The phytochemical content and the antioxidant activity in the inflorescences of six industrial hemp (Cannabis sativa L.) genotypes, four monoecious (Codimono, Carmaleonte, Futura 75, and Santhica 27), and two dioecious (Fibrante and Carmagnola Selezionata), were assessed for three consecutive years from 2018 to 2020. The total phenolic content, total flavonoid content, and antioxidant activity were determined by spectrophotometric measurements, whereas HPLC and GC/MS were used to identify and quantify the phenolic compounds, terpenes, cannabinoids, tocopherols, and phytosterols. All the measured traits were significantly affected by genotype (G), cropping year (Y), and their interaction (G × Y), although the Y effect prevailed as a source of variation, ranging from 50.1% to 88.5% for all the metabolites except cannabinoids, which were equally affected by G, Y, and G × Y interaction (33.9%, 36.5%, and 21.4%, respectively). The dioecious genotypes presented a more constant performance over the three years compared to the monoecious genotypes, with the highest and most stable phytochemical content observed in the inflorescences of Fibrante, which was characterized by the highest levels of cannabidiol, α-humulene and β-caryophyllene, which may confer on the inflorescences of this genotype a great economic value due to the important pharmacological properties of these metabolites. Conversely, the inflorescences of Santhica 27 were characterized by the lowest accumulation of phytochemicals over the cropping years, with the notable exception of cannabigerol, a cannabinoid that exhibits a wide range of biological activities, which was found at its highest level in this genotype. Overall, these findings can be used by breeders in future programs aimed at the selection of new hemp genotypes with improved levels of phytochemicals in their inflorescences, which can provide better health and industrial benefits.
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