毛细管区带电泳研究凝血酶与蛇目菊和肉苁蓉提取物的相互作用

摘要

目的采用毛细管区带电泳法研究凝血酶与蛇目菊和肉苁蓉提取物的相互作用.方法研究了在pH 7.2的Tris-醋酸缓冲溶液中,凝血酶和天然提取物于25 ℃以不同比率混合温育20 min后的相互作用情况,分离在5 min内完成.结果与阳性对照和阴性对照相比,蛇目菊中的提取物CB-1, CB-2与凝血酶有相互作用,并计算得到了CB-1,CB-2和凝血酶的结合常数;蛇目菊中的提取物CB-3和肉苁蓉中的HC-1, HC-2, HC-3与凝血酶没有相互作用.结论本实验采用的毛细管区带电泳研究天然提取物与凝血酶相互作用的方法快速、准确、可靠.%Aim A capillary zone electrophoretic method (CZE) was used to determine the interactions between natural products and thrombin. Methods Samples containing natural products and thrombin at various ratios were incubated at 25 ℃ and then were separated by CZE with Tris-acetate buffer at pH 7.2. Each run could be accomplished within 5 min. Results In CZE, the peak width broadened due to the affinity interaction between natural products and thrombin. Compared with positive and negative control, the natural products (CB-1, CB-2) from Coreopsis tinctoria Nuttt. interacted with thrombin; CB-3 from Coreopsis tinctoria Nuttt. and HC-1, HC-2, HC-3 from Cistanche deserticola Ma. did not bind to thrombin. Both qualification and quantification characterizations of the binding were determined. Conclusion The established method is capable of sensitive and fast determination of natural products and thrombin interactions, it can be employed as an alternative method.