Aim of the project was the bioassay guided optimisation of extraction methods applied to wine chain by-product to obtain extracts, fractions and biologically active biomolecules with a possible use in the nutraceutical and cosmeceutical industry. Exhausted red and white grape marc were extracted using water:ethanol 50:50 with ultrasound assisted extraction and NaviglioV R technology; and also with supercritical fluid extraction (SFE) and steam-distillation obtaining different phytocomplexes. Each extract was characterised by different molecular category: exhausted red grape marc (VCR) by anthocyanins, exhausted white grape marc (VCB) by flavonoids, and grapeseed (VIN) by proanthocyanins. SFE and steam-distillation, instead, highlighted the presence of fatty acids and their ester in every matrix, but terpenoids were not revealed at level major or equal than 0.1%, except for manoyl oxide in VCR (2.89%). VIN was the most abundant matrix in polyphenols (506.24 ± 55.91 mg gallic acid/g dried extract), and it showed the highest antioxidant activity (IC 50 of 4.30 lg/mL). Regarding the antimicrobial activity, the hydroalcoholic extracts from VCR, VCB and VIN were tested but no noteworthy activities have been recorded.
Maize ( Zea mays L.) germs are by-products from the milling industry. The objective of this work was to compare the phenolic and lipophilic antioxidant fractions of yellow and white corn varieties, provided by Corn Valley S.r.l. (Piumbega, Mantova, Italy) and among the raw materials most processed by the company. The phenolic fraction, extracted with ultrasound-assisted extraction, alone and in combination with chemical and enzymatic hydrolyses, was analyzed with high-performance thin-layer chromatography and reversed-phase high-performance liquid chromatography-diode array detector. Among the various extraction techniques used, the combination of sonication and alkaline hydrolysis proved to be an effective method for the extraction of phenolic compounds from yellow and white germs, with the highest ferulic acid concentrations (636.54 ± 3.71 and 569.23 ± 1.69 mg FA/g dried extract, respectively), total phenolic contents (844.5 ± 64.6 and 742.8 ± 15.44 mg gallic acid equivalents/g dried extract, respectively), and the best antioxidant activity (14.33 ± 0.48 and 11.41 ± 1.1 µg/mL, respectively). The lipophilic fraction, extracted using supercritical carbon dioxide was analyzed by gas chromatography-mass spectrometry. The unsaponifiable fractions were found to be 2.41% ± 0.24% in yellow corn and 1.85% ± 0.08% in white corn; β-sitosterol, campesterol, and stigmasterol were identified as the main phytosterols characterizing both lipophilic extracts which showed the most effective antioxidant activity (1.29 ± 0.26 mg/mL and 1.33 ± 0.21 mg/mL, respectively) compared with the control. Finally, the phenolic and lipophilic extracts obtained from maize by-products may be reintroduced into the health-oriented market as extracts enriched of high-added value biomolecules with antioxidant activity both as active molecules and as additives of natural origin.
Vitis vinifera L. leaves from pruning are by-products of the wine industry and represent an important source of secondary raw material, thanks to their polyphenols content. Optimization of the extraction processes is a key factor for their valorization, and Design of Experiment (DOE) could be a tool to obtain the most performing extract in terms of polyphenols quality/quantity and bioactivity. Vitis vinifera Lambrusco leaves were subjected to ultrasound-assisted extractions guided by a 23 factorial design. Three independent parameters (% solvent, time of extraction, and solvent:solid ratio) were considered to evaluate the extraction process by analyzing the extraction yield, the total phenolic content (Folin-Ciocalteu assay), and the antioxidant capacity (DPPH assay). Moreover, the content of the main molecules was identified and quantified by reversed-phase high-performance liquid chromatography coupled with diode array detection and mass spectrometry. The DOE highlighted the best extraction conditions that showed slight changes considering the different evaluating parameters. The highest extraction yield was obtained by extraction with 100% water, 60 minutes of extraction time, and 30:1 solvent:solid ratio, but it was neither the richest in polyphenols nor antioxidant capacity. The latter 2 characteristics were associated with the extraction performed using 50% ethanol, 35 minutes of extraction time, and a 20:1 solvent:solid ratio. That extract also exhibited the highest quantity of flavonols.
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