前沿色谱法对盐酸小檗碱和固定化牛血清白蛋白结合作用的研究

Investigation on Binding Interaction of Berberine Chloride with Bovine Serum Albumin Immobilized onto Chromatographic Supports by Frontal Chromatography

盐酸小檗碱(BC)是黄连的主要有效成分,它具有抗菌消炎等多种作用。采用前沿色谱法测定了不同温度下BC与固定化牛血清白蛋白(B SA)的结合常数K和结合率PPB、BC的保留因子k、BC在色谱柱上活性位点的物质的量mL,以及BC与B SA结合过程中的热力学参数。在温度为30℃时,BC与B SA的结合常数为4.79×104L/m o l。K、k和mL均随温度的升高而降低,其中以mL的变化程度最为显著,表明k的降低是由于K与mL共同作用的结果。而B SA分子构象变化可能是mL降低的主要原因。热力学分析结果表明:BC和B SA之间的作用力以静电作用力为主。

Berberine chloride （BC） is a major active constituent of coptis and can be used as an antipyrotic and antibacterial medicine. Frontal analysis was used to investigate the changes in the binding constant （K）, retention factor （k）, binding ratio （PPB） and mole of binding sites （mL） for the binding of BC on an immobilized bovine serum albumin （BSA） column at several temperatures and to obtain the thermodynamic parameters in the binding process. At 30 ℃, the binding constant was 4.79 × 10^4 L/mol. K, k and mL all decreased as the temperature was increased. Among these three parameters, the change magnitude in mL was the most significant. It could be concluded that the decrease in the retention of BC was caused by the decrease of both K and mL, and the change in the configuration of BSA was considered to be the main reason for the decrease of binding site. The thermodynamic analysis indicated that the main driving force for the interaction between BC and BSA is electrostatic force.