Biotransformation of white and black mustard grains through germination and enzymatic hydrolysis revealed important metabolites for antioxidant properties and cytotoxic activity against Caco-2 cells.

Germination and enzymatic hydrolysis are biological processes with well-recognized positive effects on phenolic composition and antioxidant potential. This study aimed to apply those processes to white (Sinapsis alba) and black (Brassica nigra) mustard grains and to analyze the influences on the total phenolic content (TPC); phenolic and peptide profile determined by ultra-performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS); antioxidant potential (DPPH, ABTS, and FRAP assays); and cytotoxicity against Caco-2, a human colorectal adenocarcinoma cell line. Enzyme combinations for hydrolysis were different for each mustard grain, but for both species, enzymatic hydrolysis and germination showed a positive effect on antioxidant properties. From UPLC-HRMS analysis and molecular network studies, 14 peptides and 17 phenolic compounds were identified as metabolites released from mustard after processes application, which were strongly correlated with increased antioxidant activity. In addition, enzymatic hydrolysis applied in germinated mustard grains for both mustards increased the cytotoxic activity against Caco-2 human colorectal adenocarcinoma cell line.

Bioaccessibility of phenolic compounds using the standardized INFOGEST protocol: A narrative review.

The INFOGEST protocol creation was a watershed for phenolic bioaccessibility studies. Because of this important initiative to standardize bioaccessibility studies, data comparisons between different laboratories are now expedited. It has been eight years since the INFOGEST protocol creation, and three from the latest update. However, the current status in terms of phenolic bioaccessibility and how far different laboratories are from reaching a consensus are still unrevealed. In this sense, this narrative review considered an evaluation of different studies that applied the INFOGEST protocol to investigate the bioaccessibility of phenolic compounds. The central objective was to compile the main findings and consensus and to identify possible gaps and future opportunities. This approach intends to further facilitate the use of this protocol by professionals in the field of food science and technology and related areas, generating a reflection on the actual level of standardization of the method. Despite the differences in phenolic compounds from diverse food matrices, and their peculiar behavior, some trends could be elucidated, in terms of phenolic release, stability, and/or transformation upon in vivo digestion. In contrast, there was no general consensus regarding sample preparation, how to report results and the form to calculate bioaccessibility, making it difficult to compare different studies. There is still a long road to effectively standardize the results obtained for phenolic bioaccessibility using the INFOGEST protocol, which is also an opportunity in terms of food analysis that can impact the food industry, especially for the development of nutraceuticals and functional foods.

Free and insoluble-bound phenolics: How does the variation of these compounds affect the antioxidant properties of mustard grains during germination?

This work aimed to investigate how the variation of free and insoluble-bound phenolics affected the antioxidant properties of mustard grains from two varieties (black - Brassica nigra and white - Sinapsis alba) during different germination parameters. The germination conditions selected for each mustard variety to improve their antioxidant properties were different, as follows: (a) for white mustard - 72 h of germination at 25 °C in the dark and (b) for black mustard - 48 h of germination at 25 °C alternating dark and light periods. At these conditions, increases of 49, 72, 80, 68, 42, 66 and 45% were detected for total phenolic compounds (TPC), total flavonoids, condensed tannins, FRAP, DPPH, ABTS, and ORAC, respectively, for soluble extracts of white mustard compared to the non-germinated white mustard. The soluble extracts from black mustard, in turn, presented increases of 44, 18, 55, 29, 3, 160 and 42% for TPC, total flavonoids, condensed tannins, FRAP, DPPH, ABTS, and ORAC, respectively, compared to the non-germinated sample. Gallic acid, 3,4-di-hydroxybenzoic acid, sinapic acid, ferulic acid, coumaric acid, and rutin were identified by UPLC-MS/MS and were the main compounds detected in mustard extracts. Given the results obtained, germinated mustard grains have the potential for application as a functional and nutraceutical food.

Alkaline conditions better extract anti-inflammatory polysaccharides from winemaking by-products.

Winemaking generates large amounts of by-products, a well recognized source of phenolic compounds. However, less attention has been paid to the polysaccharide-rich fraction (PRF) and effects of fractionation techniques on its potential bioactivity. Therefore, PRFs from Syrah and Tempranillo winemaking by-products were extracted under aqueous (neutral pH conditions), acidic and alkaline conditions. PRFs were screened for their monosaccharide composition, uronic acid content, homogeneity and molecular weight. Anti-inflammatory activity of PRFs were evaluated on stimulated RAW 264.7 macrophages. PRF obtained in water and/or under acidic conditions showed heterogeneous profiles. As like as in the others, a heterogeneous and complex profile was detected in extracts procured under alkaline conditions. A high content of uronic acid was found in aqueous extracts, thus indicating the presence of pectin. Pectin and hemicellulose were present in PRFs procured under acidic conditions. Alkaline conditions rendered extracts containing a complex mixture of monosaccharides, mainly xylose. This latter PRF was the only one exhibiting anti-inflammatory potential (at 100 µg/mL) by reducing the release of TNF-α and activation of NF-κB in LPS-activated RAW 264.7 macrophages, with no effect on cell viability. Regardless of the grape variety, PRFs obtained under alkaline conditions were the best option to obtain bioactive polysaccharides with potential application as a source of anti-inflammatory compounds. A complex mixture of polymers may be responsible for the anti-inflammatory effects. Finally, according to results procured by NMR, it is possible to suggest that bioactive fractions are composed of a chain of α-L-Araf-(1 → 3) linked, ß-D-Xylp- (1 → 4), α-D-Glcp-(1 → 4) linked, α-D-GalpA-(1 → 4), α-D-Gal-(1 → 2) forming possible RG I and RG II and xylan chains.

Should we ban total phenolics and antioxidant screening methods? The link between antioxidant potential and activation of NF-κB using phenolic compounds from grape by-products.

Free radical imbalance is associated with several chronic diseases. However, recent controversies have put in check the validity of colorimetric methods to screen the functionality of polyphenols. Therefore, in this study two antioxidant methods, based on chemical reactions, were tested for their ability in anticipating the reduction of the activation of NF-κB using LPS-activated RAW 264.7 macrophages, selected as a biological model. Grape processing by-products from winemaking showed higher total phenolic content (TPC), antioxidant capacity towards peroxyl radical (31.1%) as well as reducing power (39.5%) than those of grape juice by-products. The same trend was observed when these samples were tested against LPS-activated RAW 264.7 macrophages by reducing the activation NF-κB. Feedstocks containing higher TPC and corresponding ORAC and FRAP results translated to higher reduction in the activation of NF-κB (36.5%). Therefore, this contribution demonstrates that colorimetric methods are still important screening tools owing their simplicity and widespread application.

Phenolic acids and flavonoids of peanut by-products: Antioxidant capacity and antimicrobial effects.

Peanut skin (PS) and meal from dry-blanched peanuts (MDBP) were evaluated as sources of phenolic compounds. PS rendered the highest total phenolic content, antioxidant capacity towards ABTS radical cation, DPPH and hydroxyl radicals as well as reducing power. Phenolic acids were present in PS and MDBP whereas proanthocyanidins and monomeric flavonoids were found only in PS as identified by HPLC-DAD-ESI-MSn. Procyanidin-rich extracts prevented oxidation in non-irradiated and gamma-irradiated fish model system. Both extracts inhibited the growth of gram-positive (Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Geobacillus stearothermophilus) and gram-negative bacteria (Pseudomonas aeruginosa, Pseudomonas fluorescens, Salmonella Enteritidis, Salmonella Typhimurium, Escherichia coli). Regardless of the strain, phenolic acid-rich extracts showed the lowest minimum inhibitory capacity (MIC); therefore presenting higher antibacterial effect. The MIC of phenolic acid-rich extracts (24-49µgphenolics/mL) was higher but comparable to Ampicillin (10µg/mL). Thus, phenolics in PS and MDBP may serve as antioxidants and antimicrobial compounds.