Comparative separation of biologically active components in Rhizoma chuanxiong by affinity chromatography with α1-acid glycoprotein and human serum albumin as stationary phases

Affinity chromatography with alpha (1)-acid glycoprotein (AGP) and human serum albumin (HSA) stationary phases was applied to screen and analyze the biologically active components of Rhizoma chuanxiong. Five major peaks and a number of small peaks were resolved based on their affinity for AGP and HSA, respectively and three of them, identified as ferulic acid, chuanxiongzine and ligustilide via standard compounds, are regarded as effective components. The effects of acetonitrile concentration, pH, inorganic salt concentration and temperature on the retention behaviors of five major components on the two stationary phases were also investigated. It was observed that hydrophobicity is the major contributor to retention on both stationary phases, and ferulic acid has a weak electrostatic interaction with HSA. It demonstrated that the chromatograms of Rhizoma chuanxiong on the two stationary phases have concise and different fingerprinting characteristics. The amount of ferulic acid, chuanxiongzine and ligustilide in Rhizoma chuanxiong determined using the AGP column are as much as 0.064%, 0.021% and 2.00% by weight.