Application of waxberry flesh extract in preparation of alpha-glucosidase inhibitor

A technology of glucosidase and extract, which is applied in the field of preparing α-glucosidase inhibitors from bayberry pulp extracts, can solve the problems of reporting α-glucosidase inhibitory activity and other issues

Inactive Publication Date: 2016-06-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Studies have shown that bayberry fruit extract can effectively prevent and control diabetes, but there is no relevant literature reporting the α-glucosidase inhibitory activity of its extract

Method used

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  • Application of waxberry flesh extract in preparation of alpha-glucosidase inhibitor
  • Application of waxberry flesh extract in preparation of alpha-glucosidase inhibitor
  • Application of waxberry flesh extract in preparation of alpha-glucosidase inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1) The assay method of α-glucosidase inhibitory activity: the α-glucosidase ordered from SIGMA company is prepared into 0.2U / mL with potassium phosphate buffer (0.01mol / L, pH=6.8), adds 0.2% BSA. Among them, the enzyme activity unit is defined as: under the condition of 37°C and pH 6.8, p-nitrophenyl-α-D-glucopyranoside (PNPG) releases 1 μmol of glucose in 1 minute under the action of the enzyme, which is defined as an enzyme activity unit (U).

[0024] When measuring, mix 112 μL 0.1mol / L potassium phosphate solution and 20 μL α-glucosidase in a 96-well microtiter plate evenly, then add 8 μL inhibitor, let it stand at a constant temperature of 37°C for 15 minutes, and then add 20 μL NPG (2.5 mmol / L) , 37 ℃ constant temperature for 15min, add 80μL Na 2 CO 3 (2.5mmol / L), measure the absorbance value of 405nm with microplate reader (sample group, A 1 ), without adding α-glucosidase as a blank control (sample blank, A 2 ), without adding sample is the control (enzyme ...

Embodiment 2

[0029] Weigh 10g of freeze-dried pulp powder of bayberry variety 'Dongkui', add 200mL of 80% methanol solution containing 0.5% formic acid, wherein the ratio of solid to liquid is 1g:20mL, soak for 2h, ultrasonically extract for 30min, repeat twice, and combine the supernatant liquid, 37°C rotary vacuum concentration until only the water phase remained, centrifuged at 10,000r / min for 5min in a high-speed centrifuge, distilled into 75mL water, passed the concentrated solution through a solid phase extraction column, rinsed with 10% methanol for 100mL, then rinsed with 50% methanol 50 mL, collected the effluent, and evaporated to dryness on a rotary evaporator at 37°C to obtain 57.6 mg of 10% eluted fraction and 33.8 mg of 50% methanol eluted fraction. The results of the α-glucosidase inhibitory activity test showed that the α-glucosidase inhibitory activity IC of the crude extract was 50 IC of 2545.5μg / mL, 10% elution fraction 50 IC of 270.0 μg / mL, 50% eluted fraction 50 It i...

Embodiment 3

[0031]Weigh 8g of freeze-dried pulp powder of bayberry variety 'Konoha', add 160mL of 80% methanol solution containing 0.5% formic acid, wherein the ratio of solid to liquid is 1g:20mL, soak for 2h, ultrasonically extract for 30min, repeat twice, and combine the supernatant liquid, 37°C rotary vacuum concentration until only the water phase remained, centrifuged at 10,000r / min for 5min in a high-speed centrifuge, distilled to 70mL of water, passed the concentrated solution through a solid phase extraction column, rinsed with 10% methanol for 100mL, then rinsed with 50% methanol 50 mL, collected the effluent, and evaporated to dryness on a rotary evaporator at 37°C to obtain 72.9 mg of 10% eluted fraction and 44.5 mg of 50% methanol eluted fraction. The results of the α-glucosidase inhibitory activity test showed that the α-glucosidase inhibitory activity IC of the crude extract was 50 IC of 2083.7μg / mL, 10% elution fraction 50 IC of 307.5 μg / mL, 50% eluted fraction 50 is 30....

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Abstract

The invention provides an application of a waxberry flesh extract in preparation of an alpha-glucosidase inhibitor. The waxberry flesh extract is prepared through extraction and concentration with methanol, and extraction separation by a solid phase extraction column. Preliminary activity screening tests prove that crude flesh extracts of different waxberry varieties all show alpha-glucosidase inhibition activity. The alpha-glucosidase inhibition activity of different elution components after enrichment of the crude extracts through the solid phase extraction column is improved remarkably, and especially the activity of 50% elution components is much higher than that of positive control acarbose. The application has great significance in reasonable use of waxberry resources and increase of additional values of waxberry products.

Description

technical field [0001] The invention belongs to the field of functional food or medicine, and relates to the application of bayberry pulp extract to prepare alpha-glucosidase inhibitors. Background technique [0002] Diabetes is an absolute or relative deficiency of insulin in human blood, resulting in higher blood sugar than normal, diabetes, and a series of metabolic disorders such as fat and protein. It is an effective treatment to reduce postprandial blood sugar by inhibiting the activity of some enzymes in the body that promote carbohydrate digestion, such as α-glucosidase. [0003] Carbohydrates in the diet release glucose under the action of α-glucosidase, and the small intestine absorbs glucose into the blood, which is the main cause of postprandial blood sugar rise. On the one hand, α-glucosidase inhibitors can inhibit the activity of α-glucosidase by reversibly occupying the complex site of α-glucosidase and sugar, delaying the absorption of carbohydrates, and on ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/185A61P3/10A61K131/00
CPCA61K36/185A61K2236/17A61K2236/333A61K2236/39A61K2236/51A61K2236/53A61K2236/55
Inventor 李鲜闫淑霞孙崇德陈昆松
Owner ZHEJIANG UNIV
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