Abstract Nine different FAs were identified in TL, TG and PL fractions. The major FAs of TL, TG, PL in all Hyoscyamus species were C16:0 (palmitic acid, PA), C18:1 n−9 (oleic acid, OLA), C18:2 n−6 (linoleic acid, LA) and C18:3 n−3 (linolenic acid, ALA). Fatty acids composition range was as follows: saturated fatty acids (SFAs) 12.45–33.91%, 31.51–41.67%, 10.32–39.03% monounsaturated fatty acids (MUFAs) 7.66–16.57%, 6.24–14.67%, 11.40–26.78% and polyunsaturated fatty acids (PUFAs) 58.42–70.97%, 51.04–55.53%, 49.55–72.90% in TL, PL and TG fractions, respectively. In all Hyoscyamus species, total PUFA amounts were found to be higher than total MUFA and total SFA in TL, TG and PL fractions. The present study is a guide for biochemical and nutritional values of the Hyoscyamus species and can be useful for further investigation on industrial applications.