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.

Germination and its role in phenolic compound bioaccessibility for black mustard grains: A study using INFOGEST protocol.

Germination has been regarded as a promising natural process to improve the antioxidant properties of mustard. However, there ís one question to be solved in this area: does germination improve mustard phenolics' bioaccessibility? The aim of this study was to answer this question by using INFOGEST protocol to simulate in vitro digestion. Resveratrol, formononetin and cryptochlorogenic acid were identified for the first time as evaluated by liquid chromatography-mass spectrometry. In general, digestion positively impacted the antioxidant potential of soluble phenolics from non-germinated and germinated grains, which were probably released from cell wall matrix by digestive enzymes. Although digestion seemed to nullify the antioxidant improvement caused by germination, phenolic quantities were distinctive. The main difference was found for sinapic acid, as its concentration reached a value 1.75-fold higher in germinated digested mustard compared to non-germinated. The results obtained suggested that germination improved the phenolic bioaccessibility of mustard grains, which encourages its use and investigations.