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A rapid determination method for the molecular weight of hydrolyzed collagen powder polypeptide

A technology for rapid determination of hydrolyzed collagen, which is applied in measuring devices, material separation, and analysis of materials. It can solve the problems of complex molecular weight determination of polypeptides and many influencing factors, and achieve the effects of reasonable design, easy operation, and high accuracy.

Active Publication Date: 2019-01-11
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the determination process of the molecular weight of the polypeptide by the above method is complicated, and there are many influencing factors. Therefore, it is one of the difficult problems for scientific researchers to quickly determine the molecular weight of the polypeptide and obtain accurate results.

Method used

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  • A rapid determination method for the molecular weight of hydrolyzed collagen powder polypeptide

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Experimental program
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Effect test

Embodiment 1

[0017] Embodiment 1: a kind of rapid assay method of hydrolyzed collagen powder polypeptide molecular weight, described assay method mainly comprises the following steps:

[0018] (1) Mix hydrolyzed collagen powder and ultrapure water at a volume ratio of 1:5, stir for 30 minutes, then centrifuge at 5°C and 10,000r / min for 1 hour, take out the supernatant, and use the UF membrane for the supernatant Concentrate by ultrafiltration, the volume ratio before and after concentration is 5:1, the powder concentrate after ultrafiltration and concentration is reconstituted by ion exchange chromatography using buffer solution, and the obtained reconstituted solution is subjected to gel chromatography containing Sephadex G-100 Column, separation and purification at a flow rate of 0.3mL / min, the detection wavelength is 280nm, the peak tubes of each peak displayed on the detector are collected, eluted, and the eluate is collected at 5°C and concentrated in a vacuum environment, the volume r...

Embodiment 2

[0022] Embodiment 2: a kind of rapid assay method of hydrolyzed collagen powder polypeptide molecular weight, described assay method mainly comprises the following steps:

[0023](1) Mix hydrolyzed collagen powder and ultrapure water according to the volume ratio of 1:5, stir for 30min, then centrifuge at 11000r / min at 7.5°C for 1h, take out the supernatant, and use UF membrane for the supernatant Carry out ultrafiltration and concentration, the volume ratio before and after concentration is 5:1.5, the powder concentrate after ultrafiltration and concentration is reconstituted by using buffer solution through ion exchange chromatography, and the obtained reconstituted solution is subjected to gel chromatography containing Sephadex G-100 Column, separation and purification at a flow rate of 0.4mL / min, the detection wavelength is 280nm, the peak tubes of each peak displayed on the detector are collected, eluted, and the eluate is collected at 7.5°C and concentrated in a vacuum en...

Embodiment 3

[0027] Embodiment 3: a kind of rapid assay method of hydrolyzed collagen powder polypeptide molecular weight, described assay method mainly comprises the following steps:

[0028] (1) Mix hydrolyzed collagen powder and ultrapure water according to the volume ratio of 1:5, stir for 30min, then centrifuge at 10°C and 12000r / min for 1h, take out the supernatant, and use the UF membrane for the supernatant Carry out ultrafiltration and concentration, the volume ratio before and after concentration is 5:2, the powder concentrate after ultrafiltration and concentration is reconstituted by using buffer solution through ion exchange chromatography, and the obtained reconstituted solution is subjected to gel chromatography containing Sephadex G-100 Column, separation and purification at a flow rate of 0.5mL / min, the detection wavelength is 280nm, the peak tubes of each peak displayed on the detector are collected, eluted, and the eluate is collected at 10°C and concentrated in a vacuum ...

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Abstract

The invention discloses a rapid determination method for the molecular weight of a hydrolyzed collagen powder polypeptide. The method mainly comprises: (1) separation purification of a polypeptide: dissolving hydrolyzed collagen powder, stirring, centrifuging, carrying out ultra-filtration concentration, carrying out redissolution with ion exchange chromatography, loading onto a gel chromatography column, carrying out elution, concentrating, and drying to obtain a separated and purified polypeptide; (2) determination of the polypeptide content: carrying out mixing dissolving on the separated and purified polypeptide by using a hydrolyzed enzymolysis liquid, carrying out light transmittance detection, and detecting the proportion of the light transmittance by using a spectrophotometer so as to determine the obtained separated and purified polypeptide content; and (3) determination of the molecular weight of the polypeptide: comparing the power sample detection result to a standard map by using the linear relationship between the molecule retention time and the molecular weight logarithm value in the standard map to rapidly determine the molecular weight of the polypeptide. According to the present invention, the determination method has characteristics of reasonable, scientific and rigor design, easy operation, and high accuracy.

Description

technical field [0001] The invention belongs to the technical field of polypeptide molecular weight determination, and specifically relates to a rapid determination method for polypeptide molecular weight of hydrolyzed collagen powder. Background technique [0002] Polypeptide is a compound formed by connecting α-amino acids with peptide bonds. It is also an intermediate product of protein hydrolysis. The compound formed by dehydration and condensation of two amino acid molecules is called dipeptide. Similarly, there are tripeptides, tetrapeptides, and pentapeptides. peptide etc. Usually, compounds formed by the dehydration condensation of three or more amino acid molecules can be called polypeptides. There is a clear definition of peptides in the organic chemistry foundation of the second year of high school. [0003] Generally, the number of amino acids contained in a peptide is two to nine. According to the number of amino acids in the peptide, there are many different ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/59G01N30/02
CPCG01N21/59G01N30/02
Inventor 徐昕荣朱斌胡国成黄思静
Owner SOUTH CHINA UNIV OF TECH
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