A kind of Digupi active part and its preparation method and application
A technology of active parts and Digupi, which is applied in the field of preparation of active parts of traditional Chinese medicine, can solve problems such as insufficient stability of curative effect, and achieve clear effect mechanism, low cost and definite curative effect
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Embodiment 1
[0028] Embodiment 1. The preparation method of Digupi active site
[0029] S1. After crushing Digupi, use 10 times the amount of water, heat and reflux extraction at 100°C for 2 times, each time for 120 minutes, concentrate the extract to 1 / 4 of the original volume, add 95% ethanol for alcohol The final ethanol concentration is about 75%, and the supernatant is concentrated under pressure to obtain a stock solution.
[0030] S2. pass the stock solution through the macroporous adsorption resin, the ratio (weight) of Digupi original medicinal material to resin is 1:3, wash the resin column with deionized water 3 times the volume of the resin bed, collect the eluate, and concentrate under reduced pressure to obtain Sample 1.
[0031] S3. Exchange sample 1 with a strong-acid cation-exchange resin (a cross-linked polystyrene-based strong acid resin with a model number of 001×7), rinse the resin column with deionized water three times the volume of the resin, discard the eluate, an...
Embodiment 2
[0033] Embodiment 2. A kind of liquid phase analysis method of Cortex Dictica active site
[0034] FMOC-Cl (fluorenylmethoxycarbonyl chloride) is a relatively new amino derivatization reagent, which is mainly suitable for the derivatization of primary and secondary amines, and the derivatives after the reaction have UV absorption. The anti-diabetic active molecule of Digupi has a secondary amine structure, so FMOC-Cl can be used as a derivatization reagent for derivatization. Take 50 μl of the sample 3 solution in Example 1 in a 1.5ml centrifuge tube, add 200 μl of 0.4mol / L potassium borate buffer solution, then add 250 μl of 5 mmol / LFMOC-Cl (50% acetonitrile configuration), shake well, and place at room temperature for 30 Minutes, then add 25 μl of 1mol / L glycine (Gly) to stop the reaction, then add 80 μl of acetic acid solution with a volume fraction of 1%, add water to make up to 1000 μl, and finally filter the solution with an organic membrane to obtain the test solution. ...
Embodiment 3
[0035] Example 3 Inhibition of α-glucosidase activity by the active site of Cortex Dictica
[0036] Inhibition of α-glucosidase activity by the active part of Cortex Lycii (prepared in Example 1): get 1.85ml of pH6.8 phosphate buffer solution, add 0.1U·ml successively -1 50 μL of α-glucosidase, different concentrations of Digustifolia active site solution (0, 1, 2, 3, 4, 5, 6, 7 mg·ml -1 ) 50μL, 0.0116mol L -1 pNPG50μL, after the above reaction system was incubated in a constant temperature water bath at 37°C for 30min, 0.1mol·L -1 Na 2 CO 3 8ml of the solution was used to terminate the reaction, and three replicates were performed for each concentration, and the average value was taken.
[0037] According to the above-mentioned method and calculation formula, obtain the inhibitory action figure of Digupi active site to α-glucosidase, such as figure 2 shown. It can be clearly seen from the figure that the active part of Digupi has a strong inhibitory effect on α-glucosi...
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