Establishment of green analytical method for ultrasound-assisted extraction of azadirachtin, mevalonic acid and squalene from cell suspension culture of Azadirachta indica using response surface methodology

Azadirachta indica is used as a medicinal plant and its biological activity have been studied, but the extraction of its secondary metabolites was not clearly optimized. In this study, response surface methodology was applied for optimization of extraction of azadirachtin, mevalonic acid and squalene from cell suspension culture of A. indica. The Box-Behnken Design was used for the optimization of factors. The azadirachtin, mevalonic acid, and squalene were recognized by HPLC-DAD. Results showed that the optimal condition (solvent, temperature and ultrasonication time) obtained by response surface methodology for target compounds from cell suspension culture of A. indica were as following. Azadirachtin: water, temperature 35 degrees C and ultrasonication time 20 min, obtained 86.445 mg/g DW; mevalonic acid: 50 % ethanol, temperature 45 degrees C and ultrasonication time 30 min, provided 33.671 mg/g DW; and squalene: 50 % ethanol, temperature 55 degrees C and ultrasonication time 10 min, achieved 8.278 mg/g DW.