State Key Laboratory of NBC Protection for Civilian, Laboratory of Analytical Chemistry, Research Institute of Chemical Defence, Beijing 102205, P. R. China.
School of Chemistry and Chemical Engineering, Nanjing University of Sciences & Technology, Nanjing, 210094, P. R. China.
J Chromatogr A. 2022 Aug 16;1678:463354. doi: 10.1016/j.chroma.2022.463354. Epub 2022 Jul 19.
Sulfur mustard (HD) is a highly toxic vesicant and is prohibited by the Organisation for the Prohibition of Chemical Weapons (OPCW). HD can modify human serum albumin (HSA) to generate hydroxyethylthioethyl (HETE) adducts, which could be utilized as biomarkers for verifying HD exposure in forensic analysis. Here, five amino acid adducts generated from pronase digestion of HD-exposed human serum albumin (HD-HSA) in plasma were selected as biomarkers to retrospectively detect HD exposure. HD-HSA was precipitated from plasma with acetone, digested by pronase, derivatized with propionic anhydride (PA), and analysed with ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-TQ MS). The limits of detection (LODs) and limits of quantification (LOQs) of the HD exposure concentrations were evaluated as 1.00 ng/mL at S/N≥3 and 3.00 ng/mL at S/N≥10, respectively, which are approximately 60 times lower than those of the reported method. The approach shows good linearity (R≥0.997) from 3.00 ng/mL to 10.0 µg/mL of HD-exposed human plasma with satisfactory precision and accuracy. The developed approach was applied to analysing samples from the 6 OPCW Biomedical Proficiency Test (BioPT). The study showed that the developed approach was also suitable for analysing human plasma samples that were exposed to six of HD analogues, which were common impurities in sulfur mustard mixtures. Moreover, the method was successfully applied to plasma from other species, including rabbits, rats and cattle. This study provides a reliable and sensitive tool for the retrospective detection of vesicants exposure based on multiple biomarkers.
硫芥（HD）是一种剧毒的糜烂性毒剂，被禁止化学武器组织（OPCW）所禁止。HD 可以修饰人血清白蛋白（HSA）以生成羟乙基硫乙基（HETE）加合物，这些加合物可用于法医分析中验证 HD 暴露的生物标志物。在这里，选择从暴露于 HD 的人血清白蛋白（HD-HSA）在等离子体中经糜蛋白酶消化生成的五个氨基酸加合物作为生物标志物，以回溯检测 HD 暴露。HD-HSA 用人血浆中的丙酮沉淀，用糜蛋白酶消化，用丙酸酐（PA）衍生化，并用超高效液相色谱-三重四极杆质谱联用（UHPLC-TQ MS）进行分析。HD 暴露浓度的检测限（LOD）和定量限（LOQ）分别评估为 S/N≥3 时为 1.00 ng/mL，S/N≥10 时为 3.00 ng/mL，大约比报道的方法低 60 倍。该方法在 3.00 ng/mL 至 10.0 µg/mL 的 HD 暴露人血浆中显示出良好的线性关系（R≥0.997），具有令人满意的精密度和准确度。所开发的方法用于分析来自 6 次 OPCW 生物医学能力测试（BioPT）的样品。研究表明，该方法也适用于分析 6 种 HD 类似物暴露的人血浆样品，这些类似物是硫芥混合物中的常见杂质。此外，该方法还成功应用于其他物种的血浆，包括兔子、大鼠和牛。这项研究为基于多种生物标志物的回溯性检测糜烂性毒剂暴露提供了一种可靠、敏感的工具。
