Twelve pumpkin cultivars (Cucurbita maxima D.), cultivated in Iowa, were studied for their seed oil content, fatty acid composition, and tocopherol content. Oil content ranged from 10.9 to 30.9%. Total unsaturated fatty acid content ranged from 73.1 to 80.5%. The predominant fatty acids present were linoleic, oleic, palmitic, and stearic. Significant differences were observed among the cultivars for stearic, oleic, linoleic, and gadoleic acid content of oil. Low linolenic acid levels were observed (<1%). The tocopherol content of the oils ranged from 27.1 to 75.1 microg/g of oil for alpha-tocopherol, from 74.9 to 492.8 microg/g for gamma-tocopherol, and from 35.3 to 1109.7 microg/g for delta-tocopherol. The study showed potential for pumpkin seed oil from all 12 cultivars to have high oxidative stability that would be suitable for food and industrial applications, as well as high unsaturation and tocopherol content that could potentially improve the nutrition of human diets.