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Method for detecting fungal polysaccharide

A technology of fungal polysaccharides and polysaccharides, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of inability to strictly quantify chemical analysis methods, limit the use range of gel electrophoresis methods, and difficult to master operating techniques, and achieve a wide range of molecular weights , mild conditions and high sensitivity

Inactive Publication Date: 2011-08-10
HENAN UNIVERSITY OF TECHNOLOGY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The phenol-sulfuric acid method is not only a classic method for detecting polysaccharides, but also known as a standard method for quantitative detection of polysaccharides. However, this method is a chemical analysis method that cannot be strictly quantified. It is not only difficult to master the operation technique, but also has large errors
Gel electrophoresis and HPLC can be said to be modern methods for the analysis and detection of polysaccharides. The premise of gel electrophoresis is that the sample should be charged, and most polysaccharides are uncharged neutral compounds, which makes the gel electrophoresis method The scope of use is extremely limited
The HPLC method is indeed a good method for the analysis and detection of most macromolecular compounds. However, due to the large molecular weight and high viscosity of polysaccharides, the adsorption of polysaccharides to the stationary phase is always an insurmountable problem.
In addition, the above-mentioned methods must have a standard as a reference before they can be used.

Method used

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  • Method for detecting fungal polysaccharide
  • Method for detecting fungal polysaccharide

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Experimental program
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Embodiment 1

[0023] Embodiment 1: The method for detecting fungal polysaccharides specifically includes the following process steps:

[0024] (1) sample preparation

[0025] Weigh a certain amount (20mg) of polysaccharide samples (lentinan, Hericium erinaceus, Grifola frondosa polysaccharide, Flammulina velutipes polysaccharide, bitter gourd polysaccharide) into 5 clean test tubes or small measuring cups, add 5ml of distilled water and stir to dissolve , Store in a low temperature (4°C) refrigerator for later use. The concentration of the sample solution is 0.1mg / ml;

[0026] (2) Preparation of mobile phase

[0027] The above-mentioned solvent used for dissolving polysaccharides is degassed with a degasser and used as a mobile phase;

[0028] (3) Sampling, buffering, focusing balance

[0029] The single-phase chromatography (asymmetric field-flow separation) instrument (The Eclipse AF4), multi-angle static light scattering instrument (MALS, DAWN HELEOS II), dynamic light scattering ins...

Embodiment 2

[0035] Embodiment 2: the same as the steps of Embodiment 1, the difference is:

[0036] 1. Weigh 60mg of polysaccharide samples (lentinan, Hericium erinaceus, Grifola frondosa polysaccharide, Flammulina velutipes polysaccharide, bitter gourd polysaccharide) into a clean test tube or small measuring cup, add 30ml 0.85% sodium chloride solution and stir to dissolve it , Store in a low temperature (4°C) refrigerator for later use. The concentration of the sample solution is 0.5mg / ml;

[0037] 2. The operating process conditions are as follows: use a micropipette to draw 60 μL of the prepared polysaccharide sample and an appropriate amount of mobile phase for sample injection, buffer and focus balance. Injection, buffering, focusing balance time is 8min, injection speed: 0.5mL / min; focusing balance time is 3min, channel flow rate: 1.2mL / min, crossflow flow rate: initially run at 5mL / min for 3 minutes, then Decrease to 2.5mL / min and run for 5.5 minutes, then gradually decrease to...

Embodiment 3

[0040] Embodiment 3: the same as the steps of Embodiment 1, the difference is:

[0041]1. Weigh 100mg of polysaccharide sample into a clean test tube or small measuring cup, add 60ml of phosphate buffer (7.0mmol / L KH 2 PO 4 , 7.0mmol / L Na 2 HPO 4 , pH=6.8) stirred to dissolve, and stored in a low temperature (4°C) refrigerator for later use. The concentration of the sample solution is 0.95mg / ml;

[0042] 2. The operating process conditions are as follows: use a micropipette to draw 120 μL of the prepared polysaccharide sample and an appropriate amount of mobile phase for sample injection, buffer and focus balance. Injection, buffering, focusing balance time is 10min, injection speed: 0.95mL / min; focusing balance time is 5min, flow channel flow rate: 2.0mL / min, crossflow flow rate: initially run at 9mL / min for 1 minute, then Reduce to 4mL / min and run for 3 minutes, then gradually reduce to 0ml / min, the total running time is 25 minutes. The flow rate of the sugar sample is...

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Abstract

The invention discloses a method for detecting fungi polysaccharide. The method organically integrates MALS, high-response differential method, and DLS to carry out online, offline, and quantitative detection, with monophase chromatographic technique as core, thus, the characteristics of polysaccharide, like absolute molecular weight, dispersion degree, molecular conformation, dissolubility, and stability, can be represented and analyzed. The method comprises preparation of sample, preparation of mobile phase, sample injection, buffering, focusing balancing, elution and separation, detection,and the like. Compared with prior phenol-sulfuric acid method, gel electrophoresis method, and HPLC method, the invention has the advantages that quantitative detection and qualitative detection are integrated; the sample is not damaged through whole physical detection; the sample adsorption quantity is little; the operation is simple and rapid, and the condition is mild; the absolute molecular weight and the distribution thereof of the polysaccharide can be obtained without standard sample; the sensitivity is high and can reach 10<-9>; and the physicochemical characteristics of polysaccharide can be simultaneously detected. The invention can be widely applicable to analysis detection, quality control, research and development of polysaccharide and other related compounds.

Description

technical field [0001] The invention relates to a method for detecting fungal polysaccharides. Background technique [0002] Polysaccharides are the most abundant polymer compounds in nature. They are widely distributed in nature. They exist in higher plants, algae, fungi and animals. The various biological and pharmacological functions of polysaccharides have aroused people's attention. highly valued. Polysaccharide science, as an emerging discipline in the post-genome era, has been included in the international frontier research field. Internationally, edible and medicinal fungal polysaccharides are called "Biological Response Modifiers" (BRM for short), because they are a class of biologically active substances that can enhance the immune function of the human body and have a wide range of physiological and pharmacological functions. In recent years, edible and medicinal fungal polysaccharides have become one of the research hotspots in the fields of biochemistry, molec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
Inventor 赵永亮王卫国
Owner HENAN UNIVERSITY OF TECHNOLOGY
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