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Method for detecting quality of ossein peptide based on spectrum-activity relationship

A quality detection method, collagen peptide technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve strong objectivity, ensure accuracy and repeatability, and the effect of simple method

Active Publication Date: 2018-11-23
INFINITUS (CHINA) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on the spectrum-effect relationship mainly involves the field of traditional Chinese medicine, and the research on the spectrum effect of peptides is still in a blank stage. Therefore, the research methods for the quality control of peptides such as collagen peptides need to be further developed

Method used

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  • Method for detecting quality of ossein peptide based on spectrum-activity relationship
  • Method for detecting quality of ossein peptide based on spectrum-activity relationship
  • Method for detecting quality of ossein peptide based on spectrum-activity relationship

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0124] Example 1: Establishment of the HPLC fingerprint of the collagen peptide to be tested

[0125] 1. Experimental method

[0126] 1) Preparation method of the test solution: Weigh 0.1g of collagen peptide sample, add 0.1mol / L dilute hydrochloric acid solution to dissolve it, transfer it to a 10mL measuring bottle, shake well, filter with a 0.22μm microporous membrane, and take the filtrate As the test solution;

[0127] 2) Derivatization of the solution: Place 200 μL blank sample solution and the test solution in a centrifuge tube respectively, add 100 μL acetonitrile solution with a concentration of 140 μL / mL triethylamine and 100 μL isothiocyanate with a concentration of 12.5 μL / mL in sequence The acetonitrile solution of phenyl ester, after vortex mixing, reacted in a water bath at 40°C for 1 hour, then added 400 μL of n-hexane to it, after vortex mixing, let it stand for a period of time, took the lower layer solution, and filtered it with a 0.22 μm microporous membra...

Embodiment 2

[0131] Example 2: Determination of Pharmacodynamic Indexes of 13 Batches of Collagen Peptide Difference Samples

[0132] 1. Experimental method

[0133] 1) Preparation of sample solution:

[0134] Precisely weigh 4.0 mg of 13 batches of bone collagen peptide sample powder into a sterile EP tube, quantitatively add 2000 μL of DMEM complete medium containing 10% fetal bovine serum to the ultra-clean workbench with a pipette gun, vortex and shake, wait until After the sample was completely dissolved in the culture medium, it was filtered with a 0.22 μm sterile microporous membrane, sterilized, and used for later use. The sample concentration was 2.0 mg / mL.

[0135] 2) Anti-inflammatory model establishment and dosing regimen

[0136] Knee articular chondrocytes of C518 rats in the logarithmic growth phase were uniformly seeded on a 96-well culture plate. After 24 hours of cell attachment, they were divided into groups. The control group was cultured with complete medium, and the...

Embodiment 3

[0162] Embodiment 3: Spectrum-effect relationship analysis

[0163] This embodiment adopts the method of gray correlation degree analysis to reveal the relationship between multiple independent variables and one dependent variable, and the magnitude of the correlation degree is used to determine the influence of variables on the index.

[0164] Table 5 Normalized processing results of characteristic peak areas and anti-inflammatory activities of HPLC fingerprints of 13 batches of differential samples

[0165]

[0166] Continuation of Table 5:

[0167]

[0168]

[0169] Continuation of Table 5:

[0170] sample

IL-1β

PGE 2

TNF-α

S1

1.314

1.214

1.478

S2

2.225

1.111

2.073

S3

0.416

0.907

1.974

S4

0.247

1.021

0.972

S5

0.390

0.660

1.082

S6

0.085

1.068

1.325

S7

0.560

0.968

0.286

S8

0.909

1.214

0.448

S9

0.763

1.021

0.65...

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Abstract

The invention relates to the technical field of polypeptide quality standard, in particular to a method for detecting the quality of ossein peptide based on a spectrum-activity relationship. The method comprises the following steps: (1) establishing an HPLC (High-Performance Liquid Chromatography) fingerprint spectrum of a to-be-detected ossein peptide test sample; (2) then screening out characteristic chromatographic peaks; (3) determining an anti-inflammatory function of the to-be-detected ossein peptide test sample; (4) establishing spectrum-activity correlation analysis by adopting a greyrelational analysis method, thus evaluating the anti-inflammatory activity of a common peak. By adopting the method disclosed by the invention, a peak VII, a peak I and a peak V in ossein peptide havehigh relational degree with the anti-inflammatory activity; the method has the advantages that the method is simple, the objectivity is strong, more characteristic peaks are obtained, and the like, and the method is beneficial for carrying out quality control on the ossein peptide.

Description

technical field [0001] The invention relates to the technical field of polypeptide quality standards, in particular to a method for detecting the quality of collagen peptides based on spectrum-effect relationship. Background technique [0002] Osteoarthritis (OA), also known as degenerative bone joint disease, is the most common joint disease, especially endangering the health of the elderly and bringing a heavy burden to society. Collagen peptides mainly exist in cartilage, and the active polypeptide substances extracted from animal cartilage tissue through enzymatic hydrolysis can repair bone tissue, relieve the symptoms of osteoporosis, and promote health. Studies have shown that collagen peptides have many active functions, such as anti-hypertension, prevention and treatment of osteoarthritis and osteoporosis, etc. Effect on children: Collagen peptide combines with calcium to promote bone development and enhance bone growth. Effect on adults: Collagen peptide promotes ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 张婷梁明马忠华刘卫罗丹
Owner INFINITUS (CHINA) CO LTD
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