Nitrogen (N), phosphorous (P) and potassium (K) are the most limiting factors in crop production. N often affects the amino acid composition of protein and in turn its nutritional quality. In Brassica plants, abundant supply of N fertilizer decreases the relative proportion of glucosinolates (GSLs), thus reducing the biological and medical values of the vegetables. Hence effort was made to evaluate the influence of different proportions of nutrient solutions containing N–P–K on the GSL profiles of rocket salad (Eruca sativa Mill.). Fifteen desulpho-(DS) GSLs were isolated and identified using liquid chromatography–mass spectrometry (LC/MS) analysis. Rocket salad plants supplied with lesser amount of N, P or higher concentrations of K showed a typical improvement in total GSL contents. In contrast, total GSL levels were less at higher N supply. Furthermore, with N concentrations above 5 mM and K concentrations less than 2.5 mM, the GSL amounts were on average 13.51 and 13.75 μmol/g dry weight (DW), respectively. Aliphatic GSLs predominated in all concentrations of NPK while indolyl GSLs made up marginally less amount of the total compositions. Five and 2 mM N and P possessed much higher levels of several types of aliphatic GSLs than other concentrations, including glucoerucin, glucoraphanin and dimeric 4-mercaptobutyl GSL. From this perspective, it is contended that supply of less N results in enhancing the metabolic pathway for the synthesis of GSLs in rocket salad.