液相色谱-四极杆飞行时间质谱检测染毒动物血浆中沙林和梭曼酪氨酸加合物  

Determination of Sarin and Soman Tyrosine Adducts in Exposed Animal Plasma by Liquid Chromatography / Quadrupole-Time of Flight Mass Spectrometry

作　　者：于惠兰[1,2] 裴承新[1,2] 胡真[2] 刘石磊[1,2] 向宇[2] 

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

出　　处：《质谱学报》2015年第3期261-267,共7页Journal of Chinese Mass Spectrometry Society

摘　　要：为了追溯性检测神经性毒剂沙林(GB)和梭曼(GD)暴露染毒,以染毒白蛋白上的特异性加合位点-411位酪氨酸加合物为重要生物标志物,建立了高效液相色谱-四极杆飞行时间质谱(LC/Q-TOF MS)分析动物血液中神经性毒剂GB和GD酪氨酸加合物的检测方法。在采用不同样品处理和仪器条件的基础上,确定了最佳实验条件。血液经离心后,取50μL分离的血浆加入到50μL 10 g/L链霉蛋白酶的NH4HCO3(50 mmol/L)水溶液中,于37℃孵育2.5 h,用0.5 m L 10 ku超滤管以16 000 r/min离心15min,LC/Q-TOF MS电喷雾正离子模式分析滤液。应用本方法进行GB半致死剂量(LD50)动物施毒实验,在施毒的动物血样中检测到膦酰化酪氨酸加合物,证明了该方法快速、简单、可靠。In order to retrospective analysis of biomedical samples exposed to nerve agents sarin （GB） and soman （GD）, a method for determination of GB tyrosine adducts and GD tyrosine adducts in exposed animal plasma as a biomarker was developed by liquid chromatography/quadrupole-time of flight mass spectrometry（LC/Q-TOF MS）. After optimizing different sample preparation processes and a series of instrumental parameters, the adducts were analyzed by LC/Q-TOF MS in positive electrospray ioni- zation mode. After centrifugation, 50 μL plasma was added into 50 μL 10 g/L Pronase E in ammonium bicarbonate（50 mmol/L）and incubating at 37 ℃ for 2.5 h. The super- natant was analyzed after filtered through 0.5 mL molecular mass cutoff filters （10 ku）under centrifugation at 16 000 r/min for 15 rain. The method is rapid, simple and relia-ble, which is suitable for analyzing the phosphylated tyrosine adducts in blood samples from the rabbits injected with LD50 of GB.

关 键 词：生物标志物 神经性毒剂 沙林(GB) 梭曼(GD) 血浆 酪氨酸加合物 液相色谱-四极杆飞行时 间质谱(LC/Q-TOF MS) 

分 类 号：O657.63[理学—分析化学]