Application of gallic acid in agaricus gennadii in CDC25 phosphoprotease
A phosphoprotease and gallic acid technology, which is applied in the directions of medical preparations containing active ingredients, preparation of carboxylate esters, preparation of organic compounds, etc. The effect of simple separation and low production cost
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Embodiment 1
[0035] Preparation of the crude extract of each active component in the mushroom tortuosporum of embodiment 1
[0036] Take 10kg of fresh, undamaged Agaricus torusum fruiting body, slice it and soak it with 30L 95% ethanol for 7 days (wherein the ratio of Agaricus toricosporin to ethanol is 1:3, so that the ethanol completely soaks the fruiting body of Agaricus torisporum), filter The filtrate was obtained, the solvent was recovered under reduced pressure, and 30 L of 95% ethanol was added to the filter residue for ultrasonic extraction 3 times, each time for 2 hours. Concentrate under reduced pressure to obtain 95% ethanol extract. Operate in the same way with 65% ethanol, and concentrate under reduced pressure to obtain 65% ethanol extract. Add 30L of distilled water to the filter residue of the mushroom after alcohol extraction, and extract three times with 1800HZ ultrasound, each time for 2 hours. Concentrate under reduced pressure to obtain 538 g of water extract.
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Embodiment 2
[0041] Example 2 Petroleum ether phase component separation and purification
[0042] 10.0 g of the petroleum ether fraction obtained in the above example 1 was separated by medium-pressure silica gel column flash chromatography, petroleum ether-ethyl acetate system (100:0-0:100) gradient elution, and the eluent was detected by thin-layer chromatography. The similar fractions were combined, and the solvent was recovered to obtain a total of 9 components A to I. Component D is separated by medium-pressure silica gel chromatography to obtain D1 and D2. D1 was recrystallized to obtain 20 mg of compound 3 in the form of needles. D2 was separated by multiple medium-pressure silica gel chromatography and Sephadex LH-20 column chromatography to obtain 10 mg of compound 2 as a white solid. Component E was separated by multiple medium-pressure silica gel chromatography and Sephadex LH-20 column chromatography to obtain 5 mg of compound 1 as a yellow-green oily solid.
[0043] Among ...
Embodiment 3
[0055] Example 3 Separation and Purification of Ethyl Acetate Part Components
[0056] 6.0 g of the ethyl acetate fraction obtained in the above-mentioned Example 1 was reversed phase C by medium pressure 18 Column flash chromatography separation, water-methanol solvent system (5% to 100%) gradient elution, the eluents are combined according to the same gradient, and the solvent is recovered to obtain 6 components from I to VI. After checking by high performance liquid chromatography, it was found that components I to IV were common components in the n-butanol phase, so they were merged into the n-butanol phase. Ⅴ component through medium pressure reverse phase C 18 After detaching, use C again 18 After semi-preparative column separation, a total of 4 mg of compound 4, a total of 4 mg of compound 5 and a total of 3 mg of compound 24 were obtained. Compound 4 is a yellow granular solid; Compound 24 is a yellow granular solid; Compound 5 is a red granular solid.
[0057] whe...
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