Method for extracting antitumor active substance in fructus forsythiae, antitumor active substance and application thereof
An anti-tumor activity and anti-tumor technology, applied in the field of biomedicine, can solve the problems of high side effects and easy drug resistance of chemotherapy drugs
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Embodiment 1
[0042] Example 1: Preparation of Forsythia Water Extract (FS-1)
[0043] Take 500g of forsythia (purchased from Shiyitang pharmacy, Huangqiao, place of production in Shanxi), sieve and remove dust, wash 5 times with tap water, wash 3 times with distilled water, add distilled water to make the liquid level about 10cm higher than forsythia, soak overnight. Boil at 100°C the next day, filter while hot with absorbent cotton for 2 hours, extract the dregs repeatedly, then filter with absorbent cotton, mix the two filtrates, evaporate and concentrate at 100°C to 1.00 mg / ml, and press them out intermittently for three days Sterilize by bacterial method, 40min each time, make water extract, mark it as FS-1, and store it in a refrigerator at 4°C for later use.
Embodiment 2
[0044] Example 2: Preliminary isolation of effective anti-tumor fraction by ethanol precipitation method (FS-2)
[0045]Take 3.5ml of forsythia water extract FS-1 and place it in a plastic centrifuge tube, then slowly add ethanol solution with a mass percentage concentration of 100% to it in sections, and keep shaking until the volume of ethanol solution reaches 90% of the total volume of the solution until. Put the centrifuge tube in a 4°C refrigerator for 12 hours, centrifuge in a low-temperature centrifuge at 4°C, 4000r / min for 15min, collect the supernatant, filter the supernatant through filter paper, concentrate under reduced pressure, put it in a 4°C refrigerator, and set aside, labeled as FS- 2.
Embodiment 3
[0046] Example 3: Separation of Forsythia antitumor effective fraction (FS-3) by macroporous adsorption resin column chromatography
[0047] Take FS-2 and slowly add it to the top of the pretreated macroporous adsorption resin (X-5 type) column, the addition amount is 2% of the column volume, 8mg / time, 6 times in total. After FS-2 penetrates into the resin layer, elute with distilled water, ethanol solutions with a mass percentage concentration of 30%, 60%, and 90% in sequence at a flow rate of 5.00ml / min, collect the eluted part, and use a mass percentage concentration of 90% ethanol solution The eluate from ethanol solution was concentrated under reduced pressure at 100°C to obtain FS-3.
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