Li Dan, Yan Ling, Lam Thomas Ka-Yam, Cai Zongwei
Guangdong Key Laboratory of Environmental Catalysis and Health Risk Control, Guangdong-Hong Kong-Macao Joint Laboratory for Contaminants Exposure and Health, School of Environmental Science and Engineering, Guangdong University of Technology, Guangzhou, China.
College of Science, Eastern Institute of Technology, Ningbo, China.
Rapid Commun Mass Spectrom. 2025 Nov 15;39(21):e10117. doi: 10.1002/rcm.10117.
Investigating TCE glutathione (GSH) conjugation metabolites is essential, as these reactive intermediates play a central role in TCE bioactivation and are implicated in organ-specific toxicities, including nephrotoxicity, hepatotoxicity, and neurotoxicity. Characterizing and quantifying these metabolites enhances our understanding of TCE metabolism, supports biomarker discovery, and helps elucidate mechanisms of TCE-induced toxicity.
A robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for the detection and quantification of three major GSH-related TCE metabolites: S-(1,2-dichlorovinyl)-glutathione (DCVG), S-(1,2-dichlorovinyl)-L-cysteine (DCVC), and N-acetyl-S-(1,2-dichlorovinyl)-L-cysteine (NAcDCVC). Serum samples were obtained from male C57BL/6 mice chronically exposed to TCE (100 ppm in drinking water for 15 weeks).
Calibration curves for all three metabolites demonstrated excellent linearity (R > 0.998). The method achieved limits of detection (LOD) ranging from 0.0057 to 0.0120 nM, limits of quantitation (LOQ) from 0.0189 to 0.0401 nM, recoveries of 75.9%-115.5%, and inter-assay variation of 0.5%-11.5%. PD model mice exhibited elevated serum levels of DCVG and DCVC, while NAcDCVC levels were significantly reduced.
This study presents the first comprehensive LC-MS/MS-based quantification of TCE GSH conjugation metabolites in serum, offering high sensitivity, precision, and reproducibility. The observed elevated serum levels of the toxic metabolites DCVG and DCVC, along with the markedly reduced NAcDCVC concentrations in PD mice, provide a critical foundation for future investigations into the mechanistic links between TCE exposure and PD pathogenesis.
研究三氯乙烯（TCE）与谷胱甘肽（GSH）结合的代谢产物至关重要，因为这些反应性中间体在TCE生物活化过程中起核心作用，并与器官特异性毒性有关，包括肾毒性、肝毒性和神经毒性。对这些代谢产物进行表征和定量可增强我们对TCE代谢的理解，支持生物标志物的发现，并有助于阐明TCE诱导毒性的机制。
建立了一种可靠的液相色谱 - 串联质谱（LC-MS/MS）方法，用于检测和定量三种主要的与GSH相关的TCE代谢产物：S-(1,2-二氯乙烯基)-谷胱甘肽（DCVG）、S-(1,2-二氯乙烯基)-L-半胱氨酸（DCVC）和N-乙酰-S-(1,2-二氯乙烯基)-L-半胱氨酸（NAcDCVC）。从长期暴露于TCE（饮用水中100 ppm，持续15周）的雄性C57BL/6小鼠中获取血清样本。
所有三种代谢产物的校准曲线均显示出良好的线性（R > 0.998）。该方法的检测限（LOD）范围为0.0057至0.0120 nM，定量限（LOQ）为0.0189至0.0401 nM，回收率为75.9%至115.5%，批间变异为0.5%至11.5%。帕金森病（PD）模型小鼠的血清DCVG和DCVC水平升高，而NAcDCVC水平显著降低。
本研究首次基于LC-MS/MS对血清中TCE与GSH结合的代谢产物进行了全面定量，具有高灵敏度、精密度和重现性。在PD小鼠中观察到的有毒代谢产物DCVG和DCVC血清水平升高以及NAcDCVC浓度显著降低，为未来研究TCE暴露与PD发病机制之间的机制联系提供了关键基础。
