Pharmacokinetics of primary atractyligenin metabolites after coffee consumption
Description
Roasted coffee contains atractyligenin glycosides (ATC Glcs, 2-O-β-D-glucosyl atractyligenin "ATC Glc" and 3′-O-β-D-glucosyl-2′-O-isovaleryl-2-O-β-D-glucosylatractyligenin "ATC Glcglc"), which are not reported in any other food item or non-essential food. Consumption of roasted coffee leads to plasma appearance of metabolites of ATC Glcs. These metabolites (ATC Mets) are formed from ATC Glcs provided by the coffee brew. ATC Mets are detectable in the plasma and are excreted with the urine. Analysis of plasma and urine after coffee consumption and detection of ATC Mets indicates coffee consumption, because ATC Mets are not found in biofluids when no coffee was consumed. Consequently, ATC Mets could serve as dietary biomarkers for coffee consumption. Intact ATC Glcs in roasted coffee comprise 2-O-β-D-glucosyl atractyligenin (ATC Glc) and 3′-O-β-D-glucosyl-2′-O-isovaleryl-2-O-β-D-glucosylatractyligenin (ATC Glcglc). No traces of these intact ATC Glcs from roasted coffee were detected in plasma or urine before or after coffee consumption (human intervention study, n=12 participants), indicating ATC Glcs were not bioavailable. The analytes atractyligenin ("1"), 2β-hydroxy-15-oxoatractylan-4α-carboxylic acid ("5"), atractyligenin-19-O-β-D-glucuronide ("M1"), 2β-hydroxy-15-oxoatractylan-4α-carboxy-19-O-β-D-glucuronide ("M2"), 2β-hydroxy-15-oxoatractylan-4α-carboxylic acid-2-O-β-D-glucuronide ("M3") were quantified (UHPLC-MS/MS) in urine and plasma after coffee consumption to calculate pharmacokinetic parameters. ~10% of consumed ATC Glcs were excreted as ATC Mets in the urine ("1"~65% and "M1"~35%) within 10h after coffee. Control trial samples contained only traces of ATC Mets, probably from incomplete washout. "1" and "M1" showed two plasma maxima (0.5h, 10h). Cmax was 172.2±68.5 nM (0.5h) and 60.1±35.8 nM (10h) for "1" and 22.1±7.8 nM (0.5h) and 20.5±9.1 nM (10h) for "M1". The metabolites "M2" and "M3" appeared in negligible concentration. The findings indicate that intact ATC Glcs are not bioavailable. They are cleaved early (0.5h) and late (10h) in the GI tract, leading to the bioavailable aglycone atractyligenin ("1"), which is quickly conjugated to form "M1". The plasma profiles with two maxima suggest cleavage occurs in two different regions (prox. and distal. GI). Cleavage in distal GI could involve microbiota. The low recovery (~10%) of consumed ATC Glcs in the 0-10h urine suggests either fecal excretion, substantial metabolic breakdown and/or substantially longer excretion periods beyond the studied 10 h.
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Study participants (n=12) were recruited. They underwent a three day coffee- and caffeine free period followed by the coffee intervention (1 dose of roasted coffee brew) and the control intervention (caffeine aq. 130 mg in 100 ml). Plasma samples were collected before (0h) and after 0.5, 1.0, 1.5, 2.0, 4.0, 10.0h (additional 24h plasma was collected in a subset of n=6). Urine was collected before (0h) and after 1, 4, and 10h. See the file "Protocols" for further details. The UHPLC-MS/MS method comprised optimized ion source and ion path parameters and optimized mass transitions for the analytes (see the file "Protocols"), and chromatographic gradient elution. Calibration curves were obtained by analysis of aqueous and plasma samples spiked with known concentrations of the analytes and plotting of Area ratios analyte/IS versus concentration. Urine and plasma samples from n=12 participants who completed the control trial and the coffee intervention were analysed and concentrations were calculated based on the calibration curves. See the file "protocols" for further details and literature references. Plasma concentrations (mean±SD) were plotted versus time to derive pharmacokinetic data (Tmax, Cmax, half-life, AUC). Absolute excreted amounts of metabolites were calculated from excreted volume and metabolite concentration.