Development of ultra performance liquid chromatographic method for the analysis of dasatinib degradation products

U ovom diplomskom radu razvijena je učinkovita i osjetljiva metoda tekućinske kromatografije ultravisoke djelotvornosti (UHPLC) za analizu onečišćenja i razgradnih produkta lijeka dasatiniba. DryLab je korišten za određivanje optimalnih uvjeta temperature, vremena gradijentnog eluiranja i sastava pokretne faze kromatografskog odijeljivanja metodom UHPLC. Najprikladnija kolona bila je Acquity UPLC CSH Phenyl-Hexyl 1,7 μm; 2,1 mm 100 mm. Optimalni uvjeti su: temperatura kolone = 50 °C, pH pokretne faze = 2 (fosfatni pufer za UHPLC, mravlja kiselina za LC-MS/MS analizu) i ukupno vrijeme trajanja analize: 27,5 min. Forsirana ragradnja dasatiniba provedena je u kiselom i lužnatom mediju, uz prisutnost vodikovog peroksida te na povišenoj temperaturi. Za analizu razgradnih produkata korišten je vezani sustav LC-MS/MS. Na temelju MS i MS/MS spektara pretpostavljene su strukture razgradnih produkata i predložene odgovarajuće sheme fragmentiranja.In this diploma thesis efficient ultra-high performance liquid chromatography (UHPLC) method for determination of impurities and degradation product of dasatinib was developed. DryLab software was used to determine the optimal conditions for UHPLC separation like temperature, time of gradient elution and composition of the mobile phase. The most efficient chromatographic column was Acquity UPLC CSH Phenyl-Hexyl 1,7 μm; 2,1 mm 100 mm. The optimal condition were: column temperature = 50 °C, pH of mobile phase = 2 (phosphate buffer for UHPLC and formic acid for LC-MS/MS analysis), and total time of analysis = 27.5 min. Forced degradation of dasatinib was performed in acid and basic conditions, upon addition of hydrogen peroxide, and on the elevated temperature. The LC-MS/MS system was used for the analysis of the degradation products. Based on the MS and MS/MS spectra, the stuctures of degradation products, as well as fragmentation shemes were proposed