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Meat sample adulteration quantitative detection method based on different animal-derived meat characteristic polypeptides

A quantitative detection method and animal-derived technology, applied in the field of proteomics, can solve the problems of restricting the quality of meat products, unable to crack down on illegal traders, unable to judge the severity of adulteration, etc., so as to reduce human errors and improve desalination efficiency. and the effect of peptide concentration

Active Publication Date: 2019-12-03
NANJING AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These problems have seriously affected the meat industry worldwide, not only restricting the improvement of meat quality, but also seriously damaging the economic interests and health of consumers, and even causing some religious problems
[0004] However, the existing animal-derived detection methods in the current detection standard system can only be qualitative or semi-quantitative, unable to judge the severity of adulteration, and cannot severely crack down on illegal traders of counterfeit and shoddy products

Method used

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  • Meat sample adulteration quantitative detection method based on different animal-derived meat characteristic polypeptides
  • Meat sample adulteration quantitative detection method based on different animal-derived meat characteristic polypeptides
  • Meat sample adulteration quantitative detection method based on different animal-derived meat characteristic polypeptides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Example 1 Screening and verification of characteristic polypeptides of different animal-derived meats

[0102] 1. Standard meat sample pretreatment:

[0103] Standard meat samples are chicken, duck, pork, beef, and mutton from which animal sources have been identified.

[0104] The standard meat samples are divided into two categories: raw meat and cooked meat. The raw meat is commercially available fresh meat, which is frozen at -20°C for later use. The cooked meat is prepared by heating fresh meat in a water bath at 80°C for 1 hour, and then frozen in -20℃ for use;

[0105] (1) Extraction of total protein from meat samples: remove fat and connective tissue from standard meat samples, cut into pieces and chop, weigh 2g of meat samples, add 25mL 2% SDS-PBS extraction buffer, and homogenize in an ice-water bath at 9600rpm for 30s 2 1 time, then 13400rpm 30s, each homogenization interval is 30s, then 4000g, 4 ℃ centrifugation for 10min, the supernatant is filtered with ...

Embodiment 2

[0144] The standard curve drawing of the adulteration situation of the meat sample to be tested in embodiment 2

[0145] Using triple quadrupole high performance liquid chromatography mass spectrometry to draw the standard curve of mixed meat samples, the specific method is as follows:

[0146] Mix the two meats of chicken, duck, pork, beef, and mutton according to the mass ratio of 0:100, 20:80, 40:60, 60:40, 80:20, and 100:0, and process them before going to the machine The method is specifically:

[0147] (1) Extract the total protein of the meat sample: wash the meat sample with 70% ethanol for 30 s, then wash with pure ethanol for 15 s, then wash with 90% methanol for 30 s, and finally wash twice with deionized water, each time for 30 s;

[0148] (2) Extraction and digestion of total protein from meat samples: Take 1 g of the meat sample obtained in step (1) and add 5 mL of protein extract, grind and homogenize on ice, and centrifuge at 12000 rpm at 4°C for 10 min. Add ...

Embodiment 3

[0164] Determination of the detection limit of the adulteration situation of the meat sample to be tested in embodiment 3

[0165] Use triple quadrupole high performance liquid chromatography mass spectrometry to determine whether the method can reach the detection limit of 1%, the specific method is as follows:

[0166] Two kinds of meat in chicken, duck, pork, beef, and mutton were mixed respectively in a mass ratio of 1:99, and the pretreatment method on the machine was the same as in Example 3.

[0167] The simulation results of adding 1% chicken to mutton and adding 1% duck to beef are as follows: Figure 17 , Figure 18 As shown, it can be seen from the figure: add 1% chicken to the mutton sample, the chicken sample can be detected; add 1% duck to the beef sample, the duck sample can be detected, it can be seen that this method has good detection Out of limit.

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Abstract

Aiming at a technical problem that animal-derived detection means can only be qualitative or semi-quantitative and the adulteration severity cannot be judged in an existing meat detection standard system, the invention provides a meat sample adulteration quantitative detection method based on different animal-derived meat characteristic polypeptides. In order to achieve the purpose of the invention, standard meat sample characteristic polypeptides of chicken, duck, pork, beef and mutton are screened, and the characteristic polypeptides are applied to the quantitative detection of meat sample adulteration. The quantitative detection method comprises the following steps of preprocessing a meat sample to be detected, detecting a characteristic polypeptide in the meat sample to be detected byhigh-performance liquid chromatography-mass spectrometry combined use and the like. The invention further provides a universal screening method of a standard meat sample characteristic polypeptide. Effective technical means are provided for monitoring the adulteration condition of meat samples in a meat market.

Description

technical field [0001] The invention belongs to the technical field of proteomics, and relates to a method for quantitatively detecting adulteration of meat samples based on characteristic polypeptides of different animal-derived meats. Specifically, it is a method for determining the sources and contents of different species in mixed meat samples based on high-performance liquid chromatography-mass spectrometry, and a method for screening species-specific polypeptides by using liquid chromatography-mass spectrometry. Background technique [0002] In recent decades, the proportion of meat products in residents' consumption has increased year by year, but it has also brought some illegal activities, such as using cheap pork, duck, chicken, and even sick and dead animal meat and expired meat as high-priced cattle, The mutton incident has been exposed from time to time, which is shocking. [0003] According to relevant surveys, the adulteration of meat products is mainly manif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N33/12
CPCG01N30/88G01N33/12G01N2030/8804G01N2030/8813
Inventor 李春保冯超彦杨军周光宏胡文彦徐道坤刘真
Owner NANJING AGRICULTURAL UNIVERSITY
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