目的建立HPLC-MS/MS法同时测定人血浆中R-华法林和S-华法林浓度。方法以乙酸乙酯为萃取剂,布洛芬为内标。分析柱:Lux cellulose-4柱(4.6 mm×250 mm,5μm);流动相:5 mmol醋酸铵水溶液(含0.4%甲酸)-乙腈=45∶55,流速:0.5 m L·min~(-1);质谱条件:电喷雾离子源,负离子模式,R-华法林、S-华法林和内标(IS)的离子通道分别为m/z 307.0→m/z 161.0、m/z 306.9→m/z 106.9和m/z 204.8→m/z 161.0,考察该方法的专属性、标准曲线和定量下限、精密度和准确度、基质效应、提取回收率和稳定性。结果 R-华法林标准曲线方程为Y=2.57×10~(-3)X+3.84×10~(-3)(R2=0.997 6),在10.00~500.0μg·L~(-1)内线性关系良好,最低定量下限为10.00μg·L~(-1)。S-华法林标准曲线方程为Y=2.46×10~(-3)X+4.60×10~(-3)(R2=0.996 8),在10.00~500.0μg·L~(-1)内线性关系良好,最低定量下限为10.00μg·L~(-1)。回收率、精密度、准确度、基质效应和样本稳定性符合要求。结论本文建立的方法灵敏可靠,可用于测定人血浆中华法林对映异构体浓度。 Objective To establish a HPLC-MS / MS method for the simultaneous determination of R-warfarin and S-warfarin in human plasma. Methods Ethyl acetate as extractant and ibuprofen as internal standard. The analytical column was Lux cellulose-4 column (4.6 mm × 250 mm, 5 μm). The mobile phase consisted of 5 mmol ammonium acetate aqueous solution containing 0.4% formic acid and acetonitrile at a flow rate of 0.5 m L · min -1 ); MS conditions: m / z 307.0 → m / z 161.0, m / z 306.9 → m / z for the ion channel of electrospray ion source, negative ion mode, R-warfarin, S- warfarin and IS m / z 106.9 and m / z 204.8 → m / z 161.0. The specificity, standard curve and quantitation limits, precision and accuracy, matrix effect, extraction recovery and stability of this method were investigated. Results The standard curve equation of R-warfarin was Y = 2.57 × 10 -3 X + 3.84 × 10 -3 (R2 = 0.997 6), and within 10.00 ~ 500.0 μg · L -1 The linear relationship was good with the lowest limit of quantitation as 10.00μg · L -1. The S-warfarin standard curve equation was Y = 2.46 × 10 -3 × + 4.60 × 10 -3 (R2 = 0.996 8). The calibration curve was linear over the range of 10.00-500.0 μg · L -1 The lower limit of quantitation was 10.00μg · L -1. Recovery, precision, accuracy, matrix effects and sample stability meet the requirements. Conclusion The established method is sensitive and reliable and can be used to determine the enantiomer concentration of warfarin in human plasma.