摘要
针对神经性毒剂梭曼暴露后生物医学样品的准确性溯源分析这一迫切需求,建立了高效液相色谱-串联质谱技术定量分析染毒大鼠血浆中梭曼-酪氨酸加合物的高灵敏度高特异性新方法。本研究建立了有效的亲和分离富集及超滤提取白蛋白、链霉蛋白酶酶解和固相萃取等前处理技术方法,在反相液相色谱-三重四极杆质谱及正离子多反应监测模式下,对梭曼-酪氨酸加合物进行了定性和定量分析,检测离子对分别为m/z260/214（梭曼-酪氨酸加合物）和m/z293/247（内标）。梭曼-酪氨酸加合物在0.1～500μg/L范围内呈良好的线性关系（r2=0.999）,检出限为0.02μg/L,定量限为0.05μg/L,回收率为90.1%～103.5%,其日内和日间精密度均小于3%。对于以腹腔注射方式染毒后的SD大鼠（n=5）,考察了梭曼-酪氨酸加合物的代谢动力学。结果表明,梭曼-酪氨酸加合物与染毒的梭曼剂量之间呈现较为明显的时效、量效关系,说明梭曼-酪氨酸加合物可作为梭曼染毒后的暴露性生物标志物,可用于梭曼暴露后的溯源性检测。
Towards the urgent need of accurately retrospective analysis of biomedical samples exposed by nerve agent soman （ GD ）, we developed a exposed and high rat plasma by liquid chromatography new quantitative method of GD-tyrosine adduct （GD-Tyr） in （LC）-tandem mass spectrometry （MS/MS） with high sensitivity specificity. After a series of pretreatment procedures including extraction of albumin by affinity chromatography and ultrafiltration, enzymatic hydrolysis by pronase, and solid phase extraction, we qualitatively and quantitatively determined soman-tyrosine adduct by reversed-phase LC-triple quadrupole MS in the positive electrospray ionization and multiple reactions monitoring mode. The monitored transitions were m/z 260/214 （GD-Tyr） and m/z 293/247（ internal standard）. A good linearity was obtained in the range of 0.1-500 μg/L for GD-Tyr （ R2 = 0.999 ） with a limit of detection of 0.02 μg/L and a limit of quantification of 0.05 μg/L. The recovery of GD-Tyr was in the range of 90.1%-103.5% , and the RSD of this method was less than 3%. We then investigated the toxicokinetics of GD-Tyr for GD exposed SD rat （n = 5 ） in an intraperitoneal injection way. An obvious time and dose dependent relationship between GD-Tyr and the dose of soman was observed. We suggest that GD-tyrosine adduct can be used as an exposure biomarker to the retrospective detection of soman after exposure.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2013年第12期1801-1806,共6页
Chinese Journal of Analytical Chemistry
基金
国家"重大新药创制"科技重大专项"十二五"滚动项目(No.2012ZX09301003-001-010
2012-2015)
全军医学科技"十二五"重大研究项目(No.AWS11C004-05)资助
关键词
梭曼
梭曼-酪氨酸加合物
液质联用
大鼠血浆
毒代动力学
Soman
Soman-tyrosine adduct
Liquid chromatography-tandem mass spectrometry
Plasma
Toxicokinetics
