Nie Zhiyong, Zhang Yajiao, Chen Jia, Lin Ying, Wu Bidong, Dong Yuan, Feng Jianlin, Liu Qin, Xie Jianwei
State Key Laboratory of Toxicology and Medical Countermeasures, and Laboratory of Toxicant Analysis, Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, 27 Taiping Road, Haidian District, Beijing, 100850, China.
Anal Bioanal Chem. 2014 Aug;406(21):5203-12. doi: 10.1007/s00216-014-7916-3. Epub 2014 Jun 13.
A highly sensitive method for the determination of sulfur mustard (SM) metabolites thiodiglycol (TDG) and thiodiglycol sulfoxide (TDGO) in urine was established and validated using isotope-dilution negative-ion chemical ionization (NICI) gas chromatography-mass spectrometry (GC-MS). TDGO in the samples was reduced with TiCl3, and then determined together with TDG as a single analyte. The sample preparation procedures, including two solid-phase-extraction (SPE) clean-up steps, were optimized to improve the sensitivity of the method. The limits of detection (LOD) for both TDG and TDG plus TDGO (TDG + TDGO) were 0.1 ng mL(-1), and the limits of quantitation (LOQ) for both were 0.3 ng mL(-1). The method was used in a rabbit cutaneous SM exposure model. Domestic rabbits were exposed to neat liquid SM at three dosage levels (0.02, 0.05, and 0.15 LD50), and the urinary excretion of four species of hydrolysis metabolites, namely free TDG, free plus conjugated TDG (total TDG), free TDG + TDGO, and free plus conjugated TDG + TDGO (total TDG + TDGO), was evaluated to investigate the metabolic processes. The total urinary excretion profiles of the metabolites, including the peak time, time window, and dose-response and time-response relationships, were clarified. The results revealed that the concentrations of TDG and TDG + TDGO in the urine increased quickly and then decreased rapidly in the first two days after SM exposure. The cumulative amount of total TDG + TDGO excreted in urine during the first five days accounted for 0.5-1% of the applied dose of SM. It is also concluded that TDG and TDGO in urine existed mainly in free form, the levels of glucuronide and of sulfate conjugates of TDG or TDGO were very low, and most hydrolysis metabolites were present in the oxidized form (TDGO). The study indicates that the abnormal increase of TDG and TDGO excretion levels can be used as a diagnostic indicator and establishes a reference time-window for retrospective analysis and sampling after SM exposure.
建立了一种高灵敏度的方法，用于测定尿液中的硫芥（SM）代谢物硫二甘醇（TDG）和硫二甘醇亚砜（TDGO），并采用同位素稀释负离子化学电离（NICI）气相色谱 - 质谱联用（GC - MS）对该方法进行了验证。样品中的TDGO用TiCl₃还原，然后与TDG作为单一分析物一起测定。对包括两个固相萃取（SPE）净化步骤在内的样品制备程序进行了优化，以提高该方法的灵敏度。TDG和TDG加TDGO（TDG + TDGO）的检测限（LOD）均为0.1 ng mL⁻¹，两者的定量限（LOQ）均为0.3 ng mL⁻¹。该方法用于兔皮肤SM暴露模型。将家兔暴露于三种剂量水平（0.02、0.05和0.15 LD50）的纯液体SM中，评估四种水解代谢物（即游离TDG、游离加结合TDG（总TDG）、游离TDG + TDGO和游离加结合TDG + TDGO（总TDG + TDGO））的尿排泄情况，以研究代谢过程。明确了代谢物的总尿排泄谱，包括峰值时间、时间窗以及剂量 - 反应和时间 - 反应关系。结果表明，SM暴露后的前两天，尿液中TDG和TDG + TDGO的浓度迅速升高，然后迅速下降。前五天尿液中排泄的总TDG + TDGO累积量占SM给药剂量的0.5 - 1%。还得出结论，尿液中的TDG和TDGO主要以游离形式存在，TDG或TDGO的葡萄糖醛酸苷和硫酸盐结合物水平非常低，并且大多数水解代谢物以氧化形式（TDGO）存在。该研究表明，TDG和TDGO排泄水平的异常升高可作为诊断指标，并为SM暴露后的回顾性分析和采样建立了参考时间窗。
