Preparation and application of chromium green algae polysaccharide for preventing and curing diabetes
A technology of chlorella polysaccharides and diabetes, which is applied in the field of biotechnology applications, can solve the problems of low chromium chelation rate, preparation method has not been reported, high chromium chelation rate of chlorella polysaccharides, etc.
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Embodiment 1
[0019] Chlorella polysaccharide chromium preparation method is as follows.
[0020] (1) Add 0.02% (v / v), 0.1 mol / L EDTA to the 10 mg / mL Chlorella polysaccharide solution in a water bath at 35 °C for 1 h, then cool to room temperature.
[0021] (2) Add 0.01 mmol / L EDC and 5 μmol / L NHS to the above reaction system in sequence, the addition amount is 0.1% (v / v) and 0.01% (v / v), magnetically stirred for 0.5 h, 4 ℃ cross-linking for 3 h. After the reaction, precipitate with 3 times the volume of absolute ethanol, centrifuge at 4800 r / min for 10 min, and collect the precipitate.
[0022] (3) Add 0.01 mol / L chromium trichloride solution to the precipitate, adjust the pH to 3 with lactic acid, and incubate at 40°C for 0.5 h. Centrifuge at 10,000 r / min for 10 min after the reaction. The supernatant was dialyzed for 24 h with a dialysis bag with a molecular cut-off of 1000 Da, and freeze-dried to obtain Chlorella chromium with a chromium chelation rate of 18.9%.
Embodiment 2
[0024] Chlorella polysaccharide chromium preparation method is as follows.
[0025] (1) Add 0.03% (v / v), 0.1 mol / L EDTA to the 12 mg / mL Chlorella polysaccharide solution in a water bath at 45°C for 1 h, then cool to room temperature.
[0026] (2) Add 0.01 mmol / L EDC and 5 μmol / L NHS to the above reaction system in turn, the addition amount is 0.3% (v / v), 0.03% (v / v), magnetic stirring for 0.5 h, 4 ℃ cross-linking for 3 h. After the reaction, precipitate with 3 times the volume of absolute ethanol, centrifuge at 4800 r / min for 10 min, and collect the precipitate.
[0027] (3) Add 0.01 mol / L chromium trichloride solution to the precipitate, adjust the pH to 4 with lactic acid, and incubate at 50°C for 1 hour. Centrifuge at 10,000 r / min for 10 min after the reaction. The supernatant was dialyzed for 24 hours with a dialysis bag with a molecular cut-off of 1000 Da, and then freeze-dried to obtain Chlorella chromium with a chromium chelation rate of 19.3%.
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