Thiol precursor content in Melon B. and Sauvignon blanc grape juices obtained under vacuum was determined by quantifying cysteinylated and glutathionylated conjugates of 3-mercaptohexan-1-ol (3MH) and 4-methyl-4-mercaptopentan-2-one (4MMP). This characterization allowed the study of thiol precursor evolution during ripening of Sauvignon blanc grapes in several viticultural situations together with grape reaction product (GRP) and the main substrate of polyphenoloxidase, that is, caftaric acid. Concentration of precursors greatly increased during ripening except for the cysteinylated conjugate of 4MMP. Precursor evolution was also monitored during the oxidation of grape juice. Addition of oxygen to a grape juice set off the enzymatic oxidation of hydroxycinnamic acids but did not negatively affect precursor concentrations. Part of the glutathionylated precursor of the 3MH was produced during prefermentative operations (up to 140% in Sauvignon blanc). Consequently, this precursor naturally occurring in grapes was also formed during prefermentative operations. The proportion of biogenetic and prefermentary formation of the glutathionylated precursor of 3MH was different under industrial conditions depending on the grape variety considered. Addition of glutathione and hexenal in grape juices of Melon B. and Sauvignon induced an increase of the production of 3MH and consequently of its acetate in the resulting wines. Residual glutathione in must has to be preserved to enhance the aromatic potential of grapes.