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.
|
