茜素红S-牛血清白蛋白配合物的凝胶层析分离制备及紫外可见吸收光谱研究

UV-Vis Absorption Spectrophotometric Study on the Bovine Serum Albumin-Alizarin Red S Complex Separated and Prepared by Gel Chromatography

用葡聚糖凝胶层析分离制备牛血清蛋白(BSA)-茜素红S(ARS)配合物。在420和530 nm处用紫外可见吸收光谱法同时测定含有BSA-ARS配合物和茜素红S的联立方程为:A420=4.89×103cARS+3.06×104cBSA-ARS,A530=4.60×102cARS+2.29×104cBSA-ARS;正交试验选择了合适的分离条件:柱直径1.0 cm,柱长30.0 cm,凝胶用量1.3 g,最佳进样浓度为ARS:5×10-3mol/L、BSA:1.49×10-4mol/L,进样体积1.5 mL,洗脱流速0.33 mL/min,分离度为1.25;测定了纯BSA-ARS配合物紫外吸收光谱,最大吸收峰在530 nm。

The complex formed between bovine serum albumin（BSA） and alizarin red S （ARS） was separated and prepared by gel chromatography. The absorbances of the complex in an excess of ARS were measured at λ420nm and λ530nm respectively by UV -Vis spectrophotometer. The concentrations of ARS and BSA - ARS were then calculated according to the following simultaneous equations： A420 = 4. 89 × 10^3 CARS ＋3. 06 × 10^4 CBSA-ARS, A530 =4.60 × 10^2 CARS ＋2. 29 × 10^4 CBSA-ARS. According to the orthogonal experiment, the optimum diameter and length of the chromatographic column were 1.0 cm and 30 cm, the gel amount was 1.3 g, the sampling concentration and volume for ARS were 5 × 10^-3 mol · L^-1 and 1.5 mL and those for BSA were 1.49× 10^-3 mol · L^-1 BSA and 1.5 mL, the elution flow rate was O. 33 mL · min^-1 and the separating degree was 1.25. The wavelength of the maximum absorption peak of BSA - ARS complex was at 530 nm.