A method for detecting biological activity of marine polysaccharides

A marine polysaccharide and biologically active technology, applied in the direction of analysis by electron paramagnetic resonance, can solve the problems of heavy workload, individual animal differences, and high capital consumption, and achieve improved stability, simple detection process, and wide detection range Effect

Active Publication Date: 2017-07-28
DALIAN POLYTECHNIC UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a method for detecting the biological activity of marine polysaccharides, so as to solve the technical problems of animal individual differences, heavy workload, and high capital consumption in the method of detecting polysaccharide activity in zoological experiments

Method used

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  • A method for detecting biological activity of marine polysaccharides
  • A method for detecting biological activity of marine polysaccharides
  • A method for detecting biological activity of marine polysaccharides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Detection of Scallop Polysaccharides Regulating NO Production in Macrophages

[0040] Get the scallop polysaccharide sample, prepare 200 μg / mL scallop polysaccharide solution with DMEM medium or distilled water;

[0041] Adjust the RAW264.7 cell density to 10 with DMEM medium 5 ~10 6 a / mL;

[0042] The cells with adjusted density were placed in a 96-well cell culture plate and cultured at 37°C for 24 hours;

[0043] Add appropriate amount of scallop polysaccharide to the 96-well culture plate, so that the concentration of scallop polysaccharide is 0 μg / mL, 12.5 μg / mL, 25 μg / mL, 50 μg / mL, 100 μg / mL, 200 μg / mL;

[0044] 37°C, CO 2 Cultivate in a carbon dioxide incubator with a concentration of 5% for 48 hours;

[0045] Collect the cell supernatant;

[0046] Use the ESR method to detect the concentration of NO free radicals;

[0047] Add cell supernatant and NO radical scavenger (MGD) to the tube 2 -Fe 2+ (contains 10mmol·L -1 MGD and 2mmol·L -1 FeSO ...

Embodiment 2

[0051] Example 2: detection of wakame sporophyll polysaccharides regulating NO production in macrophages

[0052] Get the wakame sporophyll polysaccharide sample, prepare 200 μg / mL wakame sporophyll polysaccharide solution with DMEM medium or distilled water;

[0053] Adjust the RAW264.7 cell density to 10 with DMEM medium 5 ~10 6 a / mL;

[0054] The cells with adjusted density were placed in a 96-well cell culture plate and cultured at 37°C for 24 hours;

[0055] Add appropriate amount of wakame sporophyll polysaccharides to the 96-well culture plate, so that the polysaccharide concentrations are 0 μg / mL, 12.5 μg / mL, 25 μg / mL, 50 μg / mL, 100 μg / mL, 200 μg / mL;

[0056] 37°C, CO 2 Cultivate in a carbon dioxide incubator with a concentration of 5% for 48 hours;

[0057] Collect the cell supernatant;

[0058] Use the ESR method to detect the concentration of NO free radicals;

[0059] Add cell supernatant and NO radical scavenger (MGD) to the tube 2 -Fe 2+ (contains 10mmol...

Embodiment 3

[0063] Example 3: Detection of the regulation of NO production in macrophages by polysaccharides in sea cucumber boiled liquid

[0064] Take the polysaccharide sample of sea cucumber boiled liquid, and prepare 200 μg / mL sea cucumber boiled liquid polysaccharide solution with DMEM medium or distilled water;

[0065] Adjust the RAW264.7 cell density to 10 with DMEM medium 5 ~10 6 a / mL;

[0066] The cells with adjusted density were placed in a 96-well cell culture plate and cultured at 37°C for 24 hours;

[0067] Add an appropriate amount of sea cucumber boiled liquid polysaccharides to the 96-well culture plate, so that the polysaccharide concentrations are 0 μg / mL, 12.5 μg / mL, 25 μg / mL, 50 μg / mL, 100 μg / mL, and 200 μg / mL;

[0068] 37°C, CO 2 Cultivate in a carbon dioxide incubator with a concentration of 5% for 48 hours;

[0069] Collect the cell supernatant;

[0070] Use the ESR method to detect the concentration of NO free radicals;

[0071] Add cell supernatant and NO...

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Abstract

The invention discloses a method for detecting biological activity of marine polysaccharide. The method comprises the following steps: 1, compounding a marine polysaccharide sample into a polysaccharide solution with a certain concentration; 2, adjusting macrophage to certain cell density; 3, compounding a series of mixed solutions through the polysaccharide solution obtained in step 1 and cell sap obtained in step 2, and then conducting macrophage culture, wherein the mixed solutions are the same in macrophage density and different in marine polysaccharide concentration; 4, taking culture obtained in step 3, and collecting cell supernate; 5, taking the cell supernate obtained in step 4, and detecting the concentration of nitroxide free radicals NO. with an electron spin resonance spectrum method, wherein the detection value represents in-vitro immunoregulatory activity of the detected polysaccharide sample. The method has the advantages of being wide in detection range, extensive in detection object, simple in detection condition, high in detection sensitivity and the like.

Description

technical field [0001] The invention belongs to the technical field of biological activity detection of aquatic products, in particular to a method for detecting biological activity of marine polysaccharides. Background technique [0002] Plants, animals and microorganisms in nature contain polysaccharides. Polysaccharides are developing rapidly in the fields of medicine and food health care, and have good development prospects. Due to the special living environment of marine organisms, the synthesis process of polysaccharides in marine organisms is different from that of terrestrial organisms, and many active substances with novel structures and special effects are produced. high research value. [0003] Marine biological polysaccharides can be divided into three categories according to different sources, seaweed polysaccharides, marine animal polysaccharides and marine microbial polysaccharides. The biological activity of different polysaccharides is different. The acti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N24/10
CPCG01N24/10
Inventor 启航李楠董秀芳冯丁丁粱潇月董秀萍姜鹏飞
Owner DALIAN POLYTECHNIC UNIVERSITY
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