Horyn M. Development and Validation of the Methods for the Analysis of Meldonium and Metoprolol in Pharmaceuticals and Blood Plasma

The dissertation presents an experimental study of the development of analytical and bioanalytical methods for the determination of metoprolol and meldonium in substances, pharmaceuticals and blood plasma. The generalized approaches to the development of original express analytical and bioanalytical methods for the determination of API and ready-made medical forms of metoprolol and meldonium, as well as their combination, have been demonstrated. Original spectrophotometric methods for the quantitative determination of metoprolol tartrate in the substance and drug by reaction with BCG, BPB, BTB have been developed and validated. The optimal conditions for quantitative determination have been experimentally substantiated. For the first time, a methodology for the selection of a reagent for the development of spectrophotometric methods for the determination of meldonium in substances and drugs has been proposed. An original spectrophotometric method for the quantitative determination of meldonium in substances and drugs based on the reaction with alizarin has been developed and validated, and the optimal conditions for quantitative determination have been established. The chromatographic conditions have been proposed. Express and environmentally safe HPLC methods have been developed for the quantitative determination of meldonium in substances and drugs using an Agilent Zorbax C-18 SB chromatographic column (4.6 mm i.d. X 150 mm, 3.5 μm). As a result of the research, 8 HPLC variations have been obtained. Validation of analytical methods has showed their suitability for the purposes of pharmaceutical analysis. The methodology for the development of the HPLC method for the simultaneous determination of two analytes with a difference in polarities (meldonium and metoprolol) has been demonstrated, and the chromatographic conditions for the development of the HPLC method for the simultaneous determination of metoprolol and meldonium in drugs have been proposed. There have been developed and validated original HPLC methods for the simultaneous determination of metoprolol and meldonium in a binary mixture and drug using chromatographic columns (LiChrospher 100 CN of two sizes (4 mm × 250 mm, 5 μm), (4 mm × 125 mm, 5 μm), Waters Spherisorb CNRP (4.6 mm × 250 mm, 5 μm), ZORBAX StableBond CN (4.6 mm × 250 mm, 5 μm)), which are formulated in 12 variations. 9 variations (mobile phase – ACN – 0.15 % NH4H2PO4 (60:40), chromatographic column – Zorbax CN SB) and 10 variations (mobile phase – ACN – 0.15 % NH4H2PO4 (50:50), chromatographic column – Zorbax CN SB) are recommended as optimal, since there are observed low values of the limit of determination and limit of quantitative determination, expressivity (chromatography time – 3.5 min), score according to the AGREE method – 0.77. Chromatographic conditions for the development of a bioanalytical method for the determination of metoprolol and meldonium in blood plasma have been proposed. An original HPLC-MS/MS method for the quantitative determination of metoprolol and meldonium in blood plasma has been developed, which can be used by laboratories studying the pharmacokinetics and bioequivalence of drugs containing metoprolol and meldonium. Based on the proposed principles and approaches, an original HPLC-MS/MS method for the determination of metoprolol and meldonium in blood plasma has been developed (utility model patent of Ukraine No. 143684 dated August 10, 2020).