Evaluation of Apricot, Bilberry, and Elderberry Pomace Constituents and Their Potential To Enhance the Endothelial Nitric Oxide Synthase (eNOS) Activity.

Pomace, the press residue from different fruits accumulating as waste product in food industry, contains high amounts of secondary metabolites that could be utilized for health-related applications. This study aims at evaluating the potential of pomaces of apricot, bilberry, and elderberry to serve as a source for endothelial nitric oxide synthase (eNOS)-activating compounds. Five extracts obtained from the lyophilized pomace of apricot and elderberry with solvents of different polarity were found to enhance A23187-stimulated eNOS activity when tested at 50 µg/mL in an [14C]-l-arginine to [14C]-l-citrulline conversion assay in the human endothelium-derived cell line EA.hy926 (p < 0.05). The bioassay-guided fractionation of the extracts obtained with methanol/water (70:30) led to several active fractions from apricot pomace (p < 0.05) and elderberry pomace (p < 0.01). Liquid chromatography-mass spectrometry-based chemical analysis of the extracts and active fractions pointed mainly to triterpenoic acids as active compounds. One particular dihydroxytriterpenoic acid, characteristic for elderberry, was enriched as the main compound in the two most active fractions and might serve as a promising lead structure for further studies.

Triterpenoic Acids from Apple Pomace Enhance the Activity of the Endothelial Nitric Oxide Synthase (eNOS).

Pomace is an easy-accessible raw material for the isolation of fruit-derived compounds. Fruit consumption is associated with health-promoting effects, such as the prevention of cardiovascular disease. Increased vascular nitric oxide (NO) bioavailability, for example, due to an enhanced endothelial nitric oxide synthase (eNOS) activity, could be one molecular mechanism mediating this effect. To identify compounds from apple (Malus domestica Borkh.) pomace that have the potential to amplify NO bioavailability via eNOS activation, a bioassay-guided fractionation of the methanol/water (70:30) extract has been performed using the (14)C-L-arginine to (14)C-L-citrulline conversion assay (ACCA) in the human endothelium-derived cell line EA.hy926. Phytochemical characterization of the active fractions was performed using the spectrophotometric assessment of the total phenolic content, as well as TLC, HPLC-DAD-ELSD, and HPLC-MS analyses. Eleven triterpenoic acids, of which one is a newly discovered compound, were identified as the main constituents in the most active fraction, accompanied by only minor contents of phenolic compounds. When tested individually, none of the tested compounds exhibited significant eNOS activation. Nevertheless, cell stimulation with the reconstituted compound mixture restored eNOS activation, validating the potential of apple pomace as a source of bioactive components.