Detection of drug-human serum albumin binding ratios of two Chinese medicinal ingredients by high performance affinity chromatography

Abstract

The interaction of two Chinese medicinal ingredients and human serum albumin (HSA) has been investigated by high performance affinity chromatography (HPAC). HSA bounded silica-based stationary phase was prepared based on the "click chemistry" strategy, and packed in a column (named as HSA column). The drug-HSA binding ratio was calculated from the difference of the drug's retention times on the HSA column and silica column (blank column). The warfarin-HSA binding ratio determined by this method was similar to the reference reported value by ultrafiltration method. The results indicated that the new HSA column and the HPAC method can be used for the detection of binding ratio of drug and HSA. The binding ratios of puerarin and goitrin determined by the HPAC method were 10.26% and 10.20%, respectively. And the binding ratio of puerarin determined by ultrafiltration was 14.25%. All these results showed that HPAC is a useful method to investigate the interaction between drugs and protein.