血浆中米酵菌酸的高效液相色谱-串联质谱快速测定法

Fast determination of bongkrekic acid in plasma by high performance liquid chromatography-tandem mass spectrometry

目的建立高效液相色谱-串联质谱(HPLC-MS/MS)快速检测血浆中米酵菌酸含量的方法。方法于2020年11月,血浆样品经甲醇-乙腈溶液(体积比1∶1)提取净化后直接进样,经C18色谱柱分离,以5 mmol/L乙酸铵水-乙腈溶液为流动相梯度洗脱,在优化后的仪器条件下,采用电喷雾负离子多反应监测(MRM)模式,外标法定量,并对所建立的方法进行方法学验证。结果血浆中米酵菌酸在2~100μg/L线性关系良好,相关系数为0.9998,平均回收率为83.7%~112.0%,批内、批间相对标准偏差均<10%;方法的检出限为0.7μg/L,定量下限为2.0μg/L。结论该方法简便、快速、准确,灵敏度高,可满足中毒患者血样中米酵菌酸测定的要求。

Objective To establish a method for rapid determination of bongkrekic acid(BA)in plasma by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).Methods In November 2020,plasma samples were extracted by methanol and acetonitrile(1∶1)and purified directly.The samples were separated by C18 column.Gradient elution was carried out with 5 mmol/L ammonium acetate water acetonitrile solution as mobile phase.Under the optimized instrument conditions,the electrospray ionization multiple reaction monitoring(MRM)mode was used,and the external standard method was used for quantitative analysis.Results The linear relationship of BA in plasma was good in the concentration range of 2-100μg/L,the correlation coefficient was 0.9998,the average recovery was 83.7%-112.0%,the relative standard deviation within and between batches was less than 10%,the detection limit of the method was 0.7μg/L and the lower limit of quantification was 2.0μg/L.Conclusion The method is simple,rapid,accurate and sensitive,and can meet the requirements for the determination of BA in blood samples of poisoning patients.