Novel application of centella extract
A Centella asiatica extract and the technology of the extract are applied in the field of medicine, can solve the problems of large toxic and side effects, and achieve the effect of high activity of the extract
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Embodiment 1
[0021] Embodiment 1, the preparation of fatty acid synthase inhibitor Centella asiatica effective substance
[0022] Crush Centella asiatica, and extract the effective substance by the following method.
[0023] Use ethanol-water mixed solvent (the volume percentage of ethanol is 50%) and the above-mentioned pulverized Centella asiatica by weight ratio 8: 1, stir and leach for 12 hours at room temperature at 25 ° C, filter with ordinary filter paper; Then soak for another 12 hours, then filter with ordinary filter paper, and combine the two filtrates. At 0.09MPa, 40°C (reduced pressure) low-temperature evaporation to remove ethanol to obtain an aqueous phase containing effective substances, use an equal volume of petroleum ether to remove lipids, then extract three times with an equal volume of ethyl acetate, and combine acetic acid The ethyl ester layer was evaporated under reduced pressure and low temperature to remove the organic solvent to obtain an extract, which was dri...
Embodiment 2
[0025] Embodiment 2, the content determination of Centella asiatica active extract
[0026] Crush Centella asiatica, and extract the effective substance by the following method.
[0027] Accurately weigh 1.0 g of Centella asiatica extract in Example 1, place in a 100 ml volumetric flask, add water to dissolve, and set the volume to 100 ml as the test solution. Precisely measure 5 μL, and use acetonitrile (A)-0.3% acetic acid aqueous solution (B) gradient elution: 0-4min is (A) 20%, (B) 80%; 4-20min gradient gradually becomes (A) 60%, (B) 40%; 20-30min gradient gradually becomes (A) 70%, (B) 30%; Detection wavelength is 254nm; Flow velocity is 1mL / min; Column temperature is room temperature; Waters Symmetry C18 chromatographic column (4.6mm * 250mm , 5μm), the results showed that the content was 30%.
Embodiment 3
[0028] Embodiment 3, the detection of fatty acid synthase inhibitory activity
[0029] At 37°C, add the enzyme substrate to a 2mL quartz cuvette: add 1.85mL of 0.1M pH=7.0 phosphate buffer, 25μL of 0.2mM acetyl-CoA solution, 0.4mM malonyl-CoA solution 50μL and 50μL of 1.3mM NADPH solution, then add 30μL of FAS solution and mix well to start the enzymatic reaction. The spectrophotometer monitors the change of the absorbance value A at the wavelength of 340nm per unit time, and expresses the activity of FAS by (ΔA / min). Centella asiatica medicinal materials were crushed, passed through a 40-mesh sieve, added solvent (1:20), stirred at room temperature for 4 hours, centrifuged to separate the supernatant, and diluted ten times to determine the inhibitory rate of the extract to FAS. In the presence of a certain concentration of Centella asiatica extract, the activity of FAS holoenzyme reaction decreases, and when its activity decreases by half, the concentration of Centella asiat...
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