综述了定性和定量研究药物—蛋白结合作用的部分方法，包括色谱、毛细管电泳、核磁共振光谱、质谱等方法及一些传统方法如平衡透析、超滤等方法，并讨论了各自的优点和局限性。

We have used capillary electrophoresis in the frontal analysis mode(CE- FA) to determine the unbound concentration of clozapine in human serum albumin(HSA), human plasma, rabbit serum and plasma sample. The unbound clozapine concentration was directly measured from the height of the frontal peak. Samples were injected directly into an uncoated fused silica capillary (0.65 m (LC)×75 μm i. d.; LE= 0.35 m) and separation was accomplished within 11 min without extensive sample pretreatment. The most suitable running buffer to separate unbound clozapine peak from the other peaks due to endogenous substances was found to contain 1 mmol 1-1 EDTA, 0.5 mol 1-1 glycine, and 67 mmol 1-1 phosphate with pH 7.4. The concentrations of unbound clozapine agreed well with those determined by the conventional ultrafiltration method. The methodology is validated and good correlation and precision are obtained. It was found that clozapine is strongly bound to protein in human plasma, rabbit plasma, and serum, while hardly bound to HSA. The present method enables the determination of the unbound drug concentration in multiple equilibrium system with less than ultra- micro injection volume, and would be hence especially useful for the protein binding study in biological samples that are only available in minute quantities.

目的 研究格列美脲与人血清白蛋白（human serum albumin, HSA）结合作用。方法 采用高效液相色谱—迎头分析法（high performance liquid chromatography- frontal analysis, HPLC- FA）。色谱柱为内表面反相（internal- surface reversed- phase, ISRP）硅胶柱Pinkerton GFF Ⅱ- S5- 80(150 mmx 4.6 mm ID, 5 μm)；流动相为67 mmol·L-1磷酸盐等渗缓冲液(pH 7.4, I= 0.17 mol·L-1)；流速0.2 mL·min-1；检测波长230 nm；进样量900 μL；柱温37℃。结果应用非线性回归参数估算求得HSA分子上两类结合位点结合的格列美脲分子数及相应的结合平衡常数：n1= 1和K1= 5.1(μmol·L-1)-1；n2= 7和K2= 0.017(μmol·L-1)-1。结论 HPLC- FA法操作简便，适用于研究格列美脲与HSA的结合作用。

将18- 甲基炔诺酮加到人血清白蛋白—酮洛芬平衡溶液中，室温孵育达平衡后应用毛细管电泳—迎头分析法研究了18- 甲基炔诺酮和酮洛芬在人血清白蛋白分子上的置换相互作用。—大体积样品虹吸进样至未涂层毛细管(65 cm×50μm i. d., 有效长度35 cm)，进样时间80 s，毛细管两端高度差11 cm，工作电压10 kV，运行缓冲液为67 mmol/ L磷酸盐缓冲液(pH 7.4)，游离酮洛芬浓度由前沿峰的高度直接测定。当酮洛芬样品溶液中酮洛芬的总浓度分别为100 μmol/ L和200 μmol/ L时，随着18- 甲基炔诺酮加入量的增加（18- 甲基炔诺酮的浓度由0增至200μmol/ L），游离酮洛芬的浓度分别从22.4增至26.4 μmol/ L和从82.1增至106.2 μmol/ L。由上面的结果可推论：高浓度的18- 甲基炔诺酮可将酮洛芬从它的第二类结合位点上置换出来。

The EOF pump was successfully used as a means of introducing samples into a capillary system. An improved sample injection device has been developed using a TeflonTM union(TU) to link the two capillaries together. The capillary applied high voltage served as the EOF pump to pull the liquid inside while the other one served the purposes of isolating the electric field. Using the bias degree(BD) and SD of bias(SDB), it was possible to quantitatively determine the sampling bias and evaluate the effectiveness of injection approaches in bias elimination. Several related factors were evaluated, and it was found that TU approach could fully eliminate the bias under the optimal conditions. The fracture did not affect the efficiency, leak, or dilute the sample significantly. This approach was effective under both normal and reverse EOF situations and adapted to real samples. Finally, a TU method using grounded injection electrode was proposed and shown to be suitable for samples with low conductivity and high injection voltage.