Wild hemp (Cannabis sativa L. ssp. sativa var. spontanea) is a special variety of industrial hemp (Cannabis sativa L. ssp. sativa), which is believed to have originated from the acclimatization of industrial hemp to the ruderal habitat conditions of Eastern Europe. In Serbia it is found along roads, on field edges, stubble fields, garbage dumps etc., but also as a weed in fields. Due to the great popularity of industrial hemp, which can be used for various purposes (as raw material in food and pharmaceutical industry, in the production of gluten-free flour, biodiesel, detergents, in folk medicine), many researchers have analyzed its seeds, including analysing their fatty acids content. However, even though this knowledge can be of chemotaxonomic, ecological, evolutionary and nutritional significance, there is no data on the presence and composition of fatty acids in wild hemp seeds For the analysis of fatty acids in wild hemp seeds, the seed material was collected from two locations. After the extraction with hexane, determination of fatty acid esters was performed by capillary gas chromatography. Chromatographic peaks in the samples were identified by comparing the retention times with the retention times of the fatty acid esters in the analytical standard of a mixture of 37 fatty acid esters. The fatty acid composition is expressed as the relative mass fraction of the total fatty acids. The analysis of fatty acids in wild hemp seeds reveales the presence of 15 different acids, with content of 17.5% (U1) and 14.7% (U2) in relation to the total seed weight. Linoleic (45.3 and 47.5%) and a-linoleic (13.6 and 15.5%) acids were the most dominant. Of the 15 detected fatty acids, only 4 are saturated, with a representation of about 12%. Unsaturated fatty acids are present in a significantly higher proportion (about 88%), which indicates the nutritional value of these seeds.