20.500.12556/RUL-175092 Posodobitev analitske metode mikrobiološkega preizkušanja kakovosti za kalijev klavulanat in gentamicin sulfat Modernization of the analytical method of microbiological quality testing for potassium clavulanate and gentamicin sulphate Namen magistrske naloge je bil posodobiti analitski postopek mikrobiološkega preizkušanja kalijevega klavulanata in gentamicin sulfata. Posodobitev je bila potrebna zaradi optimizacije izvedbe analitske metode ter zaradi novih zahtev takrat aktualnih farmakopej: Evropske farmakopeje, Ameriške farmakopeje ter Japonske. Preverili smo vpliv nevtralizatorjev kot dodatkov gojišču in ugotovili, da le-ti doprinesejo k analizi gentamicin sulfata, ne pa tudi kalijevega klavulanata. Preverili smo možnost uporabe metode s prelivanjem pri izvedbi analize skupnega števila aerobnih mikroorganizmov in analiz skupnega števila kvasovk in nitastih gliv. Izkazalo se je, da lahko metodo uporabimo le pri analizah skupnega števila kvasovk in nitastih gliv pri kalijevem klavulanatu. Preverili smo tudi možnosti optimizacije analitske metode z uporabo manjših količin fosfatnega pufra, fosfatnega pufra z nevtralizatorjem in izpiralne tekočine, pri čemer smo uspešno znižali količine uporabljenih reagentov. Naposled smo tudi preverili uporabnost polivinilidenskih difluoridnih filtrov za metodo membranske filtracije namesto filtrov iz mešanih celuloznih estrov in ugotovili, da pri korakih, kjer se poslužujemo metode z membransko filtracijo, dobro delujejo tudi polivinilidenski difluoridni filtri. Kot testne mikroorganizme smo izbrali bakterije vrst Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa in Escherichia coli, kvasovko Candida albicans in nitasto glivo Aspergillus brasiliensis, vse pridobljene iz zbirke American Type Culture Collection (ATCC). Obe validaciji postopka sta bili uspešni, analitski metodi smo optimizirali in posodobili. The aim of this master's thesis was to update the analytical methods of microbiological testing of two active pharmaceutical ingredients, namely potassium clavulanate and gentamicin sulphate. The update was necessary due to the optimization of the implementation of the analytical method and due to the new requirements of the then current pharmacopoeias: the European Pharmacopoeia, the American Pharmacopoeia and the Japanese Pharmacopoeia. We tested the influence of neutralizers as additives to the culture medium and found that they only contribute to the analysis of gentamicin sulphate, but not potassium clavulanate. We evaluated the possibility of using the pour plate method in performing total aerobic microorganism count and total yeast and mold count analyses and demonstrated that the method can only be used in total yeast and mold count analyzes of potassium clavulanate. We also tested the possibilities of optimizing the analytical method by using smaller amounts of phosphate buffer, phosphate buffer with neutralizers and rinsing solution and successfully reduced the amounts of these reagents in the assays. Finally, we also tested the applicability of polyvinylidene fluoride filters in the membrane filtration method instead of mixed cellulose esters filters and concluded that polyvinylidene fluoride filters also work well in the steps where the membrane filtration method is used. As test microorganisms we have used bacteria Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa and Escherichia coli, fungi Candida albicans and Aspergillus brasiliensis, all purchased from American Type Culture Collection (ATCC). Both process validations were successful, the analytical methods were successfully optimized and updated. kalijev klavulanat gentamicin sulfat test sterilnosti mikrobiološke metode potassium clavulanate gentamicin sulphate sterility testing microbiological methods true false false [Z. Szabó] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2025-10-16 07:15:20 2025-10-16 07:15:30 2025-12-04 03:57:35 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 2026-10-13 579.2.083:615.014 267498 253484803 54283.pdf 54283.pdf 1 CDE6DAE6F1B42B12B5F431B610ADAFC6 75632b101274d93542a1650a6779325e59157a52997c8f2fb81fea727add09c0 de8ed2f4-aa4e-11f0-9328-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=220141 Biotehniška fakulteta 0 0 0