A sensitive, accurate, and time-saving approach was developed for the simultaneous quantification of eight sulfur compounds in the sulfur pathway, which could reflect the status of an organism, including oxidative stress, signal transduction, enzyme reaction, and so on. In order to overcome the instability of highly reactive sulfhydryl compounds, N-ethylmaleimide derivatization was adopted to effectively protect sulfhydryl-containing samples. Using isotope-labeled glutathione (GSH-¹³C₂, ¹⁵N), the validated method was demonstrated to offer satisfactory linearity, accuracy, and precision. Separation was done by UHPLC, using a BEH amide column. Accordingly, 0.1% formic acid acetonitrile was selected as the precipitant. A tandem mass spectrometer was coupled to the chromatographic system and afforded a detection limit of 0.2 ng/mL. Good linearity was maintained over a wide concentration range (r² > 0.994), and the accuracy was in the range of 86.6–114% for all the studied compounds. The precision, expressed in RSD%, ranged from 1.1% to 9.4% as intraday variability and less than 13% as interday precision for all of the analytes. The approach was applied to study the potential therapeutic mechanism of a well-known traditional Chinese medicine, Shao Fu Zhu Yu decoction. The results suggested that Shao Fu Zhu Yu decoction might protect against oxidative damage by increasing the concentrations of sulfhydryl compounds.

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开发了一种灵敏，准确且省时的方法，用于同时定量硫途径中的八种硫化合物，这可以反映生物体的状态，包括氧化应激，信号转导，酶反应等。为了克服高反应性巯基化合物的不稳定性，采用N-乙基马来酰亚胺衍生化来有效保护含巯基的样品。使用同位素标记的谷胱甘肽（GSH- ¹³ Ç ₂，¹⁵N），证明了经过验证的方法可提供令人满意的线性，准确性和精确度。使用BEH酰胺柱，通过UHPLC进行分离。因此，选择0.1％的甲酸乙腈作为沉淀剂。串联质谱仪与色谱系统相连，检出限为0.2 ng / mL。在很宽的浓度范围（r ²> 0.994），并且所有研究化合物的准确度在86.6–114％范围内。以RSD％表示的精度，所有分析物的日内变化范围为1.1％至9.4％，日间精度的变化范围小于13％。该方法被用于研究一种著名的中药“少夫逐瘀汤”的潜在治疗机制。结果表明，少腹逐瘀汤可能通过增加巯基化合物的浓度来防止氧化损伤。

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