Optimization and validation of a method for the direct determination of catechin and epicatechin in red wines by HPLC/fluorescence

Abstract

The present paper describes a direct procedure for the determination of catechin and epicatechin concentrations in red wines employing reverse-phase high performance liquid chromatography (RP HPLC) and detection by fluorescence. The method was performed using a sample volume of 10 μL without dilution. The separation process employed a Chromolith performance RP-18e (100 mm×4.6 mm) column, and the mobile phase was composed of solvent A: methanol–acetic acid–water (90:8:2) and solvent B: water–acetic acid–methanol (10:2:88) at a flow rate of 1.0 mL min−1. Linearity was observed in the range of 1 to 30 mg L−1, with limits of detection and quantification of 0.27 and 0.89 mg L−1, respectively, for catechin and 0.33 and 1.01 mg L−1, respectively, for epicatechin. The precisions estimated by the relative standard deviation were 3.34 and 1.09% for catechin concentrations of 0.5 and 20 mg L−1 respectively and 2.82 and 0.49% for epicatechin concentrations of 0.5 and 20 mg L−1, respectively. The evaluation of the accuracy was done using an addition/recovery assay. Four wine samples were used, and the recoveries varied from 105 to 108% for catechin and from 97 to 119% for epicatechin. The method was applied to the analysis of red wine samples collected from the São Francisco region, Bahia State, Brazil. Nine samples were analyzed, and the catechin and epicatechin concentrations varied from 7.51 to 73.20 and from 5.08 to 43.32 mg L−1, respectively. The concentrations found agree with data reported in the literature.