A kind of antioxidative polysaccharide extracted from Cyanosporium cyanophilus and preparation method thereof
A technique for oxidizing polysaccharides from Cyanosporium cyanophilus, which is applied in the field of antioxidant polysaccharides and its preparation, can solve the problems of few research reports on the pharmacological active ingredients of Cyanospora cyanophilus and lack of theoretical support, etc. Oxidation effect, low cost, good safety effect
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Embodiment 1
[0034] Embodiment 1, the preparation of antioxidant polysaccharide of the present invention
[0035] (1) Grind the fruiting body of Fomitiporiapunctata (P.Karst.) Murrill into powder, add 20 times the volume of distilled water and mix well, extract in a constant temperature water bath at 80°C for 5 hours, filter the obtained filter residue to extract again, and combine the filtrate, namely Crude extract of the fruiting bodies of Fomitiporiapunctata (P.Karst.) Murrill;
[0036] (2) The crude extract of the fruiting bodies of Fomitiporiapunctata (P.Karst.) Murrill was concentrated by rotary evaporation, and 4 times the volume of absolute ethanol was added to it, and vigorously stirred while adding, to produce precipitation. Place at 4°C overnight, centrifuge at 10,000rpm for 20min, collect the precipitate, wash the precipitate several times with absolute ethanol, acetone and ether in sequence, and obtain the crude extract of Fomitiporiapunctata (P.Karst.) Murrill;
[0037] (3) Re...
Embodiment 2
[0040] Embodiment 2, the purity and molecular weight determination of antioxidant polysaccharide of the present invention
[0041] (1) Prepare the standard dextran (DextranT-10, T-50, T-70, T-100, T-200) with known molecular weight into a standard solution with a concentration of 1% in double distilled water, 0.22 μm Membrane filtration, the antioxidant polysaccharide of the present invention is treated in the same way.
[0042] (2) Chromatographic conditions: chromatographic column is ShodexSUGARKS-804 sugar column (8.0×300mm); evaporative light scattering detector (ELSD), column temperature: room temperature; mobile phase: double distilled water; injection volume: 20 μL; air pressure: 3×10 5 Pa
[0043] (3) Molecular weight standard curve drawing: Inject 20 μL of standard dextran respectively, collect spectra, take molecular weight Mw as ordinate, retention time t as abscissa, draw standard curve ( Figure 4 ).
[0044] (4) Inject 20 μL of the antioxidant polysaccharide ...
Embodiment 3
[0045] Embodiment 3, the scavenging effect of DPPH free radical of antioxidant polysaccharide of the present invention
[0046] (1) Antioxidant polysaccharides were prepared into a solution with a concentration of 0.5 mg / mL using anhydrous methanol as a solvent, and DPPH was prepared with anhydrous methanol to a final concentration of 1×10 -4 mol / L solution.
[0047] (2) Take 0.06, 0.12, 0.18, 0.3, 0.6mL of the above polysaccharide solution in 1.5mL centrifuge tubes, make up 1.2mL with DPPH solution, replace the sample solution with an equal volume of anhydrous methanol as a blank control, and ascorbic acid as a positive control , placed in the dark for 30min.
[0048] (3) Measure the absorbance value of each treatment at 517nm, and make 3 parallels for each treatment.
[0049] (4) Press "clearance rate=[1-(A s -A b ) / A i ]×100%” formula to calculate DPPH clearance rate, where A i is the absorbance value of the DPPH solution without sample addition, A s is the absorbanc...
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