HPLC-MS/MS Quantitative Determination of Gallic acid and Cyanidin-3-Glucoside Content of Bilberry Fruit Extract from Turkey

Genus Vaccinium is a widespread genus with over 200 species of evergreen and deciduous woody plants varying in size from dwarf shrubs to trees. There is a great interest worldwide in the fruits of bilberry because of their high anthocyanin content. Anthocyanins are flavonoids comprising flavonol glycosides, flavan-3-ols and proanthocyanidins, whereas hydroxycinnamic acids are classified as phenolic acids. Anthocyanins are valued as pigments but are also widely used in natural health products due to their suggested positive effects on night vision, even though firm evidence from clinical trials is still lacking. [1].

In this study, quality control of Vaccinium myrtillus L. fruits have been carried out according to the European pharmacopoeia (2). Furthermore, gallic acid and cyanidin-3-glucoside contents of Vaccinium myrtillus water and EtOH (70%) extracts were investigated using with HPLC ESI/MSMS MRM method.

The assay was performed with different concentrations of gallic acid and cyanidin-3-glucoside chloride as standart solutions. The diagnostic fragmentations of gallic acid 168.7/125–79 and fragmentations of cyanidine-3-glucoside 448.7/287–150 were used for MRM quantitative determination.

Total anthocyanin content of the fruits were shown to be not less than 0.30 per cent and other features were found to conform Pharm Eur requirements.

Furthermore, the content of cyanidin-3-glucoside was shown to be 0.0538g±0.001 per cent in the EtOH (70%) extract and 0.045±0.002 per cent in the water extract, respectively. The gallic acid contents measured in the ethanolic and water extracts were 0.001 per cent and 0.136±0,001 per cent, resp.

References: 1. Riihinen K et al.(2008) Food Chem 110(1): 156–160.

2. European Pharmacopoeia 7.1 (2010) Bilbery Fruit Fresh, Myrtilli fructus recens, p 1070.