The seeds of 17 cultivars of Saskatoon berries ( Amelanchier alnifolia Nutt.) were evaluated for their seed mass, oil content, fatty acid and triacylglycerol (TAG) composition, tocopherol profile, and sterol content. The oil content of the seeds ranged from 9.4% (cv. 'Pasture') to 18.7% (cv. 'Thiessen'). The seed oils contained mainly linoleic acid in the range from 47.3% (cv. 'Success') to 60.1% (cv. 'Lee 3') and oleic acid in the range from 26.3% (cv. 'Lee 3') to 38.1% (cv. 'Success'). The total tocopherol content ranged from 1053 to 1754 mg/kg of oil. alpha-Tocopherol was the major vitamin E compound in all berry seed oils investigated, accounting for 87% of total tocopherols. The predominant sterols were beta-sitosterol, Delta(5)-avenasterol, and campesterol. The sterols content in seed oil ranged from 7357 mg/kg of oil (cv. 'Success') to 15771 mg/kg of oil (cv. 'Lee 3'). Thirteen triacylglycerols (TAG) were identified in seed oils, among which LLL, LLO, LOO, LLP, LOP (L, linoleoyl; O, oleoyl; P, palmitoyl) represented 88% of the total TAG. TAG composition suggests good oxidative stability of the Saskatoon berry seed oil, which could be suitable for food and industrial applications. Moreover, Saskatoon berry seed oil may serve as potential dietary source of tocopherols, sterols, and unsaturated fatty acids.