The effects of different processing treatments on thiol precursor concentrations have been investigated through studies involving transportation of machine-harvested Sauvignon blanc fruit and assessment of different applications of antioxidants, along with juice preparation and enzyme inhibition experiments. The influence these trials had on 3-S-cysteinylhexan-1-ol (Cys-3-MH) and 3-S-glutathionylhexan-1-ol (Glut-3-MH) concentrations in juices is discussed. Very interesting findings included the large increase in precursor concentrations after transportation, particularly for Cys-3-MH, and the limited formation of Glut-3-MH when grape proteins were precipitated during processing. The various results provided information about the ability to modulate precursor concentrations depending on the processing technique employed. Additionally, a conjugated aldehyde, which is the obvious missing link between the reaction of (E)-2-hexenal and glutathione in the formation of Glut-3-MH, has been tentatively identified for the first time. Deuterium-labeled 3-S-glutathionylhexanal (Glut-3-MHAl) was produced through the addition of labeled (E)-2-hexenal to grapes, followed by grape crushing, and detected in the juice by HPLC-MS/MS, along with the corresponding labeled Glut-3-MH.