Extraction method for optimizing alpha-glucosidase inhibitor in pomegranate flower by response surface method
A technology of glucosidase and response surface method, which is applied in the field of natural product extraction, can solve the problems that the yield of α-glucosidase inhibitors does not reach the optimal level, loss of ethanol solvent, high price of ethanol, etc., and achieves good inhibitory effect , large component content, and the effect of shortening the extraction time
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Embodiment 1
[0026] Embodiment 1 (response surface method)
[0027] a. the picked pomegranate flowers are dried, pulverized, and passed through a 100-200 mesh sieve to obtain pomegranate flower powder;
[0028] b. Utilize Design Expert 10 software to carry out Box-Behnken response surface optimization design with the pomegranate flower powder obtained in step a, use the IC of pomegranate flower extract anti-α-glucosidase activity 50 The value is the response value, using three factors and three levels of response surface analysis to optimize the ultrasonic extraction of α-glucosidase inhibitors in pomegranate flowers, the liquid-to-material ratio is 1:20g / mL, the solvent ratio is 45% ethanol water, and the ultrasonic time 3h, the extraction solution was obtained under the condition of an ultrasonic temperature of 45°C;
[0029] c. Centrifuge the extract obtained in b at 7000rpm for 10min, get the supernatant concentrated and dried and weighed to obtain the α-glucosidase inhibitor in pomeg...
Embodiment 2
[0069] A response surface method of the present invention optimizes the hypoglycemic experiment of α-glucosidase inhibitors in pomegranate flowers:
[0070] α-glucosidase activity assay:
[0071] Mix 50 μL sample solution with appropriate concentration with 50 μL 0.2 U / mL α-glucosidase, react at room temperature for 6 min, then add 50 μL 5.0 mM p-nitrophenyl-D-glucopyranoside (p-nitrophenyl-D-glucopyranoside ) (0.1MPB, pH 6.8), react at 37°C for 10min, add 100μL of 0.2M sodium carbonate to the reaction system to terminate the reaction, measure the absorbance at 405nm, acarbose is used as a positive control, and the results are measured by IC 50 The value indicates that the enzyme inhibition calculation formula is:
[0072]
[0073] Sample group As: reaction system with sample and enzyme; control group Ac: reaction system without sample; blank group Aj: reaction system without enzyme and sample;
[0074] Carry out in vitro α-glucosidase inhibition experiment on the commerc...
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