Simultaneous isolation of phyllanthin, hypophyllanthin, and niranthin from crude lignan extract of <i>Phyllanthus amarus</i> leaves by centrifugal partition chromatography (CPC)

Centrifugal partition chromatography (CPC) was successfully used for isolation of three lignan compounds such as phyllanthin, hypophyllanthin, and niranthin from <i>Phyllanthus amarus</i> by using a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (2:1:2:1, v/v/v/v) (Arizona R solvent system) in ascending mode. In CPC method, the 250 mL CPC column was filled with the lower stationary phase at a flow rate of 30 mL/min with a maintaining rotation speed at 500 rpm. Then, the upper mobile phase was pumped into the column at a flow rate of 8 mL/min and the rotation speed was set to 1600 rpm. From 0.10 (g) of the crude lignan extract, phyllanthin (<b>1</b>, 25.3 mg, 92.8% purity), hypophyllanthin (<b>2</b>, 7.4 mg, 88.9% purity), and niranthin (<b>3</b>, 14.8 mg, 94.5% purity) were obtained by a single CPC run. The isolated substances were identified by ESI-MS and comparison of retention times with standards.