Preparation method and content measuring method of macrothelypteris viridifrons total flavone with anti-tumor activity
A kind of anti-tumor activity, the technology of Pinhair fern, which is applied in the field of total flavonoids, achieves the effect of easy control, good reproducibility and high product yield
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Embodiment 1
[0027] The preparation of the total flavonoids of Pinhairia emeralds with anti-tumor activity provided by the present invention
[0028] (1) Preparation of raw materials: the emerald green needle fern (collected in Xijiujiang, Baijiang in October 2009, identified as Macrothelypteris macrothelypteris by the researcher Tan Tse-ming of the Jiujiang Forest Herbarium) The dried whole herb of viridifrons (Tagawa) Ching) is a raw material, which is cleaned, dried, pulverized, and crossed through a 40-mesh sieve to obtain the raw material coarse powder of emerald green needle hair fern for use;
[0029] (2) Solvent extraction: use organic solvent ethanol, methanol, 60-90% hydrous ethanol or 60-90% hydrous methanol to extract the raw material coarse powder of emerald green needle hair fern, heat to 70-90 ℃ for extraction, and the amount of solvent used is the same as that of emerald green The weight ratio of needle hair fern to coarse powder is 6-15:1, and the extraction is repeated 1 ...
Embodiment 2
[0036] Take the dried whole herb of Pinata emerald, crush it, sieve it, add 10 times the volume of 70% ethanol, and extract under reflux at 90°C for 60 minutes. Repeat the extraction twice, combine the extracts, concentrate under reduced pressure to recover the solvent, and dry in vacuum to obtain the total extract. Weigh a certain amount of total extract, add distilled water to dissolve it ultrasonically, extract twice with three times the amount of petroleum ether, combine the water layer, adjust the pH to 6 with 15% sodium hydroxide, filter, and filter out the precipitate. The filtrate is washed with concentrated hydrochloric acid Adjust the pH value to 1, put it in the refrigerator at 4°C overnight, filter the next day, wash the obtained precipitate with distilled water until it is neutral, and dry it in vacuum to obtain the flavonoids extracted from cloverine. After adjusting the pH to 4 with sodium hydroxide, the remaining filtrate was subjected to macroporous resin adso...
Embodiment 3
[0038] Inhibitory effect of total flavonoids of the method of the present invention on human breast cancer cell MCF-7
[0039] After subculture, human breast cancer cells MCF-7 in logarithmic growth phase were blown and dispersed in the medium containing 10% inactivated calf serum, and the concentration was measured with a cell counting plate under an inverted microscope and adjusted to 2×10 5 cells / mL. Add 100 μL of the cell suspension to each well of a 96-well plate and store at 37°C in 5% CO 2 cultured in an incubator for 24 h. Add 100 μL of sterilized sample-containing culture solution to make the final concentrations of the samples 0.1, 1.0, 10.0, and 100.0 μg / mL, respectively. Continue at 37°C 5% CO 2 cultured in an incubator for 48 h. Afterwards, 10 μL of 5 mg / mL MTT solution was added to each well to continue culturing for 4 h. Carefully aspirate the cell solution, add 200 μL of DMSO to each well to dissolve the formed formazan, shake it on a cell plate shaker for...
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