Development of a quantitative high-performance liquid chromatography-photodiode array detection measurement system for phenolic acids.

A quantitative high-performance liquid chromatography-photodiode array detection method separating 16 phenolic acids was achieved. Six columns and several mobile phases were investigated. Resolution was achieved with a high-purity silica Phenomenex Luna C18 column (150 mm x 4.6 mm, 5 microm) and a binary gradient consisting of CH3OH-water (with 0.1% formic acid) and flow rate set at 0.7 ml/min. Acids were detected and quantitation performed at wavelength representing the lowest energy lambda(max) for individual acids. Extraction procedure from wine was optimized and yields ranged from 79 to 87% based on internal standard recovery. To confirm our quantitative results, identical samples were analyzed both in-house and by a collaborating laboratory. Correlation of two data sets generated linear regression equations that approached unity (0.93-0.98) and R2 values ranging from 0.990 to 0.999.