液相色谱-电喷雾串联质谱同时检测血液中8种有毒生物碱

Simultaneous Determination of 8 Poisonous Alkaloids in Human Blood by Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry

建立了高效液相色谱-电喷雾串联质谱同时检测血液中8种有毒生物碱的方法。血液样品经过乙酸铵-氨水缓冲溶液(pH≈9)处理后,以甲醇提取,采用电喷雾电离(ESI)、多反应监测(MRM)方式,可同时对麻黄碱、毛果芸香碱、士的宁、阿托品、钩吻素子、马钱子碱、乌头碱、喜树碱8种有毒生物碱进行定性和定量分析。在优化的条件下,上述前4种组分在1～100μg/L范围内线性关系良好,后4种组分在2.5～500μg/L范围内线性关系良好。8种有毒生物碱的提取回收率在83.1%～104.0%范围内。士的宁、阿托品、马钱子碱、乌头碱和钩吻素子检出限为0.05μg/L,定量限为0.1μg/L;麻黄碱、毛果芸香碱、喜树碱的检出限为0.1μg/L,定量限为0.5μg/L。各组分的日内RSD小于9%,日间RSD小于10%。本方法操作简便快捷、选择性好、灵敏度高,适用于中毒诊断和法医毒物分析。

A novel method was developed and validated for the rapid and simultaneous determination of 8 poisonous alkaloids,ephedrine,pilocarpine,strychnine,atropine,brucine,aconitine,camptothecin,and koumine,in human blood using liquid chromatography-electrospray ionization tandem mass spectrometry in the multiple reaction monitoring（LC-ESI-MRM） mode.The alkaloids in blood samples,treated with ammonium acetate-ammonia buffer solution of pH≈9,were extracted by methanol.The conditions of extraction,chromatography and mass spectrometry were optimized.The linearity of first 4 compounds were held in the ranges of 1-100 μg/L,while others were 2.5-500 μg/L.The average extraction recoveries were in the range of 83.1%-104.0%.The limits of detection（LOD） and quantification（LOQ） of strychnine,atropine,brucine,aconitine,and koumine were 0.05 μg/L and 0.1 μg/L,respectively,while the other 3 poisonous were 0.1 μg/L and 0.5 μg/L,respectively.The intra-day relative standard deviation（RSD） was less than 9%,while the inter-day relative standard deviation was less than 10%.The method,proved to be simple,rapid,sufficiently selective and sensitive,was suitable for poisoning diagnosis and forensic toxicology analysis.