Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Liquid chromatography-mass spectrometry technology-based protein-polysaccharide complex digestibility analyzing method

Active Publication Date: 2019-03-08
DALIAN POLYTECHNIC UNIVERSITY
View PDF4 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, research in this field has not been fully carried out, and a few studies have used a relatively simple analysis method, mainly using in vitro simulated digestion combined with SDS-PAGE electrophoresis to analyze the effect of added polysaccharides on the digestibility of protein-polysaccharide beverages
The limitations of the research techniques used are: 1. SDS-PAGE can only display the digestion rate of all proteins, but cannot display the digestion rate of each protein in the protein drink; 2. SDS-PAGE can only achieve protein digestion Semi-quantitative, not accurate enough for kinetic studies of digestibility; 3. Unable to provide additional information such as protein-polysaccharide binding sites

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Liquid chromatography-mass spectrometry technology-based protein-polysaccharide complex digestibility analyzing method
  • Liquid chromatography-mass spectrometry technology-based protein-polysaccharide complex digestibility analyzing method
  • Liquid chromatography-mass spectrometry technology-based protein-polysaccharide complex digestibility analyzing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] The preparation of embodiment 1 protein solution and protein-polysaccharide complex solution:

[0074] The preparation steps of the whey protein isolate solution and the whey protein isolate-Tremella polysaccharide complex solution are as follows: dissolve the whey protein isolate in water, stir at 200rpm for 4 hours at room temperature, wherein the mass ratio of protein to water is 20%, and then stir at 5 Overnight at ℃ to fully hydrate the protein to prepare a whey protein isolate solution for later use; dissolve tremella polysaccharide in water, wherein the mass ratio of polysaccharide to water is 2%, then fully hydrate the polysaccharide at 5°C overnight to prepare a tremella polysaccharide solution, spare.

[0075] Whey protein isolate-Tremella polysaccharide complex solution preparation steps: Tremella polysaccharide solution is added to the previously prepared whey protein isolate solution, the volume ratio is 1:1, the appropriate amount of water is controlled, t...

Embodiment 2

[0076]Example 2 In vitro simulated gastric juice digestion whey protein isolate solution and whey protein isolate-Tremella polysaccharide solution:

[0077] Centrifuge tubes containing 25mL of simulated gastric juice (containing 0.05M HCl and 0.5μg / L pepsin) were placed in a constant temperature shaker, shaken for 5 minutes in a water bath at 37°C and 95r / min, divided into two groups, and added Whey protein isolate solution and 10 mL of whey protein isolate-Tremella polysaccharide complex solution were placed in a water bath at 37°C and shaken at 95 r / min for 120 minutes. After digestion, the pH of the reaction solution was adjusted to 7 with 1M NaOH solution. The two groups of solutions after the digestion treatment are respectively referred to as the whey protein solution group and the whey protein-Tremella polysaccharide complex solution group below.

Embodiment 3

[0078] Example 3 Denaturation of whey protein isolate solution and whey protein isolate-Tremella polysaccharide solution and the second step of enzymatic hydrolysis, the specific steps are as follows:

[0079] The above-mentioned whey protein isolate solution group and whey protein isolate-Tremella polysaccharide solution group degraded by simulated gastric juice were added with protein denaturant (the formula of denaturant was 6M guanidine hydrochloride, 100mM tris(hydroxymethylaminomethane), 10mM tris(2- Carboxyethyl)phosphine and 40mM chloroacetamide), then heated at 95°C for 5 minutes, and cooled on ice. The denatured protein fragments were fractionated by centrifugation using an ultrafiltration tube with a membrane pore size of 10 kDa, and the centrifugation condition was 4000 g for 40 minutes. Ammonium bicarbonate solution was added to the large fragment polypeptide remaining in the ultrafiltration tube for centrifugation washing, and the centrifugation condition was 400...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
The inside diameter ofaaaaaaaaaa
Column lengthaaaaaaaaaa
Login to View More

Abstract

The invention discloses a liquid chromatography-mass spectrometry technology-based protein-polysaccharide complex digestibility analyzing method. The liquid chromatography-mass spectrometry technology-based protein-polysaccharide complex digestibility analyzing method comprises in vitro digesting protein solution and protein-polysaccharide complex solution in simulative gastric juice; performing denaturation treatment on digested proteins and digested protein-polysaccharide complexes, removing low-molecular-weight peptide fragments through an ultrafiltration pipe, and further completely hydrolyzing large-fragment polypeptides retained inside the ultrafiltration pipe through a second endo protease with clear enzyme-digesting points except pepsase to obtain a number of polypeptide fragments;detecting the composition and abundance of the polypeptide fragments through the liquid chromatography-mass spectrometry technology; according to the abundance of the polypeptides digested by the second endo protease and the pepsase, determining the digestibility of the pepsase; further, according to the high-abundance polypeptides in the two types of peptide fragments in the protein-polysaccharide complex group, deducting the binding sites of proteins and polysaccharides.

Description

technical field [0001] The invention belongs to the technical field of analysis methods, and in particular relates to a method for analyzing the digestibility of protein-polysaccharide complexes based on liquid chromatography-mass spectrometry. Background technique [0002] Protein beverages are a large category of beverage products stipulated in my country's national standard GB / T 10789-2015 Beverage General Rules. Refers to the use of milk or dairy products, or other edible proteins derived from animals, or the fruits, seeds or kernels of plants with a certain protein content as raw materials, with or without the addition of other raw and auxiliary materials and (or) food additives, Liquid beverages made through processing and live fermentation. [0003] Protein drinks have the characteristics of low calories and rich in protein. In recent years, they have become a health drink of public concern and a leading role in the emerging beverage market. Taking whey protein drin...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N30/88
CPCG01N30/88
Inventor 纪超凡赵铁峰胡静税东宁李胜杰祁立波林心萍梁会朋董秀萍
Owner DALIAN POLYTECHNIC UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com