Peanuts (Arachis hypogaea L.) are one of the major oilseed crops of the world and are an important source of protein in many countries. In this study, some nutrients and characteristics of the seeds’ oil extracted from four peanut (Arachis hypogaea L.) varieties: Line 27r (Israel), Line 9 (Malawi), Line 4 (Brazil) and Line 18 (Israel) cultivated, for first time, in Upper Egypt were subjected to the comparative assessment with control NC variety (USA). Peanut seeds are a rich source of oil content (50.45 to 52.12 g 100 g−1 dry weight “DW”). The physicochemical properties of extracted oil were investigated in this study. The obtained data showed that the ratios of saturated fatty acids ranged from 14.24 to 17.23%, and the amounts of unsaturated fatty acids ranged from 82.77 to 85.76%. Significant variations (p ≤ 0.05) of oil content, saponification value, oleic/linoleic (O/L), and oil characteristics were found. Line 9 was found to be high in oil content, while Line 27r was said to have a high O/L ratio (3.22%) and proportion of unsaturated fatty acids (85.76%).