Method for increasing oligosaccharide ionization efficiency

An ionization and oligosaccharide technology, applied in the field of biochemical analysis, can solve problems that have not yet been seen, and achieve the effects of improving ionization efficiency, enriching fragmentation information, and inhibiting ionization

Inactive Publication Date: 2016-04-27
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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  • Method for increasing oligosaccharide ionization efficiency
  • Method for increasing oligosaccharide ionization efficiency
  • Method for increasing oligosaccharide ionization efficiency

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] 1. Mix 1 μL DP7 (1 pmol / L) with 1 μL 10 mg / mL DHB substrate solution (50% acetonitrile, 0.1% trifluoroacetic acid) and 1 μL 2 mg / mL HYNIC (98% methanol, 2% acetic acid) respectively, and perform MALDI-TOFMS analysis on the target. The result is as figure 2 shown, compared figure 2 (a) with figure 2 (b) It can be seen that using HYNIC as a matrix can obtain a higher quality DP7 mass spectrum, 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 1 μL of the mixture to spot the target for MALDI-TOFMS analysis. The results are as follows: Figure 4 As shown, when using HYNIC matrix for analysis, the signal intensity of sugar chain DP7 is higher, and the signal of standard peptides is suppressed; while the effect of DHB matrix is ​​opposite;

[0043] 3. Weigh 0...

Embodiment 2

[0045] Take 1pmol / μLDP71μL spot target for repeated experiments; randomly select 12 positions on the target spot for mass spectrometry analysis, and compare the intensities of the 12 mass spectrograms; the results are as follows Figure 6 As shown, when HYNIC is used as the matrix, the signal intensity and stability are better than DHB matrix.

[0046] In order to further verify the uniformity of matrix crystallization, 1pmol / μLDP71μL was targeted and subjected to mass spectrometry imaging analysis, the results are as follows Figure 7 As shown in a, the DHB crystallization area is mainly located on the periphery of the target point, and there is almost no crystallization in the center of the target point, resulting in low signal intensity; while the HYNIC crystallization (such as Figure 7 Shown in b) located in the entire target area and crystallized uniformly.

Embodiment 3

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

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Abstract

The invention belongs the field of biochemical analysis, and discloses a method for increasing oligosaccharide ionization efficiency, and relates to a method for increasing oligosaccharides ionization in a matrix auxiliary laser desorption flight time mass spectrum (MALDI-TOF MS). The method employs 6-HYNIC as matrix analysis oligosaccharides, and optionally increases the ionization efficiency of oligosaccharide, and inhibits ionization of other analyte such as a peptide segment. The 6-HYNIC has a structure shown in a formula (I); the 6-HYNIC as the matrix for analysis of oligosaccharide has the advantages of high sensitivity, uniform crystallization, strong salt resistance performance, and abundant tandem mass spectrometry fragment information. The method employs 6-HYNIC as the matrix for analysis of oligosaccharide, and optionally increases the ionization efficiency of oligosaccharide, and inhibits ionization of other analyte such as a peptide segment. The method has the advantages of simple operation, simple steps, economy, time saving, and no requirement of salt removal.

Description

technical field [0001] The invention belongs to the field of biochemical analysis, and relates to a method for ionizing 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) Neutral ionization method; this method uses 6-hydrazine nicotinamide (HYNIC) as a matrix to analyze oligosaccharides, selectively improves the ionization efficiency of oligosaccharides, and inhibits the ionization of other analytes such as peptides. Background technique [0002] Oligosaccharides are an important class of information substances in the body and have many biological functions. They are generally composed of 3-10 monosaccharide molecules linked by glycosidic bonds. The oligosaccharide has important physiological functions, such as activating the self-defense system of cells, determining cell recognition, agg...

Claims

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Application Information

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