title: Separation of human, bovine, and porcine insulins, three very closely related proteins, by micellar electrokinetic chromatography.
abstract: Human, bovine, and porcine insulins are small proteins with very closely related amino acid sequences, which makes their separation challenging. In this study, we took advantage of the high-resolution power of CE, and more particularly of micellar electrokinetic chromatography, to separate those biomolecules. Among several surfactants, perfluorooctanoic acid ammonium salt was selected. Then, using a design of experiments approach, the optimal BGE composition was found to consist of 50 mM ammonium acetate pH 9.0, 65 mM perfluorooctanoic acid ammonium salt, and 4% MeOH. The three insulins could be separated within 12 min with a satisfactory resolution. This method could be useful to detect possible counterfeit pharmaceutical formulations. Indeed, it would be easy to determine if human insulin was replaced by bovine or porcine insulin.
authors.affiliation: Laboratory for the Analysis of Medicines, Department of Pharmacy, CIRM, University of Liege, Liege, Belgium.
authors.forename: Caroline
authors.initials: C
authors.lastname: Lamalle
authors.affiliation: Laboratory for the Analysis of Medicines, Department of Pharmacy, CIRM, University of Liege, Liege, Belgium.
authors.forename: Diane
authors.initials: D
authors.lastname: Roland
authors.affiliation: Laboratory for the Analysis of Medicines, Department of Pharmacy, CIRM, University of Liege, Liege, Belgium.
authors.forename: Jacques
authors.initials: J
authors.lastname: Crommen
authors.affiliation: Laboratory for the Analysis of Medicines, Department of Pharmacy, CIRM, University of Liege, Liege, Belgium.
authors.forename: Anne-Catherine
authors.initials: AC
authors.lastname: Servais
authors.affiliation: Laboratory for the Analysis of Medicines, Department of Pharmacy, CIRM, University of Liege, Liege, Belgium.
authors.forename: Marianne
authors.initials: M
authors.lastname: Fillet
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D006801:Humans; D007328:Insulin / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D013552:Swine
chemical_list: D007328:Insulin
keywords: Bovine insulin; Counterfeit drugs; Human insulin; Micellar electrokinetic chromatography; Porcine insulin