超高效液相色谱串联质谱法测定人血浆中苯溴马隆的质量浓度

摘要

目的 建立一种超高效液相色谱联用质谱法(UHPLC-MS/MS)测定人血浆中苯溴马隆药物的质量浓度.方法 以苯溴马隆EP Impurity B为内标,血浆经乙腈沉淀蛋白处理后,在Waters Acquity UPLC ?BEH C18色谱柱(50 mm×2.1 mm,1.7 μm)上进行分离;流动相为分别含1 mL·L -1甲酸的2 mmol·L -1醋酸铵水溶液和乙腈;采用大气压化学电离源APCI,负离子多反应监测(MRM),检测离子对分别为422.7→250.8(苯溴马隆)和502.6→250.9(苯溴马隆EP Impurity B).结果 苯溴马隆质量浓度在50~10000 ng·mL -1范围内线性关系良好(r=0.999),最低定量限(LLOQ)为50 ng· mL -1,平均回收率为100% 4.04%,低、中、高3种质控质量浓度的批内和批间精密度实验RSD值均小于5%.结论 该实验建立的分析方法具有专属性强、灵敏度好、简便、稳定等特点,适用于苯溴马隆人体内血药质量浓度的测定及其药代动力学和生物等效性研究.%Objective To establish a UHPLC-MS/MS method for the determination of benzbromarone in human plasma.Methods The plasma samples were extracted by a simple protein precipitation method using acetonitrile,benzbromarone EP Impurity B as internal standard.Separation was achieved on a Waters Acquity UPLC ?BEH C18column(50 mm×2.1 mm,1.7 μm)with a gra-dient elution of 1 mL· L -1formic acid and 2 mmol· L -1amonium acetate in water(mobile phase A)and acetonitrile(mobile phase B).Atmospheric pressure chemical ionization(APCI)source in negative ion mode was used for multiple reaction monitoring (MRM)with transitions of benzbromarone(422.7→250.8)and benzbromarone EP Impurity B(502.6→250.9),which was used as an internal standard.Results The calibration curve showed good linearity over the range of 50-10 000 ng·mL -1(r=0.999)in human plasma.The lower limit of quantification(LLOQ)was 50 ng·mL -1.The average recovery of benzbromarone was 100% 4.04%,the intra-batch and inter-batch RSDs of low,middle and high QCs were less than 5%.Conclusion The method has been demonstrated to be selective,sensitive,simple,rapid and robust,which can be applied in the determination of benzbromarone in hu-man plasma to support pharmacokinetic and bioequivalence study.