Phosphorylated flat algae exopolysaccharide and preparation method thereof
A phosphorylation of flat algae, phosphorylation technology, applied in allergic diseases, anti-tumor drugs, drug combinations and other directions, can solve the problems of low biological activity of natural polysaccharides, poor water solubility, difficult to pharmacological research clinical preparation application research and other problems , to achieve the effect of improving biological activity and inhibiting effect
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[0028] The invention provides a preparation method of phosphorylated flat algae extracellular polysaccharide. In one embodiment, the method includes the steps of:
[0029] Step S10, dissolving sodium tripolyphosphate and sodium trimetaphosphate with water according to a preset ratio to obtain a phosphorylating reagent;
[0030] Step S20, adding the flat algae exopolysaccharide PEPs to the phosphorylation reagent to prepare a flat algae exopolysaccharide solution;
[0031] Step S30, adding a catalyst in the flat algae exopolysaccharide solution, heating to carry out a phosphorylation reaction to obtain a mixed solution, and adjusting the pH value of the mixed solution until it reaches the target pH value, to obtain a semi-finished product solution;
[0032] In step S40, the semi-finished solution is dialyzed and freeze-dried in sequence, so as to obtain phosphorylated Chrysalis exopolysaccharide pPEPs.
[0033] In the embodiment of the present invention, sodium tripolyphospha...
Embodiment 1
[0042] Example 1: Preparation of Phosphorylated Platycodonta exopolysaccharides.
[0043] Dissolve 7g of phosphorylation reagent (6g sodium tripolyphosphate + 1g sodium trimetaphosphate) and 5g of anhydrous sodium sulfate with 100ml ultrapure water, add 1g of purified flat algae exopolysaccharide PEPs after dissolving, and adjust the pH of the solution with NaHCO3 For 9, react at 80°C for 6h, then add 3 times the volume of 95% ethanol solution to the obtained semi-finished product solution, let it stand for 24h, centrifuge, collect the precipitate, and place it at 45°C for 2h to remove the residual ethanol , freeze-dried, reconstituted in a water bath at 50 °C, placed in a dialysis bag with a molecular weight cut-off of 800-1200, dialyzed for more than 24 hours, dialyzed for more than 1 time, collected the retentate, and freeze-dried to obtain phosphorylated flat algae extracellular Polysaccharide pPEPs.
Embodiment 2
[0044] Example 2: Detection of modification degree of phosphorylated flat algae exopolysaccharide
[0045] Determination of phosphate radical standard curve (molybdenum blue colorimetry):
[0046] Preparation of Tris buffer solution: Weigh 3.6g of Tris and 120mg of MgCl 2 ·6H 2 O was dissolved in ultrapure water, then diluted to 300ml by adding ultrapure water, and then adjusted to pH 7 with 1 mol / L HCl to obtain a Tris buffer solution.
[0047] Configuration of phosphorus determination reagent: take VC aqueous solution with mass fraction of 20% and H with 3 mol / L respectively. 2 SO 4 The solution and the ammonium molybdate aqueous solution with a mass fraction of 3% are mixed uniformly in equal volumes to obtain a phosphorus determination reagent.
[0048] Preparation of phosphate standard solution: Precisely weigh 1g of potassium dihydrogen phosphate, dissolve it in ultrapure water, use a 100mL volumetric flask to make up to the mark, and then dilute 100 times to obtain ...
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