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Characteristic peptide combination and method for detecting adulterate ratio of powdered milk in goat milk powder

A technology of characteristic peptides and milk powder, which is applied in the field of combination of characteristic peptides to detect the proportion of adulterated milk powder in goat milk powder, can solve the problems of few quantitative detections, achieve good linearity and ensure accuracy

Inactive Publication Date: 2017-05-31
杭州帕匹德检测技术服务有限公司
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the past ten years, many literatures have reported that this method can be used for qualitative identification of various meats such as chicken, pork and fish, but few studies involving quantitative detection
At the same time, there are few literatures to study and report on the method of species traceability analysis of milk protein

Method used

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  • Characteristic peptide combination and method for detecting adulterate ratio of powdered milk in goat milk powder
  • Characteristic peptide combination and method for detecting adulterate ratio of powdered milk in goat milk powder
  • Characteristic peptide combination and method for detecting adulterate ratio of powdered milk in goat milk powder

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Experimental program
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Effect test

Embodiment 1

[0037] The optimization of embodiment 1 sample pretreatment conditions

[0038] β-lactoglobulin has a relatively tight spatial structure, and its spatial structure is mainly maintained by disulfide bonds formed between cysteines and other non-covalent bonds. If the spatial structure of β-lactoglobulin is not destroyed, the enzyme cleavage site cannot be exposed and the enzyme cleavage cannot be completely performed. By using dithiothreitol (DTT) to hydrolyze the disulfide bond to destroy the protein spatial structure. Utilizing the specificity that trypsin acts only on arginine (R) and lysine (K), the protein is decomposed into polypeptides for detection. In the present invention, by optimizing the reducing temperature and the concentration of DTT, the purpose of the highest enzyme cutting efficiency is finally achieved.

[0039] The present invention designs a set of orthogonal experiments of DTT concentration and reduction temperature to optimize the reduction reaction condi...

Embodiment 2

[0040] Screening and determination of embodiment 2 characteristic peptides

[0041] Cattle and sheep belong to the same genus Bovine, but belong to different subgenera: Subfamily Bovidae and Subfamily Caprinae. Through the comparison of the Uniprot database, it was found that the proteins in cow's milk and goat's milk have high homology and little difference in amino acid sequence. The present invention selects whey protein β-lactoglobulin with a relatively high content in milk as a marker, and selects the specificity of the amino acid sequence difference between cow milk and goat milk through database screening and comparison, and finally achieves quantitative detection The purpose of the adulteration ratio of whole milk powder in whole goat milk powder.

[0042] The present invention simultaneously satisfies the detection of milk adulteration in goat milk and sheep milk, so the selected goat milk characteristic peptides meet the consistency of goat milk and sheep milk, and ...

Embodiment 3

[0045] The design and determination of the internal standard characteristic peptide of embodiment 3

[0046] Due to the different matrix effects of peptides in different samples, the peak area response values ​​measured by the external standard method are quite different. At the same time, the protein may be interfered by the matrix effect during the enzymatic hydrolysis process, resulting in incomplete enzymatic hydrolysis and affecting the accuracy of the results. accuracy. Therefore, it is necessary to design an internal standard so that it can correct the influence of the matrix effect on the enzymatic hydrolysis efficiency and the detection process. Therefore, according to the natural protein sequence, on the basis of the specific peptides LSFNPTQLEEQCHI and LAFNPTQLEGQCHV, the present invention synthesizes the internal standard milk characteristic peptide LSFNPTQL*EEQCHI* and the internal standard goat milk characteristic peptide LAFNPTQL*EGQCHV*, L* refers to [ 13C6,15...

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Abstract

The invention discloses a characteristic peptide combination and a method for detecting the adulterate ratio of powdered milk in goat milk powder. The characteristic peptide combination comprises milk characteristic peptides and goat milk characteristic peptides, wherein the amino acid sequence of the milk characteristic peptides is LSFNPTQLEEQCHI, and the amino acid sequence of the goat milk characteristic peptides is LAFNPTQLEGQCHV. Through the application of the characteristic peptide combination disclosed by the invention, the adulterate ratio of the powdered milk in the goat milk powder can be quantitatively detected by using a high performance liquid chromatography-mass spectrometry combined technology, the method is relatively high in linearity, sensitivity, recovery rate and accuracy rate, and characteristic peptide fragments in a protein are selected as detection substances and are applicable for detection of denatured protein, so that natured and denatured proteins in a detection sample can be detected simultaneously, and the accuracy of the method is guaranteed.

Description

technical field [0001] The invention belongs to the technical field of food detection, and in particular relates to a combination of characteristic peptides and a method for detecting the proportion of adulterated milk powder in goat milk powder. Background technique [0002] In recent years, goat whole milk powder has been sought after by the market due to its low allergenicity, high nutritional value, easy digestion and absorption, and unique flavor, and has become a hot-selling product in the dairy industry. However, the production of goat milk is low and affected by seasonal changes, and the raw material cost of goat milk is high. Therefore, driven by economic interests, the phenomenon of adulteration of milk powder with lower price and higher output mixed with goat milk powder frequently occurs. This kind of adulteration not only damages the economic interests of consumers, but also poses a threat to the health of consumers with milk allergies. [0003] At present, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47G01N33/68G01N30/02
CPCC07K14/4717G01N30/02G01N33/68
Inventor 黄棣华
Owner 杭州帕匹德检测技术服务有限公司
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