A kind of preparation method of Drupe densiflora stem extract
A technique for drupa densiflora and stem extracts, which is applied in the field of preparation of drupa densiflora stem extracts, can solve the problems of no research reports on chemical components and pharmacological activities, etc., and achieve simple process flow and good anti-tumor activity , the effect of clear chemical composition
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preparation example Construction
[0022] The preparation method steps of Drupe densiflora tree stem extract are as follows:
[0023] 1) Add 85-95% ethanol or methanol to the crushed Drupe densiflora (through a 20-80 mesh sieve), soak at room temperature, heat or ultrasonically extract, the extraction time is 60 minutes to 7 days, and the material liquid The ratio is 1:1~1:10;
[0024] 2) Filter the drupe densiflora medicinal material extract, recover under reduced pressure to obtain the total extract, add water to suspend, add water to suspend, sequentially extract with petroleum ether, chloroform, ethyl acetate and n-butanol, and recover under reduced pressure to obtain petroleum Ether part, chloroform part, ethyl acetate part and n-butanol part;
[0025] 3) Carry out repeated column chromatography on the part of petroleum ether that has been determined to have extensive anti-tumor activity: petroleum ether: ethyl acetate (20:1) silica gel column chromatography to obtain compounds (1), (3) and (10); Petrole...
example 1
[0028] The preparation of example one Drupe densiflora tree stem effective part
[0029] Drupe densiflora tree stems (20Kg), soaked in 95% ethanol at room temperature for 3 times, 7 days / time, combined the extracts, concentrated under reduced pressure to obtain the total extract (1.0Kg), added water to suspend and disperse, respectively used petroleum ether, Chloroform, ethyl acetate and n-butanol were extracted to obtain petroleum ether fraction (80g), chloroform fraction (120g), ethyl acetate fraction (15g) and n-butanol fraction (450g).
[0030] The petroleum ether part undergoes repeated column chromatography on silica gel and Sephdex LH-20 to obtain 3 triterpenoids, 7 sterols and 1 glyceride derivative, which are respectively petroleum ether: ethyl acetate Esters (20:1) silica gel column chromatography to obtain 30 mg of β-aryl resin (1), 47 mg of suberinone (3) and 356 mg of β-sitosterol (10); petroleum ether: acetone (20:1) followed by 17 mg of 3α-hydroxyoleanolic acid...
Embodiment 2
[0032] Example 2 Study on Antitumor Activity of Petroleum Ether Parts of Drupe densiflora
[0033] 1. Experimental method: Three tumor cell lines K562, SPCA-1, and SGC-7901 were used in RPMI-1640 medium containing 10% calf serum at 37°C and 5% CO 2 cultured in an incubator. The MTT method was used to carry out the cell proliferation inhibition test. The main operation was to take the tumor cell lines in the logarithmic growth phase, digest them with 0.25% trypsin, and prepare 5×10 RPMI-1640 culture medium with 10% newborn calf serum. 4 pcs / ml -1 The cell suspension was inoculated in a 96-well plate with 180 μL per well. At 37°C, 5% CO 2 Cultivate for 8-10 hours under saturated humidity conditions, and when they adhere to the wall, add sample solution prepared with PBS to each well so that the final concentrations of the samples are 1, 10, and 100 μg / mL, respectively. For each concentration, 3 wells were paralleled, and after 44 hours of continuous culture, 50 μL MTT (1 mg / ...
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