Camel milk characteristic peptide fragment combination and identification method

A technology of camel milk and peptides, applied in the field of proteomics, can solve problems such as difficulty in guaranteeing product quality and shortage, and achieve the effects of good accuracy, strong applicability, stability and accuracy

Pending Publication Date: 2022-04-05
江苏省食品药品监督检验研究院
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Problems solved by technology

my country's camel milk industry is developing rapidly and has good development prospects. However, at present, domestic camel milk, especially Bactrian camel milk products with Chinese characteristics, lacks reliable adulteration detection methods and standards, and it is difficult to guarantee product quality. Therefore, the development of camel milk The identification of adulterants in dairy products and the detection method of adulterated ratio are urgent problems to be solved

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  • Camel milk characteristic peptide fragment combination and identification method
  • Camel milk characteristic peptide fragment combination and identification method
  • Camel milk characteristic peptide fragment combination and identification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Example 1 Screening of Camel Milk Characteristic Polypeptides.

[0105] Step 1. Pretreatment of camel milk standard samples

[0106] (1) Determination of total protein content of pure camel milk powder solution

[0107] Take the pure milk powder of dromedary camel and pure milk powder of bactrian camel respectively as standard samples of camel milk, mix pure camel milk powder with pure water at a concentration of 0.01g / mL, vortex and mix well, and shake at 37°C for 30 minutes to make it After fully dissolving, dilute 20 times to prepare a working solution, and measure the total protein content; use the Coomassie brilliant blue method (Bradford protein concentration determination kit) to determine the total protein concentration, with bovine serum albumin (BSA, 5mg / mL) as the standard For protein, prepare standard solutions of 50 μg / mL, 100 μg / mL, 200 μg / mL, 400 μg / mL, 500 μg / mL, 800 μg / mL, 1000 μg / mL with PBS diluent, and then detect the total protein content of the sa...

Embodiment 2

[0152] Example 2 Screening of parameters for quantitative detection of characteristic peptides and detection of actual samples and simulated adulterated milk powder

[0153] 1. Optimize the quantitative parameters of characteristic peptides

[0154] The quantitative characteristic peptides screened in Example 1 were tuned with the MRM mode of liquid chromatography-tandem mass spectrometry described in Step 3 (5) of Example 1 to optimize important parameters such as reaction ion pairs, collision energy, declustering voltage, and retention time , all quantitative characteristic peptide mass spectrometry parameters are shown in Table 3.

[0155] Specifically, Shimadzu’s EXION AC-LC system was used for chromatographic separation, and the chromatographic column was ACQUITY UPLC BEHC8 (2.1×100mm, 1.7μm); the column temperature was: 60°C; the flow rate was 0.4mL / min; the injection volume was 5μL ; Mobile phase A selects 0.2% formic acid water for use, and mobile phase B selects 0.2%...

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Abstract

The invention relates to a camel milk characteristic peptide fragment combination and an identification method, a specific characteristic peptide marker combination is found through detection by a high performance liquid chromatography-tandem mass spectrometry technology, the characteristic peptide fragment combination comprises characteristic peptide fragments for identifying authenticity of camel milk powder as shown in SEQ ID NO.1-SEQ ID NO.8, and single-peak and double-peak camel milk can be identified by SEQ ID NO.6-SEQ ID NO.8. Through a large number of experimental screening, the peptide fragment with stable property, high response and good repeatability is screened from the specific characteristic peptide fragments of species, the influence of enzymolysis property, protein modification and matrix effect on detection is eliminated, and the established method is high in sensitivity, good in accuracy and high in operability. Besides, the characteristic peptide fragment in the protein is selected as a detection object and is not influenced by protein stability and deep processing, detection of non-denatured and denatured proteins in a sample can be met, and the method is high in applicability. And an effective technical means is provided for camel dairy product adulteration monitoring and commodity label authenticity supervision.

Description

technical field [0001] The invention belongs to the technical field of proteomics. The invention relates to a camel milk characteristic peptide combination, a screening method and an identification method thereof. It specifically relates to the combination of camel milk characteristic peptides and the authenticity of camel milk based on the combination of camel milk characteristic peptides, the detection of adulterated samples, the detection of adulterated ratios, and the identification of single-humped and double-humped camel milk. Background technique [0002] Camelus (Camelus) is a genus under Artiodactyla and Camelidae, which is divided into two species: dromedary camel (Camelus dromedarius, also known as Arabian camel) and Bactrian camel (Camelus bactrianus, also known as Asian camel). Dromedary camels are mainly distributed in West Asia, the Middle East and North Africa, accounting for 90% of the total number of camels; Bactrian camels are mainly distributed in Centra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K14/76C12P21/06G01N30/02G01N30/06G01N30/30G01N30/32G01N30/34G01N30/72
Inventor 黄青曹玲杨静张莹谭力施海蔚
Owner 江苏省食品药品监督检验研究院
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