Sample preparation affects separation of whey proteins by reversed-phase high-performance liquid chromatography

被引：8

作者：

Bobe, G ^{[1
]}

Beitz, DC ^{[1
]}

Freeman, AE ^{[1
]}

Lindberg, GL ^{[1
]}

机构：

[1] Iowa State Univ, Dept Anim Sci, Ames, IA 50011 USA

来源：

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY | 1998年 / 46卷 / 04期

关键词：

casein; whey; milk protein; reducing agent; chaotropic agent; reversed-phase high-performance liquid chromatography;

D O I：

10.1021/jf970687f

中图分类号：

S [农业科学];

学科分类号：

09 ;

摘要：

Measured concentrations of whey proteins in single milk samples often differ by more than 20% when analyzed by different separation methods. In the current study, we examined the effects of using guanidine hydrochloride or urea at zero and 8.5 h after sample preparation and the effects of using dithiothreitol or a-mercaptoethanol on the separation of bovine milk proteins by reversed-phase high-performance liquid chromatography. Treatment with guanidine hydrochloride or urea resulted in similar separation of milk proteins when samples were injected immediately after preparation. Separation was repeatable over 8.5 h for samples prepared with guanidine hydrochloride, whereas the resolution of a-lactalbumin and beta-lactoglobulin decreased for samples treated with urea. Treatment with dithiothreitol improved the resolution of alpha-lactalbumin and beta-lactoglobulin in comparison with treatment with 2-mercaptoethanol. Quantitation of whey proteins is more reliable when milk samples are treated with dithiothreitol and guanidine hydrochloride than when treated with 2-mercaptoethanol or urea.

引用

页码：1321 / 1325

页数：5

共 24 条

[1] Combined effect of pH and temperature during electroreduction of whey proteins [J].