DEVELOPMENT OF SPECTROPHOTOMETRIC METHOD OF FLAVONOIDS DETERMINATION IN THE BILBERRY SHOOTS DRY EXTRACT
DOI:
https://doi.org/10.11603/2312-0967.2019.3.10463Keywords:
bilberry shoots extract, flavonoid aglycones, quercetin, quantification, extraction, spectrophotometry, standard methodAbstract
The aim of the work was to develop a spectrophotometric method for quantifying the amount of flavonoids in the bilberry shoots dry extract using pre-hydrolysis of their glycosidic forms and extraction of the obtained aglycones.
Materials and methods. The dry extract was obtained from crushed bilberry shoots by fractional maceration method using 50-70% ethanol (v / v) as the extractant. The standard sample of quercetin (Sigma-Aldrich) was used in the work. The electronic absorption spectra and absorption measurements were recorded on a Lambda 25 spectrophotometer (PerkinElmer Ltd., USA).
Results and Discussion. To increase the selectivity of the spectrophotometric determination of flavonoids and to avoid the influence of hydroxycinnamic acids and chlorophylls, which are presented in the extract, aglycones of flavonoids first were isolated and then quantified. For this purpose, the hydrolysis conditions of flavonoids glycosides in acetone-aqueous medium in the hydrochloric acid presence and subsequent extraction of aglycones with ethyl acetate were investigated. It has been established that the obtained aglycones form a complex with aluminum chloride, which has a maximum absorption at the wavelength of 425 nm in the electronic absorption spectrum, and quercetin can be used as a standard substance to calculate the content of flavonoids. The conditions of sample preparation and spectrophotometric measurements were selected, which allowed to calculate the content by the standard method. The linearity, precision and accuracy of the method were experimentally proven in the range of 20-250% of the expected content, which indicates the suitability of the proposed method for the quantification of flavonoids in the bilberry shoots dry extract.
Conclusions. A spectrophotometric method for quantitative determination of flavonoids in the bilberry shoots dry extract has been developed, which avoids the influence of hydroxycinnamic acids presented in the extract in a significant amount. The linearity, precision and accuracy of the procedure were studied in the range of 1,232-15,400 µg of quercetin in 1 ml of the final measured solution (20-250% of the expected content), the results of which indicate the suitability of the method for determining the content of flavonoids in the bilberry shoots dry extract.
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