Determination of Active Metabolite and Secondary Metabolite of Irinotecan in Rat Liver Microsomes Incu-bation System by LC-MS/MS
- VernacularTitle:LC-MS/MS法测定肝微粒体孵育体系中伊立替康活性代谢物及其次级代谢产物浓度
- Author:
Xinlin ZHANG
;
Nanxi WANG
;
Chaoran ZHU
;
Xuejia ZHAI
;
Yongning LV
- Keywords:
7-Ethyl-10-hydroxycamptothecin;
Incubation ystem;
Metabolite;
UGT1A1 enzyme;
LC-MS/ MS
- From:
China Pharmacist
2017;20(2):238-241
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish an LC–MS/MS method for the determination of the active metabolite(SN-38) and secondary metabolite(SN-38G) of irinotecan in rat liver microsomes incubation system, and optimize the incubation conditions. Methods:Meth-anol was selected to precipitate protein in the samples, and then the concentrations were analyzed by LC–MS/MS. All the separation was carried out on a ZORBAX Eclipse XDB-C18 column(2. 1 mm × 50 mm, 3. 5 μm) with the mobile phase of acetonitrile – water (containing 0. 1% formic acid) (23 :77) at a flow rate of 0. 3 ml·min-1. The mass spectrometer was operated with multiple reac-tions monitoring ( MRM) using electrospray ionization ( ESI) . The incubation conditions were optimized by single factor design. Re-sults:SN-38 and SN-38G showed a good linearity ( r≥0. 9972) respectively within the range of 2. 3-920 ng·ml-1 and 2. 5-1000 ng ·ml-1. The intra-and inter-day RSD was below 14. 6%(n=6). The average recovery was within the range of 74. 1%-123. 4% with RSD below 13. 5% (n=6). The optimal incubation conditions were as follows:the concentration of liver microsomal protein was 0. 3 mg·ml-1 and the incubation time was 30 min. Conclusion:The method is rapid, sensitive and accurate in the quantification of SN-38 and SN-38G in the incubation system,which provides methodological basis for the activity determination of UGT1A1 enzyme in vitro.
