In 2022, 5 varieties of industrial hemp (Cannabis sativa L.) were planted to estimate grain production as an alternative source of nutritional compounds due to its fat, carbohydrate and protein content. Futura 75, Futura 83, Felina 32, Earlina 8 FC and Henola are included in the EU common catalogue of varieties of plant species and they were chosen due to its good field adaptation and productivity. Once the crop was fully developed, the seed was collected by variety, and within a representative sampling of 10 m2 the harvested seed was manually and mechanically cleaned to eliminate other plant material. The aim of this study was to characterise the chemical profile of different varieties of hemp seeds for food use. The fat content of the seeds was determined and the fatty acid profile was measured by gas chromatography. The fat content of the hemp seeds studied ranged between 23 and 31% of total composition. Earlina 8 FC was significantly (p < 0.05) the fattest. The varieties studied showed a high percentage of unsaturated fats (87% of total fatty acids). In addition, polyunsaturated acids were the main group, especially in Earlina 8 FC and Felina 32, being linoleic acid the most abundant. Thus, hemp seeds can be a valuable vegetable source of healthy fatty acids. The production of food products with a majority hemp seed base could be considered as “high polyunsaturated fat” according to Commission Regulation (EU) No 116/2010 of 9 February 2010.