A field experiment was conducted during the winter (rabi) seasons 2012-2013 at the Instructional farm of Narendra Deva university of Agriculture and Technology, Kumarganj, Faizabad (U.P.) India. Five doses of sulphur (i.e., 0, 10, 20, 30 and 40 kg S ha1) and zinc (i.e., 0, 10, 20, 30 and 40 kg S ha1) were applied on two mustard varieties namely Varuna and NDR 8501. The maximum oil content (42.60) and oil yield (10.87 Q/ha) were recorded in Varuna variety followed by variety NDR 8501 (40.33) and (10.65 Q/ha) respectively. Oil content, oil yield, refractive index were increased in response to sulphur and zinc doses in both cultivar but did not show significant results in all treatments. Maximum refractive index was observed in variety NDR 8501 (1.4663) followed by Varuna variety (1.4632). The increasing doses of sulphur decreased oleic, linoleic and eicosanoic acids and increased the undesirable erucic acid in both cultivars. Treatment S40Zn40 resulted highest iodine value over control in both cultivar. The increasing doses of sulphur and zinc decreased the saponification values while increased the iodine value in both cultivar. The highest saponification value was observed in variety NDR 8501 (174.86) followed by Varuna variety (171.86). A non-significant difference in saponification values was noticed in each cultivar. Tryptophan, methionine and protein content were increased with increasing the level of sulphur and zinc doses in both cultivars. The maximum protein content (29.93%) was found in Varuna variety followed by variety NDR-8501 (28.68%). Increasing level of sulphur and zinc did not showed significant results in all treatments. The allyl-isothiocynate content was decreased with increasing level of sulphur and zinc doses in both the cultivars.