高效毛细管电泳法测定牛乳铁蛋白的含量及杂蛋白的方法学研究

Research on HPCE Determination of Content and Related Proteins of Bovine Lactoferrin

目的:采用高效毛细管电泳法(HPCE)的毛细管区带电泳模式(CZE)分离测定牛乳铁蛋白(BLF)的含量。方法:熔融石英毛细管柱67cm(有效长度55cm)×50μm;运行缓冲液为33mmol·L^(-1)磷酸二氢钾-10mmol·L^(-1)磷酸溶液(pH=2.50);压力进样6.9 kPa×5s;运行电压25kV;检测波长λ=214nm;毛细管柱温为25℃。结果:乳铁蛋白进样浓度在100.0-600.0μg·mL^(-1)范围内线性关系良好(r=0.9995),最低检测限为25μg·mL^(-1)。结论:采用HPCE测定乳铁蛋白及杂蛋白的方法简便,结果可靠。

Objective: To determine the content and the related proteins of Bovine Lactoferrin(BLF) by HPCE. Methods: Capillary zone electrophoresis(CZE) separation, fused-silica capillary column: 67cm(efficiency length 55 cm)×50μm,running buffer;33 mmol·L^(-1) KH_2PO_4-10 mmol·L^(-1) H_3PO_4solution(pH=2. 50);eletroinjection;6. 9 kPa×5 s;running voltage;25kV;detection wavelength;214 nm;column temperature;25℃. Results; The calibration curve of BLF was linear in the range of 100. 0-600. 0μg·mL^(-1)(r=0. 9995). The detection limit is 25. 0μg·mL^(-1). Conclusion:The method of determining BLF content and related proteins of BLF sample by HPCE is easy to operate and the results of the determination are accurate.