Department of Analytical, Bioanalytical Sciences and Miniaturization (LSABM) Chemistry, Biology and Innovation (CBI), ESPCI Paris, PSL University, CNRS, 10 Rue Vauquelin, 75005, Paris, France.
Sorbonne Université, 4 Place Jussieu, 75005, Paris, France.
Anal Bioanal Chem. 2024 Apr;416(9):2173-2188. doi: 10.1007/s00216-023-04925-y. Epub 2023 Sep 13.
The objective of the present review is to list, describe, compare, and critically analyze the main procedures developed in the last 20 years for the analysis of digested alkylated peptides, resulting from the adduction of albumin by different mustard agents, and that can be used as biomarkers of exposure to these chemical agents. While many biomarkers of sulfur mustard, its analogues, and nitrogen mustards can easily be collected in urine such as their hydrolysis products, albumin adducts require blood or plasma collection to be analyzed. Nonetheless, albumin adducts offer a wider period of detectability in human exposed patients than urine found biomarkers with detection up to 25 days after exposure to the chemical agent. The detection of these digested alkylated peptides of adducted albumin constitutes unambiguous proof of exposure. However, their determination, especially when they are present at very low concentration levels, can be very difficult due to the complexity of the biological matrices. Therefore, numerous sample preparation procedures to extract albumin and to recover alkylated peptides after a digestion step using enzymes have been proposed prior to the analysis of the targeted peptides by liquid chromatography coupled to mass spectrometry method with or without derivatization step. This review describes and compares the numerous procedures including a number of different steps for the extraction and purification of adducted albumin and its digested peptides described in the literature to achieve detection limits for biological samples exposed to sulfur mustard, its analogues, and nitrogen mustards in the ng/mL range.
本综述的目的是列出、描述、比较和批判性分析过去 20 年中开发的主要方法，用于分析由于不同芥子气制剂与白蛋白加合而产生的已消化的烷基化肽，这些方法可作为暴露于这些化学制剂的生物标志物。虽然许多芥子气、其类似物和氮芥的生物标志物可以很容易地在尿液中收集，如它们的水解产物，但白蛋白加合物需要采集血液或血浆进行分析。尽管如此，白蛋白加合物在暴露于化学制剂的 25 天内，为人体提供了比尿液中发现的生物标志物更广泛的检测期。检测这些与白蛋白结合的已消化的烷基化肽是暴露的确凿证据。然而，由于生物基质的复杂性，其测定，特别是当它们存在于非常低的浓度水平时，可能非常困难。因此，在使用液相色谱-质谱法（或无需衍生化步骤）分析靶向肽之前，已经提出了许多用于提取白蛋白和在酶消化步骤后回收烷基化肽的样品制备程序，以提取和纯化文献中描述的与白蛋白结合的烷基化肽。本综述描述并比较了许多程序，包括提取和纯化与白蛋白结合的烷基化肽的许多不同步骤，以实现对芥子气、其类似物和氮芥暴露的生物样品的检测限达到 ng/mL 范围。
