液相色谱-电喷雾串联质谱同时检测尿液和胃液中12种有毒生物碱

建立了高效液相色谱-电喷雾串联质谱同时检测尿液和胃液中12种有毒生物碱的方法.优化了提取条件及色谱-质谱条件,并考察了基质效应的影响,探讨了质谱碎裂机理.尿液和预先调节至中性的胃液经硼砂-NaOH缓冲液(pH 9.6)碱化,乙酸乙酯液液萃取,采用电喷雾电离(ESI+)、多反应监测(MRM)方式,可同时对黄华碱、倒千里光碱、山莨菪碱、钩吻碱、芦竹碱、哈尔碱、吐根碱、血根碱、吴茱萸碱、吴茱萸次碱、雷公藤吉碱和雷公藤次碱12种有毒生物碱进行定性和定量分析.在优化的条件下,12种成分分别在0.5～200 μg/L、1～200 μg/L和5～200μg/L范围内线性关系良好,尿液中除黄华碱和山莨菪碱外,各生物碱的回收率为61.9％～119.1％,胃液中各生物碱回收率为61,0％～1102％,精密度RSD＜ 15％.检出限(LOD)为0.1～0 Sμμg/L,定量限(LOQ)为0.5～5.0 μg/L,时回收率不高的生物碱可通过空白基质配制标样校正,满足定量分析的要求.本方法操作简便、快捷、灵敏度高,适用于中毒患者尿液和胃液中有毒生物碱成分的检测.

Simultaneous Determination of 12 Toxic Alkaloids in Urine and Gastric Juice Using Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry

A new method has been established for the rapid and simultaneous analysis of 12 alkaloids,thermopsine,retrorsine,anisodamine,gelsemine,gramine,harmine,emetine,sanguinarine,evodiamine,rutaecarpine,wilforine and wilforgine in urine and gastric juice samples.The conditions of extraction,chromatography and mass spectrometry were optimized,and the matrix effects and fragmentation mechanism of the alkaloids were also examined.The samples were first treated with Na 2 B 4 O 7-NaOH buffer solution(pH 9.6),then extracted by ethyl acetate twice.After evaporation,the residues were reconstituted in methanol,from which 5 μL was used for detecting by liquid chromatography-electrospray ionization tandem mass spectrometry in positive ionization mode(ESI +).The linear range was 5-200 μg/L for emetine and sanguinarine,0.5-200 μg/ L for harmine and 1-200 μg / L for other alkaloids.The mean recoveries ranged from 61.0% to 119.1% in urine samples expect thermopsine and anisodamine,and 61.0% to 110.2% in gastric juice samples.The relative standard deviations(RSDs) were less than 15%,and the limits of detection(LOD) and quantification(LOQ) varied from 0.1 to 0.5 μg/L and 0.5 to 5 μg/L.The method,proved to be rapid,sensitive and easy to handle,was suitable for the determination of trace alkaloids in urine and gastric juice samples of poisonous patients.