Evaluation method of internal protein nutrition based on LC-MS-MS technology

A technology of LC-MS-MS and evaluation methods, which is applied in the direction of proteomics, peptide preparation methods, chemical instruments and methods, etc., can solve the problem of inability to faithfully reflect protein digestion and absorption, protein digestion, time, etc. long wait question

A technology of LC-MS-MS and evaluation methods, which is applied in the direction of proteomics, peptide preparation methods, chemical instruments and methods, etc., can solve the problem of inability to faithfully reflect protein digestion and absorption, protein digestion, time, etc. long wait question

CN107655985AActive Publication Date: 2018-02-02NANJING AGRICULTURAL UNIVERSITY
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  • Evaluation method of internal protein nutrition based on LC-MS-MS technology
  • Evaluation method of internal protein nutrition based on LC-MS-MS technology
  • Evaluation method of internal protein nutrition based on LC-MS-MS technology

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Experimental program
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Effect test

Embodiment 1

[0104] Taking C57BL / 6J mice as animal models, they were fed with low-salt (0.25%) and high-salt (3.5%) diets, with 5 mice in each group, and the experimental period was 8 weeks. After the end of the experiment, the contents of the duodenum, jejunum, cecum and colon were collected after neck dislocation, placed in 2ml cryopreservation tubes and quickly frozen in liquid nitrogen, and transferred to -80°C for later use. 10 mice, four types of intestinal contents, a total of 40 samples. Protein extraction was performed by adding RIPA lysate, phosphatase inhibitors and protease inhibitors (method as above). After the protein was extracted, the concentration was measured with the BCA kit, and the quality was adjusted after digestion and desalting according to the operation steps. The total amount of protein in each centrifuge tube selected in this experiment was 1.5 μg. After LC-MS-MS and database search, the identification results of proteins from three sources in four intestinal...

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Abstract

The invention discloses an evaluation method of internal protein nutrition based on an LC-MS-MS technology. The evaluation method includes steps of (1), collecting different intestinal segment contents; extracting and separating protein components; (2), testing the protein concentration; (3), mass spectrum pretreatment including digestion and desalting of full protein; (4), LC-MS-MS analysis; (5),database searching; (6), data processing. The method applies a proteomic technology to identify the protein and its digested product in different intestinal segment contents; on this basis, source ofthe proteins of the intestinal segment contents and the contents can be judged; through the bioinformatics analysis, the function of different proteins at the inner part of an organ can be learned deeply, wherein the gene expression of enzyme related to protein digestion and metabolism may be different; the method provides a scientific basis for scientific evaluation of the protein digestion anduse.

Description

technical field [0001] The invention belongs to the technical field of proteomics, and relates to a method for evaluating protein nutrition in vivo based on LC-MS-MS technology. Background technique [0002] The more authoritative method for measuring protein digestibility in vivo is fecal nitrogen balance test. The principle is that dietary protein enters the gastrointestinal tract through the esophagus, pepsin in the stomach digests dietary protein, polypeptides enter the small intestine (duodenum, jejunum, and ileum) and are acted on by trypsin and chymotrypsin. Finally, a small amount of microorganisms When cells, fecal metabolized nitrogen shed from intestinal mucosa and incompletely digested and absorbed protein pass through the colon, they are excreted in the form of feces. The ratio of the difference between intake nitrogen and fecal nitrogen to intake nitrogen is the apparent digestibility. Although 5% highly digestible casein will be added and fecal metabolic nit...

Claims

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Application Information

Patent Timeline
02 Feb 2018
Publication
CN107655985A
IPC
G01N30/02; G01N30/06
CPC
G01N30/02; G01N30/06; G01N33/6848; G16B40/10; G16B20/00; C07K1/16; C07K1/34; C07K1/36
Inventors
李春保; 王超