A kind of Anemarrhena homogeneous polysaccharide and its preparation method and application
A technology of Anemarrhena and polysaccharide, which is applied in the field of separation and purification of plant active components to achieve the effects of simple process, good antioxidant and hypoglycemic activity
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Embodiment 1
[0042] Example 1: Preparation of Anemarrhena polysaccharide homogeneous polysaccharide AABP-2B
[0043] (1) Extraction of Anemarrhena polysaccharide:
[0044] Take 1Kg Anemarrhena rhizomes, pass through a 100-mesh sieve after being pulverized by a pulverizer, add 5L of 95% ethanol to reflux at 70°C for 2 hours, repeat twice, centrifuge, and dry the precipitate at 45°C. Add 5L of distilled water to the degreased and decolorized Anemarrhena dry powder, extract in a water bath at 85°C for 3 hours, repeat the extraction 3 times, filter, centrifuge, collect the supernatant, and concentrate the supernatant under reduced pressure to the original volume. One-fifth; slowly add 4 times the amount of absolute ethanol dropwise to the concentrated solution, let it stand at 4°C for 24 hours, centrifuge, collect the precipitate and dry it to obtain 64.5g of mother crude polysaccharide.
[0045] (2) In addition to protein:
[0046] Take 30g of Anemarrhena crude polysaccharide, make a 20mg / m...
Embodiment 2
[0065] Example 2: Antioxidant activity of Anemarrhena polysaccharide
[0066] The antioxidant activity of AABP-2B was evaluated by DPPH, ABTS method and superoxide free radical test, as follows:
[0067] Detection of DPPH free radical scavenging activity: Add 100 μL of different AABP-2B polysaccharide solutions (0.1, 0.2, 0.4, 0.6, 0.8, 1.0 and 2.0 mg / mL) and 100 μL of 0.1 mM DPPH ethanol solution to a 96-well plate, mix well, and keep at room temperature Protect from light for 30 minutes, test the absorbance of the mixture at 517nm, use ascorbic acid (Vc) as a positive control, see the results Figure 9 Figure A in .
[0068] Scavenging ABTS free radical activity: Take 2 mL of ABTS (7 mmol / L) aqueous solution and 2 mL of potassium persulfate aqueous solution (2.45 mmol / L) and mix them at room temperature for 12 hours in the dark to prepare cationic ABTS free radicals. The prepared cationic ABTS radical solution was diluted with ethanol to an absorbance value at 734 nm of 0....
Embodiment 3
[0071] Example 3: Anemarrhena polysaccharide AABP-2B inhibits α-glucosidase activity test
[0072] Take 20 μL of Anemarrhena polysaccharide aqueous solution (0.1, 0.2, 0.4, 0.6, 0.8, 1.0 and 2.0 mg / mL) and 40 μL of α-glucosidase aqueous solution (0.2 U / mL) in a 96-well plate, mix well, and place at 37 ° C Incubate for 10min, then add 20.0μL pNPG aqueous solution (10mmol / L), mix well, then place at 37°C for 30min, and finally add 100μL Na 2 CO 3 (0.2mol / L) solution to stop the reaction, test the absorbance of the mixture at 405nm, take acarbose (acarbose) as positive control, see the results Figure 10 . The results showed that zhimu polysaccharide AABP-2B had inhibitory activity of α-glucosidase in the concentration range of 0.1-2.0 mg / mL, and its inhibitory activity was enhanced with the increase of concentration. When the concentration was 2.0mg / mL, the inhibition rate of AABP-2B was 63.54±1.88%. It shows that Anemarrhena polysaccharide has good activity of inhibiting α-...
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