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A method for analyzing the distribution of monosaccharide residues in polysaccharide structures

A technology of distribution law and polysaccharide, applied in the field of polysaccharide structure analysis, can solve the problems of cumbersome experimental steps, difficult analysis of experimental results, and difficult to achieve experimental conditions, and achieve the effect of simple experimental process, good repeatability, and easy analysis.

Active Publication Date: 2021-07-09
BIOLOGY INST OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The research on the structure of polysaccharides needs to use a variety of chemical and physical analysis methods for joint analysis. The structure of polysaccharides can only be inferred after the integration of various structural information, and the above analysis methods have cumbersome experimental steps, difficult to achieve experimental conditions, and difficult to analyze experimental results. , Experimental conclusions are difficult to repeat and other difficulties, resulting in the analysis of polysaccharide structures often requiring a lot of manpower and time
[0005] The Chinese patent with the publication number CN 102253167 A (a method for analyzing water-soluble polysaccharides based on hydrolysis process) discloses a method for recording the hydrolysis profiles of reducing sugars, glucose, galactose, and xylose in the polysaccharide hydrolysis process through sensors to form the polysaccharides. The standard hydrolysis map, but because the construction of the sensor requires the corresponding specific enzyme, the monosaccharide residues that are ubiquitous in various polysaccharides such as mannose, arabinose, and glucuronic acid cannot be detected by the sensor, resulting in this patent. To achieve the identification of polysaccharides, it is not possible to analyze the structure of polysaccharides as a whole

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  • A method for analyzing the distribution of monosaccharide residues in polysaccharide structures
  • A method for analyzing the distribution of monosaccharide residues in polysaccharide structures
  • A method for analyzing the distribution of monosaccharide residues in polysaccharide structures

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] (1) Hydrolysis treatment

[0069] Weigh 0.3g intracellular polysaccharides of D. spp. and place them in 7 ampoules (No. 1-7 in sequence), add 10mL of sulfuric acid solution to No. 1-6 ampoules respectively, fill with nitrogen and seal, and carry out under the condition of water bath. Hydrolysis, among them, No. 1-6 ampoules were treated according to Table 3, the hydrolysis temperature was 100°C, and they were recorded as AIPS-1~AIPS-6 respectively; as a blank control, No. 2h (recorded as IPS).

[0070] After the above treatment, centrifuge (15000r / min, 10min, 4°C), take 4.5mL supernatant and adjust the pH to 7 with NaOH solution (2mol / L), add water to 10mL, centrifuge (15000r / min, 15min, 4°C), take the supernatant, which is the step-by-step partial acid hydrolyzate samples (including a blank control sample).

[0071] Concentration and hydrolysis time of sulfuric acid solution when table 31-6 samples are hydrolyzed

[0072]

[0073] (2) Monosaccharide composition a...

Embodiment 2

[0085] (1) Hydrolysis treatment

[0086] Carry out hydrolysis treatment according to the method for step (1) in the embodiment 1, obtain step by step partial acid hydrolyzate sample (comprising a blank control sample); The difference is that, be the polysaccharide sample (respectively denoted as AEPS -1~AEPS-6, the blank control sample is recorded as EPS).

[0087] (2) Monosaccharide composition analysis of partial acid hydrolyzate products step by step

[0088] According to the method of step (a)~(d) in the embodiment 1, carry out pre-column derivation high performance liquid chromatography analysis, the result is as follows:

[0089] According to the peak time and peak area analysis of the monosaccharide species in the described step-by-step partial acid hydrolyzate samples according to the peak time and peak area of ​​the pre-column derivative high performance liquid chromatogram of the derivatization standard substance and the derivatization test sample, and obtain the pr...

Embodiment 3

[0095] (1) Hydrolysis treatment

[0096] Carry out hydrolysis treatment according to the method for step (1) in the embodiment 1, obtain step by step partial acid hydrolyzate sample (comprising a blank control sample); The difference is that the polysaccharide sample is the polysaccharide sample (respectively denoted as AIPS -1~AIPS-6, the blank control sample is recorded as IPS).

[0097] (2) Monosaccharide composition analysis of partial acid hydrolyzate products step by step

[0098] According to the method of step (a)~(d) in the embodiment 1, carry out pre-column derivation high performance liquid chromatography analysis, the result is as follows:

[0099] According to the peak time and peak area analysis of the monosaccharide species in the described step-by-step partial acid hydrolyzate samples according to the peak time and peak area of ​​the pre-column derivative high performance liquid chromatogram of the derivatization standard substance and the derivatization test ...

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Abstract

The present invention provides a method for analyzing the distribution law of monosaccharide residues in the polysaccharide structure. The present invention analyzes the hydrolysis law of polysaccharides through stepwise partial acid hydrolysis combined with pre-column derivatization high performance liquid chromatography, and draws the molar ratio of monosaccharide residues. Percentage content dissociation process diagram, according to the dissociation process diagram, the distribution of monosaccharide residues in the polysaccharide main chain and branch chain can be analyzed intuitively, quickly and accurately. Among them, the pre-column derivatization method can only detect free monosaccharides, and undegraded polysaccharides and oligosaccharides have no effect on the measurement results, so the hydrolyzate of the present invention can be directly analyzed after neutralization. In addition, according to the dissociation process diagram of the mole percentage content of monosaccharide residues in the examples, it can be found that the spectra of the same polysaccharide are basically the same, while the spectra of different polysaccharides are obviously different. Each polysaccharide map has map features closely related to its structural features, which can be used as the basis for identifying polysaccharides, and can be used to identify polysaccharides after building relevant databases and analysis software.

Description

technical field [0001] The invention relates to the technical field of polysaccharide structure analysis, in particular to a method for analyzing the distribution law of monosaccharide residues in polysaccharide structures. Background technique [0002] Polysaccharides are natural macromolecular substances formed by the condensation of more than 10 monosaccharide molecules. They are formed by aldoses and / or ketoses linked by glycosidic bonds. They are one of the four basic substances that constitute life. Polysaccharides widely exist in animals, plants, and microbial cells. However, due to the complexity of polysaccharides and the underestimation of the contribution of polysaccharides to life processes in the past, the research on polysaccharides has been lagging behind nucleic acids and proteins. It was not until Brander proved in 1958 that yeast cell wall polysaccharides have obvious anti-tumor effects that people gradually realized that polysaccharides not only have nutri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
CPCG01N30/88
Inventor 郑岚马耀宏孟庆军王丙莲杨艳杨俊慧蔡雷公维丽
Owner BIOLOGY INST OF SHANDONG ACAD OF SCI