XÂY DỰNG QUY TRÌNH ĐỊNH LƯỢNG ĐỒNG THỜI DIOSMIN VÀ QUERCETIN TRONG MICROEMULGEL DQE BẰNG PHƯƠNG PHÁP HPLC-UV

Abstract

Diosmin and quercetin have been demonstrated to possess the anti-inflammatory and vascular wall strengthening effects and have been developed as a microemulgel (DQE) to support the treatment of varicose veins. This study aimed at developing a procedure for the quantification of diosmin and quercetin in DQE using high performance liquid chromatography (HPLC). Different sample preparation methods and HPLC conditions were screened and selected. The quantification procedure was validated according to the ICH guidelines. A mixture of methanol (80%) and dimethyl sulfoxide (20%) was chosen as the solvent for sample preparation. The selected HPLC condition included a Syncronis C18 column (250 × 4.6 mm; 5 μm), a detection wavelength of 268 nm, a column temperature of 30 °C, 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 with acetonitrile ratios of 29%, 35% and 55% at 0-2 min, 3-4 min and 5-12 min, respectively. The peak areas and diosmin concentrations (or quercetin concentrations) were found in good linear correlations. The % RSD values of inter-day precision of diosmin and quercetin were 1.75% and 0.71%, respectively. The recovery percentages of diosmin and quercetin were 98.73-100.98% and 99.57 – 100.33%, respectively. The procedure for the quantification of diosmin and quercetin in DQE has been successfully developed and could be applied to evaluate the quality of DQE.