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Preparation method and application of alfalfa polysaccharide

An alfalfa polysaccharide and monosaccharide technology, applied in the digestive system, drug combination and other directions, can solve the problems of liver hemorrhage, decreased egg production rate of laying hens, and reduced economic benefits of farmers

Inactive Publication Date: 2016-06-08
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The main feature of the disease is a sharp increase in liver fat content, which generally exceeds 40% of the dry weight of the liver. Hepatic fatty degeneration is obvious. In severe cases, hemorrhage occurs in the liver, which eventually leads to a decline in laying hen egg production rate and even death. The morbidity is About 5%, thus reducing the economic benefits of farmers

Method used

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  • Preparation method and application of alfalfa polysaccharide
  • Preparation method and application of alfalfa polysaccharide
  • Preparation method and application of alfalfa polysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: the preparation of alfalfa polysaccharide APS-2a, APS-2b, APS-3a and APS-3b

[0028] 1. Preparation of alfalfa total polysaccharides: alfalfa total polysaccharides extracted by compound enzyme technology (Chinese patent, publication number CN103304678A, publication date 2013.09.18).

[0029] 2. Deproteinization of total polysaccharides from alfalfa: Mix chloroform and n-butanol at a ratio of 4:1 to make Sevag reagent, dissolve the total polysaccharides in distilled water in a beaker, add 1 / 4 volume of Sevag reagent, and use a magnetic stirrer After 30 min of vigorous stirring, centrifuge at 4000 r / min for 10 min, and collect the supernatant. Then add the same proportion of Sevag reagent, repeat the above steps several times until no white foam appears in the lower solution.

[0030] 3. Ion exchange column chromatography: the filler is DEAE-cellulose, dissolve the deproteinized alfalfa total polysaccharide (500mg) with 1-2mL distilled water, centrifuge, an...

Embodiment 2

[0042] Embodiment 2: the biological function of alfalfa polysaccharide fraction APS-2a, APS-2b, APS-3a and APS-3b

[0043] 1. Isolation of hepatic hepatic cells during peak laying period: live in situ two-step perfusion method to separate hepatic cells (Zhou Xiaodan et al., 2012)

[0044] 2. Effect of alfalfa polysaccharides on the oxidation state of liver cells: adjust the concentration of the freshly separated liver cell suspension to 10 6 cells / mL, seeded in 6-well plate, 5% CO 2 , Cultivate continuously in an incubator at 37°C for about 6 hours. After the cells are completely attached to the wall, replace with growth medium, continue to cultivate for 12 hours, and then replace the whole amount with different concentrations of polysaccharide-treated growth medium and continue to cultivate for a certain period of time, then add 4mmol / LH 2 o 2 Treat the hepatocytes for 2 hours, collect the culture medium of the sample, measure the content of lactate dehydrogenase (LDH) an...

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Abstract

The invention relates to function research on plant polysaccharide components and especially relates to a method for extraction of total polysaccharides from alfalfa by a compound enzyme technology. The method comprises the following steps of carrying out deproteinization on total polysaccharides of alfalfa by a Sevag method to obtain the deproteinated total polysaccharides of alfalfa, and carrying out grading purification on the deproteinated total polysaccharides of alfalfa by a DEAE-cellulose chromatography column and a Sephadex G-100 gel chromatography column to obtain different alfalfa polysaccharide ingredients such as APS-2a, APS-2b, APS-3a and APS-3b. An external cell test proves that the alfalfa polysaccharide ingredients have functions of improving a laying hen hepatic cell antioxidation capability and preventing hepatic cell fatty degeneration. Through the method, a theoretical basis for prevention of the laying hen fatty liver hemorrhagic syndrome by alfalfa polysaccharides is provided and creation and application of alfalfa polysaccharides as novel feed additives are promoted.

Description

technical field [0001] The invention relates to the functional research of plant polysaccharide components, in particular to the polysaccharide components extracted from alfalfa, and the function of improving the anti-oxidation ability of laying hen hepatocytes and preventing fatty degeneration of hepatocytes. Background technique [0002] Alfalfa is a perennial high-quality herbaceous leguminous forage, which is widely planted all over the world. It is known as the "king of forage" and is very rich in resources. Alfalfa polysaccharides are water-soluble polysaccharides extracted from alfalfa stems and leaves, which have multiple biological functions. At the same time, the wide range of sources of alfalfa provides sufficient and cheap raw materials for the production of alfalfa polysaccharides, so alfalfa polysaccharides have good application prospects. [0003] Fatty liver hemorrhagic syndrome of laying hens is a nutritional and metabolic disorder disease that mainly occurs...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00A61P1/16
Inventor 董晓芳佟建明王少璞
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI