20.500.12556/RUL-147543 An effective validation of analytical method for determination of a polar complexing agent the illustrative case of cytotoxic bleomycin The effectiveness of highly polar agents in cancer treatment is well recognized, but their physicochemical properties make their analytical determination a demanding task. Their analysis requires peculiar sample preparation and chromatographic separation, which heavily impacts the precision of such an analytical method. As a case study, we chose a polar cytotoxic bleomycin, which is a mixture of complexing congeners with relatively high molecular mass, a fact that creates an added challenge in regard to its detection via electrospray mass spectrometry. These issues combined lead to a deprived method performance, so the aim of this study is manifold, i.e., to optimize, validate, and establish quality performance measures for determination of bleomycin in pharmaceutical and biological specimens. Quantification of bleomycin is done at diametrically different concentration levels: at the concentrations relevant for analysis of pharmaceutical dosage forms it is based on a direct reversed-phase HPLC-UV detection, involving minimum sample pretreatment. On the contrary, analysis of bleomycin in biological specimens requires phospholipid removal and protein precipitation followed by HILIC chromatography with MS/MS detection of bleomycin A2 and B2 copper complexes being the predominant species. This study further attempts to solve the traceability issue in the absence of certified reference standards, determines measurement uncertainty, investigates BLM stability and method performance characteristics, and, last but not least, provides an explanatory example of how a method quality assurance procedure should be established in case of an exceedingly complex analytical method. liquid chromatography mass spectrometry biology pharmaceutics measurement uncertainty stability tekočinska kromatografija masna spektrometrija biologija farmacija merilna negotovost stabilnost true false true Angleški jezik Slovenski jezik Članek v reviji 2023-07-07 10:17:54 2023-07-07 10:18:00 2024-09-27 15:19:47 0000-00-00 00:00:00 2023 0 0 Str. 2737-2748 iss. 14 Vol. 415 Jun. 2023 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 543 1618-2650 10.1007/s00216-023-04675-x 148531715 s00216-023-04675-x.pdf s00216-023-04675-x.pdf 1 E3033F13BD19714C6D3937AD6B782121 6d88f86bdc29d10da3a9874d8132889b7ab5b34180f48485701029a5486f27b8 0cf2215d-1c9f-11ee-a314-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=171161 https://link.springer.com/article/10.1007/s00216-023-04675-x 1 5e33ed25-1c9e-11ee-a314-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=171160 Fakulteta za farmacijo Zdravstvena fakulteta 0 0 0