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 language | English (US) |
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Pages (from-to) | 367-380 |
Number of pages | 14 |
Journal | Biomedical Chromatography |
Volume | 18 |
Issue number | 6 |
DOIs | |
State | Published - Jul 2004 |
Keywords
- Daidzein
- Daidzin
- Genistein
- Genistin
- Kudzu-root
- Phytoestrogens
- Puerarin
- Serum
- Soy