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Method for extracting water-soluble alpha-glucosidase inhibitor

A technology of glucosidase and extraction method, which is applied in the directions of drug combination, medical raw materials derived from fungi, metabolic diseases, etc., can solve the problems of low application safety, not directly involved in the inhibitory function of α-glucosidase, etc., and achieves safety High, less toxic and side effects, good active effect

Inactive Publication Date: 2011-06-29
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Patent CN200710048738.5 is the water extract of two kinds of mixed medicinal fungi, Inonotus obliquus and Phellinus, but also uses harmful chloroform-isopropanol to remove impurities such as protein in the implementation of the example, and the application safety is low, and It does not directly involve the α-glucosidase inhibitory function. The patent CN 200810201798.0 is a mixture of alcohol extract and water extract of Phellinus Phellinus, and the water extraction is carried out after the alcohol extraction, and the function is not directly related to the α-glucosidase inhibitory function.

Method used

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  • Method for extracting water-soluble alpha-glucosidase inhibitor
  • Method for extracting water-soluble alpha-glucosidase inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Inonotus obliquus fruiting bodies → crushed and sieved to 80 mesh → weigh 100g → add 3000mL water → ultrasonic treatment (Shanghai Shengxi Ultrasonic Instrument Co., Ltd., FS1200PV ultrasonic processor, 20KHz, 360W) for 30 minutes → add 1mL liquid high temperature α- Amylase (20,000 U.mL -1 , Zaozhuang Jienuo Bioenzyme Co., Ltd.), boiling water extraction for 3h→5400g centrifugation to remove slag→vacuum concentration of the supernatant (pressure below 0.08Mpa, temperature 45℃~60℃, the same below) to the original 1 / 15 volume→add 95% ethanol to a final concentration of 80%, ethanol precipitation for 8h → precipitation plus 2000mL distilled water to redissolve → 10,000 molecular weight cut-off membrane (Millipore, USA, labscale TFF system, PXB010A50 polyethersulfone 10KD ultrafiltration membrane bag) ultrafiltration for 24h → filtrate vacuum concentration (pressure 0.08Mpa below, temperature 45 ℃ ~ 60 ℃, the same below) to 30mL → freeze-drying → 10.2g Inonotus obliquus wa...

Embodiment 2

[0036] Inonotus obliquus fruiting bodies → smashed and sieved to 60 mesh → weighed 100g → added 3000mL water → ultrasonic (20KHz, 360W) treatment for 30 minutes → added liquid high-temperature α-amylase (20,000 U.mL) with a raw material weight of 0.8mL -1 ), extract with boiling water for 3 hours→3800g centrifuge to remove slag→vacuum concentrate the supernatant to the original 1 / 10 volume→add 95% ethanol to the final concentration of 80%, ethanol precipitation for 8h→precipitate add 1500mL distilled water to redissolve→10,000 molecular weight cut-off membrane Ultrafiltration for 12 hours → vacuum concentration of the filtrate to 30 mL → freeze drying → 11.3 g of Inonotus obliquus water-soluble α-glucosidase inhibitor (polysaccharide content 21.6%) → semi-inhibitory activity IC of α-glucosidase 50 =7.1 μg / mL.

Embodiment 3

[0038] Inonotus obliquus fruiting bodies → crushed and sieved to 100 mesh → weigh 100g → add 4000mL water → ultrasonic (20KHz, 360W) treatment for 40 minutes → add 1mL liquid high temperature α-amylase (20,000 U.mL -1 ), boiling water extraction for 4 hours→5400g centrifugation to remove slag→supernatant concentrated in vacuum to 1 / 10 of the original volume→add 95% ethanol to final concentration of 80%, ethanol precipitation for 8h→precipitate plus 3000mL distilled water to redissolve→10,000 molecular weight cut-off membrane Ultrafiltration for 24 hours → vacuum concentration of the filtrate to 40 mL → freeze drying → 11.6 g of Inonotus obliquus water-soluble α-glucosidase inhibitor (polysaccharide content 23.4%) → semi-inhibitory activity IC of α-glucosidase 50 = 6.5 μg / mL.

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Abstract

The invention provides a method for extracting water-soluble alpha-glucosidase inhibitor, which comprises the following steps: crushing and screening medicinal fungal fruiting bodies (inonotus obliquus and phellinus igniarius) through a sieve with 60 to 100 meshes;, weighting and quantifying; adding water of 20 to 40 times by weight; treating through ultrasound for 20 to 40 minutes; adding liquid-state high-temperature amylase, and extracting in boiling water for 2 to 4h; centrifuging and deslagging the material of more than 3800g; vacuum condensing supernate to be 1 / 10 to 1 / 15 of the original volume; adding edible ethanol until the concentration reaches 80 percent and depositing in alcohol for over 8h; precipitating and adding distilled water for re-dissolving; ultrafiltering through a 10 thousand molecular weight cutoff film for 12 to 24h; vacuum condensing filtrate; and freezing and drying extracts (medicinal fungus water-soluble alpha-glucosidase inhibitor). By adopting the method provided by the invention, the water-soluble alpha-glucosidase inhibitor with better activity can be obtained; the inonotus obliquus and the phellinus igniarius as the raw materials are medicinal fungi having longer application history and fewer toxic or side effects. In addition, the process flow adopted in the invention does not involve toxic or harmful steps, has high safety and is especially favorable for the subsequent development of medical healthcare foods.

Description

(1) Technical field [0001] The invention relates to a method for obtaining a water-soluble α-glucosidase inhibitor by extracting fungal fruit bodies as raw materials. (2) Background technology [0002] Medicinal fungi are a class of higher fungi with high medicinal value. They generally have several or more functions such as anti-tumor, hypoglycemic, hypolipidemic, anti-viral, and immune-enhancing. [0003] With the development of modern society, unhealthy living and working styles have increased, and the aging population has continued to intensify. In the past two decades, the incidence of diabetes in my country has been on the rise. According to a new scientific report in 2010, the existing diabetes patients in my country As high as 94 million people, India has replaced India as the country with the largest number of diabetics in the world, and there are still 148 million patients with abnormal glucose metabolism (prediabetic patients) who have not yet constituted a diagnos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/06A61P3/10
Inventor 杨开顾今孙培龙季培军叶帮伟
Owner ZHEJIANG UNIV OF TECH
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