A Quality Detection Method for Collagen Peptides Based on Spectrum-Effect Relationship
A quality detection method and collagen peptide technology, which are applied in measurement devices, instruments, scientific instruments, etc., to achieve the effect of simple method, strong objectivity, and good quality control.
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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
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[0166] Continuation of Table 5:
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[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 0.652 S10 2.328 0.968 0.887 S11 1.690 1.068 0.460 S12 1.431 ...
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