Method for preparing anti-inflammatory and anti-tumor active ingredient group from liquorice dregs and application thereof
A technology of active ingredients and licorice dregs, which is applied in the preparation of anti-inflammatory and anti-tumor active ingredient groups from licorice dregs and its application field, can solve the problems of shortage of licorice resources and increasing demand for licorice, and achieve composition Clear, clear principal component content, the effect of inhibiting the growth of tumor cells
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Embodiment 1
[0027] (1) 10kg of dried licorice residue (obtained after water extraction of glycyrrhizic acid) was refluxed with 95% ethanol for three times, 2 hours, 2 hours, 1.5 hours, and the extract was recovered under reduced pressure to obtain crude ethanol extract;
[0028] (2) The ethanol extract was suspended in 5L of distilled water, and extracted four times with equal volumes of petroleum ether and ethyl acetate;
[0029] (3) The ethyl acetate extract obtained in step (2) was separated by silica gel column chromatography, and eluted with petroleum ether: acetone 10:1, 2:1, 1:1,
[0030] (4) The petroleum ether: acetone 2:1 fraction obtained in the above step (3) was separated by polyamide column chromatography, and ethanol / water 2:8, 6:4, 9:1 gradient elution, ethanol / water 6: 4 The eluate is the active ingredient group.
[0031] (5) Through HPLC analysis and standard comparison method, the composition of the obtained product was clarified (the structures of S1-S13 are shown in ...
Embodiment 2
[0042] (1) 5 kg of dried licorice residue (obtained after extracting glycyrrhizic acid by alkaline water method) was reflux extracted with methanol, and the extract was recovered under reduced pressure to obtain crude methanol extract;
[0043] (2) The methanol extract was suspended in 1L of distilled water, separated and purified by HPD100 macroporous adsorption resin, and then eluted with ethanol / water mixed solvent ratios of 3:7, 6:4, and 9:1, and the ratio of ethanol / water mixed The crude product was obtained as a 6:4 partial eluate
[0044] (3) The crude product obtained in step (2) is separated by polyamide column chromatography, and eluted successively with ethanol / water mixed solvent ratios of 2:8, 4:6, 6:4, and 8:2, and the mixing ratio of ethanol / water is 4:6~6:4 part of the eluate obtained the target active ingredient group.
[0045] (4) The composition of the obtained product was clarified through HPLC analysis and standard comparison method, and the experimental me...
Embodiment 3
[0048] Quinone reductase-inducing activity test of active ingredient group
[0049] 1. Experimental materials:
[0050] (1) Main reagents and main equipment: calf serum (purchased from Hyclone); fetal bovine serum (TBD);
[0051] Tetramethylazolium salt (MTT) American Sigma (St. Louis, MO); Glucose-6-phosphate dehydrogenase (Shanghai Sangong); 96-well cell culture plate (Costar); cell line: mouse liver cancer cell Strain Hepa 1c1c7
[0052] 2. Experimental method:
[0053] (1) Cell culture:
[0054] Seed 10 per hole 4 Cells were grown for 24 hours in a culture medium containing 10% (v / v) fetal bovine serum, 0.01% penicillin G, 0.15% sodium bicarbonate, 0.01% streptomycin sulfate, the environmental conditions were 37 ℃, containing 5% CO 2 of humid air.
[0055] (2) Preparation of drugs
[0056] The compound to be tested was dissolved in DMSO to make a mother solution with a concentration of 50mmol / L, and stored at -20°C.
[0057] (3) Crystal violet method t...
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