Establishment of Peptide Markers for Processed Pork Meat Against Various Meat-based Products (Chicken and Pork) by Shotgun Proteomics

Mansor, Muhammad Kamaruzaman (2020) Establishment of Peptide Markers for Processed Pork Meat Against Various Meat-based Products (Chicken and Pork) by Shotgun Proteomics. [Project Paper] (Submitted)

Text FPSK2 2020 34.pdf Download (1MB)

Abstract

Together with advances in meat processing technology, meat authentication has become a major concern in the food industry. Accidental or intentional mixing of meat-based products with pork is a major concern for quality control of food products, especially to consumers who have strict regulations in meat consumption such as Muslims and Jews, as well as for those with health reasons such as allergic reaction. Porcine serum albumin (PSA) is found abundance in pork meat and recognized as a major allergen. Objective: This study was to establish a sensitive method for the detection of trace amounts of pork meat using shotgun proteomics approach. Hypothesis: It was hypothesized that there were specific peptide makers for processed pork meat due to the uniqueness of peptide sequences as compared to chicken meat. Methodology: 29 processed meat samples of 2 different meat species (chicken and pork) were purchased from local supermarkets around Serdang area, Selangor. The protein extraction for each sample was executed by using trypsin enzyme, and incubated for 4 hours at 37 °C. Then, the samples were centrifuged twice and the supernatant was transferred into new 1.5µL microcentrifuge tube. Next, chromatographic separation of 20-μL injections was performed in triplicate for each sample through a reversed-phase column chromatography, followed by mass spectrometry detection. All the raw data was subjected to PatternLab software for data alignment and normalization. Lastly, the Area-proportional Venn Diagram (AVD) and TFold analysis were performed on all samples. Result: Shotgun proteomics can identify peptide and inferential protein from a complex mixture of samples. The AVD and TFold analysis of PatternLab established a series of peptide markers that was unique in both processed chicken and pork sample where out of 215 identified peptides, 99 and 83 of them were unique to pork and chicken samples, respectively. Protein with accession number of F1SS64 was chosen which contain three possible peptide markers which were VQLLHTQNTSLINTK, NLQQEISDLTEQIAEGGKR dan HADSVAELGEQIDNLQR. Conclusion: Specific peptide markers that were unique to processed pork meat samples against processed chicken meat samples can be established through shotgun proteomics coupled with PatternLab analysis.

Item Type:	Project Paper
Faculty:	Faculty of Medicine and Health Science
Depositing User:	Ms. Nor Safa'aton Saidin
Date Deposited:	23 Aug 2023 00:34
Last Modified:	23 Aug 2023 00:34
URI:	http://psaspb.upm.edu.my/id/eprint/1324

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