title: Development of analyses by high-performance liquid chromatography and liquid chromatography/tandem mass spectrometry of bilberry (Vaccinium myrtilus) anthocyanins in human plasma and urine.
abstract: Anthocyanins are potent antioxidants that may possess chronic disease preventive properties. Here, rapid, reliable, and reproducible solid-phase extraction, high-performance liquid chromatography (HPLC), and mass spectrometry techniques are described for the isolation, separation, and identification of anthocyanins in human plasma and urine. Recoveries of cyanidin-3-glucoside (C3G) were 91% from water, 71% from plasma, and 81% from urine. Intra- and interday variations for C3G extraction were 9 and 9.1% in plasma and 7.1 and 9.1% in urine and were less than 15% for all anthocyanins from a standardized bilberry extract (mirtoselect). Analysis of mirtoselect by HPLC with UV detection produced spectra with 15 peaks compatible with anthocyanin components found in mirtoselect within a total run time of 15 min. Chromatographic analysis of human urine obtained after an oral dose of mirtoselect yielded 19 anthocyanin peaks. Mass spectrometric analysis employing multiple reaction monitoring suggests the presence of unchanged anthocyanins and anthocyanidin glucuronide metabolites.
authors.affiliation: Cancer Biomarkers and Prevention Group, Department of Cancer Studies and Molecular Medicine, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester LE2 7LX, United Kingdom.
authors.forename: Darren N
authors.initials: DN
authors.lastname: Cooke
authors.forename: Sarah
authors.initials: S
authors.lastname: Thomasset
authors.forename: David J
authors.initials: DJ
authors.lastname: Boocock
authors.forename: Michael
authors.initials: M
authors.lastname: Schwarz
authors.forename: Peter
authors.initials: P
authors.lastname: Winterhalter
authors.forename: William P
authors.initials: WP
authors.lastname: Steward
authors.forename: Andreas J
authors.initials: AJ
authors.lastname: Gescher
authors.forename: Timothy H
authors.initials: TH
authors.lastname: Marczylo
mesh_terms: D000872:Anthocyanins / Q000008:administration & dosage / Q000097:blood* / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D005638:Fruit / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods*; D029800:Vaccinium myrtillus / Q000737:chemistry*
chemical_list: D000872:Anthocyanins