Quantification of monosaccharides through multiple-reaction monitoring liquid chromatography/ mass spectrometry using an aminopropyl column

Loubna A. Hammad, Dakota Z. Derryberry, Yazen R. Jmeian, Yehia Mechref

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

A simple, sensitive, and reproducible quantitative liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was designed for the simultaneous quantification of monosaccharides derived from glycoprotein and blood serum using a multiple-reaction monitoring (MRM) approach. Sialic acids and neutral monosaccharides were efficiently separated using an amino-bonded silica phase column. Neutral monosaccharide molecules were detected as their aldol acetate anion adducts [M+CH3CO2]- using electrospray ionization in negative ion MRM mode, while sialic acids were detected as deprotonated ions [M-H]-. The new method did not require a reduction step, and exhibited very high sensitivity to carbohydrates with limits of detection of 1 pg for the sugars studied. The linearity of the described approach spanned over three orders of magnitude (pg to ng). The method was validated for monosaccharides originating from N-linked glycans attached to glycoproteins and glycoproteins found in human blood serum. The method effectively quantified monosaccharides originating from as little as 1 μg of glycoprotein and 5 μL of blood serum. The method was robust, reproducible, and highly sensitive. It did not require reduction, derivatization or postcolumn addition of reagents.

Original languageEnglish
Pages (from-to)1565-1574
Number of pages10
JournalRapid Communications in Mass Spectrometry
Volume24
Issue number11
DOIs
StatePublished - Jun 2010

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