Six desulfo-glucosinolates (DS-GSLs), DS-glucoraphanin, DS-4-(β-D-glucopyranosyldisulfanyl)butyl GSL, DS-glucoerucin, DS-glucobrassicin, DS-dimeric 4-mercaptobutyl GSL and DS-4-methoxyglucobrassicin, were isolated from rocket salad (Eruca sativa Mill.) By using high-pressure liquid chromatography (HPLC) and identified by their retention times on HPLC and electrospray ionization mass spectrometry (ESI-MS) techniques. Total GSL contents were 125, 11 and 22 µmol g −1 dry weight (DW) in seeds, leaves and roots, respectively. Two DS-GSLs, namely DS-glucoraphanin and DS-glucoerucin, were the major GSLs in seeds and roots, while DS-glucoraphanin, DS-glucoerucin and DS-dimeric 4-mercaptobutyl GSL were the predominant GSLs in leaves (> 1 µmol g −1 DW or > 10% of the total GSL content). In addition, the anti-oxidant activity (n = 3) of intact leaf and root powders, whole DS-GSL eluates and purified DS-4-methoxyglucobrassicin were measured using the XYZ-dish method. DS-4-methoxyglucobrassicin showed the greatest anti-oxidant activity value (3.8 unit g −1 DW), and the value for whole DS-GSL eluates (1.6) was higher than that of intact leaves (1.5) and roots (0.87).