Extraction, isolation, and standardization of anthocyanin from Vaccinium myrtillus to evaluate their potential in glaucoma

Abstract

In order to preserve the structural integrity and bioactivity of anthocyanins while improving their extraction, identification, and isolation from Vaccinium myrtillus berries is the goal of this study. The following three goals were pursued: · Creating effective extraction procedures, · Using cutting-edge chromatographic techniques for identification, and · Isolating certain anthocyanin chemicals for possible bioactivity research. Three extraction techniques (A, B, and C) were contrasted with a fresh berry control for Objective 1. The maximum anthocyanin production (14.2 mg/g) and antioxidant activity (90%) were produced by Method C, which can be attributable to the high concentration of Petunidin-3-glucoside (55%). Method A produced anthocyanins with a concentration of 12.5 mg/g, predominantly made up of cyanidin-3-glucoside (45%), compared to Method B's 9.8 mg/g, primarily made up of delphinidin-3-glucoside (30%). The structural integrity of each approach was kept to a satisfactory level, as shown by the absorbance tests. Objective 2 was to identify anthocyanins using sophisticated chromatographic methods. Cyanidin-3-glucoside (5.21 min, 12500), Delphinidin-3-glucoside (7.46 min, 9800), and Petunidin-3-glucoside (9.83 min, 14200) were measured for retention times and peak areas. Anthocyanin molecules were identified using purification methods to work towards Objective 3. Three fractions were collected, and the anthocyanin chemicals in each fraction were identified: Fraction 1, 2.5 mg, 95% purity of Cyanidin-3-glucoside; Fraction 2, 1.8 mg, 90% purity of Delphinidin-3-glucoside; and Fraction 3, 3.2 mg, 92% purity of Petunidin-3-glucoside. Analysing UV-Vis spectra showed the purity, and chromatographic retention durations matched benchmark values. Further confirmation of chemical identities was obtained by mass spectrometry and NMR investigations, with detected peaks matching recognised references. This thorough investigation contributes to our understanding of how to extract and characterise anthocyanins from V. myrtillus berries. The findings demonstrate that Method C is a better extraction technique, producing high anthocyanin content while maintaining bioactivity.