Nutritional profiling of cold-pressed hemp seed oils: comprehensive analysis of fatty acids, fahfas, and cannabinoids

Hemp oil, derived from the seeds of Cannabis sativa, has recently gained significant attention for its nutritional and health benefits. In this study, a cold pressing extraction method for producing hemp seed oil is presented. Oils were obtained from three hemp cultivars, Felina 32, Futura 75, and Uso 31, commercially available in Italy and France. The oils were then characterized using UHPLC-QTOF/MS, and the levels of fatty acid esters of hydroxy fatty acids (FAHFA) were quantified using an LC-MS/MS platform. Furthermore, a GC-FID method was employed to measure fatty acids as methyl esters (FAME). This study is the first to systematically identify and quantify fatty acid esters of hydroxy fatty acids (FAHFAs) in cold-pressed hemp seed oil, revealing their potential nutritional and health-promoting relevance. FAHFAs, formed by the esterification of hydroxy fatty acids with long-chain fatty acids, are known to possess antidiabetic and anti-inflammatory properties making them significant from a nutritional standpoint. Across all cultivars, we identified and measured nine distinct FAHFA compounds, which primarily contain linoleic acid, oleic acid, linolenic acid and medium chain hydroxy fatty acids in their backbone. Specifically, FAHFA (18:2_6:0) and FAHFA (18:2_7:0) were the most prevalent FAHFA species found in hemp oils. Consistently, the most abundant fatty acids measured as FAME were linoleic acid, oleic acid, and palmitic acid, further supporting their role as acyl donors to esterify hydroxy fatty acids during FAHFA synthesis. These results provide valuable insights into the functional, nutritional and health-promoting properties of hemp seed oil.