Preparation method of amauroderma rude polysaccharide F212 and application thereof to prevention and/or treatment of breast cancer
A technique for the preparation of F. chinensis and F212, which is applied to the preparation of F. chinensis polysaccharide F212, and the application field of preventing and/or treating breast cancer, and can solve the problems of difficulty in reaching medicinal levels, mixing, and blurred extract components.
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[0036] A preparation method of the above-mentioned polysaccharide F212 from Pseudomonas rugosa, comprising:
[0037] (1) Dried, pulverized, and extracted with water, preferably with hot water, to obtain the water extract of F. chinensis, and adding alcohol to the water extract, preferably at a mass percentage concentration It is 95%-100% alcohol, more preferably 95% alcohol by mass percentage, adjusted to 80% alcohol mass percentage concentration, standing at room temperature, preferably standing overnight, and then centrifuging, separating and collecting the precipitate product, to obtain the crude polysaccharide F0 of Fructus Chinensis;
[0038] (2) Purify F0 using DEAE sepharose fast flow, using sodium chloride solution as eluent to carry out gradient elution, the flow rate is preferably 20-40ml / min, and the concentration of sodium chloride solution is preferably 0.1, 0.5, 1, 2Mol / l, 4 components were collected;
[0039] (3) Using breast cancer cells to screen the activit...
Embodiment 1
[0052] Example 1: Preparation of polysaccharide F212
[0053] 1. After drying and crushing 1kg of the fruiting body of False Chrysanthemum rugosa, it is extracted with hot water to obtain a water-extracted concentrated solution of False Ginger. Add 95%-100% alcohol to the concentrated solution and adjust it to 80%. Leave it overnight, then centrifuge and precipitate to separate, collect the precipitate, and obtain the crude polysaccharide F0 of False Chrysanthemum wrinkle cap, with a yield of 1%-4%;
[0054] 2. Purify F0 with DEAE sepharose fast flow, and carry out gradient elution. The eluent is eluted with 0.1, 0.5, 1, and 2Mol / l sodium chloride in turn at a flow rate of 5ml / min, and gradient collection is carried out to obtain 4 components;
[0055] 3. Activity screening experiment: breast cancer cells (4T1 and MDA-MB-231) were used for activity screening of the four components.
[0056] 4. Screen out the most active component F1, purify it with Sephaacyl S-200chromatogra...
Embodiment 2
[0058] Example 2: Composition analysis of the polysaccharide F212
[0059] After testing, the molecular weight of the polysaccharide F212 is MN (number average molecular weight): 4.5852e3g / mol, MW (weight average molecular weight): 5.8659e3g / mol.
[0060] After testing, the polysaccharide F212 monosaccharide composition of A. chinensis is: 4.307% ribose, 0.717% rhamnose, 5.265% arabinose, 4.527% xylose, 30.527% mannose, 37.284% glucose, and 17.374% galactose.
[0061] Gel chromatographic analysis and infrared spectroscopic analysis were carried out on the polysaccharide F212 of A. figure 1 As shown, the spectrogram as figure 2 shown. The attribution of the absorption peaks in the infrared spectrum is shown in Table 1.
[0062] Table 1 Interpretation of the Absorption Peaks of F212 Infrared Spectrum
[0063]
[0064] Interpret the absorption peaks of the infrared spectrum, and see Table 1 for the assignment of each peak. Combined with infrared spectroscopy figure 2 A...
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