / / / / / / / / / , #/ / / / / / / / / / 00, #++, / / / / / / / / / / # , , 11% 0' -,, -1% / / / / / / / / / /-/ / / / / / / / / / / / / / / / / / / / -/ / / / / / / / / / ,++' - . , ' / / / / / / / / / / %++0-% +- 358B/1 - *($ -; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; - BAA= B =/5@:51 - 7B 53-; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;; ; ; ; ; ; ; ; ; ; "0-1!" ! 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INTRODUCTION 46CABH$CIABCG +"G ;J$+"8G D JIHGFEDCBIGA@?=BIIBG=BCA@7GA@?=BCG =IBCG .EIG 5E0FIA FIEC6543G?EDB2BI1G4?BAIGCB2B=EF0B@4GIB/6AIBA@A@7G.A@BG5IIABI, .EI06=4AE@G EF4A0A-4AE@,G+?BG FBI.EI0@5BG E.G *! 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Krzeminska, K., et al., Suspensions of antibiotics in self-emulsifying oils as a novel approach to -,,* *%*'$ formulate eye drops with substances which (.;5 ;'/;,9=;:67)5 /.71;5915 9:5 /,;/7,769:5 %96.5 Wolska, E., et al., Microscopic and biopharmaceutical evaluation of emulsion and 6.;5 7//)92769:5 65 6.;5 9;6.927)5 ==966;;5 Self-Emulsifying Oil with cyclosporine. ;)5 4,=0)769:15 %9))5 3;5 2=/7,;$5 %96.5 Pharmaceuticals, 2023. 16. 70;015 1)069:15 72.5 4,=0)769:5 %9))5 3;5 306540,6.;,5;'/;,9=;:6157,;59:5/,,;115:5/915 Drug Deliv, 2024. 31. 2. ,;)9=9:7,#5 ,;10)615 %;,;5 /;,4,=;$5 9:5 ,7339615 undergo hydrolysis in aqueous environment. 3. Czajkowska-Kosnik, A., et.al., Comparison of 7$=9:916;,;$5 65 6.;5 ;#;5 45 ;9.65 7:9=7)15(.;5 cytotoxicity in vitro and irritation in vivo for :;'65 16;/5 %9))5 3;5 7:5 ;'/;,9=;:65 :5 :760,7))#5 aqueous and oily solutions of surfactants. Drug 9:4;26;$5;#;151969;5,;10)6154,=56.;1;56;1615 Development and Industrial Pharmacy, 2015. 41. %9))5 );65 65 7//)#5 4,5 ,;916,769:5 6.;5 5 4. Sznitowska, M., et al., Self-emulsifying 652:69:0;56.;5/,;265%96.55/,$0269:54,5 containing as the active substance a drug substance unstable in an aqueous environment. 2)9:927)5160$#59:5.0=7:1 101/;:19:5715;6;,9:7,#5=;$929:7)5/,$02657:$5 pharmaceutical composition in the liquid form, 2025. European Patent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usra Ahmed1, !0"1 1. INTRODUCTION 220 =730?470==,A+=8@;62@=*730?=70=9#(= A@?= ;:9@89@769;= ;69= 54732;61= :7;= 02= 6-#(=/2;!/2;(=/;7)=71=?12-=69=8@;6=847;= 7:/2.2-=;=,?12-=A289169;=+9-24;=*,A+)(= 6704261=6/=7=/9;2':930=;544=1/782=7;-=69= 26/2@= 0'= /96= 3246= 26@?19;= *%+$)= 9@= -552@2;6=;544=82@:2;6721=*=7;-=)#= 38@2;769;#="6/=%+$(=6/2=-@?=7;-=894'32@= 7@2=5@16=32462-=6926/2@=7;-=6/2;=26@?-2-=?1;= 2 0+ 000 7= /2762-= ;9!!42#= ;= 6/2= 96/2@= /7;-(= = @928= += 7;:976= -@?3= :9762@(= 38@2;769;= ;.94.21= 332@1;= 26/2@= *@928(= 24= 95= 54732;6= 9@= 8@;62-= 902:61= ;= 7= //4'= 87@7:267394=194?69;=9;=6/2=A=8@;62-=6704261(= :9;:2;6@762-=-@?=194?69;(=7449;=6/2=-@?=69= /42= 59@= 3:@91'@;2= 497-;(= 6/2= 7?69376:= -55?12=;69=6/2=3762@74#=A21862=61=1384:6'(= 1'@;2=1'1623=95=7=A7678/'1:1= ==-2.:2= 38@2;769;= /71= 1932= -17-.7;6721(= *A7678/'1:1(=2@37;')=71=?12-# ;:4?-;= 6/2= @2?@232;6= 59@= //= -@?= :9;:2;6@769;1(= 49;= 8@9:211= 6321(= 7;-= 6/2= 20)0000 8911046'= 95= -@?= -2@7-769;= -?@;= 6/2= 8@9:211= = 69= 9.2@:932= 6/1= -17-.7;672(= 9= -262@3;2= 6/2= -@?= 497-;(= 6/2= 6704261= 497-;= -@?= ;69= A8@;62-= 6704261= ?1;= 7;= 2@2=847:2-=;=8%=#=8/918/762=0?552@=194?69;= -289169;= 95= 7:6.2= 8/7@37:2?6:74=;@2-2;6= 7;-= 16@@2-= 9;= 37;26:= 16@@2@= 59@= 69= /9?@1= *)= 194?69;= 6/= 242:6@9;:= 1'@;2= 9@= 0'= 6/2;=6/2=7019@07;:2=71=3271?@2-=0'= = :9;6@9442-=18@7';=95=4182:6@98/9693262@=*/2@39:2;65:=2;21'1(= :976;=37'=02=;9.24(=7?693762-(=7::?@762(=7;-= "746/73(=+(=17)=76=7.242;6/=95==;3#= @28@9-?:042=7462@;76.21=59@=497-;#=/2'=7@2= /2= 32:/7;:74= 8@982@621= 2@2= -262@3;2-= //1822-(= 7?693762-= 7;-= 244= :9;6@9447042= 6/= 7= 5@7046'= 62162@= *$@27(= 7;2;(= 8@9:211(= @21?46;= ;= @2-?:2-= 3762@74= 491121#= 2@37;')#= A@?:7@@2@= ;62@7:69;1= 7;-= -@?= /1= 8@92:6= 732-= 6/2= :9387@19;= 95= 497-;= -16@0?69;=2@2=7;74'12-=6/=.767@==, 326/9-1= 69= 8@9-?:2= 87@7:267394= :9;67;;= = 182:6@93262@= 7;-= A= 737;= 3:@91:982= 894'47:6:= 7:-= *)= 0712-= ,A+= 8@;62-= *096/= /2@39:2;65:(= "746/73(= +(= )= 6704261#= ,;744'(= 39@8/949'= 16?-21= 0259@2= 7;-= 7562@= 497-;= 2@2= :9;-?:62-= ?1;= 7= 1:7;;;= 201!,*%1+"01/0,!(." 242:6@9;= 3:@91:982= *$+)= *%67:/= (= %67:/=:2;65:=6-#(=9'9(= 787;)#= 200 =54732;61=2@2=8?@:/712-=5@93=,9@34701= 0*,"#+!"01/0%")#""%./ *932@.442(=+7117:/?12661(=)(=87@7:267394= 71= ;-4'= 562-= 0'= 2-29;= :/62@= 4:#= 0 *?-78216= %?;7@')(= /'-@9'8@98'4326/'4 ,#==-28:61=6/2=-21;=95=7=847;=670426=37-2= :244?4912=*%+(=/7@37:976=(=/;$6!?(= 6/=,A+=7;-==54732;61#=/2=6704261=2@2= 787;=71=?12-=71=6/:2;;=72;6=95=6/2=87;= -21;2-= ;= 7= /9;2':930= 1/782= 6/= 69= :976;= 194?69;#= 6/2@= 3762@741= ?12-= 2@2= -16;:6=;544=82@:2;6721=*=7;-=)#= @272;6=@7-2# 217 .-,+ *)('%$ +"$ "E =67CAE ;E ?:CE BD=@?C9E ?,8C?A5E 432E ,C;9=@7E4+2E;?CDE89=@7)E *)('%$ #"$ GFE DCBDCAC@??=@E ;E ?:CE BD=@?C9E A8=9E 97CE ; "$ /@C.-6,E=@;=88) (:=AE A?'9.E A'--CAA;'88.E 9C6@A?D?CAE ?:CE (=@E?,8C?%AE =@?C7D?=@E ;E #'AC9E FCB 9=6C?CDE@9E:C=7:?E$CDCECA?=6?C9)E#F"E:AE 4#F"2E@9EB@E-?=@7E 8$E !D=,=8=?.E 4 FE E 2E =@E 9=6C?CDE @9E AE E @?C7.E ;D-C?68E :C=7:?E=@9=-?=@7E:=7:EBD=@?=@7EDCBD9'-=,=8=?.E ?,8C?A)E (:CE 'ACE ;E 3E ;=86C@?AE @9E )E :@C.-6,E =@;=88E A?D'-?'DCAE C@A'DC9E :=7:E BD=@?=@7E DCBD9'-=,=8=?.E @9E 6C-:@=-8E !"+"$ %))$$%$)%$ =@?C7D=?.)E @E -?=@7E C@,8C9E C;;=-=C@?E 9D'7E %$ 89=@7E-:=C!=@7E?=@E$=?:E "@E0E89=@7E$AE-:=C!C9E;6=@=68E 6?CD=8E 88E ?,8C?AE 9=;;CDC@?E =@;=88E BCD-C@?7CAE =@E ,9=@7E C:=,=?C9E--CB?,8CE6C-:@=-8EBD@9E ?C-:@='CA) 6C?E E ;D=,=8=?.E A?@9D9AE =@9=-?=@7E #D=,=8=?.E$AE9C?CD6=@C9E??CE?:CE?,8C?%AE A'=?,=8=?.E ;D6-C'?=-8E 'AC)E (:=AE 6C-:@=-8EDCA=A?@-CE?,DA=@E-BB=@7E@9E -6,=@C9E BBD-:E ;;CDAE E BD6=A=@7E -:=BB=@7E9'D=@7E6@';-?'D=@7EB-7=@7E@9E '?6?C9E@9EA-8,8CE6C?:9E;,D=-?=@7E ?D@AB?=@)E @E -ACE ;E B@E -?=@7E 89=@7E BCDA@8=C9E9D'7E9C8=!CD.EA.A?C6A)E ?C-:@='CE ?:CE BD-C?68E ?,8C?AE 6C?E E A?@9D9AE $=?:E E $C=7:?E 8@E 0E "$ * :$C!CDE ;9=@7E )E 3:6C9E 'AD)E C?E 8)E 39!@-CAE =@E 9=@7E ?C-:@='CE;D=,=8=?.0E$AE6@E0E$:=-:E (C-:@='CAE @9E '8=?.E ,.E FCA=7@E ; 6.E,CE9'CE?,8C?EA'D;-C)E FCBD6-C'?=-8ED9'-?=@5E 3E .A?C6?=-E C!=C$E :D6-C'?=-8AE 1 )E BBCD@-CEBBCD@-CE;EBCAE$AE 425E *)E ,ACD!C9E =@E #( E @8.A=AE =@9=-?=@7E 9D'7 B8.6CDE-6B?=,=8=?.)E 6@E6BB=@7EA:$C9E +"$ '@'C=DE 'DE 3@9D9C)E C?E 8)E 'B8=@7E ;E ?:?E ?:CE 3E $AE 9=A?D=,'?C9E 6=@8.E @E ?:CE ;'AC9E 9CB@9E =@C?E BD=@?=@7E ? A'D;-CE;E?,8C?A) BD9'-CE 9D'7E 89C9E GFE BD=@?C9E ?,8C?A)E :D6-C'?=-AE1)E 425) #=7)EE9CB=-?AE?:CE?,8C?A%EBBCD@-CE,C;@9E ;?CDE89=@7E,.EA-@@=@7EC8C-?D@E6=-D $:=-:E DC!C8C9E E A6-CE $=?:E 6=@(:CE '?:@E D=A?=@E BD=@?=@7E 9C;C-?A)E /$C!CDE ?:CACE 9C;C-?AE $CDCE @@E C.CE @9E DCE :C@-CE BD-CAA) '@8=C8.E?=DEB?=C@?E-6B8=@-C 218 4321 0/.-,+*).(+*'+0%$)%#)-."(.-+0-%*!4%* ,.-),0'-.")0/) ,."%4%-%'")+--.-")#%-),.*0)-+$'*+0%$ Pia Berglez1 1 1 1 , Barbara Sterle Zorec )%)4 , Alenka Zvonar Pobirk 7B9.1:; A; 51?162; -7@7; 8A61/78; 0@AB; 1. INTRODUCTION C; 1@B1; B,$; A:+7/+:767; *:+.A:; CBA@?A=;5 2; ; -1; 8A61/78; 0@AB; 7; 88; /7;1??@A1.7;/A;96.@717;-1/7@;; ; -1; A,/19678; 890@AB; A69; 5 7@B16+2; 7 C43;.16;,7;0A@B78;,+;96.A@?A@1/96*;1;8@=*;96/A; 4+:A98 # $; A,/19678; 0@AB; @1.7; 1; =9/1,:7; ?A:+B7@%5 7@B16+2$; 168; 7=9:96 4#; 0@AB; =9; 7*$; B7:/%,178; B7/A8; )#'; "9:7; B7:/; 7B9.1:; 68= *@16=:1/9A6; 9; 1; =9/1,:7; B7/A8; 0A@; ?@7?1@96*; 7B?:A+78; 1; C43; )!'$; 89007@76/91:; .16696*; .1:A@9B7/@+; @7:1/97;/A;4;168;/7;?A:+B7@ 534 2; 7@7; 1; 1; =70=:; /AA:; 0A@; ?@7% 0A@B=:1/9A6; .@77696*; 1; 9/; .16; .A609@B; /7; )4 )) 1BA@?A=; 7; 1./97; =,+9.1:; B9/=@7; A0; 4$; ?A:+B7@$; 168; ?A9,:7; 96/7@1./9A6; ,7/-776; 1./97; =,B7 168; 7.9?976/; )('; 77@1/@A:; 542; 9; 1; 67@961; #; ; !3; B97@; 59A76*9677@96*;C $; 61/=@1:$; ?AA@:+; -1/7@%4-9/7@:1682; ,+; B996*; 0A@; ; B96=/7; 96; /7; -9/;:A-;A@1:;,9A119:1,9:9/+$;-9.;B1+;,76709/; @1/9A;?@776/78;96;1,:7; 0@AB; 0A@B=:1/9A6; 96; C43; +8@A?9:9.; 0); AB?A9/9A6;A0;?+9.1:;B9/=@7;0A@;4; ?A:+B7@;=78;96;C43;.16;?@A/7./;16/9A9816/; -9/;?A:+B7@;168; 0@AB; @7.@+16.7; /79@; RSV Carrier 89(%) B77@; 7; HPMC AS LMP or MMP 1090 1BA@?A=; ; /7; ?A:+B7@; 168; /7; Eudragit EPO 1090 16/9A9816/;)' PEG 6000 1090 6;?@776/;-A@$;-7;19B78;/A;87BA67; PEG 6000 and Neusilin (ratio 2050 =7;A0;34 ;1;1;?@7%0A@B=:1/9A6;.@77696*;/AA:; 1:1) 96; /7; 877:A?B76/; A0; 4%:A1878; C43; PEG 6000 and Syloid (ratio 2050 ?@7?1@78; ,+; B16=1:; B7:/; *@16=:1/9A6; =96*; 1:1) 89007@76/; /+?7; A0; ?A:+B7@; 168; B7 .1@@97@ 1),)) +9.1:; B9/=@7; .A6/19696*; #; ; 168; ; ; 2. MATERIALS AND METHODS 4; :A1896*; -7@7; /@160A@B78; 96/A; ), 89 4$;1;?A:+?76A:9.;16/9A9816/$;-1;A,/19678; -1;:A-:+;18878;/A;BA:/76;?A:+B7@;96;1;BA@/1@; 0@AB; 9A+6/; 54:A1912; A@; 0A@B=:1/9A6; A7@;1;A/;-1/7@;,1/;"76;/7;ABA*767A=; .@77696*; -9/; 34 $; /@77; ?A:+B7@; 168; /-A; B9/=@7; -1; A,/19678$; /7; B7 B7.1@@97@; -1; 18878; 168; /7; 0961:; B9/=@7; -1; .1@@97@; -7@7; 7:7./78; +?@AB7::A7; 1.7/1/7; =..961/7; 5 ; C42; .AA:78;A6;1;/@1+;/A;@AAB;/7B?7@1/=@7 CC; B789=B; B7:/96*; ?A96/; 52; 168; :A-; B7:/96*; ?A96/; 52; 0@AB; 496%/=; 219 *)(' %$%#"! !#!#! /)B ,8B 325D?3/B =655G==6CB DE@FAH$D?@3B IHGFEDCB AF@AGF?G=B @AFG=6EDC7B 7B A@FGCC3/B D32B E@CG56CDFB EG=@A@F@6=B 5DFFGF=8B AH7=5DCB E4?6FG=8B D32B 2=AGF=@3B EG5HD3=E=)B IHGB FGCD?0GC7B C@1B AFGADFG2B =@C2B 2=AGF=@3B =DEACG=B 1GFGB EGC?3/B A@3?B @ G0DC6D?G2B 6=3/B DB 2DCB =5D333/B @FED?@3B @ 5DC@FEG?GFB.:-,B1?HB:I+;GB,*)('B=@?GEAGFD?6FG=8B1H5HBCGC7BGD=G2B?HGBEGC?3/B@ %G??CGFB I@CG2@8B :1?$GFCD32#) "G?1GG3 ! FG=0GFD?F@CBD32B?=B35@FA@FD?@3B3?@B?HGBA@FG=B ,' E/B @/HG2B 3B DB 'B B @GF=) DC6E36EB AD3B D32B =GDCG2)B .:-B 56F0GB 1D=B EGD=6FG2B?HBDBHGD?3/BFD?GB $%# @38B 3B D3B 3GF?B D?E@=AHGFGB 632GFB DB 3B?H=BAF@@@5@35GA?B=?6278B1GB=655G==6CC7B 3?F@/G3B3) 6?C$G2B .:-B D=B DB ?@@CB @FB =5FGG33/B FD1B ED?GFDC=BD32B@A?E$3/B?HGB5@EA@3G3?BFD?@=B '%##"#" ?@B@?D3BDE@FAH@6=B=?D?GB@3B=@C2B +=B@5DCBE4?6FG=8BB 2=AGF=@3B @FE6CD?@3=B AFGADFG2B 0DB ED36DCB *'''B 1D=B 2G3?G2B D=B ?HGB G=?B A@C7EGFB @FB EGC?B /FD36CD?@3)B IHG=GB 323/=B G=?DC=HB DB AFGADFD?@3B @?HB ;:9)B "D=G2B @3B ?HGB D==B @FB =5DC3/B 6AB ?HGB AFGADFD?@3B @2B .:-BHGD?3/B56F0G=8BD5?0GB3/FG2G3?BC@D23/=B 2=AGF=@3=B 6=3/B H@?EGC?B G4?F6=@3B EG?H@2=B @2G3?G2BD=BDAAF@AFD?GBD=B D32B @FB ?HGB 6F?HGFB G0DC6D?@3B @ 3@B @FB E3EDCB EGC?3/B AGDB 1D=B 2G?G5?G28B =@C2B2=AGF=@3=B5@3==?3/B@ 325D?3/BDB@FED?@3B@4?6FGB1?HB D32BEG=@A@F@6=B5DFFGF) B *'''B 26F3/B HGD?3/)B 3B 5@3?FD=?8B %-B +:B D32B 62FD/? %# B 2GE@3=?FD?G2B CE?G2B DAAC5DC?7B26GB?@B?HGFB?HGFEDCB2G/FD2D?@3B@FB ,)B D3B )8B G?B DC)B %$#"!" #" #" 3=?DC?7B D@0GB (*'-8B 1H5HB =B DB EGC?3/B #!$""$""# A@3?B@$!" #!)B %D?HGED?5DCB "@=5G35G=B D32B @FBDBAF@@@5@35GA?8BB*'''BC@D2G2B1?HB 3/3GGF3/8B('(()B,#', ) ('BBD32B'BB;:9B1D=BHGD?G2BD32BE4G2B1?HB ()B;6E@32@FB+)-))8BG?BDC)B #!$ G?HGFB:7C@2 # #" !""# # "#" B@FBG6=C3 )B "$" #" #" ! !$ " "$")B HDFED5G6?5DCB ;G=GDF5H8B ('')B (*,(#(! *'*) )B "H6DCB :)9)8B G?B DC)B !#" !$ " "$" " !#" ##")B +5?DB HDFED5G6?5DB :35DB "8B ('(,)B ,,#(!'! *) )B @0DG0B %)8B G?B DC)B !$" !# #" !$ " "!$ " "#" 6F@AGD3B @6F3DCB @ HDFED5G6?5DCB :5G35G=8B ('()B ('(('(#,'*') " !#%B.:-B56F0G=B@DC=BD32B+:.=)B IHGB D6?H@F=B D53@1CG2/GB 3D35DCB =6AA@F?B G/G32B B ;:9B B B B :7C@2 B B G6=C3 B D3B;G=GDF5HB+/G357B;G=GDF5HB :.(';:9 B :.(';:9: -@FGB6323/8B@)B,',#) :.';:9 B :.';:9B: .:-BD3DC7==B@@3=BFG0GDCG2B ?HGB D=G35GB @3/B AGD8B D=B =GG3B @3B 220 -,+* )('%$-#")! )'$%#)'%%#)%!!)('"('$%"## -(%"#)#%%)! #$%"!''' Mohamed Elkhashab1 2 3 1 2 1 , Ziad Sartawi , Denis Lynch , Abina Crean , Waleed Faisal , Sonja Vucen* .--,+*)-('%)$),'#"!#( *)" )+%%)+"*)+"*).)*)"%# ""# ,#(')*)")),#"*)%)%#*)+"*) )*)"%# +*)-('%)$)+'!" *)" )+%%)+"*)+"*).)*)"%# 1. INTRODUCTION /#*;395:6 -15)5=;3;95!6 216 #8!6 ,6 *0:+689=26:;=*752;:6:2;6'7*8621;+74=;7529 ?=;:998796 5546 352=1906 /?.-0,6 =;:25::+6 )98*)/?,65:860;4$95:=509:68+9026$;216 +750046 8*+6 )=;:998796 02*=2*90(6 =;)3;0986 /,(6 :=*$52;:6 526 6 5:86 (6 9:29746 ;'6 5=296 315)5=9*2=576 :+989:206 9039=2974(6';6 68540!690206:=7*89863;0296 /#7"15015$6 926 57!(6 6 5256 926 57!(6 ,!6 5:86:9+5296=;:2;706';65==*5=4!6 /.-%0,(6 9:5$796 1+16 8*+68;0:+6 :2589)57746 *:+658):0252;:(6?.-0680*32;:6;'62196 *# 0":6 $596 =5:6 3;096 56 0"6 ;'6 )=;$576 0":6 -;02029752;:(6 ?.-06 996 957*52986 ';6 :50;:6 7958:+6 2;6 56 =7:=57746 0+:'=5:26 =19)=576025$7246*0:+6-6-65:86?6 :'9=2;:6 /*76 926 57!(6 ,!6 ;6 ):)096 2106 2;6 *5:2'46 .-%6 89+5852;:6 5:86 )3*246 0"(6 02529+906 21526 =;:2;76 $;$*89:6 8*:+6 3;'7906 5:86 6 ?6 2;6 8929):96 =19)=576 ?.-6 )5:*'5=2*:+(6 ';77;986 $46 29):576 02*=2*576:29+24!6 0297052;:6 =5:6 $96 533798!6 106 02*846 ?=;:9987963140=576025$72465068929):986 :902+52906 8*+6 )=;:998796 =19)=576 5:86 $46 0*576 5:574006 ;'6 );31;7;+46 5:86 3140=576025$7246';77;:+693;0*96;'6?.-06 533955:=96 =;)$:986 216 3;=:96 0":6 2;6 =7:=57746 9795:26 029752;:6 =;:82;:0!6 39:9252;:6500900)9:2! 196 )35=26 ;'6 2;6 029752;:6 )921;806 /+5))56 5852;:6 5:86 921479:96 ;896 /#2,,6 3. RESULTS AND DISCUSSION ;:62196=19)=5765:863140=576:29+246;'6?.-06 # =;)3;0986 ;'6 +750046 25=;:5;796 /%,6 5:86 ;6 )=;$576 +;216 506 ;$09986 8*:+6 ;7)2325:6 /?,6 )=;:9987906 506 2902:+6;'6%65:86?6?.-0! :902+5298! $ # 2. $#)'%##$')(! -;026#2693;0*965:86+5))565852;:62196%6 ?.-06 925:986 2196 +9;)92=6 02*=2*9!6 $ ;99(6 ))98529746 3;026 +5))56 5852;:6 %6 5:86 ?6 996 0*337986 $46 9)3;29=6 2196 %6 ?.-06 91$2986 56 29)3;546 /,!625:85865:5742=57695+9:2065:86:9;:52576 80=;7;52;:6/+94016+99:,6/+!6 ,!6 3;=:960":6996*0986';6=155=2952;:65:86 39:9252;:602*890! A B C #5=16.-%6 506 )972986 5$;96 206 )972:+6 3;:26 5:86 );*78986 :2;6 3;748)9214707;5:96 !6%6?.-6$9';965:865'2960297052;:!6/.,6 /-?,6 5540(6 =;;7986 5:86 )=;:998796 ;:2;76%6?.-6$9';960297052;:!6/,6%6?.-6 02*=2*9069);9862;6';)6?.-0!6 5'296921479:96;8960297052;:!6/,6%6?.-65'296 +5))565852;:! 297052;:6 506 39';)986 56 +5))56 %:6 =;:2502(6 3;026 #26 93;0*96 5:86 +5))56 5852;:6 / 6 "4(6 ;,6 5:86 #26 /(6 5852;:62196?6?.-67;0262196+9;)92=576 6(661693;0*9,!65)3790699609579862;6 02*=2*96/+!6 ,! 399:26:+9006;'6929:5761*)824! A B C 297246 2902:+6 506 =;:8*=2986 :6 5==;85:=96 216 #*;395:6 -15)5=;3;956 9+*752;:06 221 0/.- ,+*) ' %$ ( L L L L L L L L L L LKJI JHG FEDCCBE A@? ADEB =EB;=A L L L L L L L L L L L L L L L L L L L L L L D L L L L L L L L L L L L L L L L L L L L L L D L L L L L L L L L L LL L L L L L L L L L L ,+*)(' -$ =@ @ ,E@EBBA00E 1IK A@? 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8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8:9;99-3: 8 8 8 8 8 8 8 8 8 8 8 8 8 8 / ?#'-2-9 -:*;?=?6 )-=*3=-90 !-,/ % 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 / @ @?10( 9'? 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 9';?='316 34 @.=( *3:=-6?;?6 = ++?; 1-@-9 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4433; +-: 8 8 8 8 8 8 8 8 8 8 8 8 8 844;;?? =2*9::?3= 6@-:-=9;99-3: 7= 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 '? 9'3;= 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 *:371?6,? 4--::*-1 =++3;99 20 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ?#*??6?6 99 9'? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8@"2 *3:*?:9;99-3: 2?97??: 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 9'? 13)?:- 8 8 8 8 8 8 8 8 8 8 8 8 8 8 :: ?=?;*' :6 $::3)99-3: ",?:*0( 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 :6 @,@ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 @1/8 8 8 8 8 8 8 8 8 8 8 8 8 8 13=?10 ;?199?6 93 )-=*3=-90 -= 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ;?=?;*' *3; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ;? 4:6-:, 3/ . 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 :6 ,;:9 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 :8 8 8 8 8 8 8 8 8 8 8 8 8 8 9'? -:?*9-3: 433;*? ;?5-;?6 433; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 : 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 48 8 8 8 8 8 8 8 8 8 8 8 8 86@-:-=9;99-3: 34 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 *3 8 8 8 8 8 8 8 8 8 8 8 8 8 8 44--::*?6 20 3);99-= 6/3/3/ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 ; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 @"2 9''?;++?9-*=/ !-,/ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 9 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 -11=9;99?= 9'? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 ;?199-3:='-+ 2?97??: )-=*3=-900 ::6 ,1-6? 4433;*? 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 9 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 9'? 433;*? ;?5-;?6 93 ??+ 9'? =0;-:,? +1:,?; 8 8 8 8 8 8 8 8 8 8 8 8 8 8 4 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 @3)-:,%(8 8 8 8 8 8 8 8 8 8 8 8 8 8 ='37-:, 9'99 -:*;?=-:, @"2 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 88 8 8 8 8 8 8 8 8 8 8 8 8 8 224 ;:: 9876795463275310/1./7516-7,57530/97+93*)/+3(*3,1+3'/57+45,/ 64764'/%358/04*,3+30 $ $ /*,#/70'/,"+13'/!'/ ;;/7,/976634, 5/1/3/,1/,/, 1/, /31, Jelena Paroj1, Ivana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INTRODUCTION DE6?=:D=2?2EA6/6@BEAF:AA1F9)A$5BAG('/@F:BA !(A"+ A4FEA79B7F9BAD=A:5BAEF;BA4F/,A4D:5A:5BA GFEEDCBA @?FD= FD:D?=A?0A)%A A"44 AG('+8G(A:?A:5BA :?A 1BA D=6?97?9F:BA D=:?A FA =F=?7F9:D6@BA E/E:B;A F2B?2EA75FEB) :5F:AF@@?4EAFA;F3D;2;A@?FF1@BAF;?2=:A?0A92 .-,+*)A G('/@F:D?=A DEA FA E:9F:BA :5BA 7#7%!#$7!7!$ 6D962@F:D?=A :D;BA ?0A 7F9B=:B9F@@/A F;D=DE:B9BA $?A B:B9;D=BA :5BA ;F3D;2;A @?FD= 0?9;2@F:D?=EA1/A2ED=9?75D@D6AG('A65FD=EA 9DE7B9D?=BAD=A:5BA!(EAF=A:5BA:D;BA9B2D9BA:?A :?A 6?F:A :5BA 7F9:D6@BE,A :5B9B1/A D=5D1D:D=F65DBCBA:5DE,A9DE7B9D?=BA4FEAFBAD=AB36BEEA:?A FE?97:D?=A ?0A EB92;A ?7E?=D=EA F=A :5BA -A;@A?0A1?:5A-AF=A+AF=A;D3BA0?9AAF/EAD=A E21EB2B=:A9F7DA9B;?CF@A?0A:5BA7F9:D6@BEA09?;A (77B=?90A :21BE,A 45D65A 4B9BA :BE:BA 0?9A :5BA6D962@F:D?=A.%*)A$5BAFD;A?0A:5DEA4?9#A4FEA:?A 9DE7B9D?=BA@?FD= B:B9;D=BA :5BA B00B6:EA ?0A G('/@F:BA +,A%,AAF=AAF/E AF0:B9A:5BA2=DEE?@CBA92A ?=A :5BA 7FEEDCBA @?FD=5FA 1BB=A 9B;?CBA 1/A 6B=:9D02 9DE7B9D?=B,A FA 7??9@/A 4F:B98E?@21@BA 21EB2B=:@/,A :5BA EB@B6:BA ?7:D;F@A F;?2=:A ?0A 7E/65?75F9;F6?@??=BA4FEAFBA:?A-A;@A?0A-AF=A+AF=A =F=?B;2@ED?=EA"!(E A0?9A7F9B=:B9F@A2EB) ;D3BA :?A F65DBCBA 6?;7@B:BA @?FD=)A $5BEBA 0?9;2@F:D?=EA 4B9BA 2EBA 0?9A 029:5B9A F=F@/EBE)A 7(-:8)6-3,7-5*7(8:90*, $5BA 6?=6B=:9F:D?=A ?0A 9DE7B9D?=BA 4FEA 7(##% B:B9;D=BA E7B6:9?75?:?;B:9D6F@@/A F:A +A =;A BD2;865FD=A :9D@/6B9DBEA "$ A 4B9BA F0:B9AD@2:D?=AD=ADE?79?7F=?@) 72965FEBA 09?;A F9?=A ';1A A 'A 7!#%7###!$ "F9E1::B@,A 'B9;F=/ )A ?/1BF=A ?D@A " ,A 9?7@B:A EDBA "8FCB ,A 7?@/DE7B9ED:/A D=B3A E?/1BF=A@B6D:5D=A" ,AE?D2;A?@BF:BA"@ AF=A "G ,AB:FA7?:B=:DF@A"G ,A7AF=A6?=26:DCD:/A G('+8G(A 4B9BA A 1/A 4B9BA;BFE29BA0?9A1?:5A-AF=A+,A1B0?9BAF=A D7?DA';1A"24D F0:B9A:5BAFD:D?=A?0A9DE7B9D?=B) 4FEA #D=@/A ?=F:BA 1/A 9FC@BA 6:FCDEA "BE#?CF6,A B91DF )A G?@/E?91F:BA A "G ,A 7!$7$#!$ 1?CD=BA EB92;A F@12;D=A " A F=A $?A FEEBEEA :5BA D=0@2B=6BA ?0A 9DE7B9D?=BA @?FD=9?3/:?@2B=BA "$ A 4B9BA 72965FEBA ?=A:5BAG('/@F:D?=AB00D6F6/,A9DE7B9D?=B8@?FBA 09?;AD;F8@9D65A?)A":)A?2DE,A,A ,A -AF=A+A4B9BAD=621F:BA0?9A+A5AD=A%A;;@A 45D@BA @/6B9?@A "'@/ A 4FEA 79?CDBA 1/A B96#A AD=AGA"7A) ,AFEA4B@@AFEA729BAGA0?9A 'FA"F9;E:F=:,A'B9;F=/ ) 6?=:9?@B)A9?7@B:AEDBA"8FCB AF=A9?7@B:AEDBA DE:9D12:D?=A "G A 4B9BA ;BFE29BA 9B2@F9@/A 7:7##!$7!7$#$!%!$ 29D=B:B6:A79?:BD=A1D=D=) $5BA !(EA 4B9BA 79B7F9BA 2ED= 79BEE29BA 5?;?76$7!7%# =?=8G('/@F:BA0?9;2@F:D?=A"- ,A:5BA?D@A75FEBA $5BA 9B@BFEBA ?0A 9DE7B9D?=BA 09?;A :5BA !(EA 4FEA "$A"-A ,A44 ,AA"A ,A44 ,AA"+A ,A FEEBEEBA2ED=D9B6:ADF@/EDEA1F 44 AF=A$A")A ,A44 AF=A:5BAF2B?2EA $5BADF@/EDEA:21BE1FA4D:5A+A;A?0A 75FEBA"'@/A"+)+A ,A44 ,AGA"+A ,A44 ,A@A :5BA !(A EF;7@BA "?=?9A 75FEB A F=A 7@F6BA D=A FA ")%A ,A44 AF=A5D5@/A729D0DBA4F:B9A:?A-A 0@FE#A6?=:FD=D=DEE?@2:D?=A;BD2;A ,A44 A4B9BA79B7F9BAEB7F9F:B@/AF=A5BF:BA:?A "GA7A)AA;B:5F=?@,A+,ACC AF:A%A) A ,A :5B=A ;D3BA F=A 0D9E:A 79?6BEEBA ?=A :5BA 5BC??;@B8*::C?* ?C098 7B8*::C?* 3. RESULTS AND DISCUSSION % $@B8%*B8@=)4@=098 /5 /, 7 6$* C+D@=*0 2 D@+:*D? ?$C-*0 ?@D?,@;DCB) % 2 EE% *B8@=)4 -*B* @00*0 @? ?9#*B0?=D*/B@=D @=0 :9+B;@=D% 8*;$@=;@: #BC#*BD*? -D$ D*=?:* ?DB*=/D$ B*?#*;DA*:) A@:9*?$/$*BD*$/$:90 :C@0? C=2-D$;@BB*B*9?:= ##/+,,/ / /5 2'@;$*A*0:* 6$* :90 #$@?* ;C=D@==/ '= :90#$@?*;C=D*=D*0-D$*B9?=/@8CBD@B @=0#*?D:*=@C#B*AC9?:)0*D*B8=*0,CB *@;$ ;@BB*B 6$* ;C@D=/ 8@D*B@: 7B@DC C, 225 @?= = = = = = = = = = = ;= = = = = = = = = = = :987:6= = = = = = = = = = = 54= = = = = = = = = = = 85:?= = = = = = = = = = = 4 45532179::05/6 9= = = = = = = = = = = @? " ! 3 = = = = = = = = = = = = = = = = = = = = = = 3617:6 6?5%. :? .06 +++93 //* 54 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 9 = = = = = = = = = = = = = = = = = = = = = = .060/:-39::. 0/ 766 :?9// 45513 20/1:6, @? 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = 4 = = = = = = = = = = = 1 = = = = = = = = = = == = = = = = = = = = = .06:0/*:0$ = = = = = = = = = = = @ @ +96 0/ :?? ; 45532179::05/6( = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = : = = = = = = = = = = = = = = = = = = = = = = 4 9= = = = = = = = = = = .060/:-39::05/ :02 9+++93. :5 8 0/4771/*. 8) 9 = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 4 = = = = = = = = = = = = = = = = = = = = = = 0/.0*9::0/- : 9 = = = = = = = = = = = :?9:: :? .311- 06 /5 75/-3 0/ 9 = = = = = = = = = = = = = = = = = = = = = = 3 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = :? 2*?9//0*97 +35+3::06 54 :?? ; :9887:6( 96 = = = = = = = = = = = 9 = = = = = = = = = = = 3 = = = = = = = = = = = = = = = = = = = = = = : = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = *3))6:9770/ 4 3 = = = = = = = = = = = 445532 3 2 9/. 06 .06657$. 0/ , = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = :? ;' = = = = = = = = = = = :987:6( %?0*? ?9$$ 9 ?0-?3 :/607 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = @? 60-/0400* = = = = = = = = = = = 499//:7) 02+35$. .066571:05/ 54 @ = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = @= = = = = = = = = = = 6:3/-:?( ?9. 9 670-?:7) 75/-3 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = .060/:-39::05/ 9= = = = = = = = = = = 443352 ; :98 = = = = = = = = = = = = = = = = = = = = = = 87:6 *9// :?16 8 9:::3081:. :5 :? = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 : 1 = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = :02 :?9// :? ;' = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = =; 45532 = = = = = = = = = = = 4 3 2179::05/( %?0*? 9 = = = = = = = = = = == = = = = = = = = = = .31- 80/- 2 = = = = = = = = = = = = = = = = = = = = = = 257*17937) .06+36. 0/ , = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = *5/:90/. :? *93303 ;)750. = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = =#= = = = = = = = = = ="! , = = = = = = = = = = = 4. CONCLU USION @? @ = = = = = = = = = = = @ ; = = = = = = = = = = = ; : 887:6 +3+93. %0:? 85:? 54 :? = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = @? 3617:6 54 .066571:05/ :6:0/- 0-, 6?5% = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = 0/$6:0-9::. 9= = = = = = = = = = = *933036= = = = = = = = = = = 6?5%.= = = = = = = = = = = 69::06499*:53)) 9 4 3= = = = = = = = = = = :?9:: @ 9 = = = = = = = = = = = @ %96 *52+7:7) 3796. 43352 :?? ; = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 4 = = = = = = = = = = = : = = = = = = = = = = == = = = = = = = = = = 2*? //0*97 9= = = = = = = = = = = 6:3/-:? 9//. .060/:-39::05/ :02, = = = = = = = = = = = 9 = = = = = = = = = = = 9 = = = = = = = = = = == = = = = = = = = = = :9887:6 0336 9 87 = = = = = = = = = = = 0 6+*:0$ 54 :?? *93303 16. 0 = = = = = = = = = = = = = = = = = = = = = = 4 ? : = = = = = = = = = = = 0 = = = = = = = = = = = . .( %?07 = = = = = = = = = = = ?07= = = = = = = = = = = @? 16 = = = = = = = = = = == = = = = = = = = = = 54 ; :*?/575-) 7. :5 9// = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9= = = = = = = = = = = 5/7) 9351/. = = = = = = = = = = = . = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = @ @ %96 3796. 433 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 4 352 ! = = = = = = = = = = == = = = = = = = = = = 02+35$2/ /: 0/ :? . = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = =066571:05/ 39:: 54 @ = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = @(= = = = = = = = = = = :9887:6, 9= = = = = = = = = = = %0:? *52+7 = = = = = = = = = = = : .311- 3796 9*?0$. 0/ 766 :?9// = = = = = = = = = = = 3 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9= = = = = = = = = = = 20/1:6, = = = = = = = = = = = 5. REFERE ENCES ,= = = = = = = = = = = 398 11( ? = = = = = = = = = = =,( : 97,( = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = , !311-= = = = = = = = = = = 3= = = = = = = = = = = .70$3) 9/. 3 = = = = = = = = = = == = = = = = = = = = = :39/679:05/97 3693*?( , ' = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = , , 760( , = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = ( : 97,(= = = = = = = = = = = ,= = = = = = = = = = = 3*?0$6 54 0 = = = = = = = = = = = = = = = = = = = = = = ?9329*)( , ', = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = ACKNOWL LEDGMENT 79876 9/. !06 66571:05/ +3540076 54 ; 9/. ! = = = = = = = = = = = 4 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = :987:6 = = = = = = = = = = == = = = = = = = = = = @?06 3693* = = = = = = = = = = = *? %96 411/.. 8) :? 0/06:3)) 54 = = = = = = = = = = = = = = = = = = = = = = 4 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 3 = = = = = = = = = = = = = = = = = = = = = = # = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = ; 0/.0*9: :?? 16 54 *93303 = = = = = = = = = = = : = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 16070/ = = = = = = = = = = = ;*0/*( @ = = = = = = = = = = = @ @*?/575-0*97 !$75+2/: 9//. = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = ; 9/. ;)75 = = = = = = = = = = = = = = = = = = = = = = 50. # = = = = = = = = = = = "! ( 36+*:0$7), = = = = = = = = = = = = = = = = = = = = = = 0 7 = = = = = = = = = = = //5$9::05/( +1870* 54 ;38809 :?351-? :%5 9 :0 = = = = = = = = = = = 870 = = = = = = = = = = = 4 ; 80 :? = = = = = = = = = = = 3 = = = = = = = = = = = ? : = = = = = = = = = = = = = = = = = = = = = = : *9/ 8 5863$$. :?9:: :? 9+++70*9::05/ 54 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 3 = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = 9 9 = = = = = = = = = = = = = = = = = = = = = = 4 :? ; = = = = = = = = = = == = = = = = = = = = = 3 //: -32/:6 %0:? /0$360:) 54 7-39.' 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = :*?/01 02+35$6 85:? :? :/: //. = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9= = = = = = = = = = =:? 39:: = = = = = = = = = = = 9= = = = = = = = = = = 9*17:) 54 ?932 = = = = = = = = = = = = = = = = = = = = = = 3 29*) 5 ''' = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = 54 @ = = = = = = = = = = = @ 3796, 53 85:? ; 9/. ! :98 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = =7:6( :? = = = = = = = = = = == = = = = = = = = = = 9/. 5 ''' , = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = *93303 ;)750. = = = = = = = = = = =#= = = = = = = = = = ="! 9++93. :5 + = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = +3525:= = = = = = = = = = = 44996:3 .311- 3796 *52+93. :5 16070/# = = = = = = = = = = = 3 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = /= = = = = = = = = = =;,= = = = = = = = = = = 53 :?9/ 54 :? .31- %96 3796 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = . 43352 = = = = = = = = = = = 4= = = = = = = = = = = ;'; 0/ :? 40036: 20/1:6, @? 4996: = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 4 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 43 .311-= = = = = = = = = = = 3= = = = = = = = = = = 3796 43352 ;' = = = = = = = = = = = 4 = = = = = = = = = = =; :9887:6 *517. 8 .1 = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = :5 :?? = = = = = = = = = = = = = = = = = = = = = = := = = = = = = = = = = 75%3 :/607 6:3/-:? 9//. 6?53::3 .060/: = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = 3 = = = = = = = = = = = -39::05/ 9= = = = = = = = = = = :02 5863$$. 44553 = = = = = = = = = = = 3 = = = = = = = = = = == = = = = = = = = = = ::??6 :9887:6, 7:?51 = = = = = = = = = = = 9 = = = = = = = = = = = 1-? :? = = = = = = = = = = == = = = = = = = = = = .31- 3796 43352 45532 = = = = = = = = = = = = = = = = = = = = = = 4 = = = = = = = = = = = 4 3 2179::05/ ;' 9 = = = = = = = = = = = ' %96 = = = = = = = = = = == = = = = = = = = = = 670-?:7) 675%3 :?9// :?9:: 43352 45532 = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = 9 = = = = = = = = = = = 4 = = = = = = = = = = = 4 3 2179:: 905/ ;' = = = = = = = = = = = ;( 253 :?9// 54 :? .31- %96 37 = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 96. 0/ = = = = = = = = = = = = = = = = = = = = = = :? 40036: 20/1:6( 0/.0*9::0/- :??9 = = = = = = = = = = = 4 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = : 99:: 85:? = = = = = = = = = = == = = = = = = = = = = 445532 3 2179::05/6 ?080:. 39++0. .31- 379 9 = = = = = = = = = = = = = = = = = = = = = = 9 = = = = = = = = = = = = = = = = = = = = = = 96 9//. = = = = = = = = = = = 9= = = = = = = = = = = 2:= = = = = = = = = = = :?= = = = = = = = = = = /0:.= = = = = = = = = = = ;:9::6 9= = = = = = = = = = = ?932 3 29 9*5+509= = = = = = = = = = = 3*522/.9:05/6 5/ .066571:05/ :6:0/-9 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = = = = = = = = = 54 @ @= = = = = = = = = = = 1/*59::. :9887:6, 9 : . : 87 : = = = = = = = = = = = 9 226 7654 32101/3.-7,0+*+,237),(+/-03,(3.,2-7-'+//,1.-.3 %$+*,012#10-.3*$-)",!-//+2!30 0*-"3%,2-2*+,21/33#+ .+)-%*+,21/3.-/+-) "371 1 1 1 2 , /3232 , % 33# 3+37 , Dijana Zadravec, Livije Kalogjera3 1 , Anita Hafner + *)('%$)#%"! 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INTRODUCTION C6ECD6D1CEBB:BC=95E=:C GFEDC EBA@?@=FE=@C ;=:?C CE=C!$C@?CEC*0*C2 E7@E=:BC6@=5C4E=F@?=:=@?EDC@B:C:;;:7=CE?BC FE=@0C1?=5C=B0-CC6EC D@A@=:BC3FE@?CB:D@2:F10C/EEDCF.=:C5EC:A:F4:BC =5:?C @?7.D=@?4C $#C :;;@7@:?=C=FE?97C;C-C$-CE?BCC6:F:C0-C0)-C ' C #* E?BC*0CA4CA -CF:9:7=@2:D10 -C @??:F2E=@?4C ECEC9F@?4CED=:F?E=@2:-C:?E3D@?4CB@F:7=CE?BC 9@FE=C ;C ?EEDC 3" 7E2@=1C +*-C )(0C '??2E=@2:C ?EEDC ;-C /EEDCB:9@=@=.B@:C6:F:C9:F;@?4C .75C EC =5:FA9@2:C #" %C 4:DD@?4C ECA.D=@#:7=@EDC7E=C3=E@?:BC 1=:A-C ;;:FC @4?@;@7E?=C 9=:?=@EDC ;EDC ;F7E?C ;C EC )#1:EF# BF.4C EBA@?@=FE=@.F:C :;;@7@:?=C 9F=7C E99F2:BC .?B:FC @B:?=@;@7E=@ B:D@2:F1C =C ;C @?=:F:=-C ?EEDC 7C !#),*#*,)C B:9@=@=.B@:C ?::BC =C EC :4@=F1C?.A3:FC)*#)##*#!0C F@ 7F.7@EDC 9EF=C ;C ;EBA@?@=FE=@EA9D:-C =5:C AC $EDC B:D@2:F1C F:D@:C :2:?D1C 7C .?@B@F:7=@9FE1-C 65@D:C 3@#B@F:7=@@?B@7E==:C =.ED@:C =5:C B:9@=@ B:D@2:F1C F:;:FC =EDC 9E==:F?C E?BC 9F:2:?=C BF@99@?40C %5:C ;FE7=@ EBA@?@=FE=@9FE1C B:9@=@C B:=:FA@?:BC :D:2E=@;=C9EDE=:C:9EFE=:C=5:C?EEDC 4FE2@A:=F@7EDD10C E75C :"9:F@A:?=C 6EC E?BC 9:F;C =5F.45C=F@DC=6EFBC=5:C?E95EF1?"CE?BC B:D@2:F:BC @?=@?4C E?C9=EFC C :"@=C=5F.45C=5:C=5:FC+!(0C 9FE1C9.A9-C:.@99:BC6@=5C=5:C))C/CE7=.E= '?C=5@C6#"%!C ?EEDC B:9@=@C ;C B=5:C C9.A9C6EC@?:F=:BC@?==F@D-C 75@=E?#3E:BC =5:FA4:DD@?4C 1=:AC .965@D:C=5:CD:;=C ?=F@DC 6EC3D7 :B0C%5:C B:2@7:C 76EC E7=.E=:BC E=C E?C E?4D:C C ;F 7F::?C =5:C @A9E7=C ;C ?EEDC B:D@2:F1C A9DE?:-CE?BCCE?4D:C;F B0C %5:C AC 7 3"1*-)+1/0312.3"-*$,.0 F:9@FE=@A.DE=:C 3F:E=5@?4-C 6@=5C #* @?9@FE= 3"3 #* CA@?0C C=5:FA4:DD@?4C75@=E?#3E:BC; ?:#=#3FE@?C B:D@2:F1C ;C BC '?C @A.DE=:C 3@#B@F:7=@ B:2:D9:BC.@?4CD6CAE?C EBA@?@=FE=@9@FE=C $C @4AE#DBF@75-C :FAE?1C E?BC # 7=F@D0C@F;D6C=5F.45C=5:C 4D17:F9595E=:CC@1?=5C=B0-CD2E @EC ?EEDC 7E2@=1C 6EC @A.DE=:BC 6@=5C =5:C; EC =5:C 4:DD@?4C E4:?=0C $5@=E?C :==@?4C 9@FE= 227 9?:?98E:F:?9A 5?;? 7)8,23:,7-5*7*6,12,,605 ;D=2:B ,A F=A 0D=F@@/A ?=A :5BA 5D5879BEE29BA 7(#7%!##% 5?;? 231 -,+* A@? =;:=9= 8768?6545:76 73 4:21?4:076? :6 122:.?/70:6'734:21?4:076?0:06750.?42?/ 5@?667?=9/2:762.4:?0-?5,??6+*)60(*) 33?855@?"! 875:6'479605@?0471/?52* #($:60:85:6'5@5"! /5:760:0675:65?43?4? 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INTRODUCTION @??6:6B3; B3; :98; C"; 2B74@:6B3; @3?; .7B.87:68=; A@=; 8@01@:8?; -; @7635; :9867; /B3/83:7@:6B3; CBA@?@=;:9878;6=;@;578@:;?84@3?;2B7;3@:17@0; A9608;:98;B60;.9@=8;/B3:83:;A@=;%8.:;@:;*;A:'' /B=48:6/; .7B?1/:=; @3?; .9@74@/81:6/@0=; .7B?1/8?;27B4;7838A@-08;-6B,78=B17/8=;+*)(';; 97%!#$!9%%#%! =:76%635;8$@4.08;B2;=1/9;.B.10@7;7@A;4@:876@0=; C"=; A878; /B326748?; 6=1@00; @3?; -; ?3@46/; 6=;/6:71=;8==83:6@0;B60=;#"!= ;A96/9;@78;%3BA3; 0659:;=/@::87635; ;;#,@87@58;?7B.08:;=68; 2B7; :9867; /9@7@/:876=:6/; 78278=9635; =/83:; @3?; @3?;.B0?6=.87=6:;63?8$;; ;.;@018;@3?; 20@B17;-1:;@78;@0=B;/17783:0;-8635;638=:65@:8?; /B3?1/:66:; 48@=178483:='; 00; =:@-08; C"=; @=;.B:83:6@0;@3:6B$6?@3:;@3:646/7B-6@0;@3?;@3:6, A878; %8.:; @:; 7BB4; :84.87@:178; # ; @3?; 6320@44@:B7; @/:68=; +*,('; 18; :B; :9867; 78:8=:8?;1.;:B;;4B3:9='; B0@:606:; @3?; =83=6:66:; :B; 98@:; 0659:; @3?; @67; :9878;6=;@;388?;2B7;633B@:68;2B7410@:6B3=;=1/9; 96%%9#!!9!98; @=;3@3B8410=6B3=;#C"= ;:B;@??78==;:98=8;6==18=; @4@3;=.8/:7@;B2;:98;/6:71=;"!=;A878;78/B7?8?; +('; 98782B78; :98; @64; B2; :96=; =:1?; A@=; :B; :B;/B32674;:9867;4@ B7;/B4.B383:=;78=.B3=6-08; ?880B.; =:@-08; 3@3B8410=6B3=; =16:@-08; 2B7; 2B7;-6B@/:66:;#A6:9;;@4@3;46/7B=/B.8; :B.6/@0; @..06/@:6B3=; A6:9; ?6228783:; /6:71=; "!=; 8$/6:@:6B3;A@80835:9;;34 ' @3?;:B;8@01@:8;:9867;@3:6B$6?@3:;@/:66:' 9 3!"%9 % 9 !9 8;9 %"9 8; 92%#%$9 AB;@3:6B$6?@3:;@==@=;A878;.872B748?;; :8=:;63;06.B.9606/;48?614;#48:9@3B0 ;@3?;; 98; 2B00BA635; ; /6:71=; 8==83:6@0; B60=; 27B4; :98; 2. MATERIALS AND METHODS $!%"#"9%!#$! " " :8=:;63;@18B1=;48?614;#;-12287 ' ;2@460;A878;:8=:8?; " ; ; -875@4B:; ; ; 084B3; 96801.0937)9)/0,100/;7 ; ; 0648; ;;=A88:;B7@358;;;4@3?@763; 9*!$%!9!%!9 B7@358;@3?;! ;;57@.82716:;.880;B60'; :;A@=;2B13?;:9@:;B.:64@0;!;@018;6=;*;@3?; B050/87B0,*;0@17@:8;#* ;A@=;1=8?;@=;:98; 46364@0;";@018;A@=;;:B;*;@3?;:98;B.:64@0; 4@63; =172@/:@3:; 50/870; /@.70@:8; # ; @=; /B3/83:7@:6B3;B2;"!;A@=;*;A:';2B7;@00;/6:71=; /B=172@/:@3:; #-B:9; /87:6268?; 2B7; 3@:17@0; "!,0B@?8?;C"=';0:9B159;:98;"!,0B@?8?;C"=; /B=48:6/= ; @3?B7; .983B$8:9@3B08:9098$0; 9@?;=653626/@3:0;0BA87;?7B.08:;=68=;#,@8; ; 50/87638; #" ; @=; @; .78=87@:68/B=172@/:@3:; ;;;34 ;:9@3;:98;-0@3%;C";#*;34 ;:98; 8:9098$0; =:8@7@:8; #" ; @=; /@77687; B60; A9608; 16/%0;=9BA8?;=653=;B2;63=:@-606:;=1/9;@=;2@=:; A@:87;.9@=8;B2;:98;C"=;A@=;10:7@,.178;A@:87'9 63/78@=8; B2; ?7B.08:; =68=; @3?; /78@4635; #8=.8/6@00; -0@3%; C"; =A88:; B7@358;4@3?@763; 9 7%!#$!9 #%%!9 %"9 B7@358; @3?; 57@.82716:; "!,0B@?8?; C"= '; !%! B7410@:6B3; B.:646=@:6B3; A@=; @/9688?; -; "!,0B@?8?;C"=;A878;.78.@78?;-;1=635;:98;0BA; @??635; '; A:'; B2; ; @3?; '; A:'; B2; "; 83875; .9@=8; 6387=6B3; /B4.B=6:6B3; # ; A96/9; =653626/@3:0; 64.7B8?; :98; =:@-606:; B2; 48:9B?; @:; 7BB4; :84.87@:178; @=; .786B1=0; C"=;0B@?8?;A6:9;-875@4B:;0648;B7;084B3;"!; ?8=/76-8?; +('; 98; 6320183/8; B2; =172@/:@3:,:B, #@-08;* ' 8410=6B3; #" ; =172@/:@3:,:B,B60; #! ; 7@:6B; @3?; :98; 6320183/8; B2; "!; @3?; /B=172@/:@3:; 233 %$% #"! JIHGFGEDCBAJ@?=CDCB9B876B !.B?DF5B!%B=5CB#D;2=FHB!.'B FD=E:;DFD5HEB=9HD;B4BF5H3EB9BEH;=2DB=HB10/ =HB7F7BHDEHB$5$D5H;=HG5/BJ5B=B$5H;=;"BH3DB HDEHDCB AJEB D;DB F:$3B ?DEEB D99D$HGDB 8B NE Z-ave PDI ;=CG$=?BE$=D52D;EBEG5$DB5?"B?GFDB=5CB?DF5B Bergamot EO .-/,+B*B)/()- )/)'B*B)/))- AJBE3 DCBFCD;=HDB=$HGGH"B=HB%B3G23D;BHDEHB $5$D5H;=HG5EB ! .(B =5CB 4-B Lime EO .%/+)B*B)/'4+ )/)-B*B)/))% ;DEID$HGD?"/B JIHGFGDCB HB ?GFDB ;B ?DF5B AJB =?EB E$=D52DCB H3DB06B Lemon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ercedes Vitek , Janez Mravljak, Andrej Jamnik3 3 , # 3 "! ! ! 1. INTRODUCTION ?D@?@5E,?AE?FF = JIHGIFEDCBA@?I= *4 ?AE DG3;A;2 D;AE ?@E C;;2E @2: FCG/E FI. 3?@4+ A@CGD@GC?E;CFIHGIFEDCBA@?I= ,I@4E@4I. /: / // G=IHGE =?=;A@CGD@GC6?C@IDBFIA: =D?:AG?@I;=E;-E4BFC;:4IID0E4BFC;:4;3ID0E?=FE 2I/4@E AD?@@ ?2:4I:4IIDEFCG/A0E;--9 @?AI@C?0E?.B@ID?0E5+ FCG/AE ?=FE :C;.IF BE .?G?3-) 2.4. 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INTRODUCTION %'=4-='=@?-=! =8;=96@;,6=836=6+7655=;2= DCBA@?= ;:98?7C65= 96:96568= = :9;@?5?4= 79;55/C??4= 468= .= 7346= 836= .?5:695?;= ::9;73= 2;9= :96869= ;2= :;;9C1= @6.?A@*= )36= 56.?@68= 05= 96.?5:6956.= ?= = 0869/5;CABC6=.9A45-=.A6=8;=836=?89?5?7=C?4./ = ;2= 5A79;56= %= %0, = .= B?.?4= .;@5= 2;9= ,9?;A5= 6.;46;A5= .= @?8;C= %= %0, -= 03?73= 0696= A56.= 5= 6+;46;A5= @;C67AC65= 0?83?= 836= = 791;:9;867885= ?= 836= 29666/.91?4= :9;7655*= @;C67AC6*= )361= 96= )36= ;B86.= .?5:695?;= 05= 29;6= % ' ! = B?;.649..= 7= 6376= 58.=29666/.9?6.=%!39?58=68='/=29666=.9169= C;.6.= .9A45= ('*= )36?9= ?89?5?7= :9?@91= .91?4= !-= *= @B9-= = 3= :55?,6= .= 78?,6= 89468?4= ,?= = 567;.91=.91?4= !-=*'=@B9-= =3 *= 9676:8;95=%6*4*=5679686.=:9;86?=7?.?7=.=9?73= )36=;:98?7C6=5?6=.=5?6=.?589?BA8?;=0696= ?= 71586?6= %$#D"! = .= 4C17;:9;86?= -= 6,@?7=C?438=578869?4=.=836?9= 03?73=96=;,696+:96556.=?=7769=.=?2C@6.= 5A9276=73946=05=@65A96.=B1=C569=;::C69= 8?55A65= (-= 2A98369= 63765= 836?9= ,= 6@;@6891= %685?69= C89-= = 894686.= .6C?,691= ;2= 8?;+?.85= .= 8?/ # ?2C@@8;91=4685=8;=5A73=964?;5=('*= 967;58?8A8?;= ;2= ;:98?7C65= 29;@= 29666/ )3A5-= 836= .9?6.= 2;9@AC8?;5= .= 836?9= :315?7 29666/.9?6.= .A9?4= .91?4= 0696= = ;2= 9:?.C1= 967;58?8A8= ;:98?7C65= C;.6.= @;9:3;C;41= ;2= 836= 29666/.9?6.= :9;.A78= 05= 0?83= ,?5A?4= 6C6789;= @?79;57;:1*= A69768?-= = 2C,;;C= 6+3?B?8?4= 8?;+?.8-= )36=718;8;+?7?81=;2=;:98?7C65=05=6,8?/?2C@@8;91= .= ,5;:9;8678?,6=:9;:698?65* ;= 836= 76CC= C?6= D*31= A5?4= = 965/ B56.=551=( =2869==3=.= =3=6+:;5A96=8;= 2. MATERIALS AND METHODS ;:98?7C6=.?5:695?;5=%*''=@4@ *= DCBA@?= ;:98?7C65= 0696= :96:96.= 29;@= 3. RESULTS AND DISCUSSION B;,?6= 569A@= = %C683@= ?26= $7?6765-= =B1=.65;C,8?;=@683;.=(*=?958-= 65;C,8?;= ;2= = 1?6C.6.= 836= C94658= ==5;CA8?;=%'=@4@ =05=:96:96.= ;:98?7C65= %#= %'= = = @ = ?= 836= ?= AC8989;CC6.= 76= ;2= A69768?*= ?55;CA8?;= ;2= A69768?= ..?8?;= ;2= 683;C=%= @@? = 03?C6= 58?99?4-= ?= 683;C= %'= @4@ = A56.= 2;9= .65;C,8?;= 03?73= 7= ;2= = .= 965AC86.= ?= = 5@;:98?7C6= 5?6= %#'= 2;9@8?;= %' =@ *=)36=:A9?2?78?;=;2=;:98?7C65= ;2= ;:98?7C65*= A69768?= %?;5183-= $C;,4= 05= :692;9@6.= B1=80;=717C65=;2=7689?2A48?;=965AC86.=?=@;96= .A9?4=.65;C,8?;=586:-=5=?8=05=.?55;C,6.=?= 3;@;46;A5= #= .?5:695?;-= 03?C6= 836= 5?6= 836= 683;C= A56.= 2;9= .65;C,8?;= ;2= *= .?589?BA8?;= ;2= #= 2;9@AC8?;5= 96@6.= ;:98?7C65= 0696= 58?4= A7346.=%?4*==' *=)3?5=7;AC.=B6=889?BA86.=8;= 836=@?6= 965?.A65= ?= = @;C67AC65= 0?83= C;5565= ;2= 5@;:98?7C65= 0?83= 836= 4CA89;:98?7C65= 96@;,6.=5A:6988-=5=35=B66=.6579?B6.=?= 0696= :A9?2?6.= B1= 80;= 717C65= ;2= 7689?2A48?;= 836= C?8698A96= (*= DCC= 29666/.9?6.= #= 239 FEDCBA@?E=; D:9E=?B?:8; =; 7; 65; ; @F?:D; ?4:; 9EC3@D@(A:;?E;?4:;/@(A?-;EF;B=?D:@?:8;9E=?DEA1; @88?E=; EF; 3BDF:8; 2@?:D1; 0/:D@.:; 0F?:D;;4;=9B(@?E=;9:AA;/@(A?-;8:9D:@:8;=; 4-8DE8-=@C9; ,:; EF; +B:D9:?=*AE@8:8; 9@:;EF;! );24A:;9:AA;=9B(@?:8;2?4;"! ; =@=E3@D?9A:; @F?:D; FD::,:*8D-=.; 2@; .4?A-; :4(?:8;=E;.=F9@=?;94@=.:;=;9:AA;/@(A?-); =9D:@:8; 9E=?D@D-; ?E; B=94@=.:8; @/:D@.:; =89@?=.; .EE8; (E9EC3@?(A?-; EF; ?4:; 3@D?9A:; ,:; =; 9@:; EF; +B:D9:?=*FD::; 3D:3@D:8;=@=E8:A/:D-;- =@=E3@D?9A:);2494;9EBA8;(:;@??D(B?:8;?E;?4:; /@(A?-; EF; 9:AA; ?D:@?:8; 2?4; @AA; =/:.@?:8; FEDC@?E=;EF;@..D:.@?:1;'4:;,:;8(B?E=); =@=E3@D?9A:;FEDCBA@?E=;2@;@(E/:;5;;@F?:D; 4E2=; @; 3EA-83:D?-; =8:); 2@; (:AE2; 516; =9B(@?E=; 2?4; 6; C.C; EF; =@=E3@D?9A:; FED; FED; @AA; =/:.@?:8; =@=E3@D?9A:; FEDCBA@?E=); ;41;'4:;8:9D:@:;=;9:AA;/@(A?-;E(:D/:8;=; =.; 4.4A-; CE=E83:D:; =@=E3@D?9A:; 9@:;EF;! ;C@-;=89@?:;@;4.4;=?D@9:AABA@D; 3E3BA@?E=;:/:=;@F?:D;D:9E=?B?E=;%$.1 6#1; +B:D9:?=; 8:A/:D-); @; 4.4; =?D@9:AABA@D; +B:D9:?=; 9E=9:=?D@?E=; 9EDD:A@?:; 2?4; 200 ns 9-?E?E9;:FF:9?1; 0.4 P oly 150 ispd ersity :; 8:/:AE3:8; A-E34A,:8); +B:D9:?=*AE@8:8; 100 @A(BC=;=@=E3@D?9A:;FEDCBA@?E=);2494;::D?; 0.2 in C=C@A; 9-?E?E9?-; @=8; 9@=; (:; D:9E=?B?:8; exd 50 D@38A-1; $BD?4:DCED:); ?4:; C:?4E8; B:8; FED; Hydrodinamic size [nm] 0 BNP QNP0.1 QNP1 4.4A-; D:3DE8B9(A:1; "@:8; E=; ?4:; D: 0.0 =@=E3@D?9A:; 3D:3@D@?E=; 2@; 4E2=; ?E; (:; E(?@=:8); EBD; FB?BD:; 2ED; 2AA; FE9B; E=; 0/:D@.:;4-8DE8-=@C9;,:;%9EABC=#;@=8; 8:?:DC==.; ?4:; @=?E8@=?; 3DE3:D?:; EF; 3EA-83:D?-;=8:;%8E?#;EF;+B:D9:?=*FD::;@A(BC=; +B:D9:?=*AE@8:8; =@=E3@D?9A:; =; /@DEB; -,+ =@=E3@D?9A:; %"! #; @=8; @A(BC=; =@=E3@D?9A:; *-)('+9:AA*(@:8;@@-1; 3D:3@D:8; 2?4; 516; C.C; %! 516#; @=8; 2?4; 6; C.C;%! 6#;+B:D9:?=;E=;(:FED:;%8;.D:-#; @=8;@F?:D;FD::,:*8D-=.;%8@4:8;24?:#1;?B8:=?;?*?: 2@; B:8; ?E; 9EC3@D:; ?2E; .DEB31; @?@; 4E2=; @D:; 61;3@8@);01);:?;@A1);%$+#,-"!$,$ +'+!-,+ ++ C:@=;;;%= ((-$(+ -,+ ,'(!+ $-*$(; EA:9BA@D; ;#);=);=E=*.=F9@=?;);3;;5151; 34@DC@9:B?9);561;6%#;6 *61; '4:; ,:?@; 3E?:=?@A; EF; =@=E3@D?9A:; =; 3BDF:8; 2@?:D;2@;AE2);2?4;/@AB:;@DEB=8; ); 1); :?; @A1); + ,+ + + !-,+ 5;C;FED; ($$)'( + $+ )($)$+ -)$ + '(+ )%$+ $-*$(+ '+ @AA; 3D:3@D:8; FEDCBA@?E=1; ?; D:C@=:8; ,)-)!'(+ (! ; $DE=?:D; =; 4@DC@9EAE.-); 51; B=94@=.:8; @A?B?E=; EF; FD::,:* 61; 8D:8;! ;FEDCBA@?E=);(B?;?4:;@(@=.:D);1);:?;@A1); !,+ $(!+!-,++ EF;,:?@;3E?:=?@A;8:9D:@:8;@F?:D;D:9E=?B?E=; ,,'()-$ + $('!---)+ '+ ($()-',+ EF; ?4:; +B:D9:?=*FD::; =@=E3@D?9A:1; '4:; ('$ + ,+ -,$)- + '+ $,)-+ $()-',; C=C@A; 94@=.:; =;@/:D@.:; =@=E3@D?9A:; ,:); =?:D=@?E=@A;EBD=@A;EF; 4@DC@9:B?9);551;%6* #;65*661; ,:; 8(B?E=; @=8; =.; FD::,:*8D-=.; =89@?:; ?4@?; ?4:; 3DE9:; B:8; ; D@.@D);1);:?;@A1);-'$*!)-',+$)%' +'(+-(', '-$ $+ ,$)-+ ,,'()-$ + =?:D=@?E=@A; ?@(A:;FED;?4:;3D:3@D@?E=;EF;8D-;FEDCBA@?E=; EBD=@A;EF; 4@DC@9:B?9);561;;651 EF;@A(BC=;=@=E3@D?9A:1 $BD?4:DCED:);?4:;3D:3@D:8;=@=E3@D?9A:;4@8;=E; @8/:D:; :FF:9?; E=; 9:AA; /@(A?-; @F?:D; @; ; 4; '4; D::@D94; 2@; FB=8:8; (-; ?4:; AE/:=@=; =9B(@?E=);@;?4:;/@(A?-;EF;9:AA);?D:@?:8;2?4; ::@D94; @=8; ==E/@?E=; @.:=9-; %3DE.D@C; +B:D9:?=*FD::;ED;+B:D9:?=;AE@8:8;=@=E3@D?9A:; 9E8:; 6*55); 6*56); *55 #1 =; 9E=9:=?D@?E=; EF; B3; ?E; 6; C.C; 2@; 240 210/ .-,-+*2)-('%(.*2'$)$#%'$*(*"-+-!' *2((%(*"$- "* .- )%+%22+$!%'$*(*",$'%)$(%%!-'%'- '2 ' , Petra Kocbek 1. INTRODUCTION ;0DIH.DC5G.=D'"DGEBDIH.DD7@?@D6?@6G?@BDIED JIHGFIED CD GEBD IHAD B@?IGHI@A=D ;::@@:9D D %D @H3GE;:.D 1AHD 6?I;?D H;D H3@D 8E;7EDGAD?@HIE;IBA=D6:G9D7@::5@AHG4:IA3@BD?;:@AD @:@ IEDA8IED639AI;:;29=DIE:1BIE2DH3@D?@21:GHI;ED;0D FI$@BD 7IH3D GED GB@1GH@D GF;1EHD ;0D IH.DC5G.D @6IB@?FG:D @::D BI00@?@EHIGHI;E=D 8@?GHIEI/GHI;E=D A;:1HI;ED7IH3D;?D7IH3;1HDGEDGEHI;$IBGEH=DEGF@:9D GEBDIFF1E@D?@A6;EA@=DH31ADH3@9DG?@D7IB@:9D1A@BD ;E:9DIH.DC5G.D.7H%D0;?F1:GHI;ED!#%=DIH.D H;6IG?I;1AD C5G.D 7IH3D '"D .7HD @G3%D 0;?F1:GHI;ED B@?FGH;:;2I!%=D GEBD IH.DC5G.D 7IH3D IH.D D .7HD @G3%D 63;H;G2IE2=D I3H39;AIA=D GEBD @?HGIED 0;?FAD ;0D 0;?F1:GHI;ED!%.D ;0D ?@HIE;I6GIE%.D G3D AGF6:@D 7GAD :;GB@BD IEH;D GD D FD 6:GAHIG8IG%D H3GHD 7GAD ?@HIE;IBD 6?@1?A;?AD A13D GAD ?@HIE;:D @AH@?A=D 6:G@BD IED H3@D A9?IE2@D 61F6=D 73I3D 7GAD ?@HIE;:=DGEBD?@HIEG:DG?@D;FF;E:9D1A@BDIEDH;6I@?=D H3@D IHGFIED CD ;1H@?D BIGF@H@?D .D FF%.D D "3@D E@@B:@D 7GAD B@?IGHI@A=D A13D GAD IHGFIEDC5G@HGH@D IH.DC5 ;EE@;:HG2@D2@E@?GH;?D) 8J%D G%=D G?@D 3@FIGH@BD 7?G66@BDIEDG:1FIE1FD0;I:.D:@@?A@D IFF@BIGH@:9D G0H@?D 6?@6G?GHI;ED 1AIE2D GED @:@I@D IED GD 3;?I/;EHG:D ?@GD A9AH@F=D I;IEI@EGHIE2D "3@D 6;:9F@?D A;:1HI;EAD 7@?@D @:@ 63;H;H;$I@?;F@DH3@A@D3G::@E2@AD ).DF3D0;?DDFIEDGHD?;;FDH@F6@?GH1?@DGEBD GEBD H;D @E3GE@D AHG4I:IH9D ;0D IH.D C5G.D IED H3@D ?@:GHI@D31FIBIH9D .D 6?;B1@:;6@BD "3@D F;?63;:;29D ;0D @:@ )%'- $%+%(.)-'*. :@ ) AH1BAD 7IH3D B;14:@5AIB@BD ;EB1@D HG6@D JIH.D C5G.D .%D GEBD H;;63@?;:D IH.D =D BIGF@H@?=D )#D FFD $0;?BD EAH?1F@EHA=D %D .%D 7@?@D 61?3GA@BD 0?;FD G?4;A9EH3D GEBDIFG2@BDGHDGEDG@:@?GHIE2D;:HG2@D;0D)D8JD @?8A3I?@=D %.D '52?GB@D G@H;EIH?I:@=D 1AIE2D GD A@;EBG?9D @:@@DDIFG2@AD1AIE2D =%D GEBD 6;:;$GF@?D D 7@?@D 61?3GA@BD FG2@D).@%DA;0H7G?@D-GHI;EG:DEAHIH1H@AD;0D 0?;FD I2FG5C:B?I3D H@IE3@IF=D @?FGE9%.D '@G:H3=D C%=D GEBD H3@D G@?G2@D -!D BIGF@H@?D :H?G561?@D7GH@?D7GAD;4HGIE@BDH3?;123DGDI::I5 G:;E2D 7IH3D H3@D AHGEBG?BD B@IGHI;ED 7GAD D 7GH@?D 61?I0IGEHG2@D ) "3@D;EH@EHD;0DIH.DC5G.DIED-!AD7GADB@H@?FIE@BD 49D GD G:IBGH@BD F@H3;BD 1AIE2D C2I:@EHD 'D "3@D IEIHIG:D 6;:9F@?D A;:1HI;ED 7GAD 6?@6G?@BD 49D @?I@AD )))#D IEAH?1F@EHD GEHGD :G?G=D BIAA;:IE2D D GEBD 6;:;$GF@?D D IED 7@I23HD C%D @1I66@BD 7IH3D GD JD B@H@%D@H3GE;:D0;?F1:GHI;ED!)%.D 3?;FGH;2?G63I@?A@B563GA@D )D ;:1FED 7IH3D GD F;4I:@D 0;?D FIEDGHD =D0;::;7@BD49DGBBIHI;EG:D# 3D 63GA@D;EAIAHIE2D;0DG@H;EIH?I:@DGEBD7GH@?DIEDGED AHI??IE2DGHD?;;FDH@F6@?GH1?@D*(.DH;8DA;:1HI;EAD 241 210/ IHGFEDCCIF A@?CIG =GF2 41.2-58.7 9 =87= 7 =I6 5ACAFCIG :DH FDEEIA G?C DCC 432 =6 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B ? B B B B B B B B B B B B B B B B B B B B B B D F3 59.0-72.9 F4 84.2-104.2 10A A;;;AAFC G; @I/0C G C0A HCD..I@IC- G; ,IC6 +*DF6 B B B B B B B B B B B ; ; B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B D B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B I C0A )EA)DEA (' :DH DHHAHHA I DFFGEFA B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B DB B B B B B B B B B B :IC0 C0A %$ #9" /?IA6 +;CCAE )EA)DEDCCIG! .-.- " , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B ; B B B B B B B B B B B D B B B B B B B B B B B C0A ('H :AEA HCGEA I D F@I=DCCIF F0 = , , , , , , , , , , , , , , , , , , , , , , % , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B D B B B B B B B B B B B D =.AE :IC0 B B B B B B B B B B B B B B B B B B B B B B D 10AB B B B B B B B B B B EAH?@C CHB B B B B B B B B B B G;B B B B B B B B B B B C0A ;GGEFA B B B B B B B B B B B ; B B B B B B B B B B B DB B B B B B B B B B B B B B B B B B B B B B @I/0C 0 B B B B B B B B B B B =G AE= )0GCGHC ..I@IC D CC-- HC?- HCEDCCA C0DCC @I/0C B B B B B B B B B B B B B B B B B B B B B B D B B B B B B B B B B B D B B B B B B B B B B BB B B B B B B B B B B E = - DCC 32 %! IIC6 +*DF6 D B B B B B B B B B B B B B B B B B B B B B B D B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B FGCAC I ('H B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B :DH A)GH?EA 0 B B B B B B B B B B BDH D HI/I;IIFDC I=)DFC G C0A B B B B B B B B B B B B B B B B B B B B B B ; D B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B B B B B B B B B B B B E =IA .- $8% =AC0G )EIGE @I/0C B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B A)GH?EA D D;CCAE 2 C6 HCD..I@IC-- G; , D C B B B B B B B B B B B B B B B B B B B B B B ,IC6 +*DF6 ,AE C0A 2* G; ('H CG @I B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B/0C ,IC6 +*DF6 FG=)@ACA@- .- +*)(') %$# #")!())" $ , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , I FGCEDHC C B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B CG C0A ,IC6 + DF6 I C0A FGCEG@ (' B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B .-- , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , ,, , , , , , , , , , , HD=)@AH! :0 B B B B B B B B B B B 0IF0 :AEA B B B B B B B B B B BB B B B B B B B B B BHCGEA )EGCAFCA ;EEG= @I/0C B B B B B B B B B B B B B B B B B B B B B B ; B B B B B B B B B B BB B B B B B B B B B B 10A I= /AH EA,AD@A 10A I= B B B B B B B B B B B B B B B B B B B B B B = /AH EA,AD@A D D B B B B B B B B B B BB B B B B B B B B B B;;GGE= ;;GGE= E E =DCIG G; =DCIG G; ? D D B B B B B B B B B B B B B B B B B B B B B B ?I;GGE= ?I;GGE= ; ; E E = =B B B B B B B B B B B ?H6 10AHA ;II/H B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B ;B B B B B B B B B B B A@AFCEGH)?B B B B B B B B B B B ('H :IC0G?C .ADC D )EG=IHI/ B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B D,AED/A (' B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 'H :DH B B B B B B B B B B BB B B B B B B B B B B D, FA D B B B B B B B B B B B 9 B B B B B B B B B B B 23 = D 394 99 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B =6 10A B B B B B B B B B B BB B B B B B B B B B B 0G:A,AE!B B B B B B B B B B B )DFD/I/ )B B B B B B B B B B B CGB B B B B B B B B B B DHH?EAB B B B B B B B B B B A;;;AAFCI,A ; ;B B B B B B B B B B B IFGE)GEDCCIG D G G; ,IC6 +*DF6 '3! ,IC6 +*D B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B DF6 :IC0 B B B B B B B B B B BB B B B B B B B B B B )EGCAFCIG G B B B B B B B B B B B B B B B B B B B B B B ; C0A ('H ;EEG= @I/0C IH AHHACID@ CG B B B B B B B B B B B B B B B B B B B B B B ; B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B "$1 '4 D B B B B B B B B B B B B B B B B B B B B B B D ,IC6 +*DF6 B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B:IC00 ,IC6 ' C B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B BGC B B B B B B B B B B BB B B B B B B B B B B DHH?EA HCD.I@ B B B B B B B B B B B @IC-- ,IC6 +*DF6 I ('H6 C B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B I=)GECC C@-E DB B B B B B B B B B B D;;;AAFC ; ;B B B B B B B B B B B C C00AB B B B B B B B B B B ('B B B B B B B B B B B =GE)) E )0G@G/-B B B B B B B B B B B FG=)DEA CG /- ! $!'( , , , , , , , , , , , ()" $) B B B B B B B B B B B B B B B B B B B B B B G C0A )?EA 7 ('H ' B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B '96 B B B B B B B B B B B 10A IFGE)G B B B B B B B B B B BEDCCIG G; ,IC6 +*DF6 D B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B BD@G/ :IC00 ,IC6 B B B B B B B B B B B C B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B ICG )G@-=A B B B B B B B B B B BE ('H :DH H0G: CG .A D )EG=IHI/ B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B HCEDCCA/- ;GG HCEDCCA/- ;GG D D B B B B B B B B B B B ; ; GE HCD..I@II/ ,IC + DF GE HCD..I@II/ ,IC6 +*DF6 / B B B B B B B B B B B D D B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B /B B B B B B B B B B B A@AFCEGH)I I/! G;;;AAEI/ D,DFA C DB B B B B B B B B B B G;B B B B B B B B B B B ,DEIG?HB B B B B B B B B B B HB B B B B B B B B B B D DH D6B B B B B B B B B B B '?EC0AE HC?H?EA C0A B B B B B B B B B B BB B B B B B B B B B B @G/*CAE= E = HCD..I@IC- G; ,IC6 +*DF6 B B B B B B B B B B B D B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B :IC0I C0A B B B B B B B B B B BB B B B B B B B B B B ('H! AIC0AE C00EG?/0 C0A ?HA G; B B B B B B B B B B B B B B B B B B B B B B C B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B D))EG)EIDCCA DB B B B B B B B B B B )DFD/I/ GE .-D B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B IFGE)GEDCCIG G; D B B B B B B B B B B B B B B B B B B B B B B DD@ HC B B B B B B B B B B B CD..I@II/ AFI)IACH6 D B B B B B B B B B B B +**+*- +**+* , , , , , , , , , , , *$!*) , , , , , , , , , , ,-, , , , , , , , , , , A)EAHACDCI,A IB B B B B B B B B B B HFDI/B B B B B B B B B B B A@AFCEEG CB B B B B B B B B B B 96 GC = B B B B B B B B B B B D =A /0AED ! B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B D B B B B B B B B B B BB B B B B B B B B B B =IFEGHFG)- I=D/AH G; C00A )?EA 7 D B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B C B B B B B B B B B B B B B B B B B B B B B B 7 B B B B B B B B B B B DG;II.AEH ;B B B B B B B B B B B EA:D@ 6 B B B B B B B B B B BB B B B B B B B B B B $#"!" "! " " # % % % % % % % % % % % # % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % '3! 7 DG;II.AEH @GD "!" B B B B B B B B B B B G G?ED@ G; %?CDAG?H AA D B B B B B B B B B B B B B B B B B B B B B B ? D B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B B '9! 7 DG;II.AEH @GD B B B B B B B B B B B B B B B B B B B B B B ; B B B B B B B B B B B B B B B B B B B B B B C B B B B B B B B B B B ,IC6 +*DF6 B B B B B B B B B B BB B B B B B B B B B B "$1 '4 D 7 DG;II.AEH @GD ?E//AE--! 33 E E B B B B B B B B B B B 336 3 96 B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B B B B B B B B B B B B 7 B B B B B B B B B B B ; B B B B B B B B B B B B B B B B B B B B B B C B B B B B B B B B B B B B B B B B B B B B B+*DF D B B B B B B B B B B BB B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 7 B B B B B B B B B B B ; B B B B B B B B B B B B B B B B B B B B B B:IC00 ,IC6 C B B B B B B B B B B BB B B B B B B B B B B +*DF6 D ,IC6 '6 36 I@GH0AH B B B B B B B B B B B D 5! GDE 6 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B "!" "! % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B " # ! % % % % % % % % % % % !"""! % % % % % % % % % % % ! % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % .-- , , , , , , , , , , , % , , , , , , , , , , ,, , , , , , , , , , , , , , , , , , , , , , " $ % % % % % % % % % % % % % % % % % % % % % % $#"!" # "! $!!"!# % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % %% % % % % % % % % % % IIC6 +*DF6 B B B B B B B B B B BB B B B B B B B B B B @GD/ A;;;IIFIAF- :DH A! ! % % % % % % % % % % % #"! #% % % % % % % % % % % +,DFAH I 10AED))-! B B B B B B B B B B B B B B B B B B B B B B DB B B B B B B B B B B .DHA G ,IC6 +*DF6 FGCAC G; C0A ('H ? B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B ?HC D;CCAE B B B B B B B B B B B ;B B B B B B B B B B B 3336 4 24 B B B B B B B B B B B B B B B B B B B B B B 429*2426 C0AIE )EA)DEDCCIG D C0A C00AGEACIFD@ ,I B B B B B B B B B B B D B B B B B B B B B B B D B B B B B B B B B B B B B B B B B B B B B B C B B B B B B B B B B B IC6 +*DF6 B B B B B B B B B B BB B B B B B B B B B B 46 5ED/DE B B B B B B B B B B B DB B B B B B B B B B B ! GDE ! GF.A 6 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B BB B B B B B B B B B B % % % % % % % % % % % FGCAC I ('H 1DD.@A 96 10A EAH?@CH E B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B 1 B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B B EA,AD@A!! 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Annual Review of Biomedical 46;53-'=59#==:9=$=#"!#2=46=3;:96=4:?= ;96:6:=3-=3;;9#(364'=4:?=:?=(35:4;2=-4= 2. Maisel, K., et al. Nanotechnologies for ;955-(96'46"=46;53-=46=3'''=24(4'-==32-9= Engineering. 2023. 8;25: 233-256. 9.-5'=3=:56'=9=46;53-46"=(35:4;2=-4=59#== 3. Rouco, H., et al. Screening strategies for surface =:9=%$=6#=4:?=3=;955-(96'46"=46;53-=46= modification of lipid-polymer hybrid nanoparticles. +=59#==#=:9==# International Journal of Pharmaceutic 2022. ,+*)+= (35:4;2= -4= 36'= 6;3(-023:496= Epub 2022 Jul 8. PMID: 35811041. 624:121973. doi: 10.1016/j.ijpharm.2022.121973. 4;3;/==95= ,= Sample Z-ave PDI EE % (nm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assiliki Papadimitriou , Georgios Sotiroudis 5 !5#, 6 5##3 4 5 1 , James M. Cook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able 1. Type of the used gelling agent for gelation CDCB85B3A6=FFCA61B50F4DCF85A469F of the nanoemulsion. D77A46F62AF5AAD=AFB7F62AFBD8A8F85?;F=?:=6DC4AF Gelling agent (1% m/m) /'-, NEG_C980_d Carbomer 980 +2AF D@F B7F 62=F =6?89F 1D=F 6BF =?D*AF 62AF @45B=65?46?5AF B7F CDCBA@?;A=F NEG_SZ_d Polyacrylate crosspolymer-6 35B8?4A8F162F625AAF877A5AC6F;A 6BF 6A=6F 2B1F 6F D77A46=F 62AF 5AAD=AF 086%"$" #$8#'#$ F5B6D6F52AB@A6A5F!2ABD:F%F.'"GFDD5F :?35B7ACFD=FDF@B8AFD46AF=?:=6DC4A, 2=4DGF A5@DC9F A?33A8F 162F DF 4?33D6AF @AD=?5AF.F 8;=56:/48218;5=014 1D=F ?=A8,F +2AF 6A=6F 1D=F 4BC8?46A8F ?C8A5F 8;#%#$ 4BC6 F$#"F1D=F3?542D=A8F DF 4BC65BA8F =2AD5F 5D6AF 75B@F "F 6BF '""F =GF 75B@F ?:5*BF !,F DC62DCF ;?@F 6534D6A, !EE F E%F 1D=F 3?542D=A8F 75B@F DC8A5:6F=GF%F!,FB9D459D6AF 8:'8"8%$%#%8%' 8:+6= 45B==3B9@A5F !A3@DF ACF 1D=F 09F +2AF 6A=6F 1D=F 4BC8?46A8F ?= 4A=F 4B?3A8F 162F DF 3B94D5:BCD6AF @A@:5DCAF 8BCD6A8F 75B@F A334F !5DC4A,F B8 162FDF3B5AF8 :AC*BD6AFDC8F65B@A6D@BF1A5AF3?542D=A8F75B@F 8B=A=F 1A5AF 4BA46A8F D6F ;@DF 8542F !,F 65D3?5AF 1D6A5F 1D=F 35A8A7 B:6D8D6A8F %F 8B%##"'8"8'#'"%$"'8 %$ @A62B8, EDCBA@?==8654-/=482181:49-44:2 CDCBA@?;A=F 85A469F :9F =1A87;8 D;AC6F162F62AFCDCBA@?=9==F35B8A8FD?D:AFD6AF., @B532BB;9F DC8F 6B3B;5D329F B7F 62AF CDCBA@?;A=,F )(F 6B3B;5D329F B7F 7B5@?D6 8"87"%8;""87; E%$#"8F !;,F .F 4AD59F =2B1=F 8AC=A9F EDCBA@?;A=F 1A5AF 8?6A8F 65D3?5AF 1D6A5F 3D40A8F 4?=6A5=F B7F 4D5:B@A5F $#"GF D=F 1AF D=F !.'"""F F DC8F BCAF 85B3F B7F =B?68F=65?46?5A=F5A35A=AC6 249 CBA@AB?A= ;:A987@6:= B:C7A5@= 4:5;663= was in agreement with the findings obtained by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igure 6. 3D topography of the formulation ;:A987@6:= 3A7@= %= 9;5B6(0= .A= 18B5.AB= NEG_C980_d. 3.2. Rheological analysis 964B:@5B8458BA= :1= 5.A@A= 4:9C7A(= 1:B987;56:@= All formulations exhibit non-Newtonian, shear- 4:926= "65.= B.A:7:364;7= 9A;@8BA9A5@#= ";@=@844A@@1877'=A@4B62A#=BA?A;763=C:5A56;7= thinning, thixotropic behavior, as evidenced by :1= 5.A= :?A7= :7';4B'7;5A= 4B:@@C:7'9AB0= flow curves and viscosity vs. shear rate curves ;B2:9AB==CB:84A@=;:A9873A7@="65.=5.A= (Fig. 2). A@A7'= C;4A= CB:84A@=C:B:8@#=7:"?6@4:@65'=3A70 0=%:5.6#=/00#=A5=;70#=-;:A9873A7==-:?A7=-;:= ;BB6AB= ;@= ;= ::7= 1:B= :C64;7= %B83= %A76?AB'0= .;B9;4A8564@#=0= =0 0=.;#=0#=A5=;70#=-;:A9873A7==B:96@63=-:?A7= :B987;56:= 1:B= BA;59A5= :1= !6= 679A5@0= :7'9AB=877A56#=0= =0 Figure 2. Flow curves (upper graph) and viscosity vs. shear rate profiles (down graph). .6@= BA@A;B4.= ";@= 18A= 2'= 5.A= /66@5B'= :1= This behavior is typical for topical semisolid !46A4A#= A4.:7:364;7= %A?A7:C9A5= ;= formulations and indicates that the formulation :?;56:#= AC82764= :1= !AB26;= 5.B:83.= 5":= could be easily squeezed out of the tube and +B;5=3BAA9A5@="65.=6?AB@65'=:1=A73B;A simply spread at the site of application. The ;4875'=:1=.;B9;4'=-:== highest viscosity was exhibited by #=-:==#=;= NEG_C980_d, while the lowest viscosity was -:=0 observed for the sample NEG_XG_d, which 250 ;:98 ;765432371043/.-/,+-*712.2)43/.-1(/.'/42,-2%-7602.$ .2/+1+-(1543'./*-;725;72.4;4+-)1.5 # -(1 1 2 1 1 , Sabina Keser-0-, 0-( , Zora Rukavina # "! !!!!! ! !! 1. 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ajafloo, R., et al., A mini-review of Thymol incorporated materials: Applications in 243D54B-3D:9.490D:BA@?=*)@(7+ antibacterial wound dressing. Journal of Drug Delivery Science and Technology, 2020. 60: 101904. 2. Pham, Q.D., et al., Chemical penetration enhancers in stratum corneum - Relation between molecular effects and barrier function. Journal of Controlled Release, 2016. 232: 175- 187. 3. Keser, S., et al., Vesicular phospholipid gels: A new strategy to improve topical antimicrobial dermatotherapy. International Journal of Pharmaceutics, 2024. 667(Pt B):124931. 4. Rukavina, Z., et al., Azithromycin-loaded liposomal hydrogel: a step forward for %')B)2.-D.24B/0DC2:9.BDCB=*)@(.+ infections enhanced treatment of MRSA-related skin . Acta Pharmaceutica, 2023. 73: 559 579. *)*)'%!''."%$'"'.$ =*)@(.B62./:96B.4D019B'D65:5.B(7B109490B 43,B:D..B'D65:5.B(7B2,62-42,1B=32.BD0 B.B.5//D0496B;B .D:26*:2 9B.405-4509BDCB;D43B)@(.B21+B7+B=39B 051B 69:290B ,,D..49'.B CD0B 4D/2-:B :2,90B 2.-D9:.42-B 0912D,B -30-490296B ;B -D,.4,4B /:495B 3909B (B ,6B (B 09'2,B ?0D42+BB 5, 252 4323 101/.-,+*)4('-%0$)4*/4+*#-+)-1.$"+1%+)-!!"1+)%+$$) ,+(0 -/"* 0) 1 1 1 2 , ) , Julijana Kristl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igure 7. The impacts of nanofiber diameter, nanofiber pore size EB6:D:BA,=FGand mat thickness on PBMC penetration and activation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evelopment of a novel in vitro cell model for 6@?=8?;5@)7 =1/.17 6=59;=8?;5@)7 8@17 evaluation of nanofiber mats immunogenicity, 4;%@;;.8@?9 795$=170?8:59;.78.?;;? 7$;?25/?7 International Journal of Pharmaceutics 650 (2024) ;@1/.;@%7 . ?5?5;.;? 7 ;@7 ,+*)7 (*')7 8@17 +7 123696. @7;@.=8417#"(7;@7?2;.!=708?4)79;!9 71/7 diameter as a critical parameter affecting skin cell response, Eur. J. Pharm. Sci. 66 (2015) 29-35. ?57 %=8?=7 ;00/@7 .997 8.?;8?;5@7 65?@?;897 @8@5;:=4-7;@708?4)725$=)708;@?8;@175=7 [2] J. Pelipenko, P. Kocbek, J. Kristl, Nanofiber 146;?7=1/.176=59;=8?;5@7;%/=7-7 of polycaprolactone, alginate, chitosan and zein 7(-7;18=)7-7/68@;)7-7=;4?9)7"-7=84)7A068.?7 9/5=4.@?7;08%;@%7.5@;=017?28?7.9971@4;? 7 nanofiber physical properties on immune cells for safe biomedical applications, Int. J. Biol. Macromol. 8@17 .9/4?=;@%7 $=7 4;%@;;.8@?9 7 =1/.17 ;@7 282 (2024) 137029. ?2;.!708?4)7$;?2705=7.99475/@1716=7;@7?27 4?=/.?/=7;%/=7-7(9457?276=;0@?7$2=7 "!! @8@5;:=478@17=;@417?207=507?27@8@5;:=7 /665=?17 : 7 95@;8@7 #48=.27 (%@. 7 $7 ./9?/=17 ?27 4807 @/0:=7 57 .9947 5@7 4/=8.7 425$17 ?28?7 05=7 .9947 =08;@17 ;@7 '=8@?7 5-7 , )7 ,)7 )7 ?2;.!=7 @8@5;:=7 08?47 4/%%4?;@%7 ?2 7 7 8@17 /=568@7 #%;5@897 9560@?7 6@?=8?1716=78@17%5?7@?=86617;@7?2708?-7 /@17'=8@?7+7(#A#A-4;- Figure 8. Proliferation of PHA-L activated PBMCs, cultured for 72 h on differently thick nanofiber mats (thickness noted in upper left corners) made of PCL, ALG/PEO2MDa, CS/PEO900kDa, and zein/PVP. Cells were 4?8;@17$;?27(,A-7.897:8=475@78.27;@1;;1/897;08%7=6=4@?770- 254 *)(' %$#"$! !$"$"#$$"$## #" Nata 1 1 2 2 1 a Holler , Bernd Gesslbauer , Ilse Letofsky-Papst , Claudia Mayrhofer , Andreas Zimmer #"! ! ! ! ! !!! 1. 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MATERIALS AND METHODS -)0:42 812 0(42 .435/94.2 9:987:665462 :32 5630,*2 0(42 6:2 9:984'-,358932 4642 )64):64.2 '4:3-64.27786.59/20820(42,5046:0-6420(42$ -359/2(5/($)6433-642(8'8/495:05892 464242 26474)08625324)64334.25920(42$274,,322 04304.2.5114649024'-,35154632#4//2,4750(5925)85.2 42 78'):64.2 0(42 -)0:42 592 0(42 $2 74,,32 2 "2 "2 2 2 :9.2 "2 ;!2 592 .511464902 )65862 082 :9.2 :10462 0(42 .511464905:05892 082 0(42 78974906:058932:9.2424642:.-3059/20(4285,*2 ':0-642 :.5)87*0432 32 789068,2 :2 )4/*,:04.2 )(:342 *2 -359/2 +:658-32 85,32 :9.2 2 9:984'-,35892 #"2 ;"!2 :32 -34.2 2 (532 6:05832 (42 4302 7:9.5.:0432 592 046'32 812 0(42 .511464.2168'20(424$2;"289,*25920(42:349742 ):6057,423542.53065-05892:9.240:2)804905:,24642 8120(42)4)05.428920(423-61:7428120(4285,2.68),403 1-60(462-34.21862)64):6:05892812)4)05.4$.564704.2 $##""$ 9:984'-,35893282.435/920(42.6-/27:6654620(42 449.59$2 :32 78-),4.2 082 0(42 "2 49.2 812 :2 ' )4/*,:04.2 )(83)(8,5)5.2 7(:592 *2 :2 7,57$ (42)64):6:058932)68+4.20824230:,428+4620(423402 7(4'5306*264:705892(53249:,4320(42:00:7('4902 )4658.2 812 05'42 32 4)4704.2 0(42 .68),402 3542 8120(42)4)05.420820(423-61:7428120(4285,2.68),4032 59764:34.2:10462789-/:05892#3442:,42!2 50(592 0(42 9:984'-,35892 (42 34,4704.2 6:2 9:984'-,35892 :32 '54.250(2 0(42 ,*8)(5,54.2 )4)05.42 789-/:042 082 )64):642 :92 449.59$2 9:984'-,35892#3442 5/-642! 255 ('% $ $##"! % BA@? =; :9=87? :654=8A36 62 199? 8=0 1 31=A $=:1A$49=..6A;A ".?= ./68 ** =6?5/.3A6 :61=A 2 78?? =; 6 6?:931 31=A "./? 48A68 16 =; =211?8 :6''/7=1A6 0A199 B%+$%+#"!))) 2 1 =; B%+$%+#"!))) $??;A$ " B Size [nm] PdI Charge [mV] raw NE - Our findin ngs demonstrate that the uptake patterns ob bserved in 3T3-L1 fibroblasts PEG NE 175,5 0,12 - 6 56,,9 were significantly diff feerent and that the ex-4 NE 0,11 - uptake rate es were higher when the peptide was presen nt on the s faace of the oil urf droplets. T This tendency is even more # pronounced d after the fibroblasts are fully ?=3/8?5?13 62 :?../.=8 /41 ? 6?8 1A5? =8? = difffeerentiated to the mature adipocytes. This 48?3?1?; A A7/8? ! 9? /41=? 0=3 suggests a specific or a receptor-mediated :62AA85 2 8 5?; 0A19 619 /;A222??8?1A=11?; =; 2 2 = uptake. Sub ubsequent studies will focus on ;A222??8?1A=11?; 2 2 = :?..3 =; 22668 619 14?3 62 = exploring t the significance of this specific =64=811A:.?3 8 60??8 9 8 A79?8 /41=? .??.3 = type of uptake using receptor antagonists 0?8? 63?8?; A :?..3 18?=11?; 0A19 19? ??;A$ 8 = and diff feeren nt endocytosis inhibitors. =6?5/.3A6 A:863:64A: A5=7?3 "3?? = A7/8? 3960A7 5=11/8? $- =;A46:1?3 = =211?8 ) 5A / = =6?5/.3A63 - 6?; B ( ?1 =. -,+*)) *(', ($$+####''"!' + % % % % % % % % % % % % ( + # # % % % % % % % % % % % %% % % % % % % % % % % % =.36 :62AA85 2 8 5 19? A1?8= =.A@=11A6 = 62 19? 6A. ;864.?13 ' +*' ## % % % % % % % % % % % % % % % % % % % % % % % % ((, *"+ ) % % % % % % % % % % % % * *'!, ,(',,) % % % % % % % % % % % % , $,,",($ % % % % % % % % % % % % $ , ( $% % % % % % % % % % % % 6.?:/.=8 = =; ?../.=8 +;6:8A6.6 !)- 7 % % % % % % % % % % % % *!"! * ! ..?8 ?1 =. "+ ) * *'!, ,(',, % % % % % % % % % % % % , % % % % % % % % % % % % % % % % % % % % % % % % ) 6.?:/.= =8 ?1=6.A35 !)- ) !-) /2==/ 2 ?1 =. % % '($$ + , ' ,,* ## % % % % % % % % % % ( % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % % * % % % % % % % % % % % % % % % % % % % % % % % % +''"(, ' " 5?8A:= 6/8=. 62 %93A6.67 $ ?.. 9 7 %93A6.6 ! 9 7 !)!- !)" *!!*- 9? ?1 =. ) )- '' +*' ' % % % % % % % % % % % % % % % % % % % % % % % %% % % % % % % % % % % %-, *+(' % % % % % % % % % % % % ## ''"(, '####,,$, ('+(' % % % % % % % % % % % % ' " % % % % % % % % % % % % ' # #% % % % % % % % % % % % + % % % % % % % % % % % % ?. ../.=8 /441=? 6?8 ! 96/83 " / B ',, ,' ' ?../.=8 %93A6.6 =; A6:9?5A3188 9 7 1 8 6199 =6?5 1 5/.3A63 0?8? ;A./11?; A 199? 3=5? / 1 ? 0= 36 = !)- !" - ---- 1199=1 199? 2AA=. 4 1 2 4?41A;? :6:?188=1A6 A3 )) 1 $ $ We would like to t annk to the Phospholipid ha Research C Center (PRC) foor sponsoring our f master stud dents (Projject number AZI-2021-o 096/1-1) and d to the Lipoid GmbH foor providing f the egg-lecit tthhins used in thhis research. t 5=7?3 61=A 2 28865 :6266:=. 2 5A:863:64? "?A:= A:86331?53 #?85= A A A ?2668? 2 2 A5=7A7 :?..3 0?8? 2AA?; A , 26685 2 2 8 5=.;?9;? =; 9 256 8765 43210/.-,+0*3)*0,-3-(80*'%)$,810*,$20/3#(/,/0".!31 (,( "014*,2.0/(, 3 3-(!'(31083,/(8,14,$0803.,(,*.2'( $1.231., (+1 1 1 1 , 4( , 1(1 , Julijana Kristl 1. INTRODUCTION Table 5. The collection parameters of three samples. NMLKJIHGFEDC DBDHA@?LIGC BILC FIH@G@D=C KBII@LIGC Movemen Movemen Sampl AHICMHKBMCCollecto t in x-axis t in y-axis e BILB8CFHIHG@J:8CBDr direction direction name ILMLBGL7C69L@ICFHJLDJ@BMC@GCLGFLK@BMM:CILML;BDJCAHIC (mm) (mm) JILBJ@D=C ;B=@DBMC @DALKJ@HDG8C 59LILC MHKBM@4L@KIH?@BMGC LD9BDKLGC LAA@KBK:C P2 Plate 110 190 0 290 BDFM@BDKL7C 3LGF@JLC L2JLDG@;LC D Drum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igure 9. Scheme of nozzle movement (red arrows) above different collectors during electrospinning process of samples (grey area) (a) plate (P1), (b) plate (P2), and (c) drum (D). Dimensions of the nanofiber layers in samples that comply with the pharmacopoeial requirements for the uniformity of mass in single-dose preparations along longitudinal sections are marked in green, while those that do not meet the criteria are shown in red. 2090;70/;3.02%'7! 6:520;70/-8/24"-4*656.8;3.02749'0:.2014';9! ?= ;:;98767 54:36210/ .-0 7,550773,9 3421;.64:437144.-+*0;/)3200:;:436*027(6.- ;'02;0-67 7.,/8 /014:7.2;.0/ .-;. :;:436*027 .-0 549905.42 .8"0! 633020:. 549905.64: 70.,"7 5;:*0"24/,50/*80905.247"6::6:.4100.-! 76:6365;:.98 ;3305.0/ .-0 0410.28 ;:/ ,2! 20,62010:.7 342 76:90)/470-4140:06.8 43 .-0 :;:436*02 1;.7! 99 "-;21;50,.65;9 34217 (-0: ;""24"26;.0 :;:436*02 1;.7 0-6*6.0/ 0/0 .-6::6: 549905.64: 7.2;.0607 ;20 ;""960/! -0 24.;.6: -4(0'02+ /2,1)549905.0/ 7;1"907 "24'6/0/ ; /2,1549905.420:7,20/*0..02-4140:06.8;:/ 9;202 ,7;*90 ;20; (6.- 1420 ,:63421 50:.2;9 ,:634216.8 43 1;77 ;52477 .-0 1;.+ (-690 2064:7!-0 50:.2;9 %!#51 705.64: 43 /2,1) 2076/,;9 749'0:.7 (020 7;3098 2014'0/ *8 549905.0/1;.7$#513,99810..-0-!,2! 54:.24990/ /286:! -070 36:/6:7 ,:/0275420 !!1;77,:634216.8526.026;!:"9;.0)549905.0/ .-0 61"42.;:50 43 *4.- 549905.64: 10.-4/ ;:/ 7;1"907+ 541"96;:. 4:07 (020 ;974 6/0:.6360/ "47.)"2450776: 6: /0'094"6: ;550".;*90 ;3.02059,/6:.-04,.02147.2064:7!99.07.0/ :;:436*02)*;70/ ,:634216.8 !!$ 20,62010:.7! 076/,;9 [1] J. Pelipenko, P. Kocbek, J. Kristl, Nanofiber (6.-54:.0:.,:634216.8 749'0:. ;:;98767 7-4(0/ $! 342165 ;:/ response, Eur. J. Pharm. Sci. 66 (2015) 29-35. diameter as a critical parameter affecting skin cell $!$ ;50.65 ;56/ 6: ,:/260/ :;:436*027+ *4.- (099*094(.-0$!-!,2!.-207-49/34259;77 %749'0:.7! 286:3,2.-0220/,50/.-07090'097 ?,""42.0/ *8 ?94'0:6;: 070;25- 0:58 *094(.-09616.432;:.4!)$+);:/:6'0276.843 2090;70/ = 1420 794(98 .-;: ,:/260/ 4:07 ,*9;:; ?.;2.)," 070;25- 242;110 ?) )$$! 258 5432 10/.-1,+.*)0+)-/*)-(('*%*1$-*#0)/"5,+")%1"','*"!* )%)(" -1/,+' Ferenc Fenyvesi 1 1 1 1 2 *)* **/-Nagy **/ *, * 3210/.-,1+-*)(*')1%$/.*/+#*"/+)0!/.,/%1$- %*+ 1. -*)(*21.1%1+*$+/. 210/.-,1+-*)(*!/.,/%1$- %/*1%!+))*+ 1. -*)(*21.1%1+*$+/. 1. 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"!. 1 .-/,/+-*0.-+.(.0/.+(+. /,/+-*0.//()/.+#.(+(. */. $#+)/,. $+-! -('/*. (. 1. 0+,. "//. )///#.%0.0/.,$))(-.)-(#/#."!.0/. ,-!.(.$$-/.+#.(+(.(. $+-!. -(.0/.+(+./,/+-*0.//()/.+#. (+(.$#.+*/#.$#/-.0/.1 .$#.,*0// 260 :987 6543210/04.-,2+5+*/)03,(24*424,:2'30/-5(*515++5+*04* 424,/5--%-,(5*$,%4+*)52-046*424,/,1:,(035 Lina Livrinska1, Marija Petrushevska1#*4"! *6! !"1#*%!*"2#*"*! "3, Beti Djurdjic1 1 2 , Maja Simonoska Crcarevska , Selestina Gorgieva 3210/.-,+*)+('1%10,$+#"!+ ,.+1+-'*/"+"-,++#*$+!+10*1 210/.-,+*)+0'101.+$+"-,+*)+1*$+#.*1 -+)*+%01.+%0**.*,$+"--/-+*)+(/.0+1.-'+*)+*-*$+*-* 1. 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yoti Verma1 1,2 2 1 , Vishal Kumar , Carl-Eric Wilen , Jessica M. Rosenholm and Kuldeep K. Bansal1,2 $#"! !!! ! !! ! ! ! ! ! ! !! ! ! 1. INTRODUCTION D:67F7.7FG 2006/?G ,' ?G ?G 0?G =ADECB68=D:67F7.647FE@DE=H 20 '*/?G IHG FEDEHCG BEAF@?G A@@=;C@G :A AFA.7FA68E:B8EG 2 /G AH8G :E!AHEG (EFEG A8EG C7G 8E ;4FD:A@E8G .F7G =3A*68F=D:? =H6AH8-G DAFF=EF@G210@/GDA;A96EG7.GE..EDC= A@=HEG 6ADC7HEG 2/G (A@G ;4FD:A@E8G .F7G 8F43@G ;FED=@E6BG C7G 8=@EA@E8G AFEA@-G ,:EG A+7FG 64D:G@@EHDE?G;A=H-G7!7F49=D=H2/GAH8G 37A6@G 7.G 10*9A@E8G DAHDEFG C:EFA;BG =HD648EG 7DECA!E6G 2/G (EFEG ;4FD:A@E8G .F7G 0G =H==5=H3G8F43G6EA)A3EG=HG7..*CAF3ECGAFEA@GAH8G A97FAC7F=E@?G-G0:67F=HEGEG20E/ 2/G @C=46AC=H3G @4..=D=EHCG 8F43G FE6EA@EG (=C:=HG C:EG (A@G;4FD:A@E8G.F7G0ABAHG0:E=DA6?G-G C47F-G'7(E7@CGDA@E@?G;FEAC4FEG8F43G E@A54F=H?G#?*=;:EHB6=@79EH57.4FAH2 /G FE6EA@EG.F7G10@G8==H=@:E8GC:E=FGA8 (EFEG ;4FD:A@E8G .F7G ,0IG 4F7;EG 76BEF%8F43G D7H+43ACE@G AFEG DA;A96EG 7.G 0,'IG 274HC=H3G E8=4G (=C: =H:=9=C=H3G ;FEAC4FEG 8F43G FE6EA@EG =HG DAHDEFG I/G (A@G ;4FD:A@E8G .F7G EDC7FG DA4@E8G 9BG D:E7C:EFA;E4C=DG A3EHC@G $#"-G A97FAC7F=E@?GIHD-G4F6=H3AE?G0-C:EFA;B?GC:EFE9BGEH@4F=H3G=H=A6G@=8EGE..EDC@G '7(E=CE8G84EGC7G C:E=FG H7H*@E6EDC=IHG C:EG ;FE@EHCG @C48B?G (EG @BHC:E@=5E8G @C=46=* 6EA8=H3GC7G;77FGE..=DADB-GIHGDAHDEFGC:EFA;B?G=CG=@G FE@;7H@= E@@EHC=A6GC7G@E6EDC=@=46CAHE74@6BG =HG AG D7HCF766E8G AHHEFG ACG C:EG =HC7G C:EG C47FG C7G ;FEC47FG FE3=7H-G EG D7H+43ACE8G 87!7F49=D=HG A@@7D=ACE8G(=C:GAHC=DAHDEFG8F43@-G1E2/?G87DECA!E6G2,/?GAH8GD:67F=HGEG20E/G C:EG =HD7F;7FAC=7HG 7.G @C=464@*@EH@=C==HACE8G ;76B2+A@=HEG D:AFADCEF=@C=D@G =HG 10@G D7468G 9EG AG ;7CEHC=A6G 6ADC7HE/G2*9**'/G;76BEF?GFE@46C=H3G @764C=7H?GEHA96=H3GHEAF*D7;6ECEG8F43GFE6EA@EG=HG =HG C:EG .7FAC=7HG 7.G *9**0E*,* FE@;7H@EGC7GAG@C=464@GA@@7D=ACE8G(=C:GAG@;ED=.=DG *,G D7H+43ACE@?G .4FC:EFG FE.EFFE8G C7G A@G 8=@EA@EG @B;C7G $ "-G ,:4@?G =HG C:=@G @C48B?G AG **,-GEG@BHC:E@=5E8G*,GAH8G @C=46=G @EH@=C=EF%8F43G D7H+43ACEG (A@G ,*,G@E;AFACE6BG9E.7FEGA88=H3GC:EGC7GC:EG @BHC:E@=5E8G.7FG@E6EDC=FEADC=7H-G ,:EG @BHC:E@=5E8G D7H+43ACE@G (EFEG =HD7F;7FACE8G=HC7G@E6.*A@@E96E8G=DE66E@G2* $"%!$$%" *,G =DE66E@/G 4@=H3G C:EG G HAH7;FED=;=CAC=7HGEC:78?GAH8GD:AFADCEF=5AC=7HG (A@G;EF.7FE8G9BGAHA6B@=H3GC:EG8F43G67A8=H3?G 76B2EC:B6EHEG36BD76/GEC:B6GEC:EFG2?GHG @=5E?GAH8G=HG=CF7G8F43GFE6EA@EG4C=6=5=H3GGAH8G G -G A/?G #??*CF=A5A9=DBD67G $--"8ED** G@;EDCF7;:7C7ECFBG$"-GEGA6@7GD7H84DCE8G EHEG 2,/G 2/?G EFDA;C7;F7;=7H=DG AD=8G # =HG =CF7G FEADC= 2/?G 8=EC:7!B* *;:EHB6ADEC7;:EH7HEG 3EHEFAC=7HGAH8G,G6=H)EFGD6EA CF=EC:B6A=HEG 2,/G 2-/?G 1*2*G =HCFADE6646AFGG3EHEFAC=7HG4@=H3G0 '?G 8=EC:B6A=H7;F7;B6/*1*EC:B6DAF978===8EG AH8G DE66G DBC7C7!=D=CBG @C48=E@G 4@=H3G C:EG6AAFG 20/ 2/?G *28=EC:B6A=H7/G ;BF=8=HEG 964EGA@@AB- 2/G 2/?G 1*:B8F7!B@4DD=H==8EG 21'/ 2-/?G '0G 3FA8E@G EC:AH76G 2-/?G ADEC7HEG 2-/?G 8E4CEFACE8G 8=EC:B6G @46.7!=8E 2*8/?G 8E4CEFACE8G 263 N =J@=?N:G=2N''NE8N3(NIG7@E=NFLEFE=J@=GLENL*N MCB/N I4N :@4N 4=@GE?N :G=2N M#.N DHA-N MLKLJIHGFGENDMCBAN@E?N?LF=@KJN :2GN MCBN GE2JE=N J?N *4FEFN :@4N I4?N =LN FLE9I8@=?N LE=LN EL7N @F=LEAN ?=F=N )MCB)M;BN 3GF4/N F@N H@J!'N 3N H@4?NFL6L53JN7G@N@N=2GL1=@J/N.EN D@4JAN@E?N('N3ND:G=2LI=N@4JA/ @==@F2?N =LN =2N 6L53J-N :2GF2N 4J7?N @4N @N @??G=GLE-N @N 62L=L4E4G=G,JN DF2+AN :@4N J@F=G7NLK58EN46FG4N8EJ@=LJN=LNF@7N=2N ;2N 4=I?5N 4IFF44*I5N FLE9I8@=?N =:LN ;0N J/N ;2N FLE9I8@=N G4N J@?G5N 4*) @E=GF@EFJN ?JI84-N MCBN @E?N M;B-N LE=LN @N @443H?N GE=LN 3GF4N 44N =2@EN (''N E3N GEN 6L5D9@43GEN@F=LEANHJNI4GE8N@EN 4G,/NGF4N?3LE4=J@=N@NEL=@HNGEFJ@4NGEN $C)4E4G=G7N ;0N J/N ;2N 62L=L4E4G=G,JN =2GJN@HGNF@=2N:2ENK6L4?N=LN +N :@4N @???N =LN @FFJ@=N ?JI8N F@7@8NGEN E@J)GE*J@J?N D%.$AN =2N6J4EFNL*N%.$NNFLE9I8@=N4*) HJ@4=NF@EFJNF4/N;2NGEFJ@4NGENF5=L=LKGFG=5N @443H?NGE=LN(''NE3N3GF4-N:2GF2N42L:?N G4N 2G82JN =2@EN =2@=N LH4J7?N :G=2N =2N 2G82JNF5=L=LKGFG=5NGENF@EFJNF4NFL36@J?N=LN FL3HGE@=GLEN L*N *JN MCBN @E?N M;BN DG8/N (A/N *JN ?JI84/N L:7J-N H@4GFN @E@5=GF@ ;2N@FFI3I@=GLENL*NMCBNGEN=2NEIFI4N@*=JN *@G?N =LN ?=J3GEN =2N K@F=N @3LIE=N L*N ?JI8N J@4N*JL3N=2N3GF4ND@4JNGJJ@?G@=GLEAN:@4N N3GF4-NGE?GF@=GE8N=2N6L=E=G@ @4LNFLE*GJ3?NH5NFLE*LF@L*N =2N ?7?N *LJ3I@=GLEN GEN J?IFGE8N 4G?N .EN=2N@H4EFNL*N4N?G?NEL=N42L:N **F=4/N I=IJN :LJ1N :G7 @E5N =LKGFG=5N :2GN =2N *JN ?JI84N :JN *LIE?N @E@5=GF@=2L?4N @E?N @66JL@F24N =LN =LKGFNGJJ46F=G7NL*N=2NK6L4IJ/N @FFIJ@=5NF@N?JI8NFLE=E=/ A B PJL-DOX-DTX Micelles 2.5 ug/mL PJL-DOX-DTX Micelles 5 ug/mL PJL-DOX-DTX Micelles 10 ug/mL DCFH-DA DOX DTX Mix 2.5 ug/mL DOX DTX Mix 5 ug/mL DOX DTX Mix 10 ug/mL Free Ce6 + DCFH-DA D0X 2.5 ug/mL D0X 5 ug/mL D0X 10 ug/mL Ce6 micelles + DCFH-DA DTX 2.5 ug/mL DTX 5 ug/mL DTX 10 ug/mL (/N ;2@1LJ-N /N =/N @J?JI8N FLE9I8@=4N Ce6 2.5 ug/mL Ce6 5 ug/mL Ce6 10 ug/mL 4 ns JFE=N @?7@EF4N @E?N *I=IJN 6J46F=G74/N ?JI8N **** 100 ns si4 n n ?G4FL7J5N=L?@5N!'!'D!AN( (( !+/ ty **** te io !/N LE=GJL-N //N =/N @?IF=GLE)J46LE4G7N rat ce In 4 n 50 sce Prolife 6L53J4N *LJN ?JI8N ?J5N GEN F@EFJN =2J@65G4N o 3 % C re ell lu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aryana Salamah1,2 3 4 4 4 , )$#" -5# , 0" Lekli , 0"-)$ , 4#$$-, 0#- $1 5,6 1 , -1-)$# , "-$$ , +*)('((%$#"$! %('$% *##$*$%(#$""')$*'%)'($#"$%%$* +*)('((%$#"$! #*')$*$'# $*'%)'($#"$%%$* '%(#(%$#"$ $)(*%$ %%#%*($')$! %(')$! %)($* %(%*($#"$! ## *'%)'($#"$ %%%*$* %(%*($#"$! %('$ %')( %%%')$*'%)'($* %*(%$"#$! ##$*$ $%)% $$ %%#%*($%%%')$*'%)'($* 1. INTRODUCTION 4/769/539=.A9?2/5;:9748A5=79A?A9A8/?/=A39 BA@?=?;:9 8;?@7A769 ;5:4@3;529 1B09 8;?@7A769 @7;529/9?=/?-98/:@@,9A8/?/=?!9;5/44-69=.A9 :/59 ;58/3A9 =.A9 :A5=?/49 5A?8@79 7-7=A,9 +*B)(9 =.;59 4;;39 ;4,9 /79 .-3?/=A39 ;=.9 .7./=A =.?@2.9 =.A9 5/7/49 A;=.A4;@,69 '43'?/;59 '@A?A397/4;5A9+%)699!(! '/??;A?9 +%%%(9 ?9 -$$ $ ,A5;52;=;79 /539 7A8A?/49 5A@?3A2A5A?/=;8A9 00)79 A?A9 :./?/:=A?;A39 ;59 =A?,9 9 3;7A/7A79 $#"!9 5=?/5/7/49 /3,;5;7=?/=;59 9 59 /539 A=/9 =A5=;/469 /5=;8;?/493?@27697@:.9/79/8;;?/8;?9+0(69;79/9 A5:/7@4/=;59 A;:;A5:-69 3?4A=9 7;A9 ,;7;529 //:.9 =9 8A?:,A9 =.A9 3;7=?;'@=;569 '*$ '(#9 /8/;4/';4;=-9 :./44A52A79 /539 ?A3@:A9 =.A9 ?/=A69/539'*$'#9 5/5 -(6+231+-4.'-'0+2++0/. /79 /7:?'-49 57;3A?A39 /9 -.$$#-4#$# 7@;=/'4A9 /5=;8;?/49 3?@29 :/??;A?9 =9 A5./5:A9 =.A9 .A9 @?9 ?,@4/=;579 7.A39 %%%959/539?A/:.9=.A97;=A99;5A:=;59 .,2A5A@795/57;A395:A5=?/=;579$6"! 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GA=2FA2=D F @ @ @ @ @E3?H7H78 B 69=E4HCHF 4EC63D= @ @ @ @ @ @ @ @ @ @ @ @ @ @ @@ @ @ @ @ @ @ @ @ @ - -% =DCDBGD ,==E3 !/15 ECDE8DC @ @ @ @ @ @ @ @ @ @ , @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ =DBFD9 4CBAADB22 B,AAD= * 3H72ADG BG AD 8DC 3BAA=H ;HA B @ @ @ @ @@ @ @ @ @ @ @ @ @ @ B CH4E4HCHF 4BGD. -DH= B??HCHA6 AE @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ B @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ B B @ @ @ @ @ , @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ =DCDBGD9 AD EHC 4BGD H33D9HBAADC6 B,AAD= ?D ,EE=3 @ @ @ @ @ , = 32CBAA B AD9 ;H @ @ @ @ @AE2A G2=,BBFAB77AG E,,,DD=G B 2 @ @ @ @ @ , B @ @ @ @ @ , , @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ B @ @ @ @ @ , @ @ @ @ @ 9HGH7AD8=BAAHE7 E, B=9 FB4G2CD FE7ABH7H78 GH87H,HHFB77A B , B @ @ @ @ @ B9IB77AB8D EID= FE7ID7AHE7BC CH4H9: B B @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ B @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ ECDE8DC $H82=D #. 7,EE=AA27BAADC6 E7C6 B44=E. ?BGD9 G6GAD3 @ @ @ @ @ 3G G2F BG 10'' 4EAD7AHBCC6 @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @@ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ , = B @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ - =D92FH78 8B @ @ @ @ @GA=EH7ADGAH7BC H==HABAAHE7 779 AEHFHA66 7 @ @ @ @ @ B @ @ @ @ @ B @ @ @ @ @ A@ @ @ @ @ @ @ @ @ @ -% ;BG =DCDBGD9 B,AAD= * 3H72ADG BG AD @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ , @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ EHC 9H8DGAHE7 92=H78 ;HCD 3BH7A @ @ @ @ @BH7H78 D,,,DDFAHID 9=228 GEC2?HCHBAHE7 @ @ @ @ @ , , @ @ @ @ @ = @ @ @ @ @ B@ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ CH4EC6GHG FB2GD9 CEGG @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ E, - B79 =DCDBGD. @ @ @ @ @ @ @ @ @ @ -% GEC2?HCHBAAHE7 FB4BFHA6. @ @ @ @ @ B @ @ @ @ @ @ @ @ @ @ 100% $ % % % % % . B7F2=B @ @ @ @ @@ @ @ @ @ 1 H4 5HDCDFB:/HE7AD . @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ 80% AdTV 60% . / /B=3BFD2AAHFG. +*++. $#. 2 @ @ @ @ @ @ @ @ @ @ asel 40% % % % % % ECE DC @ @ @ @ @ C CB CH B4D=CH7 1 IE7B= @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @@ @ @ @ @ Ol e eR eogel 1 /E?H= . = @ @ @ @ @ @ @ @ @ @ 20% 20% Ol Ol e e o o g g e e l l 2 2 @ @ @ @ @ 7AD D=7BAHE7BC E2=7BC E, /B=3BFD2AHFG. @ @ @ @ @ @ @ @ @ @ @ @ @ @ @@ @ @ @ @ 0% +* . $ # @ @ @ @ @ +:+. 0 10 20 30 Ti me [min] $# -D B22AE= @ @ @ @ @ BG BF7E;CD98D ,HH7B7FHBC G244E=AA @ @ @ @ @ @ @ @ @ @ , @ @ @ @ @ =@ @ @ @ @ % % % % % % % % % % /D=FD7AB8D E, =DCDBGD9 - @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ -% 92=H78 @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ ,,==E3 AD CE @ @ @ @ @ @ @ @ @ @ EID7HB77 DGDB=F 8D7F6 $=DGDB=F B @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ CH4EC6GHG )7(. 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omp. 1 2 3 Tetracycline $B=/81EG';K)56;5*;+5I5)*6 B. Subt. na** na na K65)6* 6;D@D';66';=1;A@6'; )%* ) +('+( )) S. Aureus 556))6K na na K65)6* %' *'+) *'++%+) *) *'+%+*') *'6; E. Coli 5+65)65 56))6K 56))6 K65)6* #?8=1808?;=.DE1';K)K56;55;K556 S. Typhi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omp. 1 2 3 Trolox #?8=?='; !=?BD@D78?A@; =,=@D./=1; AF; LP D,A18D; D0; #=E28A; :ED=?1; *5I)I Prooxidative Prooxidative inhibition K6K)6K 0.005 0.001 activity activity 5 K)K*I) K))55; AF; *5I)I5 K)K*I (IC 50 )* ;; =A;,A@-=;;#;:8 ; /7 ?/ 4;;;;;;;;;;;;;;;;;;;;; ) K))5546 ,$,$%( %% % ( "@@; ?D/.D-F; 3=E=; .E=F8?1=F; 1D; BA,=; 7DDF; DEA@; 28DA,A8@A28@81G; AF; B87B; 7A1ED81=18A@; A26; @G; ?D/.D-F; #; 3A; .E=F8?1=F; 270 /.-, /+*)('%$#$"%'!# +#!/")%'#(%$+/"%$*'!#%# /+'%"*# +#!!"%'#/*(+")#'*'"!' #1 1Department of Pharmaceutical Technology, Faculty of Pharmacy, Masaryk University Brno, Czech Republic 1. INTRODUCTION A7@?D=@=98:9=B=BD=B98A?)B@?D=9DCBD0982= 0D@?@7@?D4= " = 6= @C9= A?5= 0D@9D@= +/= EDCBAB@?D= ?= B= DD;?D:B7@9= 6= 5874= 891AB0?D4=BD=9#7?:BA9D@=B37D@=6=@C9=0B88?98=?D= B53?D?@8B@?D2= 1B8@?07AB8A/= 96690@?:9= 68= @C9= 0B9= 6= " = BD5= " = ?@= .B= 891?8B@8/= 5?9B92= BAA.?D4= 68= C?4C= A0BA= DA/= =579=@=@C9=A?3?@95=.B@98=A7+?A?@/=6= 5874= 0D09D@8B@?D= BD5= 8957095= A970?D9-=%= 5?A:9= 0C?@ 96690@-= ,909D@A/2= @C989= CB= BA= +99D= 48.?D4= .B=795=?D@9B5=6=.B@98-=%C99=A7@?D=.989= ?D@989@= ?D= 7DB8/= B53?D?@8B@?D= 68= @C9D=7+90@95=@=18B/=58/?D4- = A7D4= @?79*= AB849= 786B09=B89B2=8?0C=+A5=711A/2=B+?A?@/=@=+/1B= 18B/= 58?98= B+ABD@= ;= 9#7?1195= .?@C= A?:98=39@B+A?32=8=A?3?@95=9D)/3B@?0=B0@?:?@/= ' 33= D))A92= "'= $= = 33= 58/?D4= 0CB3+98= ('- BD5= 0/0AD9= 91B8B@8= .B= 7@?A?)95= 68= 3?081B8@?0A9= 1891B8B@?D-= %C9= 6AA.?D4= %C9=031A9$=+8BD0C?D4=6=@C9=891?8B@8/=@899= 1B8B39@98=.989=9@=?DA9@=@93198B@789='=2= 89#7?89= 190?6?0= 0D5?@?D= @= +9= 39@= 68= 6995= 8B@9= = 3A3?D2= B?8= 1995= 2= 32= D))A9= ?DCBA95= 6837AB@?D= @= 96690@?:9A/= 89B0C= @C9?8= 189789==+B8- ?D@9D595= @B849@= B89B-= %C9= 591D= 6= @C99= 6837AB@?D=?=18?3B8?A/=?D6A79D095=+/=@C9=6@98= 9:BA7B@?D2= 9A90@95= BD5= 6= @C9= 1B8@?0A92= B= .9AA= B= @C9= 1891B895=?D=@C9=D= 1B@?9D@*= ?DCBAB@?D= @90CD?#79= ("-= !.9:982= 6= @C9= 359A= 5874= 0A?@?D;7A6B@92= .C?0C= A7D4= 5969D9= 390CBD?3= A? 9= 3700?A?B8/= 0D@?@7@95=' =6=@C9=A?5=0D@9D@2=891AB0?D4= 0A9B8BD09= 8= BA:9AB8= 3B081CB49= B0@?:?@/= 0BD= BD=9#7?:BA9D@=B37D@=6=@C9=0B88?98-A?3?@=@C9= 578B@?D= 6= @C9?8=96690@= B6@98= B= 8B1?5= D9@=6=B0@?D=6=?DCBA95=6837AB@?D=(2=- #' %C9= 897A@?D4= 1.598= .989= 9:BA7B@95= 7 %C?= .8 = B?395= @= 1891B89= ?DCBAB+A9= :B8?7= 39@C5= ?D0A75?D4= 59D.= 3?081B8@?0A9= 598= 7@B?D95= 89A9B9= 18198@?9= 59@983?DB@?D2= 1@?0BA= BD5= 9A90@8D= 77=1A/398=B=0B88?98-=18B/=58/?D4= 3?0801/2=8=89?@789=39B78939D@-= .B= 931A/95= B= @C9= 39@C5= 68= 1B8@?0A9= %C9= 9/= 1B8B39@98= B995= 68= ?DCBAB+A9= 1891B8B@?D-=%C9= 897A@?D4= 1B8@?0A9= 191B8@?0A9= .989= 4939@8?0BA= 395?BD= 18198@?9= 7?@B+A9= 68= ?DCBAB@?D= .989= AB595= 59@983?D95= +/= AB98= 5?668B0@?D= BD5= 3B= .?@C= B= 359A= 5874= @= 9:BA7B@9= @C9?8= 89A9B9= 395?BD=B985/DB3?0=5?B39@98==BAD4= 186?A9-= .?@C=@C9=6?D9=1B8@?0A9=68B0@?D==59@983?D95= +/=B985/DB3?0=1B8@?0A9= 2. MATERIALS AND METHODS %C9=3B@98?BA=795=?D=@C?=.8 =.989=3BDD?@A= 5?A7@?D=@9@=78=031B8@39D@=6= 8BD)= 5?A7@?D= 09AA-= %C9= #( 874;AB595= = B= = ,@C2= 2= 5?73= BA4?DB@9= 2= 0C?@A/:?D/A= BA0CA= = 1AB095= D= @C9= 786B09= 6= "= 3A= 6= 1C1CB@9; ?43BA58?0C2= 2= 0B8+398= ,= +7669895= = = -= B48D2=2=A970?D9=/39B32=2=0A?@?D; A?#7@=6='=3A=.989=@B 9D=B@=?D@98:BA=6=-2= 7A1CB@9= '2= "2= 2= 2= BD5= = C78= B6@98= @C9= CB83B097@?0BA=-2=- B11A?0B@?D-=%C9= B37D@= 6= 5874= 89A9B95= .B= #7BD@?6?95= 783BD09= A?#7?5= #/ ## 0C83B@48B1C/=!-9689=18B/=58/?D42=B#797=A7@?D=6=9B0C= 0B88?98=.989=1891B895=?D=@.=0D09D@8B@?D=B= ?D5?0B@95=?D=%B+A9='-=970?D9=.B=B5595=@=9B0C= 271 3. RESULTS AND DISCUSSION A@? = ?;:9876 54:;? 32 9@? 1?4;=?0 /4=41 4 1?9?= 4=? /=? 87 A44..:? -, +7 139 *4 54:;? 87*=?4 *37*?79=499837) 4 ?$*?/9 2233= 9@? 4:687499? 41/:? 4 9@499 ?$@8.89?0 4 4 6=?499 4?= 9?70?7*# 233= 466=?649837 %@?7 2 4 4 4 0=8?0 2==31 4 08:;9?0 3:;9837, 2 A@? 1 4 1? 9=?70 %4 *3789?79 4*=3 4:: 41/:? 233= !! (, 2 4=98*:? %89@ 477 !! ( 32 4=3;70 ' 1 = 4 1 *47 ?222??*985?:# .? 0?/389?0 87 9@? 0??/ :;76) 2 2 8708*499876 9@499 139 *4==8?= @3% 6330 4 4 /39?7984: 233= /;:1374=# 0?:85?=# 2 , 4=66?= = /4=998*:? '- 1 = *477 98:: =?4*@ 9@? 4 ?:?4 2 49? 2==31 2 9 9=4*@?3.=37*@84: =?6837, A@? @86@ 58* @ . @8 : 8 A@ @8 @ 8 389# 32 89 2 *@3 @ 8 9@? 3:;9837 :?0 93 9@? 233=1 2 = 14 4499837 32 2288.=8: 9=;*9;=? , (? = =*:? 8? 32 ') 9@8 1 4 1/:? 0?1379=499? 4 ?0 6330 4. CONCLUSION 4?=30#741 4 18* /=3/?=998? 0;? 93 89 :3 = % .;: 0?789#, A@? ;64= *4==8?= 08/=4# 0=##876 @4 .??7 @3%7 93 .? 47 ? = 2222??*985? 2 1339@ ;=244*? 13=//@3:36#) %@8:? 9@? /3 1?9@30 233= /=30;*876 2 /4=998*:? %89@ = @ 6330 2 = :#1?=8* *4==8?= ?$@8.89?0 31? ;=244*? =3; 4?=30#741 4 18* /=3/?=998? 4770 @86@ /39?7 = 7984: 233= 2 2 ;6@7?) %@8*@ 1 ## .? 405477946?3; 233= 4?=33:8 /;:1374=## 0?/389837, = +9 %4 3. = 0 9@499 4 4 4 4 4 2 8499837, 4 @86@?= 08:;9837 32 9@? 87/;9 08 ; ; ?;:9 87 ?;:9876 /4=998*:? /=3/?=998? = = 9@? /=30;*9837 32 2887?= /4=998*:?, 13 2 = 376 9@? /3:#1?= 9?9?0) 37:# *@893 77 770 4 4 308;1 Sample 4:687499? 0?1379=49?0 9@? 4.8:89# 93 ;9 FPF 4 4 9487 9@? M =?:?4 ;:24499? %89@87 9@? 288= 2 4 2 =9 9@=?? MAN_5% (%) ',' ,' 5.29 ,- 42.02 MAN_20% @3;=, ,- ,- 7.62 , 14.83 CHIT_0.5% , , 4.86 , 54.21 5. REFERENCES CHIT_2% , , 7.25 , 22.66 ALG_1% , ,- 5.72 , 32.39 -, 498:) , ,) ?9 4:,) ALG_3% ', ,' 6.40 , 22.85 , ;76 CARB_0.5% ',' , 3.74 ,' 61.88 +7084) -, - , CARB_1% ', , 5.15 , 55.57 PVA_5% , (4=;?77?) @,) , -, ,- 5.24 ,' 42.27 3;=74: 32 ?=33: !?08*87? 470 ; :1374=# = PVA_20% -, ,- 7.77 -, 20.42 (=;6 (?:85?=#) , ---', = ,' 08 4 0#7418* , !8=4) ,) ?9 4:,) /=3/?=998?) .;9 9@? 0=;;6 40089837 =?;:9?0 = = 0 87 /33= 08, $/?=9 /87837 37 (=;;6 ( = (?:85?=#) = 4 4 2 22;;=99@?= = 9?9876, +7 9@? "@3;= 083:;9837 9? , - -, ?9) ?# 08222??=?7*? ?1?=66?0 87 9@? 288=9 @3;= 2 2 = 2 86, -, , @@47?1) ,) ?9 4:,) !477893: = 2 *4==8?=)) = 4*@8?5876 0=;;6 =?:?4 = 9? %89@ , 3;=774: 32 3 = 379==3::?0 9 73 =?94=0499837, 3:#587#: 4:*3@3: 4 @3%?0 ?:?4 818:4= =?;:9, @893477 =?:?4 4 ; 87 187;9? 4770 =?4*@?0 499;;=499837 4299?= 4 4 9 4 2 @3;=, :687499? %4 9@? :3%?9) =?:?4876 37 4 7:# ACKNOWLEDGMENT 4299?= 187;9? 470 6=40;4::# = 2 =?4*@876 *31/:?9? =?:?4 ?=30#7 1 4 18* 1?4;=?1?79 %?=? /?=233 2==1 1?0 499 4 4 @3;= 14=, = =?499?= 4 =?94=0499837 186@9 .? 4*@8?5?0 .# 8378* 6 9@? 4*;:9# 32 !?*@4778*4: 7687??=876 3 4 32 =73 4 @ 6?:499837 4 32 9@? A 272 .-,- +*)('%)($(.*#'"#! +( (.+ " (.*#'"#!+*)'#.(#( ".' (!% !#%(+!") Sonja Icheva Doneva1 2 3 , Marija Taleska Velkovski , Ljuba Karanakov ('%$#"! ""!!'#"! !" " !"!%# !"!!'# !" " '%$#"! ""!!'# !" " 1. 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" Julija Komac 1 2 1 , Brigita Mavsar Najdenov , Mojca Kerec Kos 1. INTRODUCTION ;:9876574 5:4 32150234 7:4 704 5/975./20-4 ,14 !824 1,**,+5064 37-74 +7:4 ,)-750234 1.,/4 -824 :+7**,+5064-87-4*273:4-,47047)0,./7*432*7;4504 8,:95-7*:4/2357*43,'/20-7-5,0 4-824-.70:5-4,1474*5('534,.4:,*534),*':41.,/4-824,.7*4 7%5-;4-,4-824:-,/78$4#-4704)247'-24,.48.,05"4 basic patient data (gender, age, body mass 50-2./5--20-4 ,.4 92.:5:-20-$4 !824 9.,9,.-5,04 ,14 index) 92,9*24+5-843;:987657450.27:2:4+5-84762470345-4 patient health status data (number and type 8,/2:4 :'112.4 1.,/4 ,7:5,07*4 3;:987657$4 kidney function) type of enteral feeding tube +7**,+506435:,.32.:47.247*:,4,//,0471-2.474 5:42:-5/7-234-87-4 4,14,*32.473'*-:45040'.:5064 of comorbidities, laboratory results related to :-.,2"4 504 7.50:,0:4 35:27:2"4 32/20-57"4 2-$4 number and types of prescribed oral medicines during and after hospitalisation $4 24 +2.24 50-2.2:-234 504 -824 :'5-7)5*5-;4 ,14 -824 #14 ,.7*4 12235064 %574 -824 0,./7*4 98;:5,*,657*4 9.2:.5)234,.7*43,:76241,./:41,.497-520-:4+5-84 .,'-24 5:4 0,-4 .2,//20323"4 5:4 0,-4 127:5)*24 ,.4 :+7**,+5064 35115'*-52:$4 !824 9,::5)5*5-;4 ,14 3,2:4 0,-4 /22-4 -824 ),3;:4 0'-.5-5,07*4 0223:"4 87065064 -824 3,:7624 1,./4 :9*5--506"4 .':85064 97-520-:4.225%240'-.520-:4%57470420-2.7*4-')24,.4 -7)*2-:"4 ,9205064 87.34 79:'*2:4 ,.4 35::,*%5064 :-,/74 35.2-*;4 50-,4 -824 :-,/784 ,.4 :/7**4 -7)*2-:4,.479:'*2:4+7:482234504'//7.;4,14 50-2:-502$4 #04 -82:24 97-520-:"4 /235502:4 7.24 7*:,4 .,3'-4 87.7-2.5:-54 7034 504 35112.20-4 73/505:-2.234%57420-2.7*412235064-')2:4$ 9.,12::5,07*4501,./7-5,04:,'.2:4$ !824 75/4 ,14 -824 .2-.,:92-5%24 :-'3;4 +7:4 -,4 ,.2,%2."4 -824 987./7,-82.79;4 ,14 :,/24 2%7*'7-24-824799.,9.57-202::4,149.2:.5)234,.7*4 97-520-:450*'3234504,'.4:-'3;4+7:4.2%52+234);4 /235502:4 504 73'*-4 8,:95-7*5:234 97-520-:4 +5-84 74 *5057*4 987./75:-4 7:4 97.-4 ,14 74 :+7**,+50645/975./20-$4247*:,42%7*'7-234-824 987./7,-82.79;4.2%52+$424+2.2450-2.2:-234504 .,*24 ,14 -824 *5057*4 987./75:-4 504 ,9-5/5:5064 -;924,14.2,//2037-5,0:4/7324);4-824*5057*4 987./7,-82.79;4504-85:46.,'94,1497-520-:$ 987./75:-4 -,4 ,9-5/5:24 987./7,-82.79;4 504 ! !" -82:24 97-520-:4 7034 8,+4 ,1-204 -824 .2,//2037-5,0:4+2.24729-234);4-824-.27-5064 # 98;:5570$44 24,03'-23474.2-.,:92-5%24*5057*4:-'3;4504 ! 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" ! ! $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ G? DMML 97EE K K K K K K K K D K K K K K K K K 7 E1 AB IAGFG1A;JF LB A7GB2LBDH K K K K K K K K K K K K K K K K K K K K K K K KK K K K K K K K 99ADAGBJFF@ @0 DML MA1M = LB; ::HEFJDDAGB K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K J JK K K K K K K K NML IIHGFEDL CEJBBDAD@ G? = K K K K K K K K J K K K K K K K K J K K K K K K K K K K K K K K K KK K K K K K K K LB;J::HEFJDDL9 J JK K K K K K K K L???AA;J;@ 6 ? ? K K K K K K K K $3 AH AB9A;JDDG7 G? K K K K K K K K K K K K K K K K J K K K K K K K K K K K K K K K KDML7 ::LEDA;JFF@ JK K K K K K K K 8ADMAB DML BJBBGHD7E;DE7L9 FA:A9 ;J77AL7H 654H3 K K K K K K K K K K K K K K K K J K K K K K K K K K K K K K K K K K K K K K K K KK K K K K K K K :GDLBD BK K K K K K K K ??GG72 7 2 2EFJDDAGB JK K K K K K K K 7G2 A1 ! 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M 6%3 99ADAGBJFF@0 DML K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K KK K K K K K K K EHAB1 J -GDG?AA, +* ;LBD7A?EE1L 6)LDDA;M K K K K K K K K K K K K K K K K ? K K K K K K K K K K K K K K K K ? K K K K K K K K DK K K K K K K K ??GG72 7 2EFJDDAGB JK K K K K K K K 8ADM = FGJ9L9 54/H 6*3 8JH K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K KK K K K K K K K (LBD7A?EE1LB0 'L72JB@3 JDD %$$ 7:20 *% "/ ?GG7 ? K K K K K K K K K K K K K K K K K K K K K K K K J K K K K K K K K K # K K # K K # K K # K K # K K # K K # K K # K ?K K K K K K K K ;MJ7J;DL7AL9 8ADM HFG8L7 7LFLJHL AB DML ?AA7HD M K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K ? K K K K K K K K K K K K K K K K !% 2ABEDLH0 J;7GHH ?GGE7 ;GBHL;EDA L ;@;FLH0 K K K K K K K K K K K K K K K K K K K K K K K K ? 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K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K J 6 77DG7AEH0 'L72 K K K K K K K K J 7 K K K K K K K K 7 2JBB@3 NML 7LHEFDAB1 J K K K K K K K K K K K K K K K KK K K K K K K K 1G L7BBL9 I 7 K K K K K K K K@ IGDM K K K K K K K KK K K K K K K K 9A???EEHAGB ?77G2 DML FA:A9 ? ? K K K K K K K K ? K K K K K K K K K K K K K K K KK K K K K K K K HE:L7BBJDD BBD 8JH ;GFFL;DL9 JB9 JBJF@HL9 I@ 7 J J K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K KK K K K K K K K BJBG;J77AL7 J K K K K K K K K JB9 HEIHLCELBDAJF 9A???EEHAGB DM7GE1M K K K K K K K K K K K K K K K K ? ? K K K K K K K KK K K K K K K K H:L;D7G:MGDG2LDL7 JDD *+ B2 8J LFLB1DM D M D D K K K K K K K K J D *+ K K K K K K K K K K K K K K K K K K K K K K K K F DMK K K K K K K K DML 2E;GJ9M K K K K K K K K MLHA L :GF@2L7A; ?AAF2 2JDD7A, * K K K K K K K K K K K K K K K K ? K K K K K K K K J K K K K K K K KK K K K K K K K 6J72 7 2HDJ9D0 'L72 9 K K K K K K K K 7 2JBB@3 J ' + " $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $! $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ NML ?AAF2H 8L7L :7L:J7L9 I@ 2A,AB1 :7L8LA1ML9 K K K K K K K K ? K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K KK K K K K K K K J2GEBDH G? JBB9 5J// AB J +%! 7JDAG B K K K K K K K K K K K K K K K K K K K K K K K K J K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K JK K K K K K K K 6883 AB 9AHDAFFL9 8JDDL7 EHAB1 J 2J1BLDA; K K K K K K K K K K K K K K K K K K K K K K K K J K K K K K K K K K K K K K K K K K K K K K K K KK K K K K K K K MGD:FJDDL HDA77L7 6+M0 +$$ 7:20 $ "/0 FJDDL J K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K # K # K # K # K # K # K # K # K K K K K K K K K K K K K K K K JK K K K K K K K-/N A1ADJF0 LL7L0 'L72JB@3 B;L ?EEFF@ K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K ?K K K K K K K K 9AHHGF L90 ! 1F@;L7GF0 $% 24 JBB9 ! 24 G? K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K J K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K KK K K K K K K K /J/F HGFEDAGB 8L7L J99L90 ?GGFFG8L9 I@ K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K ? K K K K K K K KK K K K K K K K ;GBDABEGEH HDA77AB1 G L7BA1MD ?DDL7 ;G2:FLDL K K K K K K K K K K K K K K K K K K K K K K K K ? K K K K K K K KK K K K K K K K 9AHHGFEDAGB G? DML /J/F0 !1 G? !$ = E K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K KK K K K K K K K HGFEDAGB 6!3 G7 54/H 6*3 8L7L AB;G7::G7JDL9 $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ K K K K K K K K 7LFLJHL :7G?AAFL G? = K K K K K K K K ? K K K K K K K K K K K K K K K K = ?77G2 K K K K K K K K ?K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K 7 J K K K K K K K K 2E;GJ9MLHA L A L ?AAF2H 6! JB9 *3 K K K K K K K K ? K K K K K K K K K K K K K K K K K K K K K K K K JB9K K K K K K K K K K K K K K K K K K K K K K K K % 1 G? LJ;M M@97G1LF ?GG72 K K K K K K K K K K K K K K K K K K K K K K K K ? 7 2EFJDDAGB 8JH J K K K K K K K KK K K K K K K K :GE7L9 ABDG LD7A 9AHMLH 8ADM J % 22 J % $ $ $ $ $ $ $ $ K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K 9A 2 2LDL7 ' In conclusio on, the controlled slow initial release $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ NML ;E2EFJDDA L 7LFLJHL G? K K K K K K K K J K K K K K K K K K K K K K K K K K K K K K K K K= ?77G2 ! JBB9 * K K K K K K K K ? K K K K K K K K K K K K K K K K J K K K K K K K K K K K K K K K K loaded NLC ffrrom mucoa adhesive fiilms embedded wit f th DEX 8 8JH JHK K K K K K K K L JFEJDDL9 L JFEJDDL9 J JK K K K K K K K EHAB1 EHAB1K K K K K K K K 9AJF@HAH 9AJF@HAHK K K K K K K K DEIAB1 DEIAB1K K K K K K K K therapeutic efffeect, especially in conditions like C can be benefiicial foor su f f ustained 6=--/=40 / $$$0 -/ K K K K K K K K K K K K K K K K K K K K K K K K /0 L7 J K K K K K K K K 7K K K K K K K K OLP, where longer exposure to the druug at the u ff r LABIAG;ML2A;J '2I)0 'L72JBB@30 8ML K K K K K K K K K K K K K K K K J K K K K K K K K L7L LJ;M K K K K K K K KK K K K K K K K mucosal site is desirable, leading to prolonged ??AAF2 8JH K K K K K K K KK K K K K K K K:FJ;L9 ABDG +$ 24 G? :MGH:MJDD K K K K K K K K K # K # K # K # K # K # K # K # K K K K K K K K K K K K K K K K JL IE???LL7 K K K K K K K K ? ?K K K K K K K K therapeutic e feects, improved ffsafety prof fiile, ;GBDABEGEH and enhanced patient compliance. K 6:) 3 JB9 AB;EIJDDL9 JDD + ! "/ K K K K K K K K K K K K K K K K K K K K K K K K J K K K K K K K K J K K K K K K K K # # # K # # # K # # # K # # # K # # # K # # # K # # # K # # # K / 8ADMK K K K K K K K K K K K K K K HDA77AB1K K K K K K K K J JDDK K K K K K K K *$$ 7::2 # # # # # # # # 7 2K K K K K K K K DK K K K K K K K :7L9LDL72 7 2ABL9 DA2L :GABDH 6!0 *0 0 0 0 K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K 0 * JBB9K K K K K K K K #### $ $ $ $ $ $ $ $ 8ADM97J8 1. Park, S.Y., et al., Factors affffecting treatment a f J 8B JB9 JBBJF@HL9 NML ;GB;LBD7J K K K K K K K K K K K K K K K K J K K K K K K K K K K K K K K K K JJDDAGB G? K K K K K K K K K K K K K K K K outcomes in patients with oral lichen planus M30 % 24 JFACEGDH G? DML 7LFLJHL 2L9AE K K # K K # K K # K K # K K # K K # K K # K K # K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K E2 8L7L K K K K K K K KK K K K K K K K CEJBDA?AAL9 EHAB1 J JFA9JJDDL9 H:L;D7G:MGD lesions: a retrosppective studyy off 113 cases. s d o = ? K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K DG2LD7A;K K K K K K K K 2LDMG9 6*+ B20 K K K K K K K K # # # # # # # #K K K K K K K KJ72HDJ9D0 'L72 K K K K K K K K 7 2JB@3 2018. 48(4): 213-223. Journal of Periodonttal and Implant Science, n # 2. Sodata, P., et al., Optimization off Mucoadhhesive o f d $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ $ Film R nfforced ein with Functionalized NML N Naanostructured Lipid Carriers (N ( N NL LCs)) for s K K K K K K K K :7L:J7L9 K K K K K K K K 54/H K K K K K K K K ? ?GG72 7 2EFJDDAGB J B K K K K K K K K 8JH K K K K K K K K DML ?JJ;AFADJDDL9 :LBLD7JDAGB G? DML 54/H via Buccal Adm d ministration: A Box B Behnken K K K K K K K K ? J K K K K K K K K J K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K ABDG DML K K K K K K K K $ $ $ $ $ $ $ $ Designn Apppproach. A p Sci, 2025. 7(22). 2E;GHJF HE7?JJ;L JBB9 :GDLBDAJFF@ 9LL:L ;MJ7J;DL7AL9 !$ B2 8MA;M AB9A; Enhanced Triamcinolone Aceton dee Deliveryy id r K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K ;JDDLH GB J K K K K K K K K K K K K K K K K K K K K K K K K ? K K K K K K K K J K K K K K K K K K K K K K K K K L7 DAHHEL K K K K K K K KK K K K K K K K FJ@@L7H NML BJ77G8 EBA2G9JF 9AHD7 J K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K 7AIEDAGBK K K K K K K K 6 6 5*3 LBHE7LH DML MG2G1LBLGE 5 *3 LBHE7LH DML MG2G1LBLGE K K K K K K K K K K K K K K K K K K K K K K K K EH 97EE1 K K K K K K K K 7K K K K K K K K 9LFA L7@ JBB9 K K K K K K K KK K K K K K K K 7L9E;L9 J117L1JDDAGB 7AH K K K K K K K K J K K K K K K K K H DMEH K K K K K K K KK K K K K K K K A2:7G AB1 DML G L7JFF J K K K K K K K K K K K K K K K K K K K K K K K K HDJIIAFAD@K K K K K K K KJBB9 :L7?GG K K K K K K K K ? G72 7 2 BB;L JK K K K K K K K 288 ;:98 76543;241650/.-6,-+6.*-4/;43)(5163,'-;4/;242163,-/%20(2163,- 2,)-.12$6061#-.1()# "! -%1 1 1 1 1 , "!-7 , !!--.-;! , 0-; Miroslava ;! 1 ! 1. INTRODUCTION #* ? DI-D7-C4ECBI-*? +EDCHG? ? /@? I.? E114I14BEC7?79I/74?8747?B99IH07?B-??/H? JIHGFHEDCBDADIA@HGDIHBD? EDB=? FJ;:=? B9? E? 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FB@/E? )-?0BC4I?47H7E97?C79CB-@?FCB/7?DI6497??EG9=?8E9? :H4BD(=*?:HH?IC(74?/EC74BEH9?8747?I.?E-EHGCBDEH? 174.I4/7? B-? E@E4I97? @7H? F#=*? ? /@? I.? @4E7? E-? 8747? 697? 8BC(I6C? .64C(74? /BD4I1E4CBDH79?8747?1HED7?B-CI?E?0BEH,?.B+7?B-? 164B.BDECBI-* E@E4I97? @7H? E-? E.C74? @7H? 9IHBB.GB-@,? ? /H? I.? -.!-!! 1(I91(EC7? 56..74? 9IH6CBI-? 1!? *? 8E9? E7*? BD4I1E4CBDH79? 8747? 1471E47? 69B-@? 8 I 8#? (7?56..74?8E9?9E/1H7?F#*?/H=?47@6HE4HG?B-?,? 7/6H9BI-*? 3I4? 8,? ? /@? I.? EHE47HB-? ED7CEC7? #,? ,? #,? ? E-? ? (49,? E-? DI/1H7C7HG? 8747?B99IH07?B-?*?@?I.?@7HECB-7?9IH6CBI-?I.? 471HED7?8BC(?.479(?/7B6/?E.C74?9E/1HB-@*?:HH? 289 $#"! HGFEDCGHGBAE ?GCG GC=504-5-LS stability 76FCG6:B 50 ,2 5 00 HGA-; ;GE5C:3G; F3,G8 +AF3:6:A2 EAD;2 ?FE @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ 1/ 0 ,2 GC=Time 0 mg( 0,1 ,)) )( '. BB; %$ -DH:;:A2# CG=CC:"GCFAA,g0) 5 @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ F @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ ion t 1 month mg FB; =CCGGGC 39 !% '.8 @ @ @ @ @ @ @ = @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ CDD" 5ar 0 ,1 nt 3 mont hs ;:EE6DA:C=::6GE ?GCG 3EGC,,G; :B # # D @ @ @ @ @ @ @ = @ @ @ @ @ @ @ @ @ @ @ @ @ @ C @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ 0 ,0 5 6 mont hs once HC 0 0 2 Tim 1e 1 0 0hr 0(00 ( s ) DC:B" CGFCFA:+AF3 3:6:A22 = (%( A @ @ @ @ @ @ @ @ @ @ @ @ @ @/+ :B H @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ A @ @ @ @ @ @ @ @ @ @ @ @ @ @ GH62G; A GBEDCG -:"-GC 2:G6;8 G6FAA:BG :B A-G @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ F @ @ @ @ @ @ @ @ @ @ @ @ @ @@ @ @ @ @ @ @ :BAGCBF6 ?FAAGC -FEG ? EGC,G; = +FH H6GE CGFCG; ?:A-- ($ "G6FA::BG FB; @ @ @ @ @ @ @ @ @ @ @ @ @ @ A @ @ @ @ @ @ @ @ @ @ @ @ @ @ A @ @ @ @ @ @ @@ @ @ @ @ @ @ @ @ @ @ @ @ @ F @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ = @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ A-G ;GBE:A22 5G; A 5CGFAAG %%% CH - @ @ @ @ @ @ @ C @ @ @ @ @ @ @ GG; 5-FCF5AGC:FA: @ @ @ @ @ @ @ @ @ @ @ @ @ @ A-G ;GBE:A22 5G; A @ @ @ @ @ @ @ A A @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ 5CGFAAG F F @ @ @ @ @ @ @ E-GC:5F6 FCAA:56GE ?:A- ;G=::BG; E-FG8 0 @ @ @ @ @ @ @ C @ @ @ @ @ @ @ @ @ @ @ @ @ @ = @ @ @ @ @ @ @ @ @ @ @ @ @ @ 0CGH:9@ @ @ @ @ @ @ Sample Yield(%) EE (%) DL (%) = ? GHD6E: 6-FEG EAD5 A A-G ,GEEG6 @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ 6 3203-5-LS 76.03 (8%8 %8%%8% ;DC:B" A-G @ @ @ @ @ @ @@ @ @ @ @ @ @ GHD6E:=::5FAA:504-5-LS 33.02 88% %8%8% A C,:;G FCAA:56GE ED:AF36G =F @ @ @ @ @ @ @ @ @ @ @ @ @ @ C @ @ @ @ @ @ @ @ @ @ @ @ @ @ = @ @ @ @ @ @ @ FCGBAGCF6 B@ @ @ @ @ @ @ F;H:B:EACFAA:653-5-LS 63.71 (8((8%( %8%8%( F 766 ) EFH @ @ @ @ @ @ @ @ @ @ @ @ @ @ F H6GE ?GCG ED55GEE=DD662 C @ @ @ @ @ @ @ @ @ @ @ @ @ @ = @ @ @ @ @ @ @ CGFCG;8@ @ @ @ @ @ @ 1:"-GC -GG; E- FCAA:56G E:G 3DA F6E EAC::B"62 C @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @@ @ @ @ @ @ @ 6ED6FAA::5CFCAA:56 C 6GE ?:A-- 5ED6FAAG; @ @ @ @ @ @ @ A @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ F@ @ @ @ @ @ @ F F @ @ @ @ @ @ @ = = @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ F A E:H:6FC @ @ @ @ @ @ @ 2:G6; ,F6DGE8 -:E -:BAE FAA ;CDD" 6B1F F6F @ @ @ @ @ @ @ FCG6:B@ @ @ @ @ @ @ F5GAFAAG ?GCG ED55GEE=DD662 F @ @ @ @ @ @ @ @ @ @ @ @ @ @ =@ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ F C @ @ @ @ @ @ @ @ @ @ @ @ @ @ C:B" A-G @ @ @ @ @ @ @ @ @ @ @ @ @ @ CGFCFAA:CGFCG;8 0F @ @ @ @ @ @ @ FCAA:56G E:G ?FE =GG;8 @ @ @ @ @ @ @ @ @ @ @ @ @ @ EHF66 =F @ @ @ @ @ @ @ = @ @ @ @ @ @ @ FCGBAGCF6 F;H:B:EACFAA: GC=FB; :A ?FE @ @ @ @ @ @ @ @ @ @ @ @ @ @@ @ @ @ @ @ @ GB; A-G @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @@ @ @ @ @ @ @ = C HG; ?:A- EG6G5AG; EFH6GE CG @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ FCG; FAA @ @ @ @ @ @ @ @ @ @ @ @ @ @ F ;"! "CIHC:32-5,65,C 2376-67+EC "10/1954:C C 254C C :=14C 7= =.C :7;15,:7C 65=6671;+C 37:C 254C 2-1;28 4015:7;274C;246328C:32-5,65,C2376-67+C27C7=/;46374C /=+:6313=06328!C /=2;023165763C 7:74C 315357;27615:C *)A.?)AC (,'0%EC $2:4C 254C71 63181,6328C/;168:!C 3 15C #")AC -289:!C 310/1954:C C 254C C =6674C ;6:C 1C ;C254C 1/760627615C 6:C 544C 71C 64576+C 25281,9:C !C =63=C :=14C 2C ;246328C :32-5,65,C 67=C60/;1-4C:27+C/;168:E 2376-67+EC=: :72542;4C $HC *#" )AC C D)E?C ,'0%!C =63=C /;1-4C 71C 310/1954:E DEC "=5,!C "E!C 7C 28E!C " "! ""! ! " "! EC 19;528C 1C 88C19;C310/1954:C "=;10271,;2/=+C$!C?AA?EC@D*D?%CF@DFEC . C2;?EC$1;,:!CE!C7C28E!C" "! ! ! ,2:7;1657:76528C "! ""! " ! !" ! 2:1;/7615EC 1 63181,6328C /;4637615:C 6546327"" "" C "9;;57C 04636528C 3=06:7;+!C 5157901;6,563!C254C5156;;67257!C97C02+C/1:?AA)ECD?C@@DE 2C=6,=C;6:C1C;GEC #86!C E!C 7C 28E!C !" ! "! "" " "!"""! " "!EC 23415625C =2;023976328C $988765!C :+:70C*;2/64C :+:70:EC #5C 7=!! "! "EC IHC,5;27615%!C:60682;C71C19;:!C 528+76328C$613=06:7;+!CD@ECD)*D%C?D)?DE =+4;1 +C * %C 254C 07=1 +F )EC 2+!C E!C 7C 28E!C !"! " 07=+831902;65C * %C :=14C 51C :6,5663257C "! " " ! " !"! 65=667615C27C DAA(!C =68 %C " " ! *D?%C)!C=1C:79464C IHC:32-5,65,C65C71C " "!"! " "! " I9;C ;:987:C /2;76288+C 286,5C 67=C 2+C 7C 28EC FEH288688!C $E!C 7C 28E!C ?A?GEC*:9//8CD%C??AEC 254C !@46=+4;1 +F07=+831902;65C *% "E $613=06328C /=2;023181,+!C D?E FF*?%C 65=6674C 41 +;61:?A)?DFE 3157;2:7!C 65C 19;C :794+C *)A.?)AC (,'0%!C 310/1954:CC254CC:=14C2376-67+C310/2;2871C $H!C =;2:C 310/1954:C C 254C C =24C 2;C 37:EC #57;:765,8+!C 65C 7==G H?I?C :+:70C *:81C 36533=5181,6328C -81/057C 254C IHC ,5;27615%!C 310/1954:C C254C ;4934C #551-27615C *;137:C F)DAGDG'?A?) IHC8-8:!C=68AG'?AADDGC254CF)DAGDG'?A?)AG'?AADDG%E C254C :=14C/;11 64257C37:C )EC=:IHC:32-5,65,C 4 28:1C15C /;605728C315467615:E #5C 19;C 2;86;C 1;C G!C 310/1954:C C 254C C 65=6674C 86/64C /;1 6427615!C =;2:C C 254C C ; 1 64257C 37:C 27C =6,=;C 315357;27615:EC 1728+!C 7=6:C 7;54C 2:C ;-;:4C 65C 7=IHC :32-5,65,C 2::2+!C =;C 254C C ; ;2463287+/4 278 ;:98 765435210/.0-.2,7.+3*05),7(167.;'0;7'217(.0-.040;525)1/7.77. )'0;(%.1/-437/*7.0-.;04+7'.*0+$1/5210/ 5#"! .' 1 2 3 , 6! .# .)##.# *)('%$#("$! !%#($!(" !"! #($ !$! !("(!"(%$! !"!! "!"! (%( " )('%$#("$! !%#!$! !("(!"(%$! ! !! !!(% )('%$#("$! !%#($!(" !"! #($ !"(%$! !(%($! !%#!(% 1. INTRODUCTION ?8B=,?7;B;=;,?8B;3C8?8=B9; 2B9E?;=;1?8)??; FEDCBA@?=C; =; B; E?:E9; 87B8?65; 8C; =437C2?; 8@?;3C954?7;B CDE9B7;B2B=9B1=9=85;15;=87CAED=6;3C954?7;=8C; .@?;4EDCBA@?=2?;87?68@;)B;4?BE7?A;)=8@; 8@?; DC43C=8=C; C:; 9=0E=A; C3@8@B94=D; B; -;8?#8E7?; B 37?3B7B8=C/; .@??; 3C954?7; =?; 8@?; E=6;4ED=;A=D/;.@?;4B#=4E4;:C7D?;7?0E=7?A; 2=DC=85; C:; 8?B7-:=94; B 8C; A?8BD@; 8@?; 4ED=; A=D; :7C4; 8@?; ?5?; A7C3; 1=CBA@?=2?; 37C3?78=?; 8C; 7?AED?; A7B= B439?; 7?37??8; 8@?; 4EDCBA@?=2?; :C7D?; 37C9C6; 7?8?8=C; 8=4?; B8; 8@?; B339=DB8=C; =8?+; "4+;)@=9?;8@?;B7?B;E B )B;?8=4B8?A;B;)C7;C:;BA@?=C;"4/ 2=DC=85; 4EDCBA@?=2?; C9E8=C; B7?; 4C7?; .' ! .!!!# DC4:C78B19?; 8@B; @=6@-2=DCE; C+; 4EDCBA@?=C; =; 37?:?77?A; C2?7; B; =439?; @?C9C6=DB9;B?;C:;8@?;2=DCE;?5?;A7C3; =?; =; 2=DC=85; ('/; %2?8=6B8=6; 8@?; )?7?;DB77=?A;CE8;15;!;F *;7@?C4?8?7; 4EDCBA@?=2?; 1?@B2=C7; C:; B; :C74E9B8=C; =; ".@?74C; *D=?8=:=D+; B797E@?+; ?74B5; D7ED=B9;:C7;E8B=4E9B8=6;8@?;"! ?0E=33?A; )=8@; B; DC ;=8?7BD8=C;)=8@;CDE9B7;8=E?/;.@?7?:C7?+;B; 4?BE7=6; 58?4/; 9C); DE72?; )?7?; "!$% +;(! +;B"! ;4?8@CA; A?8?74==6; B; DCB#=B9; D59=E7?4?8; 58?4+; :C7; 8@?; @?B7; 7B8?; :7C4; D@B7BD8?7=,?; 1=CBA@?=2?; C3@8@B94=D; ACB6?; /'; 8C; '; '; B8; $; ; B;($/;.@?;B=4;C:;8@?;37??8;8EA5;)B;8C; BAA=8=C;C:;=4E9B8?A;8?B7;:9E=A/ ?2B9EB8?; 4EDCBA@?=2?; 37C3?78=?; C:; C9C3B8BA=2 !..C43C=8=C;C:;8@?;8?8?A;?5?;A7C3/ ?5?; A7C3; DC8B==6; @5A7C#537C359; 6EB7; 6E4; "! ; BCA=E4; Composition Sample label @5B9E7C=6; (% m/V) F1 F2 F3 A=::?7?8; "!$% ;4?8@CA/ OLO 0.1 0.1 0.1 .+527'154(.5/).+72,0)( HPG 0.25 - 0.25 .+! . SH - 0.4 0.4 9C3B8BA=;7?D?=2?A; NaCl q.s. q.s. q.s. B;B;6=:8;B439?;:7C4;!?4C:B74;//;"*?71=B/; Na2HPO4 0.5 0.5 0.5 ! ;)B;3E7D@B?A;:7C4;@9B; BAC 0.01 0.01 0.01 B;=8C4BD@+; 853?; %%; )B; purified 100 ml 100 ml 100 ml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ushko Shalabalija, Natasa Draskovic, Lina Livrinska, Nikola Geskovski, Marija Glavas Dodov, Ljubica Mihailova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ilution in CS F CS+F DMEM (mmol/L) (mmol/L) (mmol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Ljubica Mihailova, Matej Nikolovski, Metodi Naumovski, Magdalina Petrushevska, Teodora Tasevska, Marija Glavas Dodov, Dushko Shalabalija ! 1. INTRODUCTION ($!$ $#(!"%# 6GG;8 0.7 6FCAA:56GE FE C @ @ @ @ @ @ @@ @ @ @ @ @ @ ?G668@ @ @ @ @ @ @ ===GG5A = - A-FAA A--G ;GGB;GB5G 3GA?GGB A-G FH EGG; BB; "G @ @ @ @ @ @ @ F @ @ @ @ @ @ @ G6FA:BG 5 "G6FAA:BG GE / :B AGCH @ @ @ @ @ @ @ C HE = EAF33:6:A2 = A-G ;:EE6DA:C=::6G8 @ @ @ @ @ @ @ @ @ @ @ @ @ @ EAF33:6:A2 = A-G ;:EE6DA:C=::6G F F @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ = = / BB; FCG F5-:G @ @ @ @ @ @ @ F ; @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ -: ,G; FAA ($ "G6FAA:BG8 ; A ($ @ @ @ @ @ @ @ @ @ @ @ @ @ @ F @ @ @ @ @ @ @ 6 A: F @@@ @@@ @@@ @@@ @@@ @@@ @@@ FF38 @ @ @ @ @ @ @ @ @ @ @ @ @ @ E-GG; F @ @ @ @ @ @ @ F @ @ @ @ @ @ @@ @ @ @ @ @ @ ED3EGA8@ @ @ @ @ @ @ 6GE @ @ @ @ @ @ @ CGFCG;@ @ @ @ @ @ @ ==CC 566G5A:62HGCE :B ,GAGC:BFC2 E5:GB5G FB; C @ @ @ @ @ @ @ @ @ @ @ @ @ @ C @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @@ @ @ @ @ @ @ @ @ @ @ @ @ @ = @ @ @ @ @ @ @ @ @ @ @ @ @ @ = ;C22:B" C @ @ @ @ @ @ @ 5GEE8 B @ @ @ @ @ @ @ @ @ @ @ @ @ @ F;H:B:EACFA: @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ A-:E CG"FC;# A-G 3GEA CGED6AE 2:G6;G; 0/ ,GCF66# A-G 2:G6; ?FE 68 :6-F6,F# 78 8# GA F68# A:6:FA: @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @@ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ EFH F H6G @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ /7 ! @ @ @ @ @ @ @@ @ @ @ @ @ @ )% EFH 76:G; 7B:HF6 GEGFC5-# )%8 * 8 @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ F H6GE !8( A 8%)$8 -G E @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ @ EA 33:6:A2 F @ @ @ @ @ @ @ $ %" % %5 3:*.,65 !$5 !5 ;08/6$/ 5 ;08/6$/5 7/;,:345 )95 .5 $.9)7;/0;$/):895% 5-(5%! 5 " 5 .'/#/05 ';95 3.5 /30:8/,5 ;5 ;365 .*9/0#/,5 1;0/.5 20.35 :95 ;5 0/27/8:.35 .25 0//5 5 ;05 5 /5 ;7 5 " % ,:22/0/35 90;% % 5 ;)0/5 /.,95 5-%(5%%!% ;22:3:65 ;95 ;775 3;3.*.,:/95 '::35 /5 ,;;9/5 % 5=.#;75 5/5;7 5% ! % ";:3;:35;55:35/59)*!":80.".7;050;34/ 5/5 ;79.5,/%%! % % % 5 =8:/38/5 5%%-(5! .25 /5 ;*.#/5 /30:8"/395 *65 ;3;769:345 ':85 :3 /5 9;"/5 ;08/6$/ 5 /5 5 ;08/6$/5 ';95 /30:8/,5 :35 ,/39:65 .25 $.7;05 8.3;895 ! /5 ;).095 ;8+3.'7/,4/5 2:3;38:;75 9)$$.05 *.3,95 ;3,5 9/8.3,;065 90)8)0/5 $0.$/39:65 20."5.#;0:955;3,5;3+58.77/;4)/95;5*.5 9.':345;5/5:3.#;0:95 ;3,5 /5 ;8)765 .25 /":9065 ;3,5 8.32.0";:.3;7765 8.390;:3/,5 ':5 ;5 40/;/":8;75/83.7.465 52.05 /:05 :3#;7);*7/5 .$:":;:.35.259$/8:2:858.3;89 5/5 ! 8.30:*):.39 298 )(' %$#"!# % ## %)%) %#"%!"# #)#% )% !"# % #1 2 1. 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")'/$#')'*'"(%%#"%('' #)/"$)%('%('#"/$)'%(+'%)$'%* %)'"'"' %! *)()" $)'$)( Aleksandra Damjanoska1,2 2 2 1 , Stefan Radevski , Cveta Dolikjoska Trajkova , Katerina Brezovska, Jasmina Tonikj-Ribarska1, Natalija Nakov1 ) ('%$%#%"! !$"!"$%!'!"#%$!'$!#%! !!!$!'!"% $#! '$"$%!$'! "!"# $! ! %!" '$ '#%#$'!"'$!"%$ '!(!!"# $! ! %!" '$ 1. INTRODUCTION ,D88-DD8#,D =BD8?4:3BD@=7ABD1?EA:AB9D?6D 7D@D,D4566BC-D8B=7E?3D7E9D71B?E:C:3BD:ED7D FED CBA@?EABD ?D =BD *?358BDC7:?D?6D/D'D***#,D =BD63?;DC7BD A5A7:E743BD 7E732:173D @C71:1BAD 0/.-D =BCBD =7AD ;7AD87:E7:EB9D7D/,D8$8:E-D;:=D7ED:EB1:?ED 4BBED 7D E?743BD A=:6D ?;7C9AD *?358BD?6DD$,DBB1:?ED;7AD@BC6?C8B9D7D7D 8B=?9?3?:BAD :ED 7E732:173D 1=B8:AC2,D ;7*B3BE=D ?6D D E8D 7E9D =BD 1?358ED ;7AD +?E*BE:?E73D =:=)@BC6?C87E1BD 3:(5:9D ?@BC7B9D 7D 784:BED B8@BC75CB,D =BD 8B=?9D 1=C?87?BED CB3:BAD ?ED ;7AD 9B*B3?@B9D :E)=?5ABD 7E9D *73:97B9D =7"7C9?5AD ?C7E:1D A?3*BEA-D ;=:1=D @?ABD 711?C9:E?DF+DD5:9B3:EBAD0., BE*:C?E8BE73D7E9D=B73=DC:A!A,D =BD79?@:?ED?6D 73BCE7:*B-D8?CBDA5A7:E743BD7@@C?71=BAD:AD=5AD '+ '' '%'' ' 4B1?8:E:73,D =:AD A:3:17)47AB9D 8B=?9D =BD1=C?87?:?ED;7AD1?E951B9D 0.-D3B*BC7:E=23D:A?A?C4:9BD'F#D7AD7D ?ED7D:3:17D@=BC:A?C4D1?358ED'D88DD,D 4:?)CBEB;743BD A?3*BE-D @CBABEAD 7ED :EE?*7:*BD 88-D D 8#,D =BD 8?4:3BD @=7ABD 1?EA:AB9D ?6D 7D A?35:?E,D FD :AD 7D *BCA7:3B-D B1?)6C:BE932D ,D71B:1D71:9DA?35:?ED7E9DFD:ED7D*?358BD A?3*BED=7DA:32DCB951BAD=BDCB3:7E1BD?ED C7:?D ?6D /D 'D **#,D =BD 63?;D C7BD ;7AD ?:1D ?C7E:1D A?3*BEA,D =BD 8B=?9?3?2D 87:E7:EB9D 7D /,D 8$8:E-D ;:=D 7ED :EB1:?ED 13?AB32D 73:=D @BCD 7(5B?5AD 3:(5:9D *?358BD?6DD$,DBB1:?ED;7AD@BC6?C8B9D7D7D 1=C?87?B1=E:(5BD =7D ;7*B3BE=D ?6D D E8D 7E9D =BD 1?358ED ;7AD @C:?C::"BAD ;7BC)47AB9D A?3*BEAD 6?CD BE=7E1B9D ?@BC7B9D7D+, BE*:C?E8BE73D A76B2D 0.,D =:AD A592D @CBABEAD =BD9B*B3?@8BED?6D7D=?9D6?CD '*)$'"('%#%( =BD(57E:7:*BD9BBC8:E7:?ED?6D1B@=73B:ED:ED =BD 7@@C?71=D 5:3:"B9D :ED =BD =?9D :AD 1B@=73B:ED17@A53BA, :EE?*7:*B-D7AD:DCB@371BAD=BD1?E*BE:?E73D?:1D '")*%"$'"(') 8?4:3BD@=7ABD1?8@?AB9D?6DD9:35B9D71B:1D E?E)@?37CD 8?4:3BD @=7ABAD ;:=D 7ED B1?)6C:BE932D .'' 71:9D ',D **#D 7E9D FD 7AD 7D 4:?)CBEB;743BD FD '-D :?BEB;743B-D B7%35A#D ?C7E:1D A?3*BE,D =:AD 1=7E=D =BD ;7AD A?5C1B9D 6C?8D :87D 39C:1=,D B=7E?3D @C:E1:@3BAD ?6D C2-D ?66BC:E?E:C:3BD '%$+D A5A7:E743BD A?35:?ED 6?CD 1=C?87?7EB)/)A536?E:1D71:9DA?9:58D 7E732A:AD;:=?5D1?8@C?8:A:E=BDB66:1:BE12D?CD A73D'$:+=C?@5CD6?CD:?ED@7:CD1=C?87?2D?6D=BDCBA53A, C:B=2378:EBD '#-D 7E9D ?C=?)%=?A@=?C:1D BC1!D 7D 7C8A79-D BC87E2,D =BD @BC6?C87E1BD 4B;BBED 1?E*BE:?E73D 7E9D 8B=?9D6?CD9BBC8:E7:?ED?6D1B@=73B:ED:ED+B673B:ED 1B673B:ED:E)=?5ABDAB1?E97C2DCB6BCBE1BDA7E97C9D 71:9DD'D6?CD7E732A:A#D;BCBD6C?8D ' .D +?8@7C:A?ED ?6D 1=C?87? 17@A53BA, 7E9D =BD 6?C8537:?ED 7E732"B9D '+B673B:ED D Conventional Green 8 Method Method D!?@B, Retention Time 6.9 5.5 '# ' !' (RT) (minutes) =BD1=C?87?:?ED;7AD1?E951B9D Total Runtime 12 8 ?ED 7ED BCC7D %)/D 1?358ED '/D 88D DDDDDDDDDDDD (minutes) 301 Capacityy t /A5.856A5?9 @8CA3-E E E E E E E E E E E EE E E E E E E E E E E EC8 36;;;88B85/8 6 @ E E E E E E E E E E E E ; ; E E E E E E E E E E E E E E E E E E E E E E E E @656@?9E E E E E E E E E E E E 3.2 1.9 ?53 3A8 5A ?;;;88/ C8 @8CA3" :::96/ Factor (k') r E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E ;; E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E ? /?4696*,E E E E E E E E E E E E Column +C6 65/B8?8 6 96#89* 378 A C8 C6=C8B ;99 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E ; 9A2 B?8 E E E E E E E E E E E E ?E E E E E E E E E E E E Backpressure 174 194 783 65 CC8 =B885 @8CA3- 47 6 B8@?65 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E 26C65 E E E E E E E E E E E EE E E E E E E E E E E E (bar) ?//8:?498 96@6 ;AAB C8 696/? /A97@ E E E E E E E E E E E E E E E E E E E E E E E E ; E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E @5, +C8 E E E E E E E E E E E EE E E E E E E E E E E E Theoretical *@@8B** ;??/AB )$0 ;AAB 4ACC @8C B E E E E E E E E E E E E ; E E E E E E E E E E E E E E E E E E E E E E E E ; E E E E E E E E E E E E E E E E E E E E E E E E CA3 6 E E E E E E E E E E E E E E E E E E E E E E E E 3251 9 9126 6@69?B- 26C C8 =B885 @8CCA3 Plaates (N) E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E ? 96=CE E E E E E E E E E E E Factor (As) Symmetryy 6@:BA.8@85 A.8B C8 /A5.856A5?9 @ E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E @8CA3-E E E E E E E E E E E E r 1.3 1.2 2C6 /C 7==8 /A@:?B 4498 :8?# C7 8%76.?985 ?//7B?/* ?553 B8:BA37/64 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E ? E E E E E E E E E E E E 4696* A; E E E E E E E E E E E E E E E E E E E E E E E E B879, +C8 @8CA3 C E E E E E E E E E E E E C 3 3 765= ? ? 46AB858 E E E E E E E E E E E E 6 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E 46 82?498-E E E E E E E E E E E E 5A5A6/ 65 /A575/6A5 26C E E E E E E E E E E E E E E E E E E E E E E E E 5 E E E E E E E E E E E EE E E E E E E E E E E E E E E E E E E E E E E E ? 696/?E E E E E E E E E E E E /A97@5E E E E E E E E E E E E 38 8@A5B?8 ?E E E E E E E E E E E E B896?4498 ?E E E E E E E E E E E E ?5 5?9*6/?9 *E E E E E E E E E E E E :8B;AAB@ ; B @?55/8 ? 8 2C698 6=56;66/ 559* B837/ E E E E E E E E E E E E E E E E E E E E E E E E ; ? E E E E E E E E E E E E65= C8 E E E E E E E E E E E EE E E E E E E E E E E E 58=?6.8 85 ? E E E E E E E E E E E E 5.6BA5@85?9 6@:?/ ?53 E E E E E E E E E E E E E E E E E E E E E E E EE E E E E E E E E E E E ??55?9*E E E E E E E E E E E E 8:A7B8 E E E E E E E E E E E E A A6/ E E E E E E E E E E E EE E E E E E E E E E E E B8?=85, 6.8 ? E E E E E E E E E E E E 85 C8 E E E E E E E E E E E EE E E E E E E E E E E E 2638:B8?3 7 E E E E E E E E E E E E 78 A; /8:C?9865 ?553 C8 65/ E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E ? E E E E E E E E E E E E E E E E E E E E E E E E /B8?65=E E E E E E E E E E E E 8@:C?6 A5 E E E E E E E E E E E E E E E E E E E E E E E E 5 =B885 /C8@6B**- C6 @8CA E E E E E E E E E E E E B E E E E E E E E E E E E E E E E E E E E E E E E3 A;;;88B E E E E E E E E E E E E ; ;E E E E E E E E E E E E ? :B?/6/?9 E E E E E E E E E E E EE E E E E E E E E E E E ??553 8/A E E E E E E E E E E E E/A58B55? E E E E E E E E E E E E B ? 6.8 ;AAB E E E E E E E E E E E E ;E E E E E E E E E E E E BA7658 :C?B E E E E E E E E E E E E : BB@ @?/876/?9 %7?96* /A5BA9, 7 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E D DCBA@?A=B?@ A; ? E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E EE E E E E E E E E E E E 6A5E E E E E E E E E E E E A4?6583 26 E E E E E E E E E E E E C1 ?0 /A5.856A5?9 @8CCA3- 4 E E E E E E E E E E E E E E E E E E E E E E E E 5 E E E E E E E E E E E E E E E E E E E E E E E E 40 =B885 E E E E E E E E E E E EE E E E E E E E E E E E (, C??4?5-E E E E E E E E E E E EE E E E E E E E E E E E !,- 8 ?9,- 82 56=C 65A E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E A 6%763 E E E E E E E E E E E EE E E E E E E E E E E E @8CA3,EE EE EE EE EE EE EE EE EE EE EE EE DCBA@?A=B?: :C** ;AAB AB8 /A'B68539* $5 C E E E E E E E E E E E E ; E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E?9*6 A; 9 E E E E E E E E E E E E E E E E E E E E E E E E C?B@?/876/ /?9,E E E E E E E E E E E E - (-E E E E E E E E E E E EE E E E E E E E E E E E +C8 A4?658 E E E E E E E E E E E E 83 B879 E E E E E E E E E E E EE E E E E E E E E E E E 38@A5B?8 6= ? E E E E E E E E E E E E =56;66/?55 ; ?E E E E E E E E E E E E )01 E E E E E E E E E E E E , 6@:BA.8@85 5 65 /CBA@?A=B?::C6/ :8B;AA E E E E E E E E E E E E E E E E E E E E E E E E ? ? E E E E E E E E E E E E ; AB@?5/8E E E E E E E E E E E E , A4?#? E E E E E E E E E E E E ?- $, ,- 8 ?9,- @8B= E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E =65= AB E E E E E E E E E E E EE E E E E E E E E E E E ?53 85.6BA5 E E E E E E E E E E E E @85?9 7?65?44696* 26C C E E E E E E E E E E E E ? E E E E E E E E E E E E E E E E E E E E E E E E C8 =B885 E E E E E E E E E E E E E E E E E E E E E E E E 538B86@? 83 696/? 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E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E ; ; ; ; E E E E E E E E E E E E :?B?6A5 ?E E E E E E E E E E E E :BA/8,E E E E E E E E E E E E C698 CC8 /A97@5 4?/#:B87B E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E B8 65 C8 E E E E E E E E E E E E E E E E E E E E E E E EE E E E E E E E E E E E =B885 @8CA3 6 96=C9* C6=C8B C?5 E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E E EE E E E E E E E E E E E E E E E E E E E E E E E 65 C8E E E E E E E E E E E E 302 %$# REALISTIC SIMULATIONS OF GASTRIC EMPTYING VARIABILITY ON A POPULATION LEVEL USING COMPASS" SOFTWARE Dorota Danielak1 2 3 1 , ! , Maciej Winiarski , Grzegorz Banach, Jadwiga Paszkowska 1 1 1 1 1 , Marcela Staniszewska ! , Justyna Dobosz , Grzegorz Garbacz 1,+*)('%$('#",')! "",()%)!('"$"-"' ",' # 2 $#$"'",+*)( ",+ *" #",+ '!(#$()",'# #"#( )($*"'"( "(#)",'# " Poland 3#)$($%$"'"'" $%" #"$%$%") +",'()+" *"'"(#)"' ",' # 1. INTRODUCTION variabilities (BSV). The algorithm creates virtual subjects with varying PK parameter The performance of the immediate-release (IR) values, and assigns them to one of the four GE oral dosage form may depend on the variability $" ! $! ""! #$! ! of gastric emptying (GE). Under fasting (Table 1). conditions, GE may be described by the rate the gastric fluid flows out of the stomach (kGE) and Table 1. COMPASS GE motility types $#"!$ !!$#"!#""" !"!! GE kGE Prevale Maximum plasma drug concentrations (C max ) or GET [h] type [1/h] nce [%] area under the time-concentration curve (AUC) may differ, depending on the kGE and GET Sprint 3 - 14 27 combinations. For example, rapid kGE = 14 1/h and early GET = 0.25 h would lead to a higher Tight 3 22 0.75 C max than when the gastric emptying was slow these combinations are more probable than Average 3 - 14 31 (kGE = 3 1/h and GET = 2 h). However, some of 0.75 others, and some distinct types of GE patterns were proposed [1]. To address this, we Lazy 2 20 developed a novel module for the COMPASS software [2]. Its novelty is introducing the Dissolution data were taken from the literature prevalence of the GE emptying patterns on a [3]. PK parameters and the whole workflow, population level, as proposed earlier [1]. including dissolution curve fitting, were the We describe the working principles of the same as described earlier by Danielak et al. [2]. module using two ritlecitinib formulations as an example: one 100 mg capsule (Reference) vs. Simulations were performed for 100 virtual two 50 mg capsules (Test). In the virtual subjects. The BSV was set to the values reported bioequivalence trial, these formulations were previously [1]. assessed as bioequivalent [3]. 2. MATERIALS AND METHODS 3. RESULTS AND DISCUSSION 2.1 Software COMPASS is a Python-based tool with an PK profiles 3.1. Influence of GE patterns on predicted intuitive user interface, and its performance was evaluated earlier [2]. It allows a straightforward $!!""!$ $!$$"!"!$! translation of the in vitro dissolution profiles to the highest predicted ritlecitinib plasma reliable PK simulations. In the interface, the concentrations for both Reference and Test user provides the values of effective products (Fig. 1). permeability (Peff), volumes of distributions (V1/F, V2/F), and clearances (CL/F, Q/F). In the novel module, the user defines the study group size and respective between-subject 303 *)(' (visualized as a 3D plane) without weighting their probabilities for a subject with typical PK parameters. The novel module focuses on the physiologically plausible kGE and GET combinations and includes the BSV of the PK parameters. Table 2. Comparison of the observed (n = 6) [4] and simulated (n=100) ritlecitinib PK parameters. For Figure 1. Influence of GE patterns on ritlecitinib Cmax and AUC, data are shown as geometric means plasma profiles for Reference and Test products (n = with a geometric CV. For Tmax data are shown as 100). medians with range. %$%#"! !## #!#!# AUC inf Source C max Tmax [h] 0.5 Observed 647.7 (24) 1085 (23) (0.5 - 0.617) Simulated 500.6 (37.4) 1404 (35.7) 1 (0.5 - 3.5) Figure 3. Cmax similarity evaluation with the standard COMPASS module vs. the novel GE variability module (n = 100). 4. CONCLUSION The novel COMPASS module realistically Figure 2. simulates the endpoint PK parameters and their Relative C max and AUC differences between Test and Reference products (n = 100). variabilities, including physiologically plausible kGE and GET combinations. Fig 2. shows the relative differences of Cmax and 43210//.-,1+/*1)..1(''1%*$#).1"#! -$"14..1 5. REFERENCES ),1"0.%$1%$/11/$%.%$1$0-"1-14-*)-1 (1/)" %11-$1).1/*-1)1)1#$1--.%4 motility type produced the highest median Combination of Physiologically Driven Dissolution 2 max, but the greatest differences (over 6%), and Pharmacokinetic Modeling as a Tool for -*-1/!"-*-,1+/*1$-10*%$1$0-1/*14321 Understanding Human Gastric Motility. Mol Pharm. the differences were negligible, due to good and 2024;21:382437. complete ritlecitinib absorption from the 1 )%-.) 1 1 -$1 ).1 2/0)*-1 ),1 41 1 )"$1 C probability with the COMPASS software. Int J max was similar to the observed one. The Pharm. 2025;670:125123. differences for AUC and T intestine. screening of oral dosage forms for bioequivalence Table 2 shows that the mean predicted max can be attributed to a small study group size. Assessment of Ritlecitinib Capsules with 3. Saadeddin A, et al. Virtual Bioequivalence * # a Physiologically Based Pharmacokinetic Model. AAPS J. 2024;26:17. Fig 3. compares how the standard COMPASS %%# ! # # # !! # Incorporation of Observed Clinical Variability Using module and the novel one evaluate the influence 215830Orig1s000, Multidiscipline review Available 4. Center for Drug Evaluation and Research. of GE. The standard module systematically from:https://www.accessdata.fda.gov/drugsatfda_do analyzes 100 combinations of kGE and GET cs/nda/2023/215830Orig1s000MultidisciplineR.pd 304 210/ .-,+*-)+,('*%'%$$%#*'-$'2-"! +,!"2!))-"+-,%'-,'*%' 2)%#+2+**+-,'% +-)'-$''.-%"')( *'$1 1 , Marija Bogataj 1. 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INTRODUCTION 7:? 6;53?@3 = = = = = = 3: 3946@1. @/ = = = = = = = = = = = == = = = = = )'++001. 02/.--' , , , , , , ., , , , , , A@? ? ;@: 98 7@965;4 2'11( ', , , , , , ..00. .= = = = = = )0= = = = = = +)(+'0, , , , , , = = = = = = = = = = = = = = = = = = = = = = = = ; 4532?@351:0 ? = = = = = = = = = = = = @/.2:?;-- 5: :@8??. 8;;94 ?;5.@?@9/51 :?5@/1::, '2.+ ., , , , , , '++0 ' 0= = = = = = ) 5/. = = = = = == = = = = = + ' )'++1, , , , , , ; = = = = = = = = = = = = 8 = = = = = = 8 = = = = = = = = = = = = :?1 +2@64--. $// . , , , , , ,21= = = = = =)(% = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 45@/1- 45. 98 615:?@3: 5/. 15:?94;:% A@1 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = 3. RESULT TS AND DISCUSSION (' :-:?4: 9888; 811$@7@1@?- 5//. 888@@3@-= = = = = = = = = = = = 8 8 = = = = = = 8 = = = = = = 5 = = = = = = 8 8 = = = = = == = = = = = . 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 92 0 0 0 0 0 00 0 0 0 0 0 51:9 @/?;9.23 /# ;@:":0 = = = = = = = = = = = = = = = = = == = = = = = 65;?@3215;1- ? ; = = = = = = ?= = = = = = 0 0 0 0 0 0 *1= %$$#" 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 69?571: )! $ 5 = = = = = = 5 = = = = = = = = = = = == = = = = = '( = = = = = == = = = = = )/22.1 ++. 2 --. / , , , , , , . , , , , , , , , , , , , , , , , , , ?9 4@*;5?? @/?9 .;22* 6;9.23?: ) .2;@/* 1@+2@. = = = = = = 5 = = = = = = = = = = = = ; = = = = = = = = = = = = = = = = = = = = = = = == = = = = = .+ , , , , , , , , , , , , 220**221.0' )(2'210.12/ '0 0 * ' , , , , , , , , , , , ,, , , , , , 6;93:: :?;54 = = = = = = 5 4:%= = = = = =!$?;53?5771: 5; :27:?5/3: 9; $ 5 = = = = = = = = = = = = = = = = = = = = = = = = )'++001. ' ., , , , , , 2, , , , , , +(2'210.12/ +, , , , , , '0, , , , , , 34@351 00.221 ., , , , , , 2 )'++001. , , , , , , ' .0= = = = = = 8883?@ 5-- 0 8 8 = = = = = = 5 = = = = = == = = = = = 5/ $?;53?@9/ 4.@24 2/.; $5**;5??. = = = = = = $ = = = = = = = = = = = = = = = = = = 5= = = = = = 0 @: = = = = = == = = = = = 32;; "- 49/9*;56 = = = = = = = = = = = = = = = = = == = = = = = 15779; 15779; 5 555??9;--55??9;--; ;= = = = = = ?:? ?:?= = = = = = 39/.@?@9/:0 39/.@?@9/:0= = = = = = @/312.@/* @/312.@/*= = = = = = :? 771@:@/* ? 71@ @ 5= = = = = = = = = = = = 5 3946; ; = = = = = = 8 = = = = = == = = = = = ?46;5??2;0 .2;5??@9/0 $??;53?@9/ 6;93:: 5//. 5 = = = = = = 5 = = = = = = $ = = = = = = = = = = = = 5= = = = = = +251@8@@35??@9/ 8 5 / 98 (' = = = = = = = = = = = == = = = = =45//28553?2;@/* 39469/ .@4571:= = = = = = 5;= = = = = = ?5?? 394 5 = = = = = == = = = = = @/?9 39/?53? #@? 65;4 / = = = = = = = = = = = = = = = = = = ; 4532?@351 2= = = = = = :27:? //3: ?5?? 53?2511- 4@*;5?? 8;;94 ? ?:? 5 = = = = = = 5 = = = = = = = = = = = = 5 = = = = = = 8 = = = = = = = = = = = == = = = = = 6;93:: :?; = = = = = =554 4:% 45@/ /91?- #@? '( = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = 5;??@31 2/.; ;51 45/28553?2;@/* 39/.@?@9/: % ; = = = = = = = = = = = = = = = = = = 8 = = = = = = = = = = = == = = = = = @: ? = = = = = = = = = = = = ? :@8?? ?9 5 :? //.5;.@.0 ;@:",75:. = = = = = = 8 = = = = = = = = = = = = = = = = = = 5 = = = = = == = = = = = 15:?@3 (' 39469/ :?;54: 5? 421?@61 45/28553?2;@/* :?5*: 5//. = = = = = = = = = = = = 5 = = = = = = 8 = = = = = = 5 = = = = = = 5= = = = = = 65;4 ; 4532?@ @351= = = = = = 45//28553?2;@/* 5 8= = = = = = @/312.@/*= = = = = = 3 // 6;:@:? ?;92*92? ? 6;93::0 15.@/* ?9 5 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 39/:@:? 53324215??@9/ @/ ? 5 = = = = = = = = = = = = = = = = = = 00 #@3 69:: 69? ;@:": ?9 = = = = = = = = = = = = 6;9.23? :?577@1@?-0 +251@?-= = = = = = 5 = = = = = == = = = = = 5//. 65??@ 49/9*; 66 @: 51;5.-= = = = = = 5 = = = = = = = = = = = == = = = = = :58 2. MATERIALS AND METHODS .1964 6;9.23?: 51; = = = = = =5.- 9/ ? 45;"@: #@11 16 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = 3946; 45/28553?2; 8 ;: 59@. ;*215??9;-- .15--: 5//. = = = = = = = = = = = = 5 ; = = = = = = 5 = = = = = = 5= = = = = = 3946 39:?1- ;#9; = = = = = = ;;"" 55. 98 @?: 39/.23?.0= = = = = = @/312.@/*= = = = = = '(= = = = = = 49/9*;56:= = = = = = ) 5//. )8883?@ 899; = = = = = = = = = = = = 5 = = = = = = = = = = = = 8 8 = = = = = = = = = = = = 8= = = = = = 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5125??@/* ! : 8;;94 615:?@3 5 * = = = = = = = = = = = = 8 = = = = = = 6= = = = = = 45/28553?2;@/* 8 *= = = = = = ! ?:?@/* = = = = = = *= = = = = =556661@: ?9 511 615:?@3 39469/ 6;93:: 39469/ : 6;9 = = = = = = = = = = = = 93:: :?; 4 = = = = = = 5 4 )@*% %% = = = = = = = = = = = = = = = = = = )= = = = = = 5..;::@/*= = = = = = ! := = = = = = 8 8;;94= = = = = = 653"5*@/* 5//. .1@;-- :-:?4:0 5: #11 5: ? 5 = = = = = = 5 = = = = = = ; = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = !14 !$?;53? 771: 5/. 53571: ) *2@.1@/:% $ 5 = = = = = = = = = = = = = = = = = = = = = = = == = = = = = - .5??5775:: 899; :3@ 39/?5@/@/* :58 ??;53?@9/ :?2.- .5?5 9/ ! :0 27 . 5//. $ = = = = = = = = = = = = 5 = = = = = = = = = = = = = = = = = = = = = = = = 5= = = = = = (3@ 311 ! 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"' = = = = = = = = = = = = = = = = = = = = = = = = = = = = -6%6:76* 0(:00 ,5+0-:,0/5+) $/0(/+ 0(6 -:+# = = = = = = = = = = = = = = : = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = #6 54 "! = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = ' ,(+6/*6-- ' 60 :7' = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = **)) #('' $# ! ! ! ! ! ! ! ! ! ! ! ! ! ! ( ' ! ! ! ! ! ! ! ! ! ! ! ! ! ! (#) ( ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! 05 ! 9:- :-6 0((6 95)0 4--66+0 $ = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 0 = = = = = = = = = = = = = = = = = = = = = = = = = = = = 4 = = = = = = = = = = = = = = $/0( 0(6 = = = = = = = = = = = = = == = = = = = = = = = = = = = '#$(#" % ! ! ! ! ! ! ! ! ! ! ! ! ! ! %%$(''#" '$$' '##$ ( ! ! ! ! ! ! ! ! ! ! ! ! ! ! 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' 59:)/ / ' 60 :7' = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = ' ) ( ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ##)% ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! #)(#) $# ##" ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! % ! ! ! ! ! ! ! ! ! ! ! ! ! ! '#)! ! ! ! ! ! ! ! ! ! ! ! ! ! )059:,( 455- 8 05 (5-) $(/76 50(6-= = = = = = = = = = = = = = 4 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = =-) */* )5 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = #$$ (# ) ) '"#)(#" ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! #)% $ ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! 4455- 5+7. 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(##( ( (= = = = = = = = = = = = = = 57' (:-= = = = = = = = = = = = = =-9' ' " ' = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 66 0(:+ 8-5 = = = = = = = = = = = = = = = = = = = = = = = = = = = = 5466))5-4= = = = = = = = = = = = = = 66-++6- 66/0),(/6) --= = = = = = = = = = = = = = = = = = = = = = = = = = = =-6)6:-,(= = = = = = = = = = = = = = #-58 :00 0(6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = : 6 +/%6-)/0. 54 3-6/4))$:7* 2 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 4 = = = = = = = = = = = = = = 2+)0/006= = = = = = = = = = = = = = 54 (:-9 = = = = = = = = = = = = = =-9:,. . 455- )(:-/+# 0(6 - $ = = = = = = = = = = = = = = 4 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = : $ *:00: 4--= = = = = = = = = = = = = = : = = = = = = = = = = = = = = 4-59 0(6 = = = = = = = = = = = = = == = = = = = = = = = = = = = 9:-00 /77 -= = = = = = = = = = = = = =0 0676960-. ,:88)76 )0*. )6* = = = = = = = = = = = = = = : = = = = = = = = = = = = = = = = = = = = = = = = = = = = * /+ 5-= = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = :+:7.)/)' 1(6 6 8-6%:76+,6 54 ,5+0--:,0/5+ /+06 = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = 0 = = = = = = = = = = = = = = 6+)/0.. . 0 = = = = = = = = = = = = = = = = = = = = = = = = = = = = 8-6))--6 )/6 = = = = = = = = = = = = = == = = = = = = = = = = = = =,7:))' - )0*. -= = = = = = = = = = = = = = = = = = = = = = = = = = = =6%6:76* 0((:00 )9:77 ,5+0-:,0/ = = = = = = = = = = = = = = 0 : = = = = = = = = = = = = = = = = = = = = = = = = = = = =5+) 8 = = = = = = = = = = = = = == = = = = = = = = = = = = = 05 " 9: = = = = = = = = = = = = = = = = = = = = = = = = = = = = :- :-6 95-6 ,5995+ 0(: = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = :++ 7:-#6 = = = = = = = = = = = = = =-= = = = = = = = = = = = = = ,5+0-:,0/5+) ) :5%6 " 9:- $/0( 0( = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = (6 95)0 = = = = = = = = = = = = = == = = = = = = = = = = = = = ,5995+ 6 = = = = = = = = = = = = = =/+# /+ 0(6 -:+#6 60$66+ "! = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = =:+* ! = = = = = = = = = = = = = = = = = = = = = = = = = = = = 9:- 6)86,/:77. /+ 0(6 )059:,(' 5+0-:,0/5+) = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = 60$66+ ! :++* ! 9:- 8-6*59/+:06 /+ 0(6 = = = = = = = = = = = = = = = = = = = = = = = = = = = = : = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = : = = = = = = = = = = = = = = = = = = = = = = = = = = = == = = = = = = = = = = = = = 314 2111 0/.-,+*)(0'%$)-/*)/-/%2,-#"0'#)0/)".+)+#20-"%!)#!#"+( ) '+/")+0*+/'+)-/*)(+".%*%,%0'-,)'.-,,+/+# Petra Party 1 1 1 1 1 , Bence Sipos , ) )0)' , Rita Ambrus 1. INTRODUCTION 1)( ,75 96+86 +5 +"+-5 -875 =;10 =6+05 71!5 85 ?=;:9876=75 4?3725 8105 181;:9876=75 4/372.5 7*76+)86=5 8::;8=,5 6;5 0+16(*5 8105 8189*7+5 -,=,5 8+5 7*16,+6=5 7;905 :86=9+75 -6,5 85 +9+"816576 0+75;151,89+05?37.5/37581056,+5 08)+6+5 ;(5 '5 )%$5 ))5 8105 #'5 ).5 +((+=675 ;15 6,+5 +7:86;*5 7*76+)5 6 0+75 +7:+=6"+9*.5 ,8"+5 +)+!1(=8165 +):9;*1!5 "8; 75 +:+)+16895 );0+975 8105 +1";1)+168958105: 9=5,+896,577 +5 )8175 =;): 686;18957) 986;175-++5+"+-+0 8+5 1+"689*5 +:;7+05 6;5 ?375 8105 /375 5 1,8986;15(;)5"8; 757; =+75+=+16576 0+75 )+#,"#)-/*)*0#'##0%/ 10=86+5 6,865 1,891!5 8;1+5 ?375 8105 /375 =; 905:;6+16899*508)8!+56,+5, )815+7:86;*5 1)) 7*76+)5 4! +5'2575 6,+5:86=9+5 7+5 81!+75 5 =+995 91+5 76 0+75 :98*5 85 = =895 ;9+5 15 8+5 "8; 7.5 6,+5 0+:;76;15 8+5 15 0((++165 10+768101!5 6,+5 =+99 985 )+=,817)75 +!;175;(56,+5+7:86;*568=658=9+75 10+9*1!5 ?375 6;=6*5 +7+8=,5 1";9"1!5 8+5 ;(6+15 +9)186+05 *5 6,+5 =9+881=+5 , )815 ;1=,895 +:6,+9895 =+9975 425 )+=,817)7.5 65 7)899+5 :86=9+75 )8*5 8105 , )815 9 1!5 +:6,+9895 $5 =+9975 ,875 8== ) 986+5 15 6,+5 9 1!75 ,+5 :)8*5 0+);17686+05 6,865 ?375 8105 /375 =8 7+5 5 )+=,817)56,; !,5-,=,5?3758105/375++65 :;0 =6;1.5 1(98))86;1.5 =+99 985 8105 677 +5 6,+5 ,8)( 95 +((+=675 75 877;=86+05 -6,5 6,+5 08)8!+5 ,75 )8*5 9+805 6;5 : 9);18*5 8+5 10 =6;15;(5;086"+576+77.51(98))86;1.58105 0*7( 1=6;15 8105 =,;1=5 ;76 =6"+5 9 1!5 : 9);18*5 (;775 '.5 ,+5 0+:;76;1.5 8105 07+87+5 4 325 %$5 ,++5 8+5 );+5 :;779+5 ,+896,5 +((+=675 ;(5 1,89+05 ?375 -++5 :,*7;9;!=899*5 +9+"8165 5 );0+975 76 0+05 15 6,75 -;.5 -6,5 85 (;= 75 ;15 6,+5 +:+7+161!5 6,+5 9 1!5 4? =925 6;5 76 0*5 8"8989+5 )+)81+5 :8778!+5 ;(5 "8; 75 ?37.5 -,=,5 5 8105 5 =,88=6+86;15 )+6,;075 6;5 0+16(*5 +7+8=,5 7,;-75 ,!,+5 5 6817986896*5 !8:756,; !,5 =;):886"+58189*775;(5(101!75 =;):8+056;5=;1"+16;1895);0+975571!585 8105)+6,;0;9;!=8959)686;1758=;77576 0+7 9 1!;18=,:56+=,1 +56,+576 =6 895(+86 +75 8105+*5( 1=6;175;(56,+589"+;989;;058+5 =815 +5 8== 86+9*5 7) 986+05 -,9+5 16+!861!5 ) 96:9+5 )) 1+5 =+9975,+5 7*76+)5899;-756,+5 89"+;98=8:998*5 8+.5 "8; 75 =+997.5 677 + 677 +5 16+(8=+7.5 8105 "87= 985 :+( 7;15 6;5 +5 16+!86+056;5);0+956,+5=,88=6+76=75;(59 1!5 5 ) 1)895 );0+975 ,8"+5 +6+10+05 6,+5 10+768101!5 ;(5 6,+5 ):8=675 ;(5 ?375 +*;105 =+99 985 +7:;17+7.5 1"+76!861!5 6,+5 :,8)8=;1+6=75 8105 7*76+)=5 +((+=675 ?=+5 8105 865 );0+975 8+5 (+ +169*5 7+05 6;5 6+765 6,+5 +:;7 +5 ;(5 ?375 1,89+05 :9876=75 =815 9+805 6;5 )1),+5+((+=675;(5/358105?35+:;7 +5;156,+5 , )815 ;0*.5 8105 6,+5 1"+76!86;15 )+6,;075 6;5 6,+58=6"86;15;(5)) 1+5:86,-8*7.59+801!56;5 10+7681056,+5)+=,817)75+,1056,+) 1=+87+059 1!51(98))86;156 0+757,;-56,865 1,8986;15;(5?3759+80756;5:86=9+58== ) 986;15 )(-"+0-,#)-/*)(+".%*# 15 9 1!5 677 +7.5 +7 961!5 15 :,*7;9;!=895 08)8!+.5 -,=,5 :;6+16899*5 +6+1075 6;5 ;6,+5 315 DCBA@? =;: 987 A@A65?4? 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ample Dosage FA content NFA content form [ng] S1 Tablets 1000 202.1 S2 Tablets 400 71.1 S3 Solution 200 LOD S4 Tablets 400 2.0* S5 Tablets 800 88.9 S6 Capsules 300 LOD 5?;=C/B1@C5;:AC?.CB5@D:?C!B5C31E!C, Figure 10. Representative chromatogram of separation between folic acid and N-nitroso-folic acid #" ! (NFA). 7C.;$.10C5:5;?;/4CBAA@DB?CB:=CDB2;=CB:B10?;AB1C $#%#"! 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'%# ""(,' %"(#!')# %#((!,%$# ,!"#('##('(% '%##(% %(#()()#%# % "'%$#(,(%' Irena Slaveska Spirevska1, 1 1 1 Katerina Kochova , Kristina Grncharoska , Elena Petrovska, 1. INTRODUCTION BCG67C;69AG:@ H)6A,BHG;@ CGD@ AG@ ;-H@ CGCEFHD@ IHGFEDCBAGH@ A?@ C@ =E@ C;AG:@ G=G?;H9=ADC@ ?CB,H@ .H9H@ 6C;HD(@ .A;-@ ;-H@ =;CAGHD@ @ $C6H?@+@:"E@ CG;A8AG7CBBC;=9E@ D96:@ 543210/@ .A;-@ =@ CGCH?;-H;A@CGD@CGC:H?A@,9=,H9;AH?+@*-H@B=?;@ + =BB=G@ ,-C9BC@ D=?C:H@ 7=9B?@ =7@ ;-H@ *-H@=H;A$H@=7@;-A?@?;6DE@.C?@;=@DH$H=,@CGD@ D96:@C9H@C@A)6AD@B=6;-.C?-(@=9=B6=?C@?,9CE@ $CADC;H@ C@ ?HG?A;A$H@ 9H$H9?HD8,-C?H@ -A:-@ CGD@,C?;AH?+ ,H97=9BCGA)6AD@ -9=BC;=:9C,-E@ 5 #8 *='A==:A@ ,9=,H9;AH?@ =7@ HGFEDCBAGH@ C9H@ #/@ BH;-=D@ 7=9@ DH;H;AG:@ HGFEDCBAGH@ .H@%G=.G@D6H@;=@A;?@H';HGDHD@;-H9C,H6;A@6?H@ 9H?AD6H?@AG@?.C@?CB,H?@C?@,C9;@=7@;-H@HCGAG:@ AG@ -6BC?@ CGD@ G=@ ?,H@ -CFC9D?@ 7=9@ -6BCG?@ $CADC;A=G@,9= -C$H@HHG@=?H9$HD+@#9HAGA@?;6DAH?@-C$H@G=;@ # '( !)# %#(') 9H,=9;HD@ CGE@ G=@ =?H9$HD@ H77H;@ H$H"G=@ =?H9$HD@CD$H9?H@H77H;@H$H@54! "@4!2 /@ +# ## $C6H?@+ -9=BC;=:9C,-E@ .C?@ ,H97=9BHD@ 6?AG:@ C@@ -HG@ 7=9B6C;HD@ C?@ C@ ,C?;AH(@ HC-@ 6GA;@ 3-ABCDF6@4H'H9C@ @#@?E?;HB@H)6A,,HD@ =G;CAG?@ @ B:@ =7@ HGFEDCBAGH@ -ED9=-=9ADH+@ .A;-@C@)6C;H9GC9E@=.8,9H??69H@:9CDAHG;@5#/@ *-H@=.H?;@9H=BBHGDHD@D=?H@=7@HGFEDCBAGH@ ,6B,(@ .A;-@ DH:C??H9(@ C6;=?HB,H9(@ =6BG@ -ED9=-=9ADH@ A?@ @ B:+@ *-H9H7=9H(@ ;-H@ =.H?;@ =$HG(@=G;9=H9@CGD@C@,-=;=DA=DH@C99CE@5#02/@ =?H9$HD@ H77H;@ H$H"=.H?;@ =?H9$HD@ CD$H9?H@ DH;H;=9(@=G;9=HD@E@C@3=6;A=G?@?=7;.C9H+@ H77H;@ H$H@ 5! "!2 /@ =7@ @ B:"DCE@ A?@ 2DDA;A=GC@ H)6A,BHG;@ AG6DHD@ C@ ?HBA8BA9=@ 6?HD@7=9@6C;A=G@=7@,H9BA;;HD@DCAE@H',=?69H@ CGCE;A@ CCGH9@ *=HD=@ #(@ 5#0 /@+ *#"@6;9C?=GA@C;-(@CGD@C@H;;H9@*=HD=@ 3@ ,"=GD6;A$A;E@ BH;H9+@ ;9C,69H@ .C;H9@ 1G@C=9DCG.C?@ ,9=D6A DH;H9BAGC;A=G@=7@#0 @DH?9AHD@AG@6ADHAGH@=G@ 3E9AG:H@7A;H9?@5+@B(@9H:HGH9C;HD@6=?H/@ 3H;;AG:@-HC;-8C?HD@H',=?69H@ABA;?@7=9@6?H@AG@ .H9H@ =;CAGHD@ 79=B@ 2:AHG;@ *H-G==:AH?@ 9A?%@ ADHG;A7A DA77H9HG;@BHDA@,9=D6;?@AG@?-C9HD@7CA;AH?@ 5 2@/(@;-H@=;CAGHD@$C6H@7=9@#0 @.C?@ #" ## +@B:"DCE@+ *-H@ 9HC:HG;?@ ;-C;@ -C$H@ HHG@ 6?HD@ C9H@ DAH;-ECBAGH(@BH;-CG=(@#=8,-=?,-=9A@C =9@ ?H;;AG:@ BC'AB6B@ C=.CH@ CGD@ CH@ ,69-C?HD@ 79=B@ C9=@ 9C+# 52!/@ ABA;@ 7=9@ HGFEDCBAGH@ 9H?AD6H?@ =G@ H7H9HG BCG67C;69AG:@ H)6A,BHG;(@ C?@ C@ GH';@ ,9=D6;@ -ED9=-=9ADH@ I# 3(@ .C?@ ?6,,AHD@ E@ ;-H@ .C?@ ?HH;HD@ ;-H@ ,9=D6;@ .A;-@ ;-H@ -A:-H?;@ 69=,HCG@ 0A9H;=9C;H@ 7=9@ ;-H@ 6CA;E@ =7@ BC'AB6B@DCAE@D=?H@C?@;-H@.=9?;8HDA *-H@=;CAGHD@2!@$C6H@=7@@B:@=9@+@ 5C9:H@ 2,-C@ 3.C/@ .H9H@ 6?HD@ 7=9@ DA9H;@ ,,B@5H=.@;-H@8,,B/(@HG?69H@;-H@?C7H@$C6H+@ ?CB,AG:+ 69;-H9B=9H(@ ;-H@ ABA;@ ,H9@ ?697C 325 #""! ?=;:98765463291068766/9.2;84=2;6-,6 26,/.6:280654601+8=067061+90:06 ?+3:6 *6 )('6 %6 $#"6 %6 '6 !6 38+ 7%6 :6 8;6 0:;97976 ;490 +6 8 7:46 86 ".%6 49769483;:936+ :987%659:26+856;:6)#)6 70976 :;6 :26 3+7976 =;830 ;6 /97#626973:987618+ 6546("6/6706 0874:;:976 4749:919:%6 +97;9:%6 33 ;36 0:3:987651+7:26546((67689+6 706 =;394987#6 26 4:+94206 +99:46 ;6 =246387494:068669: ;686("618+ 4686 =;3:93+%6 3291+%6 706 74 ;6 =;80 3:6 :278+%6 ('6 18+ 46 86 3:879:;9+6 706 ''6 +9:6706=:97:64:670697996:26;946 18+ 4686"#$611609:2+97659:26:26=6 863;8446387:97:987#6*406876:261+90:9876 0 4:06 :86 #"6 4976 =284=28;936 390#6 66 ))6 ;4 +:4%6:26:28069464 9:+68;60:;97:9876 116 9: ;6 86 :278+6 706 5:;6 =6 #"6 8670976;490 469764564=+46;86 0 4:06 59:26 =284=28;936 3906 546 406 46 7 3: ;976 9=7:%6 706 :26 =;8=8406 09+ 7:#6 +99:46 376 6 ==+906 8;6 ;8 :976 3+7976 1+90:9876=;830 ;4 6 26 1+90:9876 86 :26 -,,/.6 :2806 546 3870 3:06 976 338;0736 59:26 :26 3;9:;96 1. National Center for Biotechnology Information. PubChem Compound Summary for CID 12555, 4=39906 976 :26 7:;7:987+6 .87;736 876 Benzydamine. https://pubchem.ncbi.nlm.nih.go ;879:9876 .6 90+976 8;6 1+90:9876 v/compound/Benzydamine. Accessed May 14, 867+:93+6=;830 ;46' #6?+3:919:6:4:976 2025. 428506 786 97:;;736 ;86 :26 +76 456 2. International Conference on Harmonization, 2024. Impurities guideline for residual solvents 4=+6 :6 :26 ;:7:9876 :96 86 *7097#6 Q3C (R9). ICH, Geneva, Switzerland. -381;6 4: 0946 706 4=9976 :4:46 5;6 3. EMA/CHMP/CVMP/SWP/169430/2012. 3870 3:0687664:97+444:+6=+:6:863879;6 European Medicine Agency (EMA) (2014). :264 9:9+9:686:2609;3:64=+976=;830 ;6 Guideline on setting health-based exposure limits for use in risk identification in the 8;60:;97:987686*70976;490 46876 manufacture of different medicinal products in 9=7:64 ;3664=99764:70;0648+ :98746 shared facilities /%6('6/6706"6/#626329106 Technologies for Pharmaceutical manufacturing. Interpharm Press, Denver, 456 ;381;6 3:8;6 86 "#(6 (#6 6 546 86*70976:6:2;638737:;:9876+1+46'6 4. LeBlanc Destin A., 2000. Validated Cleaning Colorado, US. ==+906 46 6 38;;3:9876 3:8;6 976 :26 3+7976 5. International Conference on Harmonization, 1+90:9876;4 +:4# 2005. Validation of analytical procedures: Text 26+97;9:686:26:28065460874:;:06 and methodology Q2(R1). ICH, Geneva, Switzerland. :2;8 26417638737:;:9874%6976:26;76;86 $"6 /6 :86 (#'6 /6 59:26 :26 38;;+:9876 389397:61+ 6-(6546"##626+99:686 0:3:9876 /6 706 +99:6 86 7:993:9876 /6 5;6 0:;9706 6 4976 4;946 86 +97;9:6 48+ :98746 706 5;6 8 706 :86 6 "#")6 /6 706 "#"$6 /%6 ;4=3:91+#6 26 =;394987686:2644:6546+4861+ :06:6:26 /6+1+%6706:268:9706-?61+ 6546+446 :276'6# 326 6554 3210/.-,+6/0/*1)(-'-26.3-)%-6+6*1$+-,#.%)"1$+,-3)(*/1(1(!- )"1$1 +$-+*21)(1(+-1(-*2+10-,+#+(3+ Lucian- 1 1 2 3-, , Corina-3-/-!-3- -6, Cristian V.A. Munteanu2 %$#"! " ""# #!"#" " # #" # ""#"# " "#" " " #"# #" # "#" " " #"# #" 1. INTRODUCTION -26.33$- =;:98765:;:43278 318 56540678 06;%=68 :$739)468 +321#);:4328 8 :208 8 (:78 :208 ,8 :.238 :+078 :;68 :2:9*4+:98 .64=3078 :44;$)4608 438 6:+=8 354+8 73.6;8 $*8 ,8 06/693560813;8+3.53)2078(4=8:7*..64;+8' :2:9*778)72#8:874:432:;*85=:76806; :43.8 %=68 $393#+:98 :+4+699)93768 %=68 765:;:4608 1;:+43278 (6;68 746;6373.6;78.:*80116;8:2084878.53;4:248438 +3996+4608 :208 5);1608 3;8 6:+=8 5654068 06/69358354.).8.64=308318765:;:4328"! 8 ;:+6.+8 74:$94*8 (:78 /6;1608 13;8 4(38 +32043278 2834=6;8+:76784=687)95=);8:43.8.:*8$68+=;:98 -0+,#.*,-/($-$1,3#,,1)( 13;8 4=68 56540678 +324:22#8 .64=32268 648 306084387)9130678%=6768+3.53)2078=:/68 28+=;:98,8:2:9*778(683$4:2608765:;:4328 0116;6248 :+4318 73.68 ;:+6.+8 7)913068 56540678 (4=8 )23060856540678" 8$)484878234823(2818 0116;6248 76)62+678 +324:22#8 3268 30608 4=68 $393#+:98 :+4.64=32268(=628+=:34;35+8:#62478(6;68:00608 354+:9873.6;780116;8 4384=68.3$9685=:768%=6874:$94*83184=68;:+6.+8 .4);678(:78/6;16080);2#8:8.324=8318743;:#68 );8:.878438765:;:4684=6873.6;78)72#8:8+=;:98 :48'8:2088 +=;3.:43#;:5=+8 46+=2)68 13;8 1)4);68 5377$968 $393#+:98 )768 6540678 +324:22#8 :48 96:748 :28 -3)(3.#,1)( 30608 .64=32268 ;670)68 (6;68 765:;:4608 %=68 3$4:2608 ;67)9478 06.3274;:468 4=:48 4=68 )72#89)08+=;3.:43#;:5=*8+3)59608(4=8' (3;193(8 78 7)4:$968 13;8 4=68 739:4328 :208 78:208+;+)9:;80+=;37.8-80646+4328%=68 5);1+:4328318746;6373.6;78(4=8=#=8354+:98 76)62+68 318 4=68 +3996+4608 1;:+43278 (:78 5);4*8:+=83184=6.87874:$968:48'88(4=3)48 +321;.608$*8.:778756+4;3.64;*88 1);4=6;8 246;+32/6;7328 )4);68 74)0678 (998 8 646208 438 34=6;8 64'+324:22#8 56540678 8%=68 -/*+01/.,-/($-+*2)$, +3996+46081;:+43278.:*8$68)760828+6998+)94);678 !8- 28 3;06;8 438 /6;1*8 18 0116;6248 $393#+:98 6116+48 +:28$68:44;$)46084386:+=8746;6373.6;8 :;3)78 232:56540678 +324:22#8 30608 .64=32268 ;670)678 (6;68 :2:9*608 )72#8 -0+%+0+(3+, ;6/6;768 5=:768 +=;:98 9)08 +=;3.:43#;:5=*8 )72#8:8+=;3.:43#;:5=86)55608(4=8-8:208 !8988648:980/:2+67828+=;:98765:;:432783187.:998 80646+432813993(608$*88:2:9*77 56540678 $*8 +:599:;*8 696+4;35=3;6778 :208 +=;3.:43#;:5=*8 3);2:9831865:;:4328+62+68!8 8!8 8 =;3)8 8 648 :98 64=32268 30:4328 7696+4 .69:23.:8:24#628+62+6888! 327 3210/1.-,+*-)('(/(0-%$)#2.$-.)+1%-"! " -+10--+)#-+/+- 1/()*/%- #- (0/'/1(- +10- "$+) +'- 2'$+)(*%- # +1/+- #1%)+'%- 1#- "3- 210(0- - %$(- /1/*%)- #- (*(+)'$- +10- 11#+%/#1- ,/%$/1- "!- - ")#.)+ -((# (1%-#-%$(-!+%/#1+-- **%( - 2)#.)+ - 1*%/%2%/#1+- "()#) +1'(- ('((1'(- 210/1.- )#('%*- 328 8776 5434210/.30-/,-+,*45014/3.30-/,2.34,0/,1452-)(82-'4*342-/4, .43.34,0/%43.$#4,*"*84/*0-/ 4! ,1 1 1 1 ,8 ,' ,' , Ivana Mijakovac ,1 ,5 1. INTRODUCTION 3 ,7,58*.2:-725:4:2/14.*/.27,(7/1*/1+7*:36-1*! ;:987 65:43:210/.2:-7 64,+02/*7 :417 :)01,0*7 API* *0*619*.,9*7 ,(7 :7 ':/14.9*,-0%-17 :2/.$17 PSD Amount of PEG 3350 Sample .9#41+.19/7 .97 /517 641*19217,(7 1"2.6.19/*!7 51.47 Dx (90) (mg/ml) *1+.319/:/.,9725:4:2/14.*/.2*7:417,(7*.#9.(.2:9/7 ,! .36,4/:9217 .97 (,430-:/.,97 41*1:4257 %12:0*17 0 9 20 /5187+1/143.917/5172,36:2/91**7,(7/517*1+.319/7 1 30 20 +04.9#7*/,4:#17:9+7/50*7/51741*0*619+:%.-./87,(7 20 /517 *0*619*.,9!7 97 (:2/7 /517 5,3,#191./87 ,(7 :7 2 30 (1/2 the amount of *0*619*.,97:(/147*5:.9#7:((12/*7/517:3,09/7,(7 API) :9+7 519217 ./*7 1((.2:287 !7 97 /5.*7 */0+87 /517 3 30 (1/2 the amount of all the excipients) 25:4:2/14.:/.,97 ,(7*1+.319/*7 ':*71"136-.(.1+7 :2/.$17.9#41+.19/7/5:/7.*7641*19/7.97:7#.$197+,*17 10 4 30 10 %87 0*.9#7 :)01,0*7 *0*619*.,9*7 ,(7 31+4,"864,#1*/14,917 :21/:/17 ;7 :7 7 7 2/.$17 65:43:210/.2:-7 .9#41+.19/7 7 7 6:4/.2-17 *.17 5 30 40 +14.$:/.$17,(764,#1*/14,917!7 517*1+.319/:/.,97 +.*/4.%0/.,9 64,614/.1*7 ,(7 /51*17 3,+1-7 ,(7 65:43:210/.2:-7 *0*619*.,9*7'14173:9.60-:/1+7%87:+0*/.9#7/517 6:4/.2-17*.17+.*/4.%0/.,97:9+7%87/517:++./.,97,(7 ,1, 6,-81/58-1917#-82,-7!7 517-://1475:*7%1197 7,*!, ,! , ,7 416,4/1+7/,75:$17:72,9219/4:/.,97:9+73,-120-:47 1+.319/:/.,97 4:/17 ':*7 1":3.91+7 './57 :97 '1.#5/7 +1619+19/7 1((12/7 ,97 *1+.319/:/.,97 .9*/40319/7 ;. 0#17 ;7 3%7 14-.97 64,614/.1*7'5.2573:87%17:*24.%1+7/,7+16-1/.,97 13:987 '5.257 *611+*7 067 /517 *16:4:/.,97 ,(7 ,47 %4.+#.9#7 (-,220-:/.,97 !7 ,-8314.9+021+7 +.*614*.,9*7 %87 0*.9#7 219/4.(0#:-7 (,4217 519217 (-,220-:/.,97 './57 5.#57 3,-120-:47 '1.#5/7 *11*7 /517 41*0-/*7 ,(7 +1*/:%.-..9#7 64,21**1*7 (:47 6,-83147 64,211+*7 /54,0#57 6:4/.2-17 %4.+#.9#7 (:*/147/5:97$.*0:-7,%*14$:/.,9!7 5171"614.319/7 3125:9.*3!74.+#.9#7(-,220-:/.,97,2204*7'5197 ':*72,9+02/1+7,977(,430-:/.,9*7:36-1777 5.#57 3,-120-:47 '1.#5/7 6,-83147 3,-120-1*7 7 :9+7 7 :/7 7 +.((1419/7 4,/:/.,9:-7 *611+*7 7 :+*,4%7 *.30-/:91,0*-87 ,97 3,417 /5:97 ,917 7 :9+7 7 463!7 :*1+7 ,97 1"614.319/:-7 6:4/.2-17:76519,319,97/5:/72:9764,+0217-:4#17 $:-01*7 ,(7 $1-,2./87 :9+7 41-:/.$17 219/4.(0#:-7 (-,2*7!7 5.*76519,319,97.*7+1*.41+7.97/143*7 :221-14:/.,97 7 :7 -.91:47 1)0:/.,97 ':*7 ,(71:*.14741*0*619+:%.-./87,(7*0*619*.,9*! ,%/:.91+7 :9+7 *1+.319/:/.,97 4:/17 :/7 #7 ':*7 1"/4:6,-:/1+!7 ,1.3420.#*,./5,143-5* ,*!,,! , 7,1 , .((1419/7 (,430-:/.,9*7 ,(7 ;7 .912/:%-17 /517 *1+.319/7 '1417 1":3.91+7 (,47 7 +.((1419/7 1+.319/:/.,974:/17:/774637:9+7/51751.#5/7,(7 *0*619*.,9*7'14171$:-0:/1+7:*7*1197.97 :%-17! (,430-:/.,9*7:*7*1197.97 :%-177(,430-:/.,977 7:9+7! 329 2110 /.-,+*)('*-%$-$#*")**#!% /..-* - - 341;968-,9;98((96;6=26;,.;6=;0:9.;45;2; /.1.-* - - --1 =;:98769;54:;32107;/;2:;0:9.6-;,.;2+7; 0:4726,.(;96:868:;,9;0499,+7;4+62,.-;%,6=; 72:(;2148.6;45;=,(=;147872:;%,(=6;" ;2; *; 2.-; ,.; ),(8:; /'; =; 9-,1.626,4.; :269; 6:204726-; 64; /(; 54:; 277; 696-; 921079; 2:; 9,(.,5,2.6;.=2.1.6;64;5748726,4.;/';=; 6=; 0471:; 54:1,.(; 2; +2+4. ; 0:4,-,.(; 9=4%.;,.;2+7;$' =414(.489; 5748726-; .6%4:; +419; 32107; #; %,6=; 91277:; "3!; 9=4%-; 974%:; 974%7; 410:99-; 8.-:;6=; 556; 45; (:2,6; 9-,1.626,4.; :26; 6=2.; 32107; /; %,6=; =,(=:; /';.;24:-2.;%,6=;6=26;6=4: ;6=;:98769; "3! ; %=,=; ,9; ,.; 24:-2.; %,6=; 6=; 72%; 45; 0:9.6-; ,.; 2+7; '; 9=4%; 6=26; :-86,4.; 45; (:2,6';32107;*;=2-;91277:;,949,6;-8;64; 2148.6; 45; "; :9876-; ,.; 74%:; 9-,1.6; 6=; :-8-; 2148.6; 45;"; +; =275 ; 6=:54:; =,(=6 ;%=,=;,107,9;6=26;";02:6,79;%:.6; 9-,1.6-;5296:;6=2.;32107;/';32107;$;=2-; 6=26; 5748726-; 2.-; 9-,1.6; %29; 14:; =275; 6=; 2148.6; 45; 277; 4104..69 ; =.; 410:99-;6=2.;,.;32107; ;%=,=;4.62,.-; 9=4%-;6=;=,(=96;9-,1.626,4.;:26;-8;64;6=; 14:; "; 2.- ; 6=:54: ; 799; 410:99-; 74%96;,949,6'; 9-,1.6; -8; 64; =,(=:; 5748726,4.; 45; "; 02:6,79'; 98769; 2794; ,107; 6=26; 6=:9; 2; ' - .-74,6; 2.-; ; :98769; 26; -,55:.6; .(26,; 4::726,4.; +6%.; 9-,1.626,4.; :4626,4.27;90-9;54:;32107;/' 74,6;2.-;9-,1.6;=,(=6'; speed (rpm) Rotational RCF Velocity RCA ' -.-74,6;26;##;:01;2.-;9-,1.6;=,(=6 (g) (g) 200 6 2.3 5.1 Sediment 500 36 Sample height Velocity at 500 15.1 31.4 - (mm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ample *'; 4(( ; ' ; Sedimentation rate (mm/h) 0 0.7 ;"4%-:;2.-;"2:6,7;48:.27;4'$# ;*#/$' 1 1.4 "%!(+$+%' 2 3.1 30,27;:-,69;64;6=;%=47;!;";621' 3 3.8 330 8776 54321014/.1-,1+341-*)(80+.')3-30/41+30)%$4(#,)"#!)"#034$5)1-) %$,1+1-$5)3-,)"##,)5'880$%$-45 )4)!, !)! 1. 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C254B6DAA?@3AB5B@6 ECBA?B@54 @ 8A3A4CEA8 2CD E45?5D 1D256A0 /A C2 B:A EC@B@3A49 6:5D*A8 E5B6: /@B: B:A @8A?B@291D256A5EDA8C,@?5?B49 EC@B@3A49 6:5D*A8 5?B@7C89 C2 B:A E5B6: BC B:A 7AB-2@B E45?A1D256A0 544C/@?* 2CD B:A :98DCE:C7@6 BD5?45BA8 BC B:A ,5+@,1, EC@?B 54C?* B:A @?BAD56B@C?A3@6C@B9= ?CD,543A6BCD)(@*=' :@ A4A6BDCB5B@6 *15D8 :@*:4@*:B B:A 6C1E4A8@?241A?6A C? ? ! E:9@6-75A8 6C,E1B5B@C?54 ,AB:C8 2CD %A );7AB-2@B )*DAA? :@2BA8 BC E5 B:DC1*: B:A E5B6:" :@*:AB EDA8@6B@?* B:A :98DCE:C7@6 6C?BD@71B@C? BC EC@?B54C?*@8A3@A/ EDCBA@? C41B@C? 3@6C@B9= A@*?A8 2CD A5D49 E5B6:0:C/@?*1D256A)9A44C/= 5EE4@65B@C? @? B:A 8D1* DAA5D6: 5?8 8A3A4CE,A?B E@EA4@?A0 C1D 5EEDC56: A?574A ,CDAC1D6A@?3AB,A?B5?8 A C7AD3A8 5 BDC?* 6CDDA45B@C? 7AB/AA? B:A 5 *A?AD54@574A 54BAD?5B@3A BC ,5?9 A+@B@?* 6546145BA875A8 ,C8A40 ,5@?* @B B:ACDAB@65449 A?AD*@A 5?8 B:A ,A51DA8 3@6C@B@A 5EE4@6574AB5B@6 @?BAD56B@C? 6DAA?A8 1@?* ', 5#4= ;?B@7C8@A /@B: @?BAD56B@C? A?AD*@A BDC?*AD B:5? 5EEDC+@,5BA49 ! 6C?@BA?B49 A+:@7@BA8 :@*: 3@6C@B@A ),CDA '=/5,@==0 B:5?:C48 ;?B@7C8@A0'=) B:CA-= B:@ B:DA:C48 )(@*= = CB57490 B:@ 61BC22 #:A??5,ABB9 =0 AB 54=0 54@*? /A44 /@B: EDA3@C149 AB574@:A8 4@,@B 2CDAB@74A5B@C?54 ;658A,9 C2 6@A?6A0 = ') $DC6AA8@?* @?BAD56B@C?= ''-''= ;*D5/54 =0 AB 54=0 ;70'=)' -= C?548C? ==0 AB 54=0 5?*,1@D0 '= ') '-= = ! @6C@B9 E4CBBA8 /@B: DAEA6B BC B:A @?BAD56B@C? A 2C1?8 B:5B 64CA49 7CD8AD@?* A4A6BDCB5B@6 E5B6:A ,C8145BA :C/ :98DCE:C7@6 E5B6:A @?241A?6A3@6C@B9=C,A5?B@7C8@A0588@B@C? 292 8766 543210/.-30,32+/223*5)(,52(0'2(53)2%)1032/)+$2 #3"+ .32#+.1032+02#1024')(51+02(0'2%())13)2/-0,51+0 Alenka Zvonar Pobirk1 1 1 1 2 , Mercedes Vitek 2$2%2 , Katarina Bolko Seljak , 02, Eylem Atak3, Daniel Bosch3, Katja Klun3, Ana Rotter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ornneometer Table 1. S Skin hyydration values measurred as h u r CM 825 arbitraryy unnits (a.u.) befoore r u f 98BB@7 67C@ 8D BEC: +*C,) (') E :EDC3@7866@ 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8? ? ? ? ? ? ? ? ? ? and aftter 1 an f nd 2-week(s) usage of unloaded HC and :% ,@ C B%@ D5C .$ E7 @7#B%@48 C3@" 9@7@ 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? LLCs, and the eir Dunaliella e ttract loaded countterparts x n E6D@7AA@3! 9%C:% CD C @ used alone (H HCa; LLCa) or aftter prior application of f 7 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 9CB% B%%@ C4.7EA@4@B ? ? ? ? ? ? ? ? ? ? B ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? water brine (H HCa+b, LLCa+b). E= B%%@ D5C D 6877C@7 =;;:BCE) ? ? ? ? ? ? ? ? ? ? B ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? = ? ? ? ? ? ? ? ? ? ? @A@7BB%@@DD! 7 ? ? ? ? ? ? ? ? ? ? A8;@D 9@7@ 3@B@:B@3 9%@ 84. B ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? B ( 9@@5 B ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? -? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 98BB@7 67C@ 98D 8...:87@ @ 9@@5D? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 7@,C4@) $ ( (! ! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ! ! ! -! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? $ $8 !- -! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?-! -!- ! -! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? // ( (! -!-? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? (! (! -!- (! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? //8 ! -! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? -! -!( -! (! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? $86 ( (! !-? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ! -!- !( (! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? //86 ( (! -! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? -! (! -! (! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 2 2EE =;;# @". ? ? ? ? ? ? ? ? ? ? = ? ? ? ? ? ? ? ? ? ? .@=CC:C8 @===@@:B E= 98B@7 ? ? ? ? ? ? ? ? ? ? B ? ? ? ? ? ? ? ? ? ? = ? ? ? ? ? ? ? ? ? ? = = ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8? ? ? ? ? ? ? ? ? ? 67C@ E B%@ 67C@ E B%@ ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ?D5C D 6877C@7 =;;:BCE! CB D%E;3 6@ ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? = ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 6@DB ;D@3 ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? BE,@B%@7 9CB%% @4EBD ? ? ? ? ? ? ? ? ? ? befoore (t=0) a :EA@BCE8 8 :7@84 ? ? ? ? ? ? ? ? ? ? 8 4D E7 EA@ 3@f Figure 1. Th he inflluence of water brine on f n TEWL and aftter 1 and 2 week(s) of its f usage as skin tonic on lower foorearm f r m. 8D //D! 9C ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? CB% E7 9CB%E;B 8,8@ @"B78:B! .EDC, ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? " ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? D5C %#378BBC ? ? ? ? ? ? ? ? ? ? 8 C, @===@@:B) ? ? ? ? ? ? ? ? ? ? = = ? ? ? ? ? ? ? ? ? ? B%@ =;;B;7@ B%@ ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? = ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ED@;@B#! B%@ ;D8,@ E= 98B@7 67C ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? @ 9CB% ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? .7EB@:BCA@ 8 ? ? ? ? ? ? ? ? ? ? 8:BCACB# E= :ED4@BC: .7E3;:BD 9CB% ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? D;6D@;@B? ? ? ? ? ? ? ? ? ? 8.. E= $ 3 // 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? D 9CB% ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? %$#"! !!"" ! @"B78:B D%E;3 8 83 9CB%E;B ? ? ? ? ? ? ? ? ? ? B? ? ? ? ? ? ? ? ? ?%$#"! !!" !? ? ? ? ? ? ? ? ? ?@"BB78:B 98D 8 ; :87@ .EB@BC8) ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? 2%@7@6# B%@ ? ? ? ? ? ? ? ? ? ? @ 83A8B8,@D ? ? ? ? ? ? ? ? ? ? 8 8? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? E= 98BB@7 67C@ 8 ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? ;D@3 8D ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? B C BEC: 9@7@ 4@7,,@3 9CB% D5C %#378BCE @ ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 7 3 CB% 5C % 3 BC ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? @===@@:B E= ==== B = = = ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? D5C :87@ =EE74 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? = 7 4;8BCED +2866@ (' 8 ? ? ? ? ? ? ? ? ? ? 2 8 ? ? ? ? ? ? ? ? ? ? ') 2%@ ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? 4ECDB;7CC, 8:BCACB# E= =EE74 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? = 7 4;8BBCED 98 8 ? ? ? ? ? ? ? ? ? ? 8D B@DB@3 ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? ()? ? ? ? ? ? ? ? ? ? CCB@5! ) 83 ED@:8 8B8! )! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? B ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? ! # "! ! 833CBCE BE 210/! D5C .$! 83 @7 !! ! $ ""! " $$" # " ! ! ! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 7#B%@48 # ? ? ? ? ? ? ? ? ? ? # # " $$# # ? ? ? ? ? ? ? ? ? ? :B8 F%874! --) +-' ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? C3@") 6# 4@8D;7C, DB78B;;4 :E7@;4 %#378 ""! " # !!" !" " ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 B ? ? ? ? ? ? ? ? ? ? 7 ? ? ? ? ? ? ? ? ? ? 88BBCE C ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? .. E= $! 9CB% 83 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8? ? ? ? ? ? ? ? ? ? 9CB%E;B? ? ? ? ? ? ? ? ? ? (() 8,8@ @"BB78:B! 7@D; DC,C=CC: B# C ? ? ? ? ? ? ? ? ? ? " ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? = 8 ? ? ? ? ? ? ? ? ? ? :7@8D@3? ? ? ? ? ? ? ? ? ? DE;:@3 9%@ 98BB@7 67 # $ # $ ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? 7C@ 98D ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? C:C@:@! --) (- ((() ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ==CC7DB 8... E= DB78BB;4 -)? ? ? ? ? ? ? ? ? ? EBBB@7! )! @B 8)! B ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? "" # "# $$ ! 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? :E7@;4) 2%CD @=== ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? = ==@@:B 98D ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? = @3 6# CBD ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 210/ , @===@@:BD) E DC,C=CC:8B @ )? ? ? ? ? ? ? ? ? ? $8A8D! *)! @B 8)! 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? %$#"! !!" "! #" " ! ! ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? = = ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? = 8 ? ? ? ? ? ? ? ? ? ? @===@@:B E = = ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? D5C %#378BBCE 98D E6D@7AA@3 =EE7 //D D $#" # # $# ### !" ? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 7 ? ? ? ? ? ? ? ? ? ? = ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 84 4.@D) ? ? ? ? ? ? ? ? ? ? " " # "# # !!" # "# # ! 2%CD 98D :EB787# BE E;7 @".@:B8BCE ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 ED! 8D ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? ##!""" # ! ) 87) 7;,D! ---) -+-' ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?? ? ? ? ? ? ? ? ? ? .7@87## 7? ? ? ? ? ? ? ? ? ? # B@DBC, E .C,D @87 D ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?5C 8() 7@D; C4.7EA@3 D5C %#378BBCE 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? 83 210/ A8;@D! 9%C:% 9@7@ C B%@ D 4 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 8 4@ 7 ,@ ? ? ? ? ? ? ? ? ? ? 8? ? ? ? ? ? ? ? ? ? ;.E 8...#C, @CB%@7 //D E7 $ 9 ? ? ? ? ? ? ? ? ? ? 8 ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? 9CB% 83 ? ? ? ? ? ? ? ? ? ? 8? ? ? ? ? ? ? ? ? ? 9CB%E;B C %? ? ? ? ? ? ? ? ? ? % %$#"! !!"" ! !! ! @"B78:B) 833CBCE! 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