title: A proteomic-based approach for detection of chicken in meat mixes.
abstract: A proteomic-based method has been developed for the detection of chicken meat within mixed meat preparations. The procedure is robust and simple, comprising the extraction of myofibrillar proteins, enrichment of target proteins using OFFGEL isoelectric focusing, in-solution trypsin digestion of myosin light chain 3, and analysis of the generated peptides by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). Using this approach, it was possible for example to detect 0.5% contaminating chicken in pork meat with high confidence. Quantitative detection of chicken meat was done by using AQUA stable isotope peptides made from the sequence of previously selected species-specific peptide biomarkers. Linearity was observed between the amount of the peptide biomarker and the amount of chicken present in the mixture; further independent replication is required now to validate the method. Apart from its simplicity, this approach has the advantage that it can be used effectively for the detection of both raw and cooked meat. The method is robust, reliable, and sensitive, representing a serious alternative to methods currently in use for these purposes. It is amenable to highly processed foods which can be particularly problematic, as the tertiary protein structure is often affected in processed food precluding immunoassays. In addition, this proteomic analysis will permit the determination of definitive discriminatory sequence, unlike the DNA PCR based methods used presently. The present article also demonstrates the translation of the technology to routine mass spectrometry equipment, making the methodology suitable for public analysts.
authors.affiliation: Center for Systems and Synthetic Biology, School of Biological Sciences, and Specialist Bioanalytical Services Ltd., Royal Holloway, University of London, Egham TW20 0EX, UK.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Sentandreu
authors.forename: Paul D
authors.initials: PD
authors.lastname: Fraser
authors.forename: John
authors.initials: J
authors.lastname: Halket
authors.forename: Raj
authors.initials: R
authors.lastname: Patel
authors.forename: Peter M
authors.initials: PM
authors.lastname: Bramley
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis / Q000737:chemistry; D002645:Chickens; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005504:Food Analysis / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D007553:Isotope Labeling; D008461:Meat Products / Q000032:analysis*; D018994:Myosin Light Chains / Q000032:analysis / Q000737:chemistry / Q000145:classification; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000145:classification; D040901:Proteomics / Q000379:methods*; D013045:Species Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D014357:Trypsin
chemical_list: D015415:Biomarkers; D018994:Myosin Light Chains; D010446:Peptide Fragments; D014357:Trypsin