Optimization of ultrasound-assisted extraction of flavonoid compounds and their pharmaceutical activity from curry leaf (Murraya koenigii L.) using response surface methodology

BACKGROUND Extraction prior to component analysis is the primary step in the recovery and isolation of bioactive phytochemicals from plant materials. METHODS Response surface methodology was applied to optimize ultrasound-assisted extraction conditions followed by ultra high performance liquid chromatography (UHPLC) to achieve high catechin, myricetin, and quercetin contents, and high antioxidant and anticancer activities in the curry leaf extracts. The antioxidant and anticancer activities of the leaf extracts were determined by the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, respectively. The central composite experimental design (3-level, 3-factorial) was employed to consider the effects of ultrasonic power (80-150 W), temperature (40-80°C), and methanol dilution (40-80%) on the properties of the curry leaf extracts. RESULTS It was found that ultrasonic power of 145.49 W at 55.9°C with 80% methanol was the most appropriate set of conditions for the extraction of catechin, myricetin, and quercetin from curry leaves with the consequent high antioxidant activity. Using the optimum extraction conditions, the extraction yields of catechin, myricetin, and quercetin were 0.482, 0.517, and 0.394 mg/g DW, respectively, and the antioxidant activity was enhanced to 83%. The optimized extract showed more distinct anticancer activity against HeLa cancer cells in a concentration of 67.2 μg/mL (P < 0.01) without toxicity to normal cells. CONCLUSIONS The results indicated that the pharmaceutical quality of curry leaves could be improved significantly by optimizing the extraction process using response surface methodology.