微透析取样-流动注射电化学检测测定表柔比星与牛血清白蛋白体外结合参数

建立了微透析取样-流动注射电化学检测法测定表柔比星与牛血清白蛋白体外结合参数的方法。流动相为pH 3.0的0.1 mol/L H3PO4-NaH2PO4缓冲液(含3.4×10﹣5mol/L Na2EDTA),流速为0.3 mL/min。电化学检测的工作电极为玻碳电极,工作电位为0.52V。表柔比星浓度在5.0×10-5～5.0×10-7 mol/L范围内与峰电流呈良好的线性关系,其相关系数为0.9997,检出限为2.0×10-7mol/L。结合微透析取样,实现了对表柔比星与牛血清白蛋白结合参数的测定。表柔比星与牛血清白蛋白的体外结合符合Scatchard曲线,结合常数和结合位点数分别为2.41×104L/mol和4.02。

Determination of the binding parameters of epirubicin to BSA in vitro by microdialysis coupled with flow-injection electrochemical detection

Microdialysis sampling coupled with flow injection-electrochemical detection was developed and applied to determine the binding parameters of epirubicin with bovine serum albumin(BSA) in vitro.The mobile phase was 0.1 mol/L H3PO4-NaH2PO4 buffer solution with pH 3.0 containing 3.4×10-5 mol/L Na2EDTA in flow injection.The mobile phase flow rate was 0.3 mL/min.The glassy carbon electrode was chosen as the working electrode,and the applied potential was 0.52 V.The results showed that the peak current of epirubicin was linear with its concentration ranging from 5.0×10-5to 5.0×10-7 mol/L,and the correlation coefficient of the compound was 0.9997.The detection limit of epirubicin was 2.0×10-7 mol/L.Coupled with microdialysis sampling,the binding of epirubicin with BSA in vitro was studied,and the result showed that the values for the association constant and the number of the binding sites on one BSA molecule were 2.41×104(mol/L)-1 and 4.02,respectively.