Quantitative analysis of phytoestrogens in kudzu-root, soy and spiked serum samples by high-pressure liquid chromatography

Elhabib Benlhabib, John I. Baker, Daniel E. Keyler, Ashok K. Singh

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

A sensitive and reliable HPLC method that allows simultaneous quantification of phytoestrogens extracted from kudzu-root and soy preparations, and serum samples has been developed. Kudzu-root and soy preparations were mixed with 5 μg flavone and 15 μg rutin (internal standards) and the phytoestrogens were extracted by using solid-phase (C18) extraction cartridges. Blank or spiked serum samples were extracted by using either C18 cartridges or trichloroacetic acid-methanol extraction. The extracts were analyzed by the HPLC equipped with a reverse-phase (250 × 4 mm, C18 column and UV, diode-array or MS detector. A linear gradient of acetic acid and acetonitrile provided excellent separation of glycoside and aglycone-phytoestrogens from kudzu root and soy preparations. The C18 cartridge extraction of serum yielded excellent recovery of both glycoside- and aglycone-phytoestrogens, while the trichloroacetic acid-methanol extraction yielded excellent recovery of glycoside but poor recovery of aglycone compounds. UV and MS detectors were suitable for phytoestrogen analysis in plant and serum samples, while the diode-array detector was suitable for generating the UV absorbance curve for phytoestrogens.

Original languageEnglish (US)
Pages (from-to)367-380
Number of pages14
JournalBiomedical Chromatography
Volume18
Issue number6
DOIs
StatePublished - Jul 1 2004

Keywords

  • Daidzein
  • Daidzin
  • Genistein
  • Genistin
  • Kudzu-root
  • Phytoestrogens
  • Puerarin
  • Serum
  • Soy

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