Phenolic compounds including phenolic aldehydes, acids and flavonoids are separated by high-performance liquid chromatography (HPLC) with analysis time shorter than described in the literature. The use of a fluorescence detector in series with absorbance detector allowed increasing selectivity and sensitivity for the determination of catechin, vanillic acid, syringic acid, epicatechin and trans-resveratrol in wine samples. An optimised sample preparation method using liquid-liquid extraction with diethyl ether at pH 2.0 was used. The optimised method was applied to analyse wine samples with good results.