LC-MS/MS法同时检测生物检材中钩吻素子、钩吻素甲和钩吻素己  

Simultaneous Quantitative Analysis of Koumine,Gelsemine and Gelsenicine in Biological Samples by LC-MS/MS

作　　者：姬圣洁 刘伟[2] JI Sheng-jie LIU Wei(Department of Forensic Medicine, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China Shanghai Key Laboratory of Forensic Medicine, Shanghai Forensic Service Platform, Institute of Forensic Science, Ministry of Justice, PRC, Shanghai 200063, China)

机构地区：[1]复旦大学基础医学院法医学系,上海200032 [2]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市司法鉴定专业技术服务平台,上海200063

出　　处：《法医学杂志》2017年第2期141-147,共7页Journal of Forensic Medicine

基　　金：十三五国家重点研发计划项目(2016YFC0800706);上海市法医学重点实验室资助项目(17DZ2273200);上海市司法鉴定专业技术服务平台资助项目(16DZ2290900)

摘　　要：目的建立准确、灵敏的生物检材中钩吻素子、钩吻素甲及钩吻素己的液相色谱-串联质谱(LCMS/MS)检测方法,并进行方法学验证。方法以士的宁为内标,血液、尿液及肝组织样品经1%氢氧化钠溶液碱化后用乙酸乙酯提取,采用ZORBAX SB-C_(18)柱(150 mm×2.1 mm,5μm)分离,以甲醇-20 mmol/L乙酸铵缓冲溶液(含0.1%甲酸和5%乙腈)为流动相进行梯度洗脱。定性定量分析采用电喷雾正离子化(ESI+)、多反应监测模式。结果血液、尿液及肝组织中钩吻素子、钩吻素甲和钩吻素己在相应的线性范围内线性良好,相关系数(r)>0.995 0,检出限分别为0.1 ng/m L(或0.1 ng/g)、0.1 ng/m L(或0.1 ng/g)及0.01 ng/m L(或0.01 ng/g),各生物碱提取回收率为61.9%~114.6%,准确度为92.4%~114.3%,日内、日间精密度的相对标准偏差均不超过11.0%。结论本方法选择性好、灵敏度高,适用于同时检测生物体液和组织中的钩吻素子、钩吻素甲和钩吻素己,可为钩吻中毒的临床诊治和法医学鉴定提供有效的技术支撑。Objective To establish a LC-MS/MS method which is accurate and sensitive for determina-tion of koumine, gelsemine, and gelsenicine in biological samples and to verify the method. Methods Strychnine was used as internal standard. Analytes in blood, urine and liver with 1% sodium hydroxide solution were extracted by ethyl acetate. Chromatographic separation was achieved on a ZORBAX SB- C18 column （150 mm×2.1 mm,5μm）, and gradient elution was performed with the buffer solution of methanol-20mmol/L ammonium acetate （including 0.1% formic acid and 5% acetonitrile） as mobile phase. Qualitative and quantitative analysis was performed in the multiple reaction monitoring mode coupled with an electrospray ionization source under positive ion mode （ESI＋）. Results The linearity of koumine, gelsemine and gelsenicine in blood, urine and liver was good within corresponding linear limitation and the correlation coefficients （r） 〉0.995 0. The limits of detection were 0.1ng/mL （0.1ng/g）, 0.1 ng/mL （0.1ng/g） and 0.01 ng/mL （0.01 ng/g）, respectively. The extraction recovery and accuracy of the alkaloids ranged from 61.9% to 114.6% and 92.4% to 114.3%, respectively. The relative standard deviations of the intra-day and inter-day precisions were not more than 11.0%. Conclusion The method is selective, sensitive and suitable for simultaneous determination of koumine, gelsemine and gelsenicine in body fluids and tissues, which offering technical support for clinical diagnosis and treatment and forensic toxicological analysis of Gelsemium elegans poisoning.

关 键 词：法医毒理学 液相色谱-串联质谱法 钩吻素子 钩吻素甲 钩吻素己 生物检材 

分 类 号：R286.95[医药卫生—中药学]