Application of astaxanthin to preparation of aldose reductase inhibitor
A reductase inhibitor, astaxanthin technology, applied in the field of application of astaxanthin in the preparation of aldose reductase inhibitors, to achieve the effect of reducing the occurrence of diabetic complications
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Embodiment 1
[0028] The preparation of embodiment 1 astaxanthin
[0029] The Phaffia fermented liquid was 5534×g, centrifuged for 8 min, the supernatant was discarded, washed with distilled water and centrifuged again, and the operation was repeated twice to obtain Phaffia cells.
[0030] Phaffia yeast cell wall breaking: Use dimethyl sulfoxide to break the Phaffia yeast cell wall, the breaking condition is: the cell and dimethyl sulfoxide are based on the mass-volume ratio, that is, g / ml, and the ratio is 1 / 2. At 55°C, break the wall for 5 minutes; using dimethyl sulfoxide has a high breaking rate and good extraction effect; the short treatment time (5min) and low treatment temperature (55°C) are conducive to the stability of the astaxanthin structure .
[0031] Crude extraction of astaxanthin with acetone: after breaking the wall, extract with acetone according to the ratio of material to liquid 1:15, the ratio of material to liquid is the mass-volume ratio of Phaffia thallus to aceton...
Embodiment 2
[0036] The measurement of the inhibitory rate of embodiment 2 astaxanthin to aldose reductase activity
[0037] Preparation of reagents:
[0038] Preparation of phosphate buffered saline (PBS): accurately weigh 6.36g NaH 2 PO 4 , add distilled water to dissolve, transfer to a 250mL volumetric flask to obtain liquid a; take another 3.31g Na 2 HPO 4 , add distilled water to dissolve, transfer to a 250mL volumetric flask to constant volume to obtain liquid b; then, pipette the two solutions of a and b to a 500mL volumetric flask to constant volume, use a pH meter to calibrate the phosphate buffer solution to pH=6.2 .
[0039] Preparation of aldose reductase solution: Take 100 U of aldose reductase powder and add 1 mL of PBS, that is, the enzyme activity of aldose reductase is 100 U / mL. Take 20 μL of 100 U / mL aldose reductase solution and add 980 μL of PBS, that is, the concentration of aldose reductase is 2 U / mL, and the aldose reductase solution should be prepared and used im...
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