A fast, sensitive and reproducible method using LC-MS/MS for simultaneous quantification of glutathione (GSH), glutathione disulfide (GSSG) and glutathione-S-sulfonate (GSSO3H) was developed, optimised and applied in analysis of grape juice and wine samples. The results show that only GSH (10–60 mg⋅L− 1) and GSSG (2–11 mg⋅L− 1) are found in grape juice when SO2 is not added. GSSO3H was detected in must samples treated with SO2 but only at a low concentration (<1 mg L− 1). In the wine samples, the dominant form of glutathione was GSSO3H (5–11 mg L− 1), followed by GSH (0–5 mg L− 1) and GSSG (0–6 mg L− 1), underscoring the importance of GSSO3H quantification. GSSO3H formation in wine was correlated with the total SO2 level in the wine. We believe this is the first report on GSSO3H quantification in wine