Food Chem. Fast LC-MS/MS method for detecting 7 kinds of animal meat in meat products

• By optimizing the steps of reduction, alkylation, digestion and purification, a rapid sample preparation method for identifying peptide markers has been established
• During rapid sample preparation, most proteins that provide species-specific peptides can be detected
• Established a fast LC-MS/MS method for detecting 35 species-specific peptides in 7 animals

As many food frauds and bad behaviors have occurred worldwide in the past few years, the authenticity of food has gradually become the focus of food safety and quality

For highly processed meat products containing multiple mammalian and poultry species, the use of mass spectrometry (MS) to detect specific peptides is a powerful, high-throughput meat certification method

The detection strategy of peptide markers for species identification is based on bottom-up proteomics, which is characterized by digestion of specific peptide bonds in proteins

In this study, Mingyue Zhang, Shouwei Wang, etc.

The protein concentration in the 7 kinds of raw meat shows that the protein extraction of this method is sufficient

Without reduction and alkylation, the protein extracted from meat samples can be directly digested by trypsin

After confirming the steps of rapid sample preparation, the amount of supernatant and trypsin and the digestion time were evaluated to minimize time and reagent consumption

After verifying the ability of the RSP method to generate enough peptides, the protein data of seven species of raw meat samples obtained by the traditional sample preparation method and the RSP method were compared to study the quality of the protein obtained by the RSP method

As shown in Figure 2, the identified proteins of the seven animal species are distributed in three cases, namely the only discovery in the RSP method, the only discovery in the traditional method, and the discovery in both methods

RSP method to screen and verify the specific peptides of 7 species

After confirming the effectiveness of the RSP method, a rapid LC-MS/MS method involving the RSP method to detect peptides of specific species was established

Screening of species-specific peptides

Screen the peptides extracted from the QE analysis in the raw meat samples of 7 species, and select the species-specific peptides according to the following parameters: 1) Each species has a unique test result; 2) The length is 6-25 amino acid residues Between; 3) There are trypsin-specific cleavage sites at both ends, no cleavage is missed; 4) It is abundant in muscle

China Meat Research Center, 100068 Beijing, China

Correspondence to: cmrc_wsw@126.

Abstract

The adulteration of meat products has been reported worldwide, and detection of specific peptides through mass spectrometry (MS) is a reliable method for meat species identification.
However, the practical application of this method is limited by complicated steps and long reaction time of the traditional sample preparation.
Therefore, this paper introduced a convenient and time-saving sample preparation by optimizing the steps of reduction, alkylation, digestion, and purification.
With the rapid sample preparation, 35 species-specific peptides for seven species (pig, cattle, sheep , deer, chicken, duck, and turkey) were screened using high-resolution MS, and a rapid LC-MS/MS method was established.
The method only takes 3 h from sample receipt to results.
The meat species of 20 processed meat products were detected,and three samples were found potentially adulterated.
The method is proved to have high sensitivity, specificity, practicability with respect to rapid identification of meat species in meat products.

ZHANG MY, LI YY, ZHANG YY, et al.
Rapid LC-MS/MS method for the detection of seven animal species in meat products[J].
Food Chemistry, 2021, 371: 131075.
DOI:10.
1016/j.
foodchem.
2021.
131075.