title: The use of label-free mass spectrometry for relative quantification of sarcoplasmic proteins during the processing of dry-cured ham.
abstract: The aim of this work was to quantify changes in the abundance of the major sarcoplasmic proteins throughout the ham dry-curing process by using a label-free mass spectrometry methodology based on the measurement of mass spectral peak intensities obtained from the extracted ion chromatogram. For this purpose, extraction of sarcoplasmic proteins was followed by trypsin digestion and analysis by nanoliquid chromatography coupled to tandem mass spectrometry (Q/TOF) for the identification and relative quantification of sarcoplasmic proteins through individual quantification of trypsinised peptides. In total, 20 proteins, including 12 glycolytic enzymes, were identified and quantified. The accuracy of the protocol was based on MS/MS replicates, and beta-lactoglobulin protein was used to normalise data and correct possible variations during sample preparation or LC-MS/MS analysis. Mass spectrometry-based proteomics provides precise identification and quantification of proteins in comparison with traditional methodologies based on gel electrophoresis, especially in the case of overlapping proteins. Moreover, the label-free approach used in this study proved to be a simple, fast, reliable method for evaluating proteolytic degradation of sarcoplasmic proteins during the processing of dry-cured ham.
authors.affiliation: Instituto de Agroquímica y Tecnología de Alimentos (CSIC), Avenue Agustín Escardino 7, 46980, Paterna, Valencia, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Gallego
authors.affiliation: Instituto de Agroquímica y Tecnología de Alimentos (CSIC), Avenue Agustín Escardino 7, 46980, Paterna, Valencia, Spain.
authors.forename: Leticia
authors.initials: L
authors.lastname: Mora
authors.affiliation: Instituto de Agroquímica y Tecnología de Alimentos (CSIC), Avenue Agustín Escardino 7, 46980, Paterna, Valencia, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Concepción Aristoy
authors.affiliation: Instituto de Agroquímica y Tecnología de Alimentos (CSIC), Avenue Agustín Escardino 7, 46980, Paterna, Valencia, Spain. Electronic address: ftoldra@iata.csic.es.
authors.forename: Fidel
authors.initials: F
authors.lastname: Toldrá
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005511:Food Handling; D007782:Lactoglobulins / Q000032:analysis*; D008461:Meat Products / Q000032:analysis*; D011506:Proteins / Q000032:analysis; D059748:Proteolysis; D040901:Proteomics; D012519:Sarcoplasmic Reticulum / Q000737:chemistry*; D013552:Swine; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000032:analysis
chemical_list: D007782:Lactoglobulins; D011506:Proteins; D014357:Trypsin
keywords: Dry-cured ham; Label-free; Mass spectrometry; Peak intensity; Peptides; Quantification; Sarcoplasmic proteins