高效亲和色谱法测定2种中药成分与人血清白蛋白的结合率

摘要

采用高效亲和色谱技术(HPAC)对中药成分与人血清白蛋白(HSA)的相互作用进行了研究.首先采用点击化学的方法制备了表面键合有HSA蛋白的硅胶固定相并装填成亲和色谱柱,根据药物在该色谱柱上与空白硅胶柱上的保留时间差计算得到药物与蛋白的结合率.利用该方法测得模型化合物华法令与HSA的结合率与文献中采用超滤法测得的结果基本一致,表明该方法可用于测定药物与HSA的结合率.在此基础上用该方法测定了葛根素和告依春两种中药成分与HSA的相对结合率分别为10.26%和10.20%.同时用超滤的方法测定了葛根素与HSA的结合率为14.25%.结果表明,HPAC可以作为研究药物与蛋白相互作用的一种简便可行的方法,其测定结果与超滤方法一致.%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.