Composition of anticancer agent using extract or active substance thereof
A kind of technology of Aphrodisiac sativa and its composition, which is applied in the field of anticancer agent composition
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Embodiment 1
[0058] Manufacture of extracts and fractions of A.
[0059] Put 1 kg of finely chopped Azalea (whole herb) into 80% ethanol, soak for 48 hours, extract repeatedly three times, filter to obtain the extract, concentrate the extract under reduced pressure to remove the solvent, and freeze-dry to obtain a powder extract.
[0060] In addition, after suspending and dissolving the obtained ethanol extract in 10 wt. 2 Cl 2 ), ethyl acetate (EtOAc) butanol (BuOH) for fractional distillation to obtain fractions of a hexane layer, a dichloromethane layer, an ethyl acetate layer, a butanol layer and the remaining aqueous layer.
Embodiment
[0062] Hereinafter, the present invention will be described with reference to examples and experimental examples. However, the scope of the present invention is not limited by these Examples and Experimental Examples.
[0063] Manufacture of the extract and fraction of A. sativa and the separation of its effective substances
[0064] Manufacture of extracts and fractions of A.
[0065] Put 1 kg of finely chopped Azalea (whole herb) into 80% ethanol, soak for 48 hours, extract repeatedly three times, filter to obtain the extract, concentrate the extract under reduced pressure to remove the solvent, and freeze-dry to obtain a powder extract.
[0066] In addition, after suspending and dissolving the obtained ethanol extract in 10 wt. 2 Cl 2 ), ethyl acetate (EtOAc) butanol (BuOH) for fractional distillation to obtain fractions of a hexane layer, a dichloromethane layer, an ethyl acetate layer, a butanol layer and the remaining aqueous layer.
Embodiment 2
[0067] Separation and identification of effective substances
[0068] 8 g of the above-mentioned ethyl acetate fraction was mixed with Celite 545, and the resulting mixture was concentrated under reduced pressure, followed by open column chromatography. Using n-Hex., MC (methylenechloride, dichloromethane), DE (Diethylether, diethyl ether), EA (Ethylacetate, ethyl acetate), Methanol (MeOH, methanol) as mobile phases, a total of five small fractions were obtained. However, silica gel column chromatography was performed on the DE (800 mg) fraction. Using chloroformate / methanol (9 / 2) as mobile phase at a flow rate of 2.5 ml / min, a total of 18 small fractions were obtained. The No. 14 fraction (DE-14) was simply purified to obtain a single compound (compound1).
[0069] To analyze the structure of the isolated substance, 10 mg of a single compound was dissolved in methanol-d to measure NMR spectrum. In 1HNMR spectrum ( figure 1 and image 3) in which three methyl groups we...
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