Anti-tumor saponin in anemone plants and preparation method thereof as well as application
Anemone genus, anti-tumor technology, applied in the field of medicine, to achieve the effect of high yield
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preparation example 1
[0030] Pulverize 500g of anemone rhizome, pass through a 10-mesh sieve, add 8 times the amount of 0.2M 65% ethanol sodium hydroxide alkali solution, soak for 6 hours, reflux and extract three times (8 times, 8 times, 8 times the amount of alcohol added each time) ), 1.5 hours each time, combined the extraction solution, recovered until there was no alcohol smell, diluted the medicinal solution with distilled water, and adsorbed it with HPD-400 macroporous resin for 5 hours. The amount of resin used was 1kg. Elution removes impurities, and then elute with 12L 80% ethanol at a rate of 0.8L / h, collect the eluted part of 80% ethanol, spin dry, put on a silica gel column, and wash with ethyl acetate:methanol:water=65:30:8 The elution process of this step was monitored by TLC. The AR17 fraction was collected, spin-dried, and recrystallized with ethanol:methanol=6:1. The resulting product AR17 was 4.7g with a purity of 95.6%.
preparation example 2
[0032]Pulverize 500g of Anemone anemone medicinal material, pass through a 10-mesh sieve, add 6 times the amount of 0.3M 50% ethanol sodium hydroxide alkali solution, soak for 10 hours, reflux extraction three times (6 times, 6 times, 6 times), each time 2h , combine the extraction solutions, neutralize them with 2% hydrochloric acid, recover until there is no alcohol smell, dilute the medicinal solution with distilled water appropriately, and use D101 macroporous resin for adsorption for 3 hours. The amount of resin used is 2kg. h speed elution to remove impurities, washed until colorless, then eluted with 12L 80% ethanol at a speed of 0.5L / h, collected the eluted part of 80% ethanol, spin-dried, put on a silica gel column, and used ethyl acetate:methanol:water =70:30:7 elution, the elution process of this step was monitored by TLC, the AR17 fraction was collected, spin-dried, and recrystallized with ethanol:methanol=3:1, and the obtained product AR17 was 6.0 g with a purity o...
preparation example 3
[0034] Pulverize 500g of black mercury rhizomes, pass through a 16-mesh sieve, add 5 times the amount of 80% ethanol solution, soak for 12 hours, reflux and extract three times (5 times, 5 times, 5 times), each time for 1h, combine the extraction solutions, and recover to no Alcoholic taste, add NaOH to make 1.5M NaOH extract, heat at 80°C for 3h, and keep stirring. Dilute the medicinal solution with distilled water, and absorb it with HPD-300 macroporous resin for 8 hours. The amount of resin used is 1.5kg. Elution at a speed of 80% ethanol was collected, spin-dried, put on a silica gel column, eluted with ethyl acetate: methanol: water = 6: 0.5: 1, collected the eluted part, and the obtained product AR17 was 5.8 g, with a purity of was 91.6%.
[0035] Preparation Example 4 of Saponin AR17
[0036] Pulverize 500g of Anemone anemone medicinal material, pass through a 10-mesh sieve, add 10 times the amount of 0.5M KOH aqueous solution, soak for 10 hours, heat and extract thre...
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