Department of Anthropology, University of Florida, 1112 Turlington Hall, Gainesville, FL 32611.
Defense POW/MIA Accounting Agency, 590 Moffet Street, Joint Base Pearl Harbor-Hickam, HI 96853.
J Anal Toxicol. 2020 May 18;44(4):391-401. doi: 10.1093/jat/bkz118.
A recently proposed model for the incorporation of xenobiotics of forensic interest into the human skeleton suggests nerve agent metabolites may incorporate into bone at relatively elevated concentrations based on their unique chemical properties. To test the hypothesis that nerve agent metabolites interact with bone, methods for the extraction, isolation and semi-quantitative detection of nerve agent metabolites (MPA, EMPA, IMPA, iBuMPA, CMPA and PMPA, corresponding to the nerve agents VX, Russian VX, sarin, cyclosarin and soman, respectively) from osseous tissue were developed using liquid chromatography-mass spectrometry with both quadrupole time-of-flight and triple quadrupole (QqQ) instruments. The optimized methods were validated on the QqQ instrument. Despite high ion suppression, the achieved limits of detection (5-20 pg/g for four analytes; 350 pg/g for the fifth analyte) were lower than many of those published for the same analytes in other biomatrices, including serum and urine. These methods were tested on the skeletal remains of minipigs exposed to the chemical weapon VX in vivo. The VX metabolite was detected in multiple minipig bone samples; to the authors' knowledge, this is the first time in vivo nerve agent exposure has been detected from bone. Further, detected concentrations and diaphyseal-to-epiphyseal area count ratios reflect animal exposure history. Although the results are limited, they are promising, indicating that nerve agent metabolites may interact with bone as a pharmacokinetic compartment and can be extracted from bone postmortem. Additional studies, assessing the effects of different agents, exposure pathways and taphonomic variables, are needed; however, these results suggest the method may be used with human bone to detect use of chemical weapons from postmortem biomatrices even well after a suspected attack. More general implications for both nerve agent toxicology and skeletal toxicology are also discussed.
最近提出的一个关于将法医感兴趣的外源化学物纳入人体骨骼的模型表明，根据其独特的化学性质，神经毒剂代谢物可能以相对较高的浓度纳入骨骼。为了测试神经毒剂代谢物与骨骼相互作用的假设，开发了从骨骼组织中提取、分离和半定量检测神经毒剂代谢物（MPA、EMPA、IMPA、iBuMPA、CMPA 和 PMPA，分别对应 VX、俄罗斯 VX、沙林、环沙林和梭曼神经毒剂）的方法，采用液相色谱-质谱法，同时使用四极杆飞行时间和三重四极杆（QqQ）仪器。优化后的方法在 QqQ 仪器上进行了验证。尽管存在高离子抑制，但达到的检测限（四种分析物为 5-20 pg/g；第五种分析物为 350 pg/g）低于许多在其他生物基质（包括血清和尿液）中发表的相同分析物的检测限。这些方法在体内暴露于化学武器 VX 的小型猪的骨骼遗骸上进行了测试。在多个小型猪骨样本中检测到 VX 代谢物；据作者所知，这是首次从骨骼中检测到体内神经毒剂暴露。此外，检测到的浓度和骨干到骨骺区域计数比反映了动物暴露史。尽管结果有限，但很有希望，表明神经毒剂代谢物可能作为药代动力学隔室与骨骼相互作用，并且可以从死后骨骼中提取。需要进行更多的研究，评估不同试剂、暴露途径和埋藏学变量的影响；然而，这些结果表明，该方法可以用于检测从死后生物基质中使用化学武器的人类骨骼，即使在疑似袭击后很久。还讨论了神经毒剂毒理学和骨骼毒理学的更普遍意义。
