[Comparison of photometric, electrochemical and post-column fluorescence detection for the determination of flavonoids by HPLC].

For the analysis of flavonoids by HPLC, we compared three different detection methods, namely UV, electrochemical detection and post-column chelation with aluminum followed by fluorescence detection. Ten flavonoids were used: apigenin, myricetin, luteorin, taxifolin, quercetin-3-O-sulfate, kaempferol, isorhamnetin, isoquercitrin, quercetin and rutin. Nine flavonoids except apigenin were efficiently detected by the electrochemical method with a detection limit of 0.025-0.05 pmol. Flavonols having free 3-hydroxyl and 4-keto oxygen formed a fluorescent complex by post-column chelation and were detected by the fluorescence method. The detection limit of the fluorescence method was 0.05-0.5 pmol. Nine flavonoids except taxifolin were detected by the UV method (absorbance at 370 nm), but the detection level was poor (5-10 pmol). Flavonols added to human plasma were recovered by solid phase extraction, and were analyzed using the three detection methods. Most of the flavonols were efficiently detected by the electrochemical and fluorescence methods, and the detection limits were similar to those of standard samples.