TY - JOUR
T1 - Elucidation of (-)-epicatechin metabolites after ingestion of chocolate by healthy humans
AU - Actis-Goretta, Lucas
AU - Lévèques, Antoine
AU - Giuffrida, Francesca
AU - Romanov-Michailidis, Fedor
AU - Viton, Florian
AU - Barron, Denis
AU - Duenas-Paton, Montserrat
AU - Gonzalez-Manzano, Susana
AU - Santos-Buelga, Celestino
AU - Williamson, Gary
AU - Dionisi, Fabiola
PY - 2012/8/15
Y1 - 2012/8/15
N2 - After absorption in the gastrointestinal tract, (-)-epicatechin is extensively transformed into various conjugated metabolites. These metabolites, chemically different from the aglycone forms found in foods, are the compounds that reach the circulatory system and the target organs. Therefore, it is imperative to identify and quantify these circulating metabolites to investigate their roles in the biological effects associated with (-)-epicatechin intake. Using authentic synthetic standards of (-)-epicatechin sulfates, glucuronides, and O-methyl sulfates, a novel LC-MS/MS-MRM analytical methodology to quantify (-)-epicatechin metabolites in biological matrices was developed and validated. The optimized method was subsequently applied to the analysis of plasma and urine metabolites after consumption of dark chocolate, an (-)-epicatechin-rich food, by humans. (-)-Epicatechin-3′-β-d-glucuronide (Cmax 290±49 nM), (-)-epicatechin 3′-sulfate (Cmax 233±60 nM), and 3′-O-methyl epicatechin sulfates substituted in the 4′, 5, and 7 positions were the most relevant (-)-epicatechin metabolites in plasma. When plasmatic metabolites were divided into their substituent groups, it was revealed that (-)-epicatechin glucuronides, sulfates, and O-methyl sulfates represented 33±4, 28±5, and 33±4% of total metabolites (AUC0-24 h), respectively, after dark chocolate consumption. Similar metabolites were found in urine samples collected over 24 h. The total urine excretion of (-)-epicatechin was 20±2% of the amount ingested. In conclusion, we describe the entire metabolite profile and its degree of elimination after administration of (-)-epicatechin-containing food. These results will help us understand more precisely the mechanisms and the main metabolites involved in the beneficial physiological effects of flavanols.
AB - After absorption in the gastrointestinal tract, (-)-epicatechin is extensively transformed into various conjugated metabolites. These metabolites, chemically different from the aglycone forms found in foods, are the compounds that reach the circulatory system and the target organs. Therefore, it is imperative to identify and quantify these circulating metabolites to investigate their roles in the biological effects associated with (-)-epicatechin intake. Using authentic synthetic standards of (-)-epicatechin sulfates, glucuronides, and O-methyl sulfates, a novel LC-MS/MS-MRM analytical methodology to quantify (-)-epicatechin metabolites in biological matrices was developed and validated. The optimized method was subsequently applied to the analysis of plasma and urine metabolites after consumption of dark chocolate, an (-)-epicatechin-rich food, by humans. (-)-Epicatechin-3′-β-d-glucuronide (Cmax 290±49 nM), (-)-epicatechin 3′-sulfate (Cmax 233±60 nM), and 3′-O-methyl epicatechin sulfates substituted in the 4′, 5, and 7 positions were the most relevant (-)-epicatechin metabolites in plasma. When plasmatic metabolites were divided into their substituent groups, it was revealed that (-)-epicatechin glucuronides, sulfates, and O-methyl sulfates represented 33±4, 28±5, and 33±4% of total metabolites (AUC0-24 h), respectively, after dark chocolate consumption. Similar metabolites were found in urine samples collected over 24 h. The total urine excretion of (-)-epicatechin was 20±2% of the amount ingested. In conclusion, we describe the entire metabolite profile and its degree of elimination after administration of (-)-epicatechin-containing food. These results will help us understand more precisely the mechanisms and the main metabolites involved in the beneficial physiological effects of flavanols.
KW - (-)-Epicatechin
KW - Chocolate
KW - Cocoa
KW - Free radicals
KW - Humans
KW - Metabolites
UR - http://www.scopus.com/inward/record.url?scp=84864397350&partnerID=8YFLogxK
U2 - 10.1016/j.freeradbiomed.2012.05.023
DO - 10.1016/j.freeradbiomed.2012.05.023
M3 - Article
C2 - 22664313
AN - SCOPUS:84864397350
VL - 53
SP - 787
EP - 795
JO - Free Radical Biology and Medicine
JF - Free Radical Biology and Medicine
SN - 0891-5849
IS - 4
ER -