Gao Xia, Liu Xuehan, Zheng Xinping, Liu Yaxin, Lin Jinming, Zhang Pengbo
School of Public Health, Xinxiang Medical University, Xinxiang, 453003, P. R. China.
J Fluoresc. 2025 Apr 30. doi: 10.1007/s10895-025-04350-7.
Biothiols play essential roles in various biological processes and are closely associated with diseases such as cancer, neurodegenerative disorders, and cardiovascular diseases. To monitor biothiols in organisms and living cells, we developed a novel intramolecular charge transfer (ICT)-based probe, hemicyanine-2,4-dinitrobenzenesulfonamide(HCD), by coupling a hemicyanine dye with 2,4-dinitrobenzenesulfonyl chloride.The HCD probe exhibits remarkable sensitivity, with low detection limits of 0.32 µM for Cys, 0.72 µM for Hcy, and 0.27 µM for GSH, and demonstrates high selectivity for biothiols in the presence of various interfering species. The detection mechanism was thoroughly validated using high-resolution mass spectrometry (HRMS), UV-Vis spectroscopy, fluorescence spectroscopy and Density Functional Theory (DFT) calculation. Furthermore, the applicability of HCD was successfully demonstrated in human urine samples and in vivo settings using RAW264.7 cells, confirming its potential as a powerful tool for monitoring endogenous biothiols in biological systems.
生物硫醇在各种生物过程中发挥着重要作用，并且与癌症、神经退行性疾病和心血管疾病等病症密切相关。为了监测生物体内和活细胞中的生物硫醇，我们通过将半花菁染料与2,4-二硝基苯磺酰氯偶联，开发了一种基于分子内电荷转移（ICT）的新型探针——半花菁-2,4-二硝基苯磺酰胺（HCD）。HCD探针具有显著的灵敏度，对半胱氨酸（Cys）的检测限低至0.32 μM，对同型半胱氨酸（Hcy）为0.72 μM，对谷胱甘肽（GSH）为0.27 μM，并且在存在各种干扰物质的情况下对生物硫醇表现出高选择性。使用高分辨率质谱（HRMS）、紫外可见光谱、荧光光谱和密度泛函理论（DFT）计算对检测机制进行了全面验证。此外，通过使用RAW264.7细胞，成功证明了HCD在人尿液样本和体内环境中的适用性，证实了其作为监测生物系统中内源性生物硫醇的有力工具的潜力。
