Ultra-Performance Liquid Chromatography/Mass Spectrometry Study of Metabolism of 5-Methylpyranopelargonidin
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Anthocyanin dyes represent an important group of plant polyphenols significantly affecting human diet. Their stability in solution is low and it depends on pH, presence of sulfur dioxide and other parameters. Many stable pyranoanthocyanins are formed by condensation of anthocyanins with small reactive compounds, i.e. pyruvic acid, acetaldehyde, acetone and others, commonly during maturation of food products (e.g. wines and juices). To date, little is known about their metabolism. This communication deals with the study of in vitro metabolism of 5-methylpyranopelargonidin (5-MePPl) as a simple member of the pyranoanthocyanin family and its direct comparison with common flavonoids quercetin and pelargonidin. Human colon adenocarcinoma cells LS174T and human hepatocellular carcinoma cells HepG2 were used for experiments. Metabolites were analyzed by UPLC/MS/MS. Quercetin underwent extensive biotransformation with a metabolic profile typical for flavonoids, corroborating that the used cells were metabolically active in parallel experiments with pelargonidin and 5-MePPl (condensation of two and three quercetin molecules was tentatively proposed in addition). Biotransformation of 5-methylpyranopelargonidin proceeded with higher yield of metabolites compared to pelargonidin. Processes related to accumulation of 5-MePPl metabolites in pellets exhibit significant differences compared to quercetin and pelargonidin. The fact can be ascribed to the presence of an additional ring (D) in the flavonoid skeleton and probably to a higher hydrophobicity of pyrano-dye compared to both other studied flavonoids. Hydroxylation, glucuronation and glucosidation are the main metabolic processes observed during in vitro metabolization of 5-MePPl providing several chromatographically resolved isomers of metabolites. Moreover, some combined metabolites were found and the site of metabolization in the 5-MePPl structure was specified based on collision spectra.
KeywordsUPLC/MS Anthocyanin Pyranoanthocyanin Metabolite
Authors thank to the support of Grant Agency of the Czech Republic (GACR 303/12/G163) and Operational Program Research and Development for Innovations—European Regional Development Fund (project CZ.1.05/2.1.00/03.0058) of the Ministry of Education, Youth and Sports of the Czech Republic.
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