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Updated: Mar 17, 2026
07:16
Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation
Published on: June 21, 2021
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Simultaneous assessment of endogenous thiol compounds by LC-MS/MS
Yao Sun1, Tong Yao1, Xiucai Guo1
1School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, Liaoning 110016, PR China.
Summary
This study presents a rapid LC-MS/MS method for quantifying key biological thiols like glutathione (GSH) and cysteine (Cys). The approach offers high sensitivity and selectivity for biomarker discovery in pathological changes.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Biomarker Research
Background:
- Biological thiols are crucial for physiological processes.
- Altered thiol levels can indicate early pathological changes.
- Accurate quantification of thiols is vital for disease detection.
Purpose of the Study:
- To develop a sensitive and selective method for simultaneous determination of multiple biological thiols.
- To establish a high-throughput assay for thiol analysis in biological samples.
- To validate a derivatization and LC-MS/MS approach for thiol quantification.
Main Methods:
- Chemical derivatization of thiol compounds with monobromobimane (mBrB).
- Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis.
- Gradient elution on a C18 reversed-phase column for separation.
Main Results:
- Simultaneous quantification of glutathione (GSH), cysteine (Cys), N-acetyl cysteine (NAC), homocysteine (Hcy), and cysteinylglycine (CysGly).
- Baseline separation achieved within 10 minutes.
- Excellent linearity (1-400μM) and high sensitivity with low limits of detection (0.31-4.98 fmol).
Conclusions:
- The developed LC-MS/MS method is sensitive, selective, and efficient.
- This approach is well-suited for high-throughput quantitative determination of biological thiols.
- The method facilitates the use of thiols as early biomarkers for pathological conditions.
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