Method for extracting mangosteen pericarp anthocyanin and application thereof in preparing anti-inflammatory drugs
A technology of mangosteen peel and extraction method, which is applied in the field of natural medicinal chemistry, can solve the problems of extraction and its medicinal use that have not been reported, and achieve the effect of good application prospects
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Embodiment 1
[0016] Separation and extraction of anthocyanins from mangosteen peel:
[0017] (1) Separation of the crude extract of anthocyanins. Crush the completely air-dried mangosteen peel, add 10-40 times the volume of 20%-90% ethanol under the condition of 20-70°C, adjust the pH value to 1-5, and repeatedly extract 3-8 times, each extraction 1 ~5h, after filtering, combine the extracts. The obtained extract is distilled under reduced pressure at 30-70°C and a vacuum of -0.03--0.09MPa to recover the alcohol solvent, so that the alcohol concentration of the filtrate is reduced to below 10%, centrifuged and filtered to obtain anthocyanin leaching from mangosteen peel liquid.
[0018] (2) Macroporous resin to separate anthocyanins. The XAD-7 macroporous resin is fully washed with ethanol / water and pretreated, then packed into a column, and the anthocyanin extract from mangosteen peel is loaded onto the resin column. Slowly carry out the adsorption of anthocyanins on the macroporous r...
Embodiment 2
[0020] Inhibitory effect of mangosteen peel anthocyanins on acute peritonitis in mice: 1-5 mL of 1-10% Thioglycollate broth (Sigma) or sterile saline was injected intraperitoneally into male mice. After 5 to 30 minutes, sterile saline, 0.01, 0.1, and 1 mg / ml saline solution of anthocyanins from mangosteen peel were injected into the tail vein of the mice. After 1-5 hours, the mice were sacrificed, lavaged with 1-5 mL of normal saline, and the peritoneal fluid was collected. After centrifugation at 500-2000rpm for 1-10min, a part of the cells was double-blindly counted the total peritoneal cells with a hemocytometer; the other part was fixed, smeared and stained, and the number of neutrophils per mL was counted. The above experiment was repeated three times, and the average value was taken. The results showed that compared with the negative control group, mangosteen peel anthocyanins could significantly inhibit the exudation of leukocytes in the abdominal cavity of model mice,...
Embodiment 3
[0022] Mangosteen peel anthocyanins inhibit the binding of neutrophils to CHO-P under laminar flow: the adhesion of neutrophils to firmly adherent CHO-P was determined by laminar flow experiment in vitro to simulate capillaries. All single-cell suspensions used in laminar flow experiments had a density of 1–5×10 6 cells / mL. A 35 mm culture dish full of CHO-P was assembled in a parallel plate flow chamber, and then placed on the stage of an inverted microscope (before interacting with neutrophils, CHO-P cells expressing P-selectin were mixed with 0.01, 0.01, 0.1, 1mg / ml mangosteen peel anthocyanins act for 10-120 minutes). The inverted microscope is connected with a color video camera device, which can transmit video images to a video recorder, and can be simultaneously observed with a connected computer monitor. First, use a syringe pump (Cole-Parmer Instrument Co.) to inject PBS to wash the parallel plate flow chamber for 1-5 minutes, and inject neutrophil single-cell suspe...
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