The paper presents a novel multi-purpose enzymatic system and procedures for fluorescent determination of several flavonoids in herbal pharmaceuticals and plant materials after their enzyme-catalyzed oxidation by hydrogen peroxide and further derivatization with meso-1,2-diphenylethylenediamine. This system may be used for rapid (15-30min/20 samples) simultaneous screening of samples containing a certain flavonoid in a standard microplate, or as a HPLC detection system for analyzing plant extracts with uncertain composition. In the first case, this indicator system provides sensitive and reproducible microplate determination of quercetin, epicatechin, caffeic acid, and taxifolin in the ranges 0.1-5, 1-10, 0.1-10, 0.5-5μM, respectively. In the second case, quercetin, epicatechin, and caffeic acid can be determined in the ranges 0.05-0.75, 0.05-0.75, and 0.01-0.75µg/ml (0.16-2.5, 0.17-2.6, 0.06-4.2μM), respectively. We have demonstrated the application of the system for the analysis of 3 pharmaceuticals and 3 types of plants.
Keywords: Enzymatic derivatization; Flavonoids; Fluorescence; HPLC; Peroxidase; Simultaneous screening.
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