Phellinus polysaccharide II having antineoplastic activity, extraction and separation method thereof
A technology of polysaccharide and anti-tumor activity of P. acicularis, which is applied in the directions of organic active ingredients, anti-tumor drugs, and medical preparations containing active ingredients, etc., can solve problems such as large blindness, loss of active ingredients, and toxicity to normal cells , to achieve good anti-tumor activity and reduce blindness
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Embodiment 1
[0031] Example 1: Get the fruiting body of the fungus, pulverize it, pass through a sieve with a diameter of 260 μm, get 1 kg of dry fruiting powder, add 10 kg, 5 kg and 2 kg of distilled water respectively, extract 3 times at 100° C. for 2 hours each time, combine the extracts, and use Rotary evaporator concentrated to 3kg. Slowly add 12kg of 90% ethanol to the concentrated solution, put the solution in a 4°C refrigerator, let it stand overnight (about 10 hours), then centrifuge at a speed of 4000r / min for 10min, take the precipitate, and dry it at a low temperature of 40°C. get crude polysaccharides.
[0032] Dissolve the crude polysaccharide in 0.01mol / L acetic acid-sodium acetate buffer solution with a pH value of 4.5, prepare a solution with a polysaccharide content of 10mg / ml, centrifuge at 4000r / min for 10min, discard the precipitate, and filter the centrifugate through a 0.45μm microporous membrane To remove impurities, the filtrate was loaded on a 2.6×20cm DEAE-Sepharo...
Embodiment 2
[0036] Example 2: Take the fruiting body of Phytophthora aciculatus, crush it, add 5 times the amount of water to the fruiting body powder, boil and extract 4 times, each time for 1 hour, combine the extracts, add 3 times the amount of 90% ethanol after concentration, and place at 4 °C In the refrigerator, let it stand overnight (about 7 hours), centrifuge at a speed of 3500r / min, and dry the precipitate at a low temperature of 40°C to obtain the crude polysaccharide.
[0037] Dissolve the crude polysaccharide in 0.01mol / L acetic acid-sodium acetate buffer solution with a pH value of 5.0, centrifuge at 3500r / min for 10min to discard the precipitate, filter the centrifugate through a 0.45μm microporous membrane to remove impurities, and load the filtrate on a 2.6×20cm DEAE-Sepharose Fast Flow column, 0.01mol / L acetic acid-sodium acetate buffer solution with a pH value of 5.0 elutes 2 column volumes, and the eluate is discarded or collected for use in antitumor experiments (the e...
Embodiment 3
[0039] Example 3: Take the fruiting body of Phylophora acutiformis, pulverize, add 10 times, 2 times, 5 times, and 3 times the amount of water to the fruiting body powder and boil for extraction 4 times, each time for 1.5 hours, combine the extracts, and concentrate Add 5 times the amount of 90% ethanol, put it in a refrigerator at 4°C, let it stand overnight (about 14 hours), centrifuge at a speed of 4500r / min, and dry the precipitate at a low temperature of 45°C to obtain crude polysaccharide.
[0040] Take the crude polysaccharide, dissolve it in the acetic acid-sodium acetate buffer solution with a pH value of 4.0, centrifuge at a speed of 4500r / min, discard the precipitate, filter the centrifugate through a 0.45 μm microporous membrane, put the filtrate on an ion exchange column, and use a pH value of 4.0 acetic acid-sodium acetate buffer was used to elute 2 column volumes, and the eluate was discarded or collected for anti-tumor experiments (the eluate was collected by a ...
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