超高效液相色谱-串联质谱法快速同步测定人血中35种毒(药)物及主要代谢产物  

Rapid and simultaneous determination of 35 drugs and their main metabolites in human blood by grinding scroll oscillation extraction coupled with ultra-high performance liquid chromatography-tandem mass spectrometry

作　　者：陈顺琴[1] 王爱民[1,2] 黄江[1] 王杰[1] 夏冰[1] 朱发泽 刘玉波[1] 罗鹏[1] 王莎莎 张秋娜 CHEN Shunqin;WANG Aiming;HUANG Jiang;WANG Jie;XIA Bing;ZHU Faze;LIU Yubo;LUO Peng;WANG Shasha;ZHANG Qiuna(School of Forensic Medicine,Guizhou Medical University,Guiyang 550004,China;Provincial Key Labratary of Pharmaccutics in Guizhou Province,Guizhou Medical University,Guiyang 550004,China;School of Pharmaceutical Sciences,Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学法医学院,贵阳550004 [2]贵州医科大学贵州省药物制剂重点实验室,贵阳550004 [3]贵州医科大学药学院,贵阳550004

出　　处：《分析试验室》2022年第6期691-697,共7页Chinese Journal of Analysis Laboratory

基　　金：贵州省高层次“百”层次创新型人才项目(黔科合平台人才[2020]6012);贵州省科技计划项目(黔科合支撑[2019]2825号、黔科合支撑[2020]4Y057号);贵州省高技术产业化示范工程项目(黔发改高技[2016]1345号)资助。

摘　　要：建立了超高效液相色谱-串联质谱法(UPLC-MS/MS)快速测定人血中常见35种毒(药)物及主要代谢产物的方法。取1 mL血液样品,加入5颗研磨珠、40 mg(NH_(4))_(2)SO_(4)、1.5 mL乙腈进行提取,涡旋振荡60 s,静置60 s,冷冻离心后取上清液体过0.22μm有机滤膜后进行分析。结果表明,目标化合物在10~1000 ng/mL质量浓度范围呈良好的线性关系(R^(2)为0.9921~0.9998),检出限(S/N≥3)为0.05~2.00 ng/mL,目标毒(药)物定量限(S/N≥10)为20 ng/mL占比为17%,其余目标毒(药)物定量限均为10 ng/mL。在20,50和100 ng/mL加标浓度水平下,目标化合物回收率为71.5%~119.8%,日内相对标准偏差(Intra-RSD)为1.1%~13.8%。在50 ng/mL加标水平下,日间相对标准偏差(Inter-RSD)为2.6%~13.4%。该方法适用于血液中毒(药)物及代谢产物的快速分析测定。A method for the rapid determination of 35 common toxins and metabolites in human blood by grinding vortex oscillation extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was established.The following pretreatment steps were ruquired in this method.The first step was taking 1 mL blood sample,then 5 grinding beads,40 mg salting-out agent(NH_(4))_(2)SO_(4) and 1.5 mL acetonitrile were added for extraction.The next steps followed by grinding the sample and vortex for 60 s,put it for 60 s,refrigerated centrifugation and taking the supernatant liquid for filtration with 0.22 μm organic filter membrane for detection employed by UPLC-MS/MS.The results showed that the 35 kinds of toxicants and drugs presented good linear relationships in the mass concentration range of 10 ng/mL to 1000 ng/mL.The correlation coefficients(R~2)were between 0.9921 and 0.9998.Meanwhile the limits of detection(S/N≥3)of the method were in the range of 0.05-2 ng/mL.Among 35 kinds of toxicants and drugs,17% of the target compounds had limit of quantification(S/N≥10)of 20 ng/mL,while the rest of 10 ng/mL.The average recoveries of the 35 kinds of toxicants and drugs ranged from 71.5% to 119.8% with the intra-day relative standard deviations(Intra-RSDs)ranging from 1.1% to 13.8% by taking blank blood samples at three spiked concentration levels of 20,50 and100 ng/mL.And the inter-day relative standard deviations(Inter-RSD)ranged from 2.6% to 13.4% at the spiked concentration level of 50 ng/mL.The method is suitable for rapid simultaneous determination of 35 kinds of commonly found toxicants and drugs in human blood.

关 键 词：蛋白沉淀 研磨 乙腈 UPLC-MS/MS 人血 毒(药)物 

分 类 号：R991[医药卫生—毒理学]