The potential of near-infrared spectroscopy (NIRS) for screening the sinigrin, gluconapin, 4-hydroxyglucobrassicin, and total glucosinolate contents of Indian mustard (Brassica juncea L. Czern. & Coss.) seed was assessed. Intact seed samples of this species were analyzed by NIRS and their reference values regressed against different spectral transformations by modified partial least-squares (MPLS) regression. The coefficients of determination (r (2)) for sinigrin, gluconapin, 4-hydroxyglucobrassicin, and total glucosinolate contents were, respectively, 0.86, 0.95, 0.33, and 0.82. The standard deviation to standard error of prediction (SEP) ratio, and SEP to standard error of laboratory ratio were for these constituents as follows: sinigrin, 2.59 and 2.70; gluconapin, 4.16 and 2.08; 4-hydroxyglucobrassicin, 1.18 and 1.40; and total glucosinolates, 2.18 and 1.60. By comparison of commercial sinigrin spectrum with the first MPLS loadings of the sinigrin equation, it can be concluded that the molecule of sinigrin has a specific signal in the seed spectrum of Brassica.