The application of the extract of chrysanthemum in the preparation of anti-oxidation and anti-tumor drugs
An anti-tumor drug, the technology of the yellow hairy ear grass, which is applied in the field of biomedicine and achieves the effects of simple preparation method, induction of apoptosis and easy operation.
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Embodiment 1
[0033] Embodiment 1: the preparation of Fructus japonica extract
[0034] Grind the dried whole grass (1.0Kg), and extract with 80% (v / v) ethanol aqueous solution under continuous reflux for 3 times, each time for 2 hours, combine the extracts, concentrate to dryness under reduced pressure to obtain crude ethanol extract total extract (crude, 123.0g). The total extract (100.0g) was dispersed in water, extracted successively with an equal amount of petroleum ether, ethyl acetate and n-butanol, separated and concentrated under reduced pressure to constant weight to obtain petroleum ether extracts (PE fraction , 2.8g), ethyl acetate extract (EtOAc fraction, 3.58g) and n-butanol extract (BuOH fraction, 3.24g).
Embodiment 2
[0035] Example 2: Determination of Antioxidant Activity of Fructus Fructus Extract
[0036] Dissolve 1,1-diphenyl-2-[(2,4,6)-trinitro]phenylhydrazine (DPPH) (3.94 mg) in absolute ethanol to prepare a 0.4 mmol / L DPPH ethanol solution . The ethanol crude extract, petroleum ether extract, ethyl acetate extract and n-butanol extract were prepared into ethanol solutions with different concentration gradients (concentrations of ethanol crude extract were 50, 100, 150, 200, 250, 300 μg / mL ; The concentration of petroleum ether extract is 100, 150, 200, 250, 300, 350 μg / mL; the concentration of ethyl acetate extract is 100, 200, 300, 400, 500, 600 μg / mL; , 200, 300, 400, 500, 600 μg / mL). Take 1.5mL of each extract ethanol solution with different concentration gradients and the above prepared 0.4mmol / L DPPH ethanol solution and mix them in equal amounts at room temperature, react in the dark for 30min, and measure the concentration of each mixed solution with a UV spectrophotometer a...
Embodiment 3
[0041] Example 3: Determination of the inhibitory activity of the extract of Fructus japonica on the proliferation of tumor cells
[0042] MTT method was used to determine the effects of ethanol crude extract, petroleum ether extract, ethyl acetate extract and n-butanol extract on five kinds of human tumor cell lines (human breast cancer cell MDA-MB-231, colon cancer cell HCT-116, lung cancer cell line). Inhibitory effect on the proliferation of cell A549, cervical cancer cell Hela and oral epidermal cancer cell KB). DMSO was used as blank control, and 5-fluorouracil (5-FU) was used as positive control.
[0043] The above five tumor cells were divided into 3×10 3 Cells / well were seeded in 96-well plates in 5% CO 2Incubate at 37° C. for 24 hours in the presence of constant temperature. Then, the DMSO solutions of the above extracts (ethanol crude extract, petroleum ether extract, ethyl acetate extract and n-butanol extract) with concentrations of 6.25, 12.5, 25, 50, 100, 200...
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