液相色谱-串联质谱法测定生物检材中乌头生物碱  

Determination of aconitum alkaloids in biological samples by liquid chromatography-tandem mass spectrometry

作　　者：刘伟[1] 沈敏[1] 吴何坚[1] 向平[1] 

机构地区：[1]司法部司法鉴定科学技术研究所,上海市法医学重点实验室,上海200063

出　　处：《药物分析杂志》2009年第12期2023-2028,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：中央级公益性科研院所专项基金资助项目(GY0702);上海市科委资助项目(072512019)

摘　　要：目的:建立检测生物检材中乌头生物碱的液相色谱-串联质谱(LC-MS/MS)法。方法:取血液、尿液或肝组织0.5mL(g),以丁丙诺啡为内标,经pH9.2的硼砂溶液碱化后,用乙醚进行提取;采用Capcell Pak MG-ⅡC18柱(250mm×2.0mm,5μm),以乙腈-pH4的20mmol·L-1乙酸铵和0.1%甲酸的缓冲液(70∶30)为流动相,流速0.2mL·min-1,采用电喷雾正离子模式离子化、多反应监测模式检测乌头生物碱,内标法定量。结果:血液、尿液和肝中乌头碱、次乌头碱、新乌头碱与内标丁丙诺啡色谱分离良好,乌头生物碱在0.1～50ng·mL-1(ng·g-1)内均具有良好的线性,相关系数>0.993,最低检出限为0.01～0.1ng·mL-1(ng·g-1),日内与日间精密度RSD均小于13.3%,血液中乌头生物碱提取回收率不小于64.8%。结论:所建LC-MS/MS方法灵敏度高、操作简便、快速、准确,适用于血液、尿液及组织等生物检材中痕量乌头生物碱成分的检测。Objective：To establish a method for determination of aconitum alkaloids in biological samples by LC-MS/MS.Methods：After 0.5 mL（g） of biological sample was extracted using a liquid-liquid extraction with diethyl ether at pH 9.2 and buprenorphine as an internal standard,aconitum alkaloids were separated on a Capcell Pak MG-Ⅱ C18 column （250 mm×2.0 mm,5 μm） using acetonitrile-buffer solution（20 mmol·L-1 ammonium acetate and 0.1% formic acid,pH 4） （70∶30） as the mobile phase at a flow-rate of 0.2 mL·min-1 with LC-MS/MS in the positive ionization mode.Quantitative results were obtained by the internal calibration procedure.Results：Aconitine,hypaconitine,mesaconitine and buprenorphine in blood,urine and liver were separated well.Calibration curve was linear within the range of 0.1-50 ng·mL-1（ng·g-1） with the LOD range from 0.01 to 0.1 ng·mL-1（ng·g-1）.The inter-day and intra-day RSDs were less than 13.3%.The extraction recoveries of aconitum alkaloids in blood were more than 64.8%.Conclusion：This method is sensitive,fast and accurate for determination of trace aconitum alkaloids in biological samples.

关 键 词：法医毒理学 液相色谱-串联质谱法 乌头生物碱 生物检材 

分 类 号：R917[医药卫生—药物分析学]