巯基化衍生结合分散液相微萃取-气相色谱/串联质谱-选择反应监测技术测定尿样中超痕量氯乙烯基胂酸  

Determination of ultra-trace 2-chlorovinylarsonic acid in human urine by GC/MS/MS(SRM)after thiolation and dispersive liquid-phase microextraction

作　　者：吴姬娜 刘石磊[1,2] 杨旸[1,2] 梁龙辉[1] 刘景全[1,2] 

机构地区：[1]防化研究院分析化学实验室,北京102205 [2]国民核生化灾害防护国家重点实验室,北京102205

出　　处：《国际药学研究杂志》2016年第5期909-914,921,共7页Journal of International Pharmaceutical Research

摘　　要：目的建立路易氏剂染毒尿样中超痕量氯乙烯基胂酸(CVAOA)的分析检测方法。方法采用正交实验,优化分散液相微萃取(DLPME)条件,优化结果为:将250μl甲醇(分散溶剂)、250μl乙酸乙酯(萃取溶剂)、3,4-二巯基甲苯(DMT)(衍生试剂,用量满足摩尔比CVAOA∶DMT=1∶100)混合液,加入到p H=1的1 ml尿样中,90℃衍生并萃取60 min,使用气相色谱/串联质谱-选择反应监测模式〔GC/MS/MS(SRM)〕对衍生产物进行定性定量分析。结果本法在50 pg/ml^1μg/ml浓度范围内呈现良好的线性(r2=0.9999),相对标准偏差(RSD)<10%,检出限可达18 pg/ml,定量限为56 pg/ml,远低于已报道的DLPME分析方法。对低、中、高3种浓度(0.5、5和50 ng/ml)的路易氏剂染毒尿样进行检测,准确度为98.2%~104%,RSD为6.9%~8.9%。结论本文所建方法灵敏度高,专属性好,准确度和精密度良好,且操作简单,适用于真实染毒样品分析。Objective To develop a method for determination of ultra-trace 2-chlorovinylarsonic acid（CVAOA）in human urine specific to lewisite exposure. Methods The conditions for dispersive liquid-phase microextraction（DLPME）were optimized by orthogonal experiments with results as follows：a mixture of 250μl methanol（dispersive solvent）,250μl ethyl acetate（extraction sol-vent）,and 3,4-dimercaptotoluene（DMT）（derivatization reagent,using 1∶100 mole ratio of CVAOA to DMT）was injected into 1 ml urine when pH was adjusted to 1. In the next 60 min,the CVAOA was derivated and the CVAOA-DMT derivative was extracted simul-taneously at 90℃. The derivative was then analyzed by gas chromatography/tandem mass spectrometry-select reaction monitoring〔GC/MS/MS（SRM）〕. Results The linear calibration extended from 50 pg/ml to 1μg/ml（r2=0.9999）with the relative standard deviations （RSD）less than 10%. The limit of detection was 18 pg/ml and the limit of quantitation was 56 pg/ml,which was much lower than that of the reported DLPME methods. When detecting human urine samples with low,middle and high spiked concentrations（0.5,5 and 50 ng/ml）of lewisite,the analysis accuracies ranged from 98.2%to 104%and the RSD ranged from 6.9%to 8.9%. Conclusion The method developed in this study has high specificity and sensitivity as well as good precision and accuracy. It is s imple and can be readi-ly applied to the exposed sample analysis.

关 键 词：氯乙烯基胂酸 路易氏剂 超痕量 分散液相微萃取 尿样 

分 类 号：R446.12[医药卫生—诊断学]