Large scale isolation and purification of salvianolic acid b in high purity from roots of dansham (Salvia miltiorrhiza bunge)

Salvianolic acid B (Sal B) is the most abundant representative compound in roots of dansham (Salvia miltiorrhiza) which is beneficial in treatment of cardiovascular disease. The present study was performed to establish a large scale isolation method of Sal B in high purity from dansham roots. Water suspension of dansham roots’ (100 g d.w.) methanol extract (13.16 g) was partitioned with chloroform, and the water layer was adjusted to pH 3.0 by triflouroacetic acid, because the Sal B partition coefficient in acidic condition was predominately higher than that in neutral condition, and then continuously extracted with ethyl acetate (EtOAc). The EtOAc-soluble acidic fraction was subjected to Sephadex LH-20 column chromatography. The fractions (Ve/Vt 1.75–2.71, 1.82 g) containing Sal B were finally purified by preparative high performance liquid chromatography using an octadecylsilane column. The isolated Sal B fraction (1.08 g) represented a 75.0% recovery, with a purity exceeding 99.2%.