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Analytic method of polysaccharide topological structure based on mass spectrometry

A technology of topology and mass spectrometry analysis, applied in the field of polysaccharide topology analysis based on mass spectrometry, to achieve good universality and credibility, ensure credibility, and facilitate the unification of standards

Active Publication Date: 2017-02-15
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing polysaccharide topological structure analysis methods need to perform "de-isotope" approximate preprocessing on the original experimental data, which has large deviations and has many limitations in application.

Method used

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  • Analytic method of polysaccharide topological structure based on mass spectrometry
  • Analytic method of polysaccharide topological structure based on mass spectrometry
  • Analytic method of polysaccharide topological structure based on mass spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A polysaccharide topology analysis method based on mass spectrometry mainly includes five specific analysis steps, such as figure 1 shown; each step is briefly described as follows:

[0035] (1) The standard polysaccharide NA2 is prepared into a solution, and after electrospray ionization and mass spectrometry detection, a first-order mass spectrogram containing the experimental isotope profile information of the precursor ion is obtained. The concentration of the standard polysaccharide NA2 is 1 μM, and the solvent includes NH 4 HCO 3 , methanol and water, where NH 4 HCO 3 The concentration is 10mM, the quality of methanol and water is the same; the voltage of electrospray ionization treatment is 2.5kV, and the temperature is 300°C; the primary mass spectrogram is as follows figure 2 shown;

[0036] (2) Compare the fingerprints of the experimental isotope profile of the precursor ion with each polysaccharide in the theoretical database of the precursor ion one by ...

Embodiment 2

[0044] Using the same standard polysaccharide and analysis steps as in Example 1, the difference is that the conditions used are different:

[0045] (1) During electrospray ionization, the concentration of standard polysaccharide NA2 is 0.5 μM, and the solvent includes NH 4 HCO 3 , methanol and water, where NH 4 HCO 3 The concentration is 8mM, the quality of methanol and water is the same; the voltage of electrospray ionization treatment is 2kV, and the temperature is 280℃.

[0046] (2) The normalized collision energy for high-energy collision-induced dissociation is 15%.

[0047] The conclusion of the final polysaccharide topology remains unchanged.

Embodiment 3

[0049] Using the same standard polysaccharide and analysis steps as in Example 1, the difference is that the conditions used are different:

[0050] (1) During electrospray ionization, the concentration of standard polysaccharide NA2 is 2 μM, and the solvent includes NH 4 HCO 3 , methanol and water, where NH 4 HCO 3 The concentration is 12mM, the quality of methanol and water is the same; the voltage of electrospray ionization treatment is 3kV, and the temperature is 350℃.

[0051] (2) The normalized collision energy for high-energy collision-induced dissociation is 20%.

[0052] The conclusion of the final polysaccharide topology remains unchanged.

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Abstract

The invention relates to an analytic method of a polysaccharide topological structure based on mass spectrometry. The method comprises the steps of obtaining a first grade mass spectrum containing experimental isotope outline information of precursor ions after conducting electrospray ionization and mass spectrometry on polysaccharide samples ready to be analyzed, and conducting cascade stage mass spectrum dissociation on the precursor ions of the first grade mass spectrum to obtain a second grade mass spectrum of corresponding fragment ions; conducting comparison between the experimental isotope outlines of the precursor ions and the theoretical database of the precursor ions to obtain multiple candidate polysaccharides which correspond to the precursor ions; and conducting comparison between the theoretical isotope outline of each theoretical fragment ion of each candidate polysaccharide and the experimental isotope outlines of the fragment ions observed in the second grade mass spectrum which corresponds to the candidate polysaccharides, and then the candidate polysaccharide which has the highest matching rate is the final polysaccharide ID. Compared with the prior art, the analytic method of polysaccharide topology is simple and clear, efficient and accurate, and is applicable to qualitative identification and quantitative analysis of the polysaccharide topological structure based on high resolution cascade stage mass spectrometry.

Description

technical field [0001] The invention relates to the field of molecular structure identification, in particular to a method for analyzing the topological structure of polysaccharides based on mass spectrometry. Background technique [0002] Glycosylation modification is one of the most common and important post-translational modifications on proteins, which has extremely important physiological and pathological functions. Glycoomics based on tandem mass spectrometry has become one of the main analytical methods for the identification and analysis of glycosylated polysaccharide modification sites, monosaccharide composition and topological structure. The topological structure of existing polysaccharides is described by a tree. However, the existing polysaccharide topological structure analysis methods need to perform "de-isotope" approximate preprocessing on the original experimental data, which has large deviations and has many limitations in application. Contents of the i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62
CPCG01N27/62
Inventor 田志新
Owner TONGJI UNIV