Isotope dilution gas chromatographic-mass spectrometric method for the determination of lignans and isoflavonoids in human urine, including identification of genistein

Clin Chim Acta. 1991 Jul 15;199(3):263-78. doi: 10.1016/0009-8981(91)90120-2.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 4-Butyrolactone / analogs & derivatives
  • 4-Butyrolactone / urine
  • Butylene Glycols / urine
  • Chromans / urine
  • Equol
  • Female
  • Furans / urine
  • Gas Chromatography-Mass Spectrometry
  • Genistein
  • Humans
  • Indicator Dilution Techniques
  • Isoflavones / chemistry
  • Isoflavones / urine*
  • Lignans
  • Lignin / analysis*
  • Magnetic Resonance Spectroscopy
  • Male


  • 4',7-dihydroxy-3,4-dihydroisoflavone
  • Butylene Glycols
  • Chromans
  • Furans
  • Isoflavones
  • Lignans
  • Equol
  • matairesinol
  • daidzein
  • 2,3-bis(3'-hydroxybenzyl)butane-1,4-diol
  • Lignin
  • Genistein
  • 4-Butyrolactone
  • O-desmethylangolensin
  • 2,3-bis(3'-hydroxybenzyl)butyrolactone