A kind of method to improve the ionization efficiency of oligosaccharide

An ionization and oligosaccharide technology, applied in the field of biochemical analysis, can solve the problems that have not yet been seen, and achieve the effects of improving ionization efficiency, uniform crystallization, and beneficial to sugar chain structure analysis.

Inactive Publication Date: 2018-07-24
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] But so far, there is no report on the use of 6-hydrazine nicotinamide (HYNIC) as a matrix to analyze oligosaccharides and improve the ionization method of oligosaccharides in matrix-assisted laser desorption time-of-flight mass spectrometry.

Method used

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  • A kind of method to improve the ionization efficiency of oligosaccharide
  • A kind of method to improve the ionization efficiency of oligosaccharide
  • A kind of method to improve the ionization efficiency of oligosaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] 1. Take 1μL of DP7 (1pmol / L) and mix with 1μL of 10mg / mL DHB matrix solution (50% acetonitrile, 0.1% trifluoroacetic acid) and 1μL of 2mg / mL HYNIC (98% methanol, 2% acetic acid) respectively, and click the target for MALDI- TOF MS analysis, the results are as figure 2 Shown, contrast figure 2 (a) and figure 2 (b) It can be seen that using HYNIC as a matrix can obtain higher quality DP7 mass spectra, and its signal-to-noise ratio is increased by 10 times; the specific implementation process of other analytes is similar to that of DP7;

[0042] 2. Take the standard peptide and DP7 and mix them according to the molar ratio of 1:1 and 1:10 respectively, and take 1μL of the target in the mixture for MALDI-TOF MS analysis. The results are as follows Figure 4 As shown, when the HYNIC matrix is ​​used for analysis, the signal intensity of sugar chain DP7 is higher, and the standard peptide signal is inhibited; while the DHB matrix has the opposite effect;

[0043] 3. Weigh 0.5mg ...

Embodiment 2

[0045] Take 1pmol / μL DP71μL point target for repeatability experiment; randomly select 12 positions on the target point for mass spectrometry analysis, and compare the intensity of 12 mass spectra; the results are as follows Image 6 As shown, when HYNIC is used as the matrix, the signal strength and stability are better than DHB matrix.

[0046] In order to further verify the uniformity of the matrix crystallization, 1pmol / μL DP71μL was spotted on the target and subjected to mass spectrometry imaging analysis. The results are as follows Figure 7 As shown in a, the DHB crystal area is mainly located at the periphery of the target point, and there is almost no crystal in the center of the target point, resulting in low signal intensity; while HYNIC crystals (such as Figure 7 b) It is located in the entire target area and has uniform crystallinity.

Embodiment 3

[0048] Take DP7 mixed with different concentrations of non-volatile salt NaCl (25mM to 200mM) and then take 1μL of the spot target for MALDI-TOF MS analysis. The results are as follows Figure 8 As shown, compared with the DHB matrix, the HYNIC matrix has stronger salt tolerance and DP7 signal strength is higher.

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Abstract

The invention belongs to the field of biochemical analysis, and relates to a method for improving the ionization efficiency of oligosaccharides, especially a method for improving the ionization of oligosaccharides in matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS); the method adopts 6 ‑Hydrazine nicotinamide (HYNIC) is used as a matrix to analyze oligosaccharides, which selectively improves the ionization efficiency of oligosaccharides and inhibits the ionization of other analytes such as peptides. The 6-hydrazine nicotinamide in the present invention has the structure of formula (I); when the 6-hydrazine nicotinamide is used as a matrix to analyze oligosaccharides, it has high sensitivity, uniform crystallization, strong salt tolerance, and tandem mass spectrum fragmentation The advantages such as abundant information; The method of the present invention, adopts described 6 hydrazine nicotinamide as matrix analysis oligosaccharide, can selectively improve the ionization efficiency of oligosaccharide, and suppressed other analyte such as peptide segment Ionization has the advantages of simple operation, simple steps, economical and time-saving, and no desalination is required.

Description

Technical field [0001] The invention belongs to the field of biochemical analysis, and relates to a method for ionization of oligosaccharides, in particular to a method for improving ionization efficiency of oligosaccharides, in particular to a method for improving oligosaccharides in matrix assisted laser desorption time-of-flight mass spectrometry (MALDI-TOFMS) Medium ionization method: This method uses 6-hydrazine hydrazine amide (HYNIC) as a matrix to analyze oligosaccharides, which selectively improves the ionization efficiency of oligosaccharides and inhibits the ionization of other analytes such as peptides. Background technique [0002] Oligosaccharides are an important type of information substance in the body and have many biological functions. They are generally composed of 3-10 monosaccharide molecules connected by glycosidic bonds. The oligosaccharide has important physiological functions, such as activating the cell's self-defense system, determining cell recognitio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/64
Inventor 陆豪杰焦竞张莹
Owner FUDAN UNIV
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