New method for separation and determination of denatured caseins by hydrophobic interaction chromatography

• Published in: Talanta (Oxford) Vol. 54; no. 2; pp. 343 - 349
• Main Authors: Bramanti, Emilia, Ferri, Fabrizio, Raspi, Giorgio, Lampugnani, Leonardo, Carla Spinetti, Maria, Miller, Keith E., Synovec, Robert E.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 12.04.2001; Oxford Elsevier
• Subjects: Biological and medical sciences; Caseins; Denaturation; Food industries; Fundamental and applied biological sciences. Psychology; Guanidine thiocyanate (GdmSCN); Hydrophobic interaction chromatography; Milk and cheese industries. Ice creams; Protein; Urea; Urea; Hydrophobic interaction chromatography; Caseins; Guanidine thiocyanate (GdmSCN); Denaturation; Protein; Separation method; Milk protein; Casein; Hydrophobic chromatography; HPLC chromatography; Dairy product; Chemical denaturation; Quantitative analysis
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/S0039-9140(00)00662-7

A new method for the separation of denatured α-, β- and κ- caseins by hydrophobic interaction chromatography (HIC) is proposed. The method is based on an easy solubilization of commercial and real samples by 4.0 M guanidine thiocyanate (GdmSCN) and elution on a TSK-Gel® Phenyl-5PW column (TosoHaas) in the presence of 8.0 M urea in the mobile phase. The procedure, applied to commercial caseins and to real, raw samples (whole milk powder and fat-free yoghurt) is not expensive, it requires common high performance liquid chromatography (HPLC) instrumentation and allows the separation of caseins also in the presence of whey proteins. Quantitative results on the analysis of α-, β- and κ-caseins in real samples are also reported.