IDENTIFICATION AND QUANTITATIVE DETERMINATION OF FLAVONOID IN AGRIMONIA EUPATORIA
DOI:
https://doi.org/10.11603/2312-0967.2017.2.7905Keywords:
Agrimonia eupatoria, herb, flavonoids, identification, quantitative determination, thinlayer chromatography, spectrophotometry.Abstract
The aim of the work. Determination of the qualitative composition of flavonoids in local samples of Agrimony herb according to the requirements of the State Pharmacopoeia of Ukraine (SPU) for flowering tops of Agrimony and development of the methods of quantitative determination of flavonoid content in it is the urgent matter.
Materials and Methods. To study the qualitative composition of flavonoids, four local samples of Agrimony were used; the evaluation of quantitative content of flavonoids was carried out for five samples. A thinlayer chromatography was used to study the qualitative composition and spectrophotometry - to determine the quantitative content of flavonoids.
Results and Discussion. The research proved that qualitative composition of flavonoids in Agrimony complied with the requirements of the State Pharmacopoeia of Ukraine for Agrimony flowering tops. To identify the Agrimony, it was suggested to detect flavonoids by thinlayer chromatography, changing the sample preparation of raw materials. Into the reference solution, hyperoside should be added to identify it completely at the chromatogram of the test solution. In electronic absorption spectra of alcohol extracts (70% V/V) of Agrimony in conditions of complexation together with aluminum chloride, the absorption maximum was observed ranging 400-403 nm. Since, the method of quantitative determination of flavonoids was suggested, equivalent to luteolin or luteolin-7-O-glucoside using the value of speciflc absorption rate of complex flavonoids compounds with aluminum chloride.
Conclusions. Rutin, hyperoside, isoquercitrin were chosen to be identification markers of Agrimony; quercitrin and other flavonoids with nonquercetin aglycones and phenolcarboxylic acids may be evidenced as zones of orange, green and blue fluorescence at the chromatogram of the test solution. The content of flavonoids in flve samples was determined within 0.90-1.46 % equivalent to luteolin or 1.47-2.39 % as luteolin-7-O-glucoside. The difference between the values of content for different samples is associated with various conditions of raw materials growth as well as with the ambiguousness of the notion "herb”. Therefore, there is a need for implementation of the national part into the monograph of SPU on Agrimony; the use of Agrimony should be provided and the concept "related substance” with criteria of largest possible quantity of gross stems should be introduced. Taking into account the high content and biological activity of flavonoids, the definition for "fl avonoids content” should be adopted in the monograph on Agrimony.
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