While methods for the identification and quantification of total plant glucosinolate content typically utilize desulfation of glucosinolates followed by reversed-phase chromatography, the analysis of intact glucosinolates has been problematic. Hydrophilic interaction chromatography offers a novel method for analyzing intact glucosinolates and when performed along with ion-pair reversed-phase chromatography offers a powerful and complementary method for glucosinolate analysis.