D.A. Borisova, D.M. Popov

Flowers, leaves and rhizomes with roots of Primula officinalis were studied. Rutin (Sigma R 5143) and quercetin (Sigma Q 0125) standards were used. Qualitative composition of flavonoids was determined by TLC and HPLC. Sorbfil PTLC-AF-A-UV plates, N-butanol - glacial acetic acid - water (5-1-1) solvent system were used for the analysis of the flavonoids. 0.05 % rutin in 95 % ethanol and 0.05 % quercectin in 95 % ethanol solutions were used as standards. The chromatogram was treated by 5 % ethanol solution of aluminum chloride. Two yellow-green adsorption zones corresponding to rutin and quercetin were detected. Two more yellow-brown unidentified adsorption zones were also detected. Gradient chromatograph Agilent 1100 (USA) was used for the HPLC analysis of the solutions. The chromatograph consists of a 4-chamber pump, autosampler, a column thermostat and a photo diode array detector. Chromatograms were obtained and processed by using Chemstation and Autochrom 1200 ACDlabs programs. The solutions were analyzed by reverse-phase HPLC method in gradient mode on Germini C18 (5 µm) 4,6*150 mm column with a mobile phases: A  water, B  acetonitrile and C  1 % solution of TFA in water (рН=2) and gradient program: 55 % B (for 4 min), 5-70 % B (for 8 min), 5 % C - constantly at a flow rate of 1.5 ml/min, temperature 25C and UV-detection at 272-360 nm. Presence of rutin and quercetin in primula flowers and leaves was determined by retention time and UV-spectra. In rhizomes and roots flavonoid compounds were not detected. Differential spectrophotometry after flavonoid complexation with aluminum chloride was used for quantification. Reaction conditions and optimal conditions for flavonoid extraction from the raw material were selected. A technique of quantitative determination of flavonoids in primula flowers and leaves was developed. This technique determined flavonoid content in flowers (4,15  4,67 %) and leaves (1,021,57 %). Grass but not leaves is more appropriate as a raw material, because flavonoid content in flowers is 3-4 times more than in leaves. Combined application of such raw material will be a good source of flavonoid compounds for the development of both individual and complex phytopreparations.