摘要
目的建立超高效液相色谱-串联质谱法检测中毒生物样本尿液中23种生物碱,为生物碱中毒检测和治疗提供可靠依据。方法试样加入硼砂-氢氧化钠缓冲液(pH=9.6)碱化,再加入乙酸乙酯提取,离心后取上清液用氮气吹至近干,再用初始流动相含0.1%乙酸和2 mmol/L乙酸铵的水溶液-含0.1%乙酸和2 mmol/L乙酸铵的乙腈溶液(90∶10)复溶,过0.22μm的微孔滤膜,待测物经Dikma leapsil C18柱(2.1 mm×50 mm×2.7μm)分离,用液质ESI源、多反应监测模式检测。结果在优化的条件下,该方法线性范围为0~100μg/L,相关系数R在0.9945~0.9998,尿液的检出限(LOD,S/N=3)为0.1~0.2μg/L,相对标准偏差(RSD)0.8%~10%,样品加标回收率在60%~115%之间。结论该方法快速、灵敏、准确,可应用于突发应急事件中检测人尿中多种生物碱,为临床救治提供可靠的依据。
Objective A method for the determination of 23 alkaloids in urine by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was established to provide a reliable basis for the alkaloid poisoning detection and treatment.Methods The samples were alkalized by adding borax-sodium hydroxide buffer(pH=9.6),and then extracted with ethyl acetate.After centrifugation,the supernatant was taken up to near dryness with nitrogen,and then was reconstituted by using the initial mobile phase contained 0.1%acetic acid and 2 mmol/L ammonium acetate aqueous solution,which was the acetonitrile solution(90∶10)containing 0.1%acetic acid and 2 mmol/L ammonium acetate,and passed through a 0.22μm micro-porous membrane.The analytes was subjected to a separation by the Dikma leapsil C18 column(2.1 mm×50 mm×2.7μm)and detected by the liquid ESI source and positive ionization multiple reaction monitoring mode.Results Under the optimized conditions,the method showed good linearity in the range of 0-100μg/L,with correlation coefficient of R in the range of 0.9970~09998.The limit of detection(LOD,S/N=3)were in the range of 0.1~0.2μg/L,and the relative standard deviation(RSD)was about0.8%~10%.The Sample spiked recovery rate was ranged from 60%to 115%.Conclusion This method is fast,sensitive and accurate,it could be used to detect a variety of alkaloids in human urine in emergency situations,providing a reliable basis for clinical treatment.
作者
罗兰
刘红河
王甫
任燕
林启辉
LUO Lan;LIU Honghe;WANG Fu;REN Yan;LIN Qihui(Longhua District Center for Disease Control and Prevention,Shenzhen 518109,Guangdong Province,China;Shenzhen Center for Disease Control and Prevention,Shenzhen 518055,Guangdong Province,China)
出处
《预防医学情报杂志》
CAS
2019年第8期832-839,844,共9页
Journal of Preventive Medicine Information
基金
深圳市卫生和计划生育委员会研究项目(项目编号:201507081)
深圳市龙华区科技计划重点实验资助项目“公共卫生应急检测重点实验室”(项目编号:20160825A041006)。
关键词
超高效液相色谱-串联质谱法
生物碱
尿液
ultra performance liquid chromatography-tandem mass spectrometry
alkaloids
urine
