天津医科大学刘阳平团队报道了使用三烷基自旋探针通过硫交换反应，实现过硫化物、生物硫醇和硫化氢的同时定量检测。相关研究成果于2024年10月21日发表在《美国化学会杂志》。

活性硫物种（RSS），包括过硫化物（RSSH）、生物硫醇和硫化氢（H₂S），是各种生理过程中的关键调节因子。为了更好地理解这些RSS的共生关系和相互转化，开发能够检测和量化它们的分析技术是非常理想但具有挑战性的。

该文报道了新型三苯甲基自由基电子顺磁共振（EPR）探针CT02-TNB和OX-TNB的合理设计和合成。CT02-TNB与两种反应性RSSH（PS1和PS2）进行了快速硫交换反应，这两种RSSH从其相应的供体PSD1和PSD2释放出来，得到了特定的偶联物。

所得共轭物表现出特征性的EPR光谱，从而能够区分检测和定量两种RSSH。此外，CT02-TNB对其他RSS也表现出良好的反应，包括谷胱甘肽（GSH）、半胱氨酸（Cys）、H₂S和亚硫酸盐。重要的是，基于更新的EPR光谱模拟程序，还实现了CT02-TNB对多种RSS（例如，PS1/GSH/Cys或PS1/GSH/H₂S）的同时定量。最后，通过亲水性OX-TNB测量分离的小鼠肝脏中PSD1释放的PS1和内源性GSH的水平。

该项工作代表了第一项通过光谱方法实现不同过硫化物和其他RSS的鉴别和定量检测的研究。

附：英文原文

Title: Simultaneous Quantitation of Persulfides, Biothiols, and Hydrogen Sulfide through Sulfur Exchange Reaction with Trityl Spin Probes

Author: Xiaoli Tan, Jiaxin Zhou, Luhua Yang, Qi Chang, Shao-Yong Li, Antal Rockenbauer, Yuguang Song, Yangping Liu

Issue&Volume: October 21, 2024

Abstract: Reactive sulfur species (RSS) including persulfides (RSSHs), biothiols, and hydrogen sulfide (H2S) are key regulators in various physiological processes. To better understand the symbiotic relationship and interconversion of these RSS, it is highly desirable but challenging to develop analytical techniques that are capable of detecting and quantifying them. Herein, we report the rational design and synthesis of novel trityl-radical-based electron paramagnetic resonance (EPR) probes dubbed CT02-TNB and OX-TNB. CT02-TNB underwent fast sulfur exchange reactions with two reactive RSSHs (PS1 and PS2) which were released from their corresponding donors PSD1 and PSD2 to afford the specific conjugates. The resulting conjugates exhibit characteristic EPR spectra, thus enabling discriminative detection and quantitation of the two RSSHs. Moreover, CT02-TNB showed good response toward other RSS including glutathione (GSH), cysteine (Cys), H2S, and sulfite as well. Importantly, based on the updated EPR spectral simulation program, simultaneous quantitation of multiple RSS (e.g., PS1/GSH/Cys or PS1/GSH/H2S) by CT02-TNB was also achieved. Finally, the levels of released PS1 from PSD1 and endogenous GSH in isolated mouse livers were measured by the hydrophilic OX-TNB. This work represents the first study achieving discriminative and quantitative detection of different persulfides and other RSS by a spectroscopic method.

DOI: 10.1021/jacs.4c10266

Source: https://pubs.acs.org/doi/abs/10.1021/jacs.4c10266