Facile and sensitive measurement of GSH/GSSG in cells by surface-enhanced Raman spectroscopy

• Published in: Talanta (Oxford) 2021-03, Vol.224, p.121852, Article 121852
• Main Authors: Zhu, Yingjie, Wu, Jianfeng, Wang, Kai, Xu, Hua, Qu, Minmin, Gao, Zhongcai, Guo, Lei, Xie, Jianwei
• Format: Article
• Language: English
• Subjects: Cellular detection; Glutathione; Glutathione Disulfide; Metal Nanoparticles; Oxidation-Reduction; Reduction-sensitive reaction-type Raman probe; SERS; Silver; Spectrum Analysis, Raman
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/j.talanta.2020.121852

Reduced glutathione (GSH) and the oxidized glutathione (GSSG) are well-known biomolecules in the main constituents of intracellular redox homeostasis system. A rapid, accurate measurement of cellular GSH and GSSG is quite needed in investigating important biochemical events. In this work, we present a novel and sensitive method to monitor intracellular GSH and GSSG concentrations by a portable surface-enhanced Raman spectroscopy (SERS) technique. We introduced a reduction-sensitive reaction-type Raman probe, 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB) to initiate GSH reduction, itself concomitantly converts to 2-nitro-5-thiobenzoic acid (TNB) to release a strong SERS signal. In a convenient way of inorganic salt MgSO4 induced aggregation of silver nanoparticles substrate, we easily implemented a good discrimination between DTNB and TNB, and a quantitative measurement of GSH and GSSG with a high sensitivity of 10 nM. This SERS method proved its feasible applicability in rapidly and sensitively monitoring GSH depletion behaviors of some notorious alkylating agents, i.e., sulfur mustard and nitrogen mustards in ex vitro or in vitro (cellular response). This SERS method may be very worthwhile in cellular detoxication event via the GSH approach and other GSH involved biomedical researches. [Display omitted] •A reduction-sensitive reaction-type SERS sensor is developed for detection of GSH and GSSG in cells.•This method reaches a high sensitivity of 10 nM for GSH and GSSG, so that cellular determination can be readily achieved.•Rapid monitoring of GSH depletion behaviors of typical alkylating agents in ex vitro or in vitro is realized.•This SERS-based method shows potential applicability in GSH involved biomedical researches.