Determination of isoflavones using high-performance liquid chromatography with electrochemical detection (HPLC- ED)

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Authors

KLEJDUS Bořivoj PETRLOVÁ Jitka POTĚŠIL David TRNKOVÁ Libuše MIKELOVÁ Radka ZEHNÁLEK Josef KIZEK René KUBÁŇ Vlastimil

Year of publication 2004
Type Article in Proceedings
Conference Book of Abstracts - 55th Annual Meeting of the ISE
MU Faculty or unit

Faculty of Science

Citation
Field Analytic chemistry
Keywords HPLC-ED; square wave voltammetry; isoflavones; daidzein; genistein; soybean products
Description A highly sensitive HPLC-ED was developed for the determination of isoflavones. The electrochemical behaviour of daidzein and genistein was studied on a carbon paste electrode by square wave voltammetry. The electrochemical results obtained were used for the development of the HPLC-ED method. The HPLC-ED system used consisted of two solvent delivery pumps and an eight-channel CoulArray electrochemical detector. Isoflavones were separated on an Atlantis dC18 column using a mobile phase consisting of acetonitrile (solvent A) and 0.15 M acetate buffer of pH 5.5 (solvent B) at a flow rate of 0.4 mL/min. The full scan of multi-channel coulometric detection was tested and an optimal potential at 450 mV was chosen for our purposes. The calibration curves were linear (daidzein R2 = 0.9993 and genistein R2 = 0.9987). The relative standard deviation (R.S.D., n = 5) was 2.5 %. The detection limits for daidzein and genistein were 480 and 394 pg/mL (1.8 and 1.5 nM), respectively. Isoflavones extracted from soybean products (farina, meat, milk) by the pressurised solvent extraction procedure and isoflavones contained in human urine were determined by this HPLC-ED method.
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