title: Isotope dilution gas chromatographic-mass spectrometric method for the determination of lignans and isoflavonoids in human urine, including identification of genistein.
abstract: We describe an isotope dilution gas chromatographic-mass spectrometric method for the quantitative determination of the lignans enterolactone, enterodiol and matairesinol and the isoflavonoids daidzein, equol, O-desmethylangolensin and genistein in urine. Furthermore we present the gas chromatographic/mass spectrometer identification of genistein. Urine samples were extracted on Sep-Pak cartridges, conjugated fractions were isolated by chromatography on the acetate form of DEAE-Sephadex and deuterated internal standards of all seven compounds were added to the samples before hydrolysis. The hydrolysate was extracted on a Sep-Pak cartridge and following chromatography on the acetate form of QAE-Sephadex two fractions were obtained: Fraction 1 contained equol, enterolactone, enterodiol, matairesinol and all estrogens and fraction 2 contained O-desmethylangolensin, daidzein and genistein. The latter was ready for gas chromatography/mass spectrometry, but the first one was further purified to eliminate the estrogens by chromatography on the carbonate form of QAE-Sephadex. Following silylation, the samples were analyzed by combined capillary column gas chromatography/mass spectrometry in the selective ion monitoring mode. The within-assay imprecision varied from 0.8-15.2% (mean 8.7%) and the between-assay imprecision from 4.1-13.9% (mean 9.3%), depending on compound and concentration level. The mean recovery of authentic standards added to urine extracts before hydrolysis varied from 96.6 to 105.5%. Values obtained from 10 Finnish omnivorous men are presented. Individual values for matairesinol (excretion range 3.3-59.9 nmol/24 h) and genistein (range 21.8-1180 nmol/24 h) in human urine have never been published before.
authors.affiliation: Department of Clinical Chemistry, University of Helsinki, Finland.
authors.forename: H
authors.initials: H
authors.lastname: Adlercreutz
authors.forename: T
authors.initials: T
authors.lastname: Fotsis
authors.forename: C
authors.initials: C
authors.lastname: Bannwart
authors.forename: K
authors.initials: K
authors.lastname: Wähälä
authors.forename: G
authors.initials: G
authors.lastname: Brunow
authors.forename: T
authors.initials: T
authors.lastname: Hase
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000652:urine; D002072:Butylene Glycols / Q000652:urine; D002839:Chromans / Q000652:urine; D060754:Equol; D005260:Female; D005663:Furans / Q000652:urine; D008401:Gas Chromatography-Mass Spectrometry; D019833:Genistein; D006801:Humans; D007201:Indicator Dilution Techniques; D007529:Isoflavones / Q000737:chemistry / Q000652:urine*; D017705:Lignans; D008031:Lignin / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy; D008297:Male
chemical_list: C007610:4',7-dihydroxy-3,4-dihydroisoflavone; D002072:Butylene Glycols; D002839:Chromans; D005663:Furans; D007529:Isoflavones; D017705:Lignans; D060754:Equol; C068935:matairesinol; C004742:daidzein; C029498:2,3-bis(3'-hydroxybenzyl)butane-1,4-diol; D008031:Lignin; D019833:Genistein; D015107:4-Butyrolactone; C050052:O-desmethylangolensin; C029497:2,3-bis(3'-hydroxybenzyl)butyrolactone