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Quantification of monosaccharides through multiple‐reaction monitoring liquid chromatography/mass spectrometry using an aminopropyl column
Author(s) -
Hammad Loubna A.,
Derryberry Dakota Z.,
Jmeian Yazen R.,
Mechref Yehia
Publication year - 2010
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.4536
Subject(s) - chemistry , monosaccharide , chromatography , selected reaction monitoring , derivatization , sialic acid , mass spectrometry , electrospray ionization , tandem mass spectrometry , glycan , glycoprotein , biochemistry
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 + CH 3 CO 2 ] − 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. Copyright © 2010 John Wiley & Sons, Ltd.