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
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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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