Simultaneous assessment of endogenous thiol compounds by LC-MS/MS

Biological thiol compounds are very important molecules that participate in various physiological events. Alteration in levels of endogenous thiols has been suggested as a biomarker of early stage of pathological changes. We reported a chemical derivatization- and LC-MS/MS-based approach to simultaneously determine thiol compounds including glutathione (GSH), cysteine (Cys), N-acetyl cysteine (NAC), homocysteine (Hcy), and cysteinylglycine (CysGly) in biological samples. Thiol-containing samples were derivatized with monobromobimane (mBrB) at room temperature, followed by LC-MS/MS analysis. Assessment of the analytes with baseline separation was completed within 10 min, using a gradient elution on a C-18 reversed-phase column. Excellent linearity was observed for all analytes over their concentration ranges of 1-400 mu M. The lowest limits of detection (S/N = 3) in a range from 0.31 fmol (for NAC) to 4.98 fmol (for CysGly) were achieved. The results indicate that this approach was sensitive, selective, and well suited for high-throughput quantitative determination of the biological thiols. (C) 2016 Elsevier B.V. All rights reserved.