Determination of Bovine Lactoferrin in Milk by Cation Exchange Chromatography and Dynamic Coating Capillary Electrophoresis[J]. 2014,33(3):339-343.DOI:
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 capillary coated by quaternized celluloses(QC).Effects of QC concentrations and pH values of the running 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 adsorption of bLF on the inner surface of fused silica capillary.QC can not only cover the silanol groups on the capillary inner surface,but also 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 extraction efficiencies of bLF were also investigated.The recoveries of bLF in milk at three spiked levels 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.