Comparative separation of biologically active components in Rhizoma chuanxiong by affinity chromatography with alpha(1)-acid glycoprotein and human serum albumin as stationary phases

Affinity chromatography with alpha (1)-acid glycoprotein (AGP) and human se rum albumin (HSA) stationary phases was applied to screen and analyze the b iologically active components of Rhizoma chuanxiong. Five major peaks and a number of small peaks were resolved based on their affinity for AGP and HS A, respectively and three of them, identified as ferulic acid, chuanxiongzi ne and ligustilide via standard compounds, are regarded as effective compon ents. The effects of acetonitrile concentration, pH, inorganic salt concent ration and temperature on the retention behaviors of five major components on the two stationary phases were also investigated. It was observed that h ydrophobicity is the major contributor to retention on both stationary phas es, and ferulic acid has a weak electrostatic interaction with HSA. It demo nstrated that the chromatograms of Rhizoma chuanxiong on the two stationary phases have concise and different fingerprinting characteristics. The amou nt of ferulic acid, chuanxiongzine and ligustilide in Rhizoma chuanxiong de termined using the AGP column are as much as 0.064%, 0.021% and 2.00% by we ight.