Extraction method and application of tobacco anti-tumor active components
A technology of anti-tumor activity and extraction method, which is applied in the field of extraction of tobacco anti-tumor active components, can solve the problems of toxicity, flammability, and volatility, etc., achieve broad application prospects, simple preparation method, and increase the effect of extraction content
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[0038] Example 1: Extraction of Tobacco Antitumor Active Components
[0039] 1. The extraction steps of tobacco anti-tumor active components are as follows:
[0040] (1) Pretreatment.
[0041] Weigh 30Kg of tobacco raw material, place it in an oven for drying, the drying temperature is 60°C, the drying time is 4h, and take out until the moisture content of the tobacco is less than 5%; take the dried tobacco raw material, and use a 50-mesh screen After being pulverized by a pulverizer, it is put into a superfine pulverizer for superfine pulverization treatment to obtain tobacco superfine powder.
[0042] (2) extraction with normal temperature water.
[0043]Weigh 30Kg of tobacco superfine powder into a 200L extraction tank, add 100Kg of purified water, stir and extract at room temperature for 0.5h; of sediment.
[0044] (3) High temperature water extraction.
[0045] Weigh 25Kg of the precipitate in step (2) and add it to a 100L concentrated extraction tank; add 60Kg of pu...
Example Embodiment
[0054] Embodiment 2: the experiment that tobacco active component suppresses the growth of human tumor A549 cell
[0055] Blank control wells: no drug was added, and DMSO with a final concentration of 1% was added;
[0056] Experimental hole: add the antitumor active component that embodiment 1 step (6) (b) gains
[0057] Cell culture:
[0058] at 5% CO 2 , 37° C., under saturated humidity, cultured human lung tumor A549 cells in RPMI 1640 (10% FBS+1% PS) medium, and selected the cells growing in the logarithmic phase as experimental cells. After counting the cells, dilute it with medium to obtain a cell suspension of about 58,000 per milliliter.
[0059] Cell growth monitoring:
[0060] Place the cell real-time monitor in 5% CO 2 , 37 ℃ saturated humidity incubator. Take an 8-well plate, add 150 μL of RPMI 1640 medium to each well, put it into the real-time cell monitor for baseline, take out the eight-well plate after the baseline, and add 345 μL of diluted A549 cell s...
Example Embodiment
[0061] Embodiment 3: the test of suppressing the growth of human liver cancer Hep-G2 cells
[0062] Blank control wells: no drug was added, and DMSO with a final concentration of 1% was added;
[0063] Experimental hole: add the antitumor active component that embodiment 1 step (6) (b) gains
[0064] Cell culture:
[0065] at 5% CO 2 , 37° C., under saturated humidity, cultured human lung cancer Hep-G2 cells with DMEM (10% FBS, 1% PS) medium, and selected the cells growing in the logarithmic phase as experimental cells. After counting the cells, dilute it with medium to obtain a cell suspension of about 58,000 per milliliter.
[0066] Cell growth monitoring: put the cell real-time monitor in 5% CO 2 , 37°C saturated humidity incubator. Take an eight-well plate, add 150 μL of LDMEM medium to each well, put it into a real-time cell monitor for baseline, take out the eight-well plate after the baseline, and add 345 μL of diluted Hep-G2 cell suspension to each well to reach t...
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