A general gas-assisted three-liquid-phase extraction method for separation and concentration of puerarin, 3′-methoxydaidzin, puerarinxyloside, daidzin and daidzein from puerariae extract

In this work, a general and novel separation technique gas-assisted three-liquid-phase extraction was established and applied in separating and concentrating isoflavonoids from the actual sample of puerariae extract by one step. For the gas-assisted three-liquid-phase extraction method, optimal conditions were selected: polyethylene glycol 2000 and ethyl acetate as the flotation solvent, pH 5, (NH4)(2)SO4 concentration 350 g/L in aqueous phase, N-2 flow rate 30 mL/min, flotation time 50 min, and flotation twice. Five isoflavonoids compounds puerarin, 3 '-methoxydaidzin, puerarinxyloside, daidzin and daidzein were separated with recoveries of 82, 84, 80, 88 and 89%, respectively. The separated products were purified by preparative high-performance liquid chromatography, and the purity of the final products was >96%. The established general gas-assisted three-liquid-phase extraction was used to separate anthraquinones from Cassiae Semen under the optimal conditions, and the recoveries were >75%. The experimental results showed that the established gas-assisted three-liquid-phase extraction method is a general technique for separating active compounds from herb extract.