Simultaneous Quantification of Biothiols and Deciphering Diverse GSH Stability in Different Live Cells by ¹⁹F-Tag

GSH, Cys, Hcy, and H₂S are important biothiols and play important roles in the living systems. Quantitative and simultaneous determination of these biothiols under physiological conditions is still a challenge. Herein, we developed an effective ¹⁹F-reactive tag that readily interacts with these four biothiols for the generation of stable thioether products that have distinguishable ¹⁹F-chemical shifts. These thioester compounds encode the characteristic fingerprint profiles of each biothiols, allowing one to simultaneously quantify and determine these biothiols by 1D ¹⁹F NMR spectroscopy. The intra-/extracellular GSH in live cells was assessed by the established strategy, and remarkable variations in the GSH stability were determined between the normal mammalian cells and cancer cells. It is notable that GSH hydrolyzes efficiently in the out-membrane of the cancer cells and the lysates. In contrast, GSH remains stable in the tested normal cells.