title: Development of an ultra-high performance liquid chromatography analytical methodology for the profiling of olive (Olea europaea L.) pulp proteins.
abstract: Ultra-high performance liquid chromatography (UHPLC) constitutes an interesting proposal to speed protein separations but it is almost not explored. In this work UHPLC is proposed, for the first time, to separate olive pulp proteins. An important difficulty in the analysis of proteins is related to their extraction. The difficulty in the extraction of proteins from the olive pulp is derived from its high content in lipids and phenolic compounds. Eight different methods for the extraction of pulp proteins were designed and evaluated. The optimized extraction procedure consisted of a cleaning step to remove interfering compounds, followed by the extraction of proteins with a Tris-HCl buffer containing sodium dodecyl sulphate (SDS) and dithiothreitol (DTT), precipitation of proteins with acetone, and solubilization in the Tris-HCl buffer. This methodology yielded the most successful isolation of pulp proteins and enabled the optimization of a UHPLC methodology for their separation. The method was applied to the profiling of olive pulp proteins from different olive cultivars observing in all cases a protein that had never been described before.
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, University of Alcalá, Ctra. Madrid-Barcelona, Km. 33.600, E-28871 Alcalá de Henares, Madrid, Spain.
authors.forename: Clara
authors.initials: C
authors.lastname: Esteve
authors.forename: Carmen
authors.initials: C
authors.lastname: Del Río
authors.forename: M Luisa
authors.initials: ML
authors.lastname: Marina
authors.forename: M Concepción
authors.initials: MC
authors.lastname: García
mesh_terms: D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004229:Dithiothreitol / Q000737:chemistry; D031658:Olea / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D020543:Proteome / Q000032:analysis; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry
chemical_list: D010940:Plant Proteins; D020543:Proteome; D012967:Sodium Dodecyl Sulfate; D004229:Dithiothreitol