Determination of warfarin enantiomers in human plasma by HPLC-MS/MS and its application
10.13699/j.cnki.1001-6821.2017.18.021
- VernacularTitle:HPLC-MS/MS法测定人血浆中华法林对映异构体浓度及应用
- Author:
Da MIAO
1
;
Wan-Ying YU
;
Hong-Yi TAN
;
Qi PEI
;
Jun CHEN
;
Xiang-Jun LIU
;
An YAO
;
Guo-Ping YANG
Author Information
1. 中南大学湘雅三医院临床药理中心
- Keywords:
HPLC-MS/MS;
warfarin;
enantiomer;
drug concentration in plasma
- From:
The Chinese Journal of Clinical Pharmacology
2017;33(18):1806-1809
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a HPLC-MS/MS method to simultaneously determinate R-warfarin and S-warfarin concentrations in human plasma.Methods Warfarin enantiomers were extracted with ethylacetate,ibuprofen was used as internal standard (IS).R-warfarin and S-warfarin were separated on column Lux cellulose-4 (4.6 mm× 250 mm,5 μm) with mobile phase 5 mmol ammonium acetate solution containing 0.4％ formic acid acetonitrile =45 ∶ 55,at flow rate of 0.5 mL · min-1.Warfarin enantiomers were protonated with electroapry ionization (ESI),the multiple reaction monitoring (MRM) transitions were preformed at m/z 307.0→m/z 161.0 for R-warfarin,m/z 306.9→m/z 106.9 for S-warfarin and m/z 204.8--→m/z 161.0 for internal standard,the specificity,lower limit of quantitation and standard curve,precision and accuracy and recovery rate as well as stability were investigated.Results The standard curves of R-warfarin and S-warfarin were Y=2.57 ×10-3X+3.84 ×10-3(R2 =0.997 6),Y=2.46 × 10-3X+ 4.60 × 10-3(R2 =0.996 8),had a good linear relationship in 10.00-500.0 μg · L-1,the limit of quantification was 10.00 μg · L-1.The method was validated by recovery,precision,accuracy,matrix effect and sample stability.Conclusion The developed HPLC-MS/MS method in our study was sensitive and accurate,and the method was able to determinate the R-warfarin and S-warfarin concentrations in human plasma.
