液相色谱-串联质谱法测定血液和尿液中秋水仙碱  

Determination of colchicine in urine and blood by liquid chromatography-tandem mass spectrometry with multiple-reaction monitoring

作　　者：刘伟[1,2] 沈敏[2] 沈保华[2] 严慧[2] 

机构地区：[1]复旦大学上海医学院法医学系,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200063

出　　处：《中国法医学杂志》2009年第1期4-7,共4页Chinese Journal of Forensic Medicine

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

摘　　要：目的建市检测血液和尿液中秋水仙碱的液相色谱．串联质谱法。方法0．5mL血液或尿液以丁丙诺啡为内标，经pH9．2硼酸盐缓冲溶液碱化后，用乙酸乙酯进行提取，在ZORBAXSB—C18液相柱（150mm×2．1mm05μm）上以V（甲醇）：V（20mmol／L乙酸铵和0．1％甲酸缓冲溶液）=80：20为流动相，流速为0．2mL／min，采用电喷雾正离子模式离子化、多反应监测模式检测秋水仙碱，内标法定量。结果血液、尿液中秋水仙碱与内标丁丙诺啡色谱分离良好，秋水仙碱在0．1—50ng／mL内均具有良好的线性，相关系数〉0．9990，最低枪出限为0．05ng／mL，方法回收率为94％～116％，日内与日间精密度（RSD）均小于8．5％。结论所建LC—MS—MS方法灵敏度高、操作简便、快速、准确，适用于血液及尿液等生物检材中痕量秋水仙碱成分的检测。Objective To establish a method of liquid chromatography-tandem mass spectrometry for determination of colchicine in blood and urine. Methods 0.5mL of blood or urine sample was extracted usinga liquid-liquid extraction （ethyl acetate ） at pH 9.2 with buprenorphine as internal standard. The extract was separated on a ZORBAX SB-C18 column （150mm x2. lmm ~51xm） using 20 mmol/L ammonium acetate with 0.1% formic acid （pH 4 ）/methanol （20/80） as mobile phase with a flow-rate of 0.2mL/min, confirmed and quantified by MS-MS in the multiple reaction monitoring（MRM） mode via positive electrospray ionization mode（ ＋ ESI）. Results Colchicine and buprenorphine in blood and urine were separated well. Calibration curves were linear within 50ng/mL（ r 〉 0. 9990） , and the the range of 0.1 limits of detection were 0.05ng/mL. The recoveries were between 94% 116%, the inter-day and intra-day precisions were less than 8.5%. Conclusion This method is sensitive,fast and accurate, and suitable for determination of trace colchicine in blood and urine.

关 键 词：法医毒物分析 液相色谱-串联质谱法(LC-MS-MS) 秋水仙碱 尿液 血液 

分 类 号：D919[医药卫生—法医学]