Methods were developed for the analysis of natural antioxidants including phenolic compounds and flavonoids in beverages and plant extracts using gradient HPLC with multi-channel electrochemical coulometric detection. Suitability of various reversed-phase columns for this purpose was compared; pH and mobile phase gradients were optimized with respect to the separation selectivity and sensitivity of detection. Because of different target compounds in various sample types, the overlapping resolution maps and the normalized resolution product approaches described earlier were used to select optimum columns and gradients to suit the analysis of the individual sample types. The methods were applied to the analysis of phenolic compounds and flavonoids in beer, wine, tea, and yacon extracts. 32 phenolic compounds were identified and determined, including derivatives of benzoic and cinnamic acids, flavones, and a few related glycosides. Eight-channel CoulArray detection offers high selectivity and sensitivity with limits of detection in the low microg L(-1) range, at least an order of magnitude lower than single-channel coulometric detection using the Coulochem detector. No special sample pretreatment is necessary and, because of the compatibility of the CoulArray detector with gradient elution, phenolic antioxidants of different polarities can be determined in a single run. In addition to the retention times, the ratios of the areas of the pre-dominant and post-dominant peaks to the area of the dominant peak can be used for improved identification of natural antioxidants.

A method was developed for simultaneous analysis of natural antioxidants in beer using multichannel electrochemical detection with a CoulArray detector, which enables selective and sensitive antioxidant detection in gradient HPLC and facilitates the identification of analytes based on the ratios of signals recorded at different potentials applied to the detection cells arranged in series. The separation conditions were optimised for 27 phenolic compounds including derivatives of benzoic and cinnamic acids, flavones, and a few related glycosides identified in beer samples. Separation selectivities of 11 columns with different stationary phase chemistries were compared, and the pH and gradient programs were optimised for the individual columns to provide best resolution and high number of resolved peaks, using the window-diagram approach. The effects of pH on the sensitivity of electrochemical coulometric detection were considered in the optimisation approach. The optimised conditions were applied to the analysis of real beer samples.