Influence of sample and mobile phase pH on HPLC-DAD-MS analysis of anthocyanins and other phenolic compounds in wine

Ivanova, Violeta and Meudec, Emmanuelle and Souquet, Jean-Marc and Stefova, Marina and Cheynier, Veronique (2010) Influence of sample and mobile phase pH on HPLC-DAD-MS analysis of anthocyanins and other phenolic compounds in wine. In: XXVth International Conference on Polyphenols, August, 24-27, 2010, Montpellier, France.

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In this study, the influence of pH of the wine sample (pH 1.2 and 3.3) and of the HPLC mobile phase (1 and 2 % formic acid in the solvent) on anthocyanin and phenolic acid analysis was explored. Wine samples acidified at pH 1.2 were analyzed immediately, after 30 min, 2h and 3.5h, in order to check if any degradation of phenolic compounds occurred during the storage. Maximum response of the flavylium cation of peonidin-3-glucoside (at 520 nm), statistically significant (p < 0.001), was observed in the sample with pH 1.2, and the hydrated forms were not detected under these conditions. The peak area of the flavylium cation was also slightly higher when elution was performed by 2 % formic acid in the HPLC solvent, but the differences were not significant (p > 0.05), meaning that both 1% and 2% formic acid in the mobile phase are suitable for anthocyanin analysis. Regarding phenolic acids, lower peak intensities, statistically significant (p < 0.001), were obtained in the negative ion mode with 2 % formic acid in the mobile phase, presumably due to the lower proportion of acids in the anionic form under more acidic conditions. Thus, 1% formic acid in the mobile phase and acidification of the sample at pH £ 1.2 were selected as the best compromise for simultaneous analysis of anthocyanins and phenolic acids. Acidification of the wine samples at pH 1.2 followed by a 30 min delay enabled total transformation of chalcone and carbinol forms into the corresponding flavylium cations, without causing degradation of the other wine phenolics. The optimized conditions were applied for MS identification of 69 compounds from different phenolic groups present in Vranec wine.

Item Type: Conference or Workshop Item (Paper)
Subjects: Natural sciences > Chemical sciences
Divisions: Faculty of Agriculture
Depositing User: Violeta Ivanova Petropulos
Date Deposited: 06 Nov 2012 15:00
Last Modified: 20 Nov 2012 11:40

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