Bioactive compounds from hemp (Cannabis sativa L.) seeds: optimization of phenolic antioxidant extraction using simplex lattice mixture design and HPLC-DAD/ESI-MS2 analysis

Abstract

The extraction of phenolic compounds from defatted hempseeds was optimized using a simplex lattice mixture design with three solvents (water, methanol, and acetone). The response variables were total phenolic content (TPC) and antioxidant activity evaluated by different spectrophotometric tests. The results showed that the binary acetone-water mixture in equal proportions is the optimal combination to achieve the maximum TPC (53.65 mg GAE per g extract) with higher antioxidant activities (265.53, 36.25, 119.03, 69.46, and 68.91 mg TE g⁻¹ extract for the TAC, DPPH, ABTS, FRAP, and CUPRAC tests respectively). In addition, the phenolic profile analysis of defatted hemp seeds by HPLC-DAD/ESI-MS² techniques showed the predominance of hydroxycinnamic acid amides and lignanamides. It allowed visualizing the effect of each solvent mixture on the relative extracted amount of each identified phenolic compound. This study suggests that N-trans-caffeoyltyramine, cannabisin A, and cannabisin B might contribute strongly to the potent antioxidant activity of hempseed extracts. Thus, it encourages the use of defatted hemp seeds as a source of antioxidants with added value for pharmaceutical and cosmetic applications.