Complex of marine oligosaccharides and collagen peptides, preparation method thereof and application thereof
A collagen peptide and compound technology is applied in the field of the compound of marine characteristic oligosaccharide and collagen peptide to achieve important economic and social benefits and strong antioxidant effects
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Embodiment 1
[0040] Embodiment 1, preparation method
[0041] 1. Preparation of mannuronic acid with antioxidant activity and a degree of polymerization of about 2-8
[0042]Use the HCL solution with a concentration of 0.1-0.3M to make a suspension solution with a weight percent concentration of 1-10% by β-D-(1,4)-connected polymannuronic acid (average molecular weight 7000Da), and then, Hydrolyze in a high-pressure reactor at 120°C for 1 hour, adjust to pH 7 with 0.1% sodium hydroxide after cooling, filter through a 0.45 μm membrane, concentrate by rotary evaporation, separate with a Bio-Gel-P6 column, and use 0.1M ammonium bicarbonate The buffer solution is used for elution to obtain oligosaccharides with a degree of polymerization of 2-8, which are desalted with Sephadex G10 and freeze-dried to obtain a brown powder, which is the mannuronic acid product required by the present invention.
[0043] 2. Preparation of tilapia collagen peptide with antioxidant activity and molecular weight ...
Embodiment 2
[0048] Embodiment 2, oxygen free radical scavenging experiment
[0049] The purpose of the experiment: to determine the effect of the compound of mannuronic acid and collagen peptide on the oxygen free radical DPPH·, O at the molecular level 2 - ·As well as the scavenging ability of OH., and compare the antioxidant effect of the compound with its single component, in order to determine the effect of the compound on the improvement of the antioxidant activity of its single component.
[0050] Experimental method: Weigh 20 mg of the compound, tilapia fish skin collagen peptide with a molecular weight of 2800-3000 Da and mannuronic acid with a polymerization degree of 2-8, and prepare a sample solution of 0.25-20 mg / ml. The oscillator is fully oscillating and ready to be tested. Using DPPH method, AP-TEMED spectrophotometry and o-phenanthroline-Fe 2+ The antioxidant activity of the complex and its single components was detected by the oxidation method.
[0051] Experimental r...
Embodiment 3
[0053] Embodiment 3, MTT cell viability experiment
[0054] Purpose of the experiment: The protective effect of the compound of mannuronic acid and collagen peptide on skin cells damaged by ultraviolet rays was detected by MTT method at the cellular level, and the antioxidant effect of the compound and its single component was compared in order to determine Compounding can improve the anti-ultraviolet activity of its single component.
[0055] Experimental method: Fetal mouse fibroblasts (MEF) and human immortalized keratinocytes (HaCaT) were randomly divided into control group (normal culture cells), model group (ultraviolet irradiation group) and drug-dosing group (different sample treatment + ultraviolet irradiation group) irradiation). When the cell growth was close to 80% confluence, under sterile conditions, the final concentration of 10, 20, 50, 100, 200, 500 and 1000 μg / ml of the complex, tilapia fish skin collagen peptide was measured by the degree of polymerization ...
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Abstract
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