Determination of Warfarin and Its Metabolite in Human Plasma by Ultra Performance Liquid Chromatog-raphy Tandem Quadrupole Mass Spectrometry (UPLC-MS/MS)
- VernacularTitle:超高效液相串联质谱法检测人血浆中华法林及其代谢产物的浓度
- Author:
Xiaohua ZOU
;
Shuanghu WANG
;
Yunfang ZHOU
- Publication Type:Journal Article
- Keywords:
Warfarin;
Plasma concentration;
UPLC-ESI-MS/MS;
7-Hydroxywarfarin
- From:
China Pharmacist
2015;(3):364-367
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish an ultra performance liquid chromatography-tandem quadrupole mass spectrometry method for the determination of warfarin and its metabolite 7-hydroxywarfarin in human plasma. Methods: An ACQUITY UPLC? BEH C18 (50 mm × 2. 1 mm, 1. 7 μm) column was used as the stationary phase at 40℃. The mobile phase consisted of acetonitrile and water (con-taining 0. 1% formic acid) with gradient elution at a flow rate of 0. 4 ml·min-1 . Warfarin-d5 was used as the internal standard. The analytes were detected on a triple-quadrupole mass spectrometer equipped with an ESI interface in a positive mode. Results:The reten-tion time of warfarin and 7-hydroxywarfarin was 1. 8 min and 1. 5 min, respectively. Excellent linear calibration curve of warfarin and 7-hydroxywarfarin was obtained within the concentration range of 25-2 000 ng · ml-1 ( r =0. 999 3 ) and 5-500 ng · ml-1 ( r =0. 999 6), respectively. The lower limit of quantification of warfarin and 7-hydroxywarfarin was 5 ng·ml-1 and 2. 5 ng·ml-1 with the average recovery of 96. 9%-105. 3% and 97. 1% -103. 3%, respectively. The intra-and inter-day standard deviations were both less than 10%. Conclusion: The method is accurate and simple, and suitable for the determination of warfarin and its metabolite 7-hydroxywarfarin in human plasma.
