To evaluate seasonal variation in fatty acid composition in broccoli, 12 commercial cultivars of broccoli were grown in spring and fall season at the field of NIHHS, and their floret, leaf and stem parts were used for the fatty acid composition analyses. Among 14 fatty acids detected in broccoli, linolenic, palmitic and linoleic acids were major fatty acids comprising more than 80% of total fatty acids in both the seasons and all the parts. Likewise, stearic and oleic acids were also present in considerable amount while remaining fatty acids; caproic, lauric, myristic, pentadecanoic, palmitoleic, heptadecanoic, arachidic, behenic and lignoceric acids showed their minor compositional ratio. Among the three parts, stem exhibited highest SFAs (49.681% in spring and 50.717% in fall season) compared to MUFA and PUFA, while highest compositional ratio of PUFAs were observed in leaves (62.588% in spring and 68.931% in fall season), which indicates leaves as a good source of health beneficial fatty acids. In contrast, floret part exhibited highest SFA (48.786%) and PUFA (57.518%) in spring and fall seasons, respectively. Major fatty acids; palmitic, linoleic and linolenic acid showed lowest cultivar dependent variation (below 10%) and leaf showed least variation in both the seasons compared to floret and stem. Our results suggest that all the fatty acids are significantly influenced by genotype of cultivars (C), plant parts (P) and growing seasons (S). Among the 14 fatty acids, myristic and palmitic acid showed highest positive or negative correlationship with oleic (r=$0.912^{**}$) and linolenic acid (r=-$0.933^{**}$), respectively. The most abundant fatty acid, linolenic acid, showed either negative or no correlation ship with other fatty acids while palmitic acid, a second major fatty acid, exhibited either positive or negative correlation ship.