A standard addition method to assay the concentration of biologically interesting polyphenols in grape berries by reversed-phase HPLC.

A reversed-phase HPLC method which allows the simultaneous assay of (+)- catechin, (-)-epicatechin, trans-resveratrol, quercetin and quercetin glycosides in grape berries is described. Kromasil 100 served as stationary phase and a gradient of acetic acid, water and methanol was used. The analytical run requires 42 min for complete sample elution. Satisfactory peak resolution was achieved following a novel extraction process and direct injection of a 20 microL sample. The method was used for the analyses of eighteen samples. Linearities were in the range of 0.98 to 0.999 regression coefficient, for all phenolics, while detection limits ranged from 30 microg mL(-1) for trans-resveratrol to 1.5 mg mL(-1) for (+)-catechin. Recoveries ranged from 95.1 to 98.7% while the method provided good precision, with standard deviations between 3.5 and 6.1%, n=5.