Quantitative analysis of phytoestrogens in kudzu-root, soy and spiked serum samples by high-pressure liquid chromatography.
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 microg flavone and 15 microg 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 x 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.
Department of Veterinary Diagnostic Medicine, College of Veterinary Medicine, University of Minnesota, St Paul Campus, 1333 Gortner Ave, St Paul, MN 55108, USA.
Elhabib
E
Benlhabib
John I
JI
Baker
Daniel E
DE
Keyler
Ashok K
AK
Singh
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods; D048789:Phytoestrogens / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D018517:Plant Roots / Q000737:chemistry*; D029908:Pueraria / Q000737:chemistry*; D021281:Quantitative Structure-Activity Relationship; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013025:Glycine max / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
D048789:Phytoestrogens