title: Development of a perfusion ion-exchange chromatography method for the separation of soybean proteins and its application to cultivar characterization.
abstract: A perfusion ion-exchange chromatography method has been designed, for the first time, for the separation of soybean proteins and its application to the characterization of soybean cultivars. For that purpose, the gradient, the mobile phase composition (buffer concentration, buffer pH, and elution salt), and the temperature were optimized. The method consisted of a two-step gradient (0% B for 2 min and from 0 to 50% B in 10 min) being mobile phase A a 2 0mM borate buffer (pH 9) and mobile phase B a 20 mM borate buffer (pH 9) containing 1M sodium chloride. The procedure used for the preparation of sample solutions was significantly simpler than that proposed by other authors and basically consisted of dissolving in water. This method enabled the separation of soybean proteins from a soybean protein isolate in 11 peaks in about 9 min. The method was used to separate soybean proteins in different commercial soybeans. In general, the 11 peaks yielded by the soybean protein isolate were also observed in the chromatograms of all soybeans. However, the area percentages of every peak in every soybean enabled the differentiation between soybeans. Moreover, the method was also used to separate soybean proteins in the proteic fractions obtained from every soybean. Multivariate methods were used for patterns recognition and the classification of samples.
authors.affiliation: Departamento de Química Analítica, Facultad de Química, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871 Alcalá de Henares, Madrid, Spain.
authors.forename: J M
authors.initials: JM
authors.lastname: Heras
authors.forename: M L
authors.initials: ML
authors.lastname: Marina
authors.forename: M C
authors.initials: MC
authors.lastname: García
mesh_terms: D002852:Chromatography, Ion Exchange / Q000379:methods*; D010477:Perfusion; D010940:Plant Proteins / Q000302:isolation & purification*; D013025:Glycine max / Q000737:chemistry* / Q000254:growth & development
chemical_list: D010940:Plant Proteins