盐酸小檗碱与牛血清蛋白结合率的测定

Determination of Plasma Protein Binding Rate of Berberine Hydrochloride

目的:建立牛血清中盐酸小檗碱浓度的分析方法,测定不同浓度盐酸小檗碱与牛血清蛋白结合率。方法:采用HPLC法,流动相乙腈(A)-0.02mol/L磷酸二氢钾溶液(p H=2.8)(B)梯度洗脱(0~32 min,20%~28%A),检测波长345 nm;采用平衡透析法测定盐酸小檗碱与牛血清蛋白结合率。结果:当盐酸小檗碱浓度在10~80μg/m L时,牛血清蛋白结合率在20%~30%之间。当盐酸小檗碱浓度在10~20μg/m L时,牛血清蛋白结合率随着浓度升高而降低;当盐酸小檗碱浓度在20~80μg/m L时,牛血清蛋白结合率随着浓度升高并无明显变化。结论:盐酸小檗碱与牛血清蛋白属于中等程度的蛋白结合率,且当药物浓度在10~20μg/m L时,具有浓度依赖性。

Objective To Establish an HPLC method for the concentration of berberine hydrochloride in bovine serum and determine the protein binding rate of bovine serum protein and berberine hydrochloride at different concentrations. Methods By using HPLC method,mobile phase acetonitrile（ A）-0. 02 mol/L potassium dihydrogen phosphate（ p H = 2. 8）（ B） gradient elution（ 0 ~ 32 min,20% ~ 28% A）,the detection wavelength of 345 nm. Equilibrium dialysis method was used to investigate binding rates of berberine hydrochloride with plasma proteins. Results When the concentration of berberine hydrochloride at 10 ~ 80 μg/m L,the protein binding rate of bovine serum was between 20% ~ 30%. When the concentration of berberine hydrochloride at 10 ~ 20 μg/m L,the binding rate of bovine serum protein decreases with the increase of concentration. When the concentration of berberine hydrochloride at20 ~ 80 μg/m L,the protein binding rate of bovine serum was not significantly changed with the increase of concentration. Conclusion Berberine hydrochloride and bovine serum protein belonged to a moderate protein binding rate,and when the drug concentration is 10 ~20 μg/m L,it had a concentration dependence.