Metabolomic investigation of inflorescences from Cannabis sativa L. variety Earlina 8FC cultivated in southern Italy: an NMR and LC-MS/MS integrated approach

Abstract Earlina 8FC is a monoecious hemp variety developed for seed production characterized by early flowering, a short development period, a high seed yield, and good pest resistance. This work, to the best of our knowledge, reports the first metabolomic investigation of Earlina 8FC inflorescences. For comparison, the same analytical protocol was applied to the well-characterized Futura 75 variety. Both highly polar and less polar/nonpolar compounds were extracted simultaneously as distinct hydroalcoholic and organic (chloroform) extracts. The extracts were analyzed by NMR after being dried and reconstituted in four different solvents for better fractionation. In the hydroalcoholic extract 23 key primary metabolism markers, such as sugars, cyclitols, amino acids, and organic acids were identified by NMR. The six most abundant flavonoids in hydroalcoholic extract, including apigenin and luteolin glycosides, orientin and vitexin were identified and quantified by LC-MS/MS. The most abundant components of organic extracts, including cannabinoids, fatty acids, phospholipids, galactolipids, sterols, and chlorophyll degradation products, were identified and quantified by NMR. According to ANOVA, Earlina 8FC had significantly higher levels of acetic acid and formic acid, a similar level of chlorogenic acid, as well as significantly lower levels of flavonoids, cannabidiolic acid, cannflavin A, myo-inositol, and quebrachitol, compared to Futura 75.