XÂY DỰNG QUY TRÌNH ĐỊNH LƯỢNG ĐỒNG THỜI DIOSMIN VÀ QUERCETIN TRONG VI NHŨ TƯƠNG DQM BẰNG PHƯƠNG PHÁP HPLC-UV

Abstract

Diosmin and quercetin, two active ingredients possess the antioxidant, anti-inflammatory, and vascular wall strengthening effects, have been simultaneously developed as a microemulsion (DQM) for a topical product. This study was carried out to develop a method for the simultaneous determination of diosmin and quercetin in DQM by high performance liquid chromatography (HPLC). The sample preparation methods were screened and selected. The HPLC method was validated according to the ICH guidelines for its specificity, system suitability, linearity, precision, and accuracy. The sample preparation method was selected using a mixture of methanol (80%) and dimethyl sulfoxide (20%) as solvent for the simultaneous extraction of diosmin and quercetin. The best HPLC condition for the simultaneous determination of diosmin and quercetin employed a Syncronis C18 column (250 × 4.6 mm; 5 μm), a detection wavelength of 268 nm, a column temperature of 30 °C, a flow rate of 0.8 ml/min and an injection volume of 20 μl. The mobile phase was a mixture of acetonitrile and 0.2% formic acid in water in gradient mode. The ratios of acetonitrile were  31%, 35% and 55% at 0-2 min, 3-4 min and 5-15 min, respesctively. The good correlations were found between peak areas and diosmin concentrations (r² = 0.9991) or quercetin concentration (r² = 0.9988). The % RSD values of inter-day precision of diosmin and quercetin were 1.60% and 0.60%, respectively. The recovery percentages of diosmin and quercetin were 98.47-103.40 % and 99.66 – 101.53%, respectively. The HPLC-UV method for simultaneous quantification of diosmin and quercetin in DQM microemulsions has been reported for the first time. The quantitation method met the requirements of validation and could be useful for quality control of DQM and its related products.