Pistacia atlantica, which belongs to the Anacardiaceae family, is an important species for rural people in arid and semi-arid areas. The fruit, rich in oil, is used in traditional medicine for the treatment of various diseases. The oil extracted from this species growing in a northern area of Algeria and its fatty acid composition were previously studied. However, the largest areas where this species is present (traditional cultivation) is located in southern Algeria. Moreover, studies on oil fatty acid composition and essential oil were always conducted separately. This study was performed in order to assess the fatty acid and volatile organic compound composition of P. atlantica vegetable oil. The seeds were collected randomly from Djelfa (300 km South of Algiers, Algeria). Oil content and fatty acid composition were determined by Soxhlet extraction. The seeds contained high concentrations of oil (32–67%). The major fatty acids were oleic (39–49%), linoleic (23.6–31%), and palmitic (21.3–26.6%) acids. The ratio of polyunsaturated fatty acids (PUFA) to saturated fatty acids (SFA) indicated that the content of unsaturated fatty acids was approximately three times higher than that of SFA. This ratio is widely used in epidemiological studies and research on cardiovascular diseases, diabetes, and metabolic syndrome. The ratios of -acids, i.e., -9/-6 and -6/-3, were 1.3–2 and 18.5–38.3, respectively. Crushed seeds were analyzed by headspace solid-phase microextraction (SPME) coupled with gas chromatography–mass spectrometry. More than 40 compounds were identified, mainly monoterpenes (C10H16), such as α-terpinene and terpinolene, but also sesquiterpenes (C15H24) at lower levels. The value of this species as a source of healthy oil rich in -3 acid and its effects on cardiovascular disease risk are discussed.