阳离子交换色谱及动态涂层毛细管电泳法检测牛奶中乳铁蛋白含量  

Determination of Bovine Lactoferrin in Milk by Cation Exchange Chromatography and Dynamic Coating Capillary Electrophoresis

作　　者：赵凌国 尤俊[2] 梁肇海 李可 丘汾 曾胜波 

机构地区：[1]深圳市福田区疾病预防与控制中心,广东深圳518040 [2]武汉大学化学与分子科学学院,湖北武汉430072

出　　处：《分析测试学报》2014年第3期339-343,共5页Journal of Instrumental Analysis

基　　金：国家自然科学基金(50973085);深圳市福田区卫生公益性重点科研项目资助(FTWS201304)

摘　　要：建立了阳离子交换色谱及动态涂层毛细管电泳检测牛奶中乳铁蛋白含量的新方法.通过purolite C115-E弱酸性离子交换树脂对牛奶中的乳铁蛋白进行提取,运用季铵化纤维素（QC）对毛细管动态涂层并测定乳铁蛋白含量.实验结果表明,该动态涂层能有效抑制毛细管内壁对乳铁蛋白的吸附.通过对QC浓度、运行缓冲液pH值、孵育时间等萃取和电泳条件的优化,实现了对牛奶样品中乳铁蛋白的定量分析.结果显示,低、中、高3个浓度组的加标回收率为87％～ 112％,相对标准偏差（RSDs）为4.4％～12.1％.该方法分析速度快、分离效率高、所需牛奶样品少,适用于牛奶中乳铁蛋白的分离分析.A dynamic coating capillary electrophoresis method with cation exchange chromatography was developed for the determination of bovine lactoferrin （bLF） in milk. The bLF was isolated from milk by using purolite C115 -E cation exchange resin, and then quantitatively analyzed in the capil- lary coated by quaternized celluloses（QC）. Effects of QC concentrations and pH values of the run- ning buffer on the separation of bLF by capillary electrophoresis were investigated. It was shown that the use of QC coating in CE could drastically reduce the adsorptio＂n of bLF on the inner surface of fused silica capillary. QC can not only cover the silanol groups on the capillary inner surface, but al- so repel the basic proteins electrostatically to prevent bLF protein adsorption. After the optimization of analytical conditions, calibration with bLF protein was carried out and linear regression data were presented. Furthermore, the influences of extraction conditions such as incubation time on the ex- traction efficiencies of bLF were also investigated. The recoveries of bLF in milk at three spiked lev- els ranged from 87% to 112% with relative standard deviations （RSDs） of 4. 4% - 12. 1%. This method had the advantages of rapidness, high efficiency, low sample and reagent consumption, and was suitable for the determination of bLF in milk.

关 键 词：阳离子交换色谱 毛细管电泳 动态涂层 乳铁蛋白 

分 类 号：O657.7[理学—分析化学] O629.73[理学—化学]