Determination of Five Polyphenols by HPLC/DAD and Discrimination of Apple Varieties

Abstract

The objective of this study was to set up a method to detect five compounds in fresh smashed apples by HPLC/DAD simultaneously. Different methods have been tested to control browning and ascorbic acid with ultrasonication was adopted. Methanol–water–acetic acid (30:69:1, v/v) containing 2.0 g of ascorbic acid L⁻¹ was chosen as the extract solvent. The method effectively simplified the sample treatment compared with the traditional ways. And primarily, the results were used to identify between different varieties. The chromatographic separation was performed on an Atlantis C18 (250 mm × 4.5 mm, particle size 5 μm) with a gradient elution program using a mixture of acetonitrile and 2% aqueous acetic acid (v/v) as mobile phase within 20 min at 270 nm wavelength. The variation of the content of five compounds was gallic acid (ND ~1.81 μg g⁻¹), protocatechuic acid (ND ~1.79 μg g⁻¹), chlorogenic acid (13.81–189.4 μg g⁻¹), caffeic acid (6.82–45.02 μg g⁻¹) and rutin (0.96–18.55 μg g⁻¹). The results could successfully be used to discriminate between different apple varieties (Gala, Fuji, Delicious, 8th Apple US, Golden Apple, Green Apple and Red Rose); chlorogenic acid and rutin being the polyphenols that contribute most to the differentiation.