Application of chitosan oligosaccharide natural polyphenol compound

Abstract

The invention relates to the field of preparation of natural active substances, and particularly discloses application of a chitosan oligosaccharide natural polyphenol compound, the chitosan oligosaccharide natural polyphenol compound is used for inhibiting the activity of alpha-glucosidase, and the chitosan oligosaccharide natural polyphenol compound is obtained by compounding chitosan oligosaccharide and natural polyphenol according to the molar ratio of 1: 1; the natural polyphenol is selected from any one or more of tea polyphenol, apple polyphenol, quercetin, dihydromyricetin, epicatechin, curcumin or ellagic acid. According to the invention, polyphenol natural substances and chitosan oligosaccharide are compounded to obtain a novel product, the inhibition activity of the product to alpha-glucosidase is measured, and an activity experiment result shows that the enzyme inhibition activity of the novel compound is greatly improved compared with that of single chitosan oligosaccharide and polyphenol substances, so that the compound is proved to have better enzyme inhibition activity; the effect of reducing blood sugar can be achieved.

Description

Technical Field

[0001] The invention relates to the field of preparation of natural active substances, in particular to application of a chitosan oligosaccharide natural polyphenol complex. Background Art

[0002] Diabetes is a chronic disease characterized by disordered carbohydrate metabolism in the human body. There is currently no cure. Diabetic patients suffer from pain and face great difficulties in diet and daily life. At this stage, it is very necessary and urgent to explore and develop a diet suitable for diabetic patients and new drugs to delay the progression of the disease. Chitosan oligosaccharide is a biological oligosaccharide with great research value. Its unique biological structure and excellent biological activity provide new raw materials for the research and development of many new foods, daily necessities and drugs. Natural polyphenols are also a kind of high-quality natural product with rich biological activity. Studies have shown that there are many types of natural polyphenols, which have antioxidant activity, anti-inflammatory activity, anti-tumor activity, hypoglycemic activity, hypolipidemic activity, etc. Since chitosan oligosaccharides are alkaline sugars and the phenolic hydroxyl groups of phenols are acidic, the complexes of chitosan oligosaccharides and phenols can form effective ionic bonds in a suitable solution system, thereby enhancing the biological activity of each other. Currently, patients with hyperglycemia and diabetes use drugs such as metformin, acarbose, and voglibose, and those with severe conditions take insulin orally or by injection. However, these drugs have drug resistance and side effects. Therefore, the development of safe hypoglycemic drugs or functional foods that are both medicine and food is of great significance. Summary of the invention

[0003] The purpose of the present invention is to provide an application of a chitosan oligosaccharide natural polyphenol complex in view of the defects of the prior art, so as to solve the problems raised by the above-mentioned background technology.

[0004] To achieve the above object, the present invention provides the following technical solution: an application of a chitosan oligosaccharide natural polyphenol complex in inhibiting the activity of α-glucosidase, wherein the chitosan oligosaccharide natural polyphenol complex is obtained by compounding chitosan oligosaccharide and natural polyphenol in a molar ratio of 1:1;

[0005] The natural polyphenols are selected from any one or more of tea polyphenols, apple polyphenols, quercetin, dihydromyricetin, epicatechin, curcumin or ellagic acid.

[0006] As a preferred technical solution of the present invention, the molecular weight of the chitosan oligosaccharide is 500-5000Da.

[0007] The invention discloses an application of a chitosan oligosaccharide natural polyphenol complex in preparing a product for reducing blood sugar.

[0008] As a preferred technical solution of the present invention, the product is a functional food, a health food, a food for special medical purposes or a new drug.

[0009] A blood sugar lowering product, comprising the chitosan oligosaccharide natural polyphenol complex according to claim 1 at a pharmacologically effective concentration.

[0010] Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention selects polyphenol natural substances and chitosan oligosaccharides to compound to obtain a new product, and determines the inhibitory activity of the product on α-glucosidase. The activity experimental results show that the enzyme inhibitory activity of the new compound is greatly improved compared with that of single chitosan oligosaccharides and polyphenol substances (the total amount of the complex is the same as the amount of single chitosan oligosaccharides or polyphenol substances, and in fact, the amount of a single component in the complex is actually halved, and the enzyme inhibitory activity effect is improved when the amount is reduced), which proves that the compound has good enzyme inhibitory activity, that is, it can play a role in lowering blood sugar; the preparation process of this new compound is simple, the raw materials used are rich in sources, and it is a natural product, so it has good development prospects and is expected to be used in the diet and daily health care of diabetic patients. DETAILED DESCRIPTION

[0011] The preferred embodiments of the present invention are described in detail below so that the advantages and features of the present invention can be more easily understood by those skilled in the art, thereby making a clearer and more definite definition of the protection scope of the present invention.

[0012] The following examples are used to prepare the target composite products of the present invention according to the following methods.

[0013] Example 1: Accurately weigh 0.5 g of chitosan oligosaccharide (mW=320-900 Da) and 0.56 g of food-grade tea polyphenols, respectively, and mix them evenly to obtain a chitosan oligosaccharide tea polyphenol complex.

[0014] Example 2: 0.5 g of chitosan oligosaccharide (mW=900-1500 Da) and 3.97 g of food-grade apple polyphenol were accurately weighed and mixed evenly to obtain a chitosan oligosaccharide apple polyphenol complex.

[0015] Example 3: 0.5 g of chitosan oligosaccharide (mW=1500-2000 Da) and 0.60 g of food-grade quercetin were accurately weighed and mixed evenly to obtain a chitosan oligosaccharide-quercetin complex.

[0016] Example 4: 0.5 g of chitosan oligosaccharide (mW=2000-2500 Da) and 0.58 g of food-grade epicatechin were accurately weighed and mixed evenly to obtain a chitosan oligosaccharide epicatechin complex.

[0017] Example 5: 0.5 g of chitosan oligosaccharide (mW=2500-3200 Da) and 0.63 g of food-grade dihydromyricetin were accurately weighed and mixed evenly to obtain a chitosan oligosaccharide dihydromyricetin complex.

[0018] Example 6: Accurately weigh 0.5 g of chitosan oligosaccharide (mW=2000-3000 Da) and 0.73 g of food-grade curcumin and mix them evenly to obtain a chitosan oligosaccharide-curcumin complex.

[0019] Example 7: Accurately weigh 0.5 g of chitosan oligosaccharide (mW=1000-2000 Da) and 0.60 g of food-grade ellagic acid, respectively, and mix them evenly to obtain a chitosan oligosaccharide ellagic acid complex.

[0020] Application examples:

[0021] α-Glucosidase is a biological enzyme widely present in organisms. Its main function is to decompose polysaccharides ingested by the body to produce glucose, thereby increasing the glucose content in the body and raising the blood sugar concentration. Therefore, inhibiting the activity of α-glucosidase can reduce the glucose content in the body and lower the blood sugar concentration.

[0022] Based on the above experimental principle, the experimental raw materials used in the present invention and the prepared novel chitosan oligosaccharide complex were selected, and acarbose was selected as the positive control of this experiment, and their inhibitory activities on α-glucosidase were determined through the following experimental process.

[0023] First, accurately weigh 80 mg of the sample to be tested and dissolve it in 8 mL of PBS buffer to prepare a sample solution with an initial concentration of 10 mg / mL. Then, the sample solution was diluted stepwise with PBS buffer to obtain sample solutions with concentrations of 0.125, 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, and 10 mg / mL, respectively.

[0024] Use PBS buffer to prepare 1 U / mL α-glucosidase solution, pNPG solution and 0.1 mol / L sodium carbonate solution respectively and set aside.

[0025] Experimental group: The test solutions were added to 96-well plates, 50 μL / well for each concentration, and then 50 μL of α-glucosidase solution was added to each well and incubated at 37°C for 10 minutes. After the time was up, 50 μL of pNPG solution was added to each well and incubated at 37°C for 20 minutes. Finally, 50 μL of sodium carbonate solution was added, mixed evenly, and the absorbance of the solution was measured at 405 nm.

[0026] Negative control group: the α-glucosidase solution in the experimental group was replaced with an equal amount of PBS buffer, and the other reagents and operations were consistent with those of the experimental group.

[0027] Blank control group: replace the test samples in the experimental group with an equal amount of PBS buffer, and keep the other reagents and operations the same as those of the experimental group.

[0028] The absorbance of each well is obtained through the above experimental operation, and then the α-glucosidase inhibition rate of each test compound at the corresponding concentration can be calculated by the following formula:

[0029]

[0030] in, A 样品 Refers to the absorbance of the sample solution in the experimental group. A 对照 Refers to the absorbance measured by the negative control group without α-glucosidase solution. A 空白 Refers to the absorbance measured by the blank control group.

[0031] Table 1: Inhibition rate of each sample (raw material) on α-glucosidase

[0032]

[0033] Table 2: Inhibition rate of each sample (chitosan oligosaccharide complex) on α-glucosidase

[0034]

[0035] By comparing the data in Table 1 and Table 2, the following conclusions can be drawn:

[0036] In the range of 0.125-10 mg / mL of the measured concentration, the inhibition rate of the chitosan oligosaccharide raw material used in this experiment on α-glucosidase is very low, even at the highest test concentration, the inhibition rate is only 2.4347%, and the selected natural polyphenol raw materials generally have good enzyme inhibition activity. After compounding with chitosan oligosaccharide, the enzyme inhibition activity of each chitosan oligosaccharide complex is significantly improved compared with chitosan oligosaccharide, especially the chitosan oligosaccharide curcumin complex and the chitosan oligosaccharide ellagic acid complex, which are significantly improved compared with the single chitosan oligosaccharide, curcumin and ellagic acid raw materials. When the test concentration is 4 mg / mL, the enzyme inhibition rate reaches 100%. In summary, the experimental results show that the chitosan oligosaccharide natural polyphenol compound product prepared by the present invention has good α-glucosidase inhibition activity and obvious synergism, so it can be considered that they have the effect of lowering blood sugar, and they are edible products of natural origin, so they have great development and application value in the development of medicine, food and health products for hyperglycemia control.

[0037] The above embodiments only express the implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention.

Claims

1. An application of a chitosan oligosaccharide natural polyphenol complex in inhibiting α-glucosidase activity, characterized in that: The chitosan oligosaccharide-natural polyphenol complex is obtained by compounding chitosan oligosaccharide and natural polyphenol in a molar ratio of 1:1; The natural polyphenols are selected from any one or more of tea polyphenols, apple polyphenols, quercetin, dihydromyricetin, epicatechin, curcumin or ellagic acid.

2. The use of the chitosan oligosaccharide natural polyphenol complex according to claim 1 in inhibiting the activity of α-glucosidase, characterized in that: The molecular weight of the chitosan oligosaccharide is 500-5000Da.

3. Use of the chitosan oligosaccharide natural polyphenol complex as claimed in claim 1 in the preparation of a product for lowering blood sugar.

4. The use according to claim 3, characterized in that: The product is a functional food, a health food, a food for special medical purposes or a new drug.

5. A product for lowering blood sugar, characterized in that: The product contains the chitosan oligosaccharide natural polyphenol complex according to claim 1 in a pharmacologically effective concentration.

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