HPLC-MS/MS法检测血液中甲卡西酮及其代谢物

Determination of the methcathinone and its metabolites in blood samples by HPLC-MS/MS method

目的建立同时检测血液中新精神活性物质甲卡西酮及其代谢物卡西酮、麻黄碱和伪麻黄碱含量的高效液相色谱-串联质谱方法,验证甲卡西酮在大鼠体内的代谢物。方法血液样品中加入内标物甲卡西酮-D3,经甲醇提取后采用InfinityLab Poroshell 120 Chiral-V型色谱柱分离,以甲醇和乙腈混合流动相恒比洗脱,采用电喷雾离子源多反应监测模式,检测腹腔注射染毒大鼠血液中甲卡西酮及其代谢物。结果血中甲卡西酮及其代谢物10~1000ng/mL浓度范围内线性关系良好(r>0.999),检出限均小于2ng/mL,定量限为10ng/mL,方法准确度为87.06%~112.62%,批间及批内精密度均小于15%;腹腔注射染毒大鼠血中检出甲卡西酮、卡西酮、麻黄碱和伪麻黄碱。结论本研究建立了血液中甲卡西酮及其代谢物的HPLC-MS/MS定性、定量检测方法,初步验证卡西酮、麻黄碱和伪麻黄碱为甲卡西酮的代谢物。

Objective To establish a HPLC-MS/MS method for determination of new psychoactive substance that methcathinone and its metabolites cathinone,ephedrine and pseudoephedrine in blood,which provides a method of validation metabolite of methcathinone in rats.Methods The blood sample was added with the internal standard Methcathinone-D3,and after being extracted with methanol,it was separated on an Infinity Lab Poroshell 120 Chiral-V column.Methanol and acetonitrile mixed solution was used as the mobile phase.The flow rate was 0.5 mL/min.The electrospray ion source was used for detection in multiple reaction monitoring(MRM)mode.Results The calibration curves of Methcathinone and its metabolites were linear in the concentration range of 10~1000 ng/mL,the correlation coefficient(r)were all more than 0.999,the limit of detection were 2 ng/mL,the limit of quantification was 10 ng/mL the accuracy were 87.06%~112.62%,the intro-batch and intrabatch precision were less than 15%.Methcathinone,cathinone,ephedrine and pseudoephedrine were detected in the blood of infected rats after intraperitoneal injection.Conclusion In this study,an HPLC-MS/MS method for the determination of Methcathinone and its metabolites in blood samples was established.It was preliminarily verified that cathinone,ephedrine and pseudoephedrine were metabolites of methcathinone.