目的建立高效液相色谱串联质谱法(HPLC-MS/MS)测定人血浆中华法林对映异构体的浓度。方法以乙酸乙酯为萃取剂,采用MS Chiral○R MS-OD(50×2.1mm,3μm)为手性柱,流动相为甲醇∶水∶甲酸=85∶15∶0.05,流速为0.18mL/min。采用负离子模式,电喷雾离子源(ESI),以萘普生为内标定量,华法林和内标的检测离子对质荷比分别为(m/z)307.2→160.9和(m/z)228.9→185.1。结果 R-华法林批内精密度,其相对标准偏差(RSD)为3.2%~5.8%,批间RSD为2.5%~5.1%;方法回收率(96.1±5.6)%~(105.4±4.7)%,提取回收率80.7%~84.4%,内标基质效应校准后的基质效应RSD<10%;S-华法林批内精密度RSD为3.7%~5.2%,批间RSD为3.2%~4.8%;方法回收率(98.3±5.1)%~(103.7±3.8)%,提取回收率81.3%~84.6%,内标基质效应校准后的基质效应RSD<10%。最低定量限均为0.1μg/mL。结论本文建立的方法准确、灵敏、可靠,可用于人血浆中华法林对映体的拆分和血药浓度的测定。 Objective To establish a method for the determination of warfarin enantiomers in human plasma by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS / MS). Methods The mobile phase consisted of methanol: water: formic acid = 85:15:0.05, the flow rate was 0.18 mL / min, the mobile phase consisted of ethyl acetate as extractant and MS Chiral MS-OD (50 × 2.1 mm, min. Negative ion mode and electrospray ionization (ESI) were used to quantify naproxen as internal standard. The mass-to-charge ratios of warfarin and internal standard were (m / z) 307.2 → 160.9 and (m / z) 228.9 → 185.1. Results The R-warfarin intra-assay precision showed a relative standard deviation (RSD) of 3.2% -5.8% and inter-assay RSD of 2.5% -5.1%. The method recovery was 96.1 ± 5.6% to 105.4 ± 4.7, %, And the recovery of extraction was 80.7% ~ 84.4%. The matrix effect RSD after internal standard matrix effect calibration was less than 10%. The intra-assay RSD of S-warfarin was 3.7% -5.2% %. The recoveries were 98.3 ± 5.1% and 103.7 ± 3.8%, respectively. The recoveries were between 81.3% and 84.6%. The matrix effect of the internal standard matrix was calibrated with RSD <10%. The minimum limit of quantification was 0.1μg / mL. Conclusion The method established in this paper is accurate, sensitive and reliable and can be used for the separation of warfarin enantiomers and the determination of plasma concentration in human plasma.