Negative ion graphitised carbon nano-liquid chromatography/mass spectrometry increases sensitivity for glycoprotein oligosaccharide analysis

Rapid Communications in Mass Spectrometry : RCM
Niclas G KarlssonNicolle H Packer

Abstract

Negative ion nano-liquid chromatography/mass spectrometry (nano-LC/MS) and tandem mass spectrometry (nano-LC/MS(2)), using graphitised carbon as separating medium, were explored for analysing neutral and acidic O-linked and N-linked oligosaccharide alditols. Compared to the sensitivity of capillary LC/MS (flow rate of 6 microL/min) coupled with a conventional electrospray ionisation source, the nano-LC/MS (flow rate of 0.6 microL/min) with a nanoflow ion source was shown to increase the sensitivity ten-fold with a detection limit in the low-femtomole range. The absolute signals for the [M-nH](n-) ions of the oligosaccharides were increased 100-fold, enabling accumulation of high-quality fragmentation data in MS(2) mode, in which detection of low abundant sequence ions is necessary for characterisation of highly sialylated N-linked oligosaccharides. Oligosaccharides with high numbers of sialic acid residues gave dominant fragments arising from the loss of sialic acid, and less abundant fragments from cleavage of other glycosidic bonds. Enzymatic off-line desialylation of oligosaccharides in the low-femtomole range prior to MS(2) analysis was shown to increase the quality of the spectra. Automated glycofragment mass fingerprintin...Continue Reading

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Citations

Feb 28, 2009·Analytical and Bioanalytical Chemistry·L Renee RuhaakManfred Wuhrer
Apr 26, 2012·Glycoconjugate Journal·Manfred Wuhrer
Sep 29, 2011·Journal of the American Society for Mass Spectrometry·Roisin A DoohanNiclas Göran Karlsson
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Dec 29, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Joomi AhnMartin Gilar
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Jun 20, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Yehia Mechref, Milos V Novotny

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