TY - JOUR
T1 - Quantification of monosaccharides through multiple-reaction monitoring liquid chromatography/ mass spectrometry using an aminopropyl column
AU - Hammad, Loubna A.
AU - Derryberry, Dakota Z.
AU - Jmeian, Yazen R.
AU - Mechref, Yehia
N1 - Copyright:
Copyright 2010 Elsevier B.V., All rights reserved.
PY - 2010/6
Y1 - 2010/6
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=77952907264&partnerID=8YFLogxK
U2 - 10.1002/rcm.4536
DO - 10.1002/rcm.4536
M3 - Article
C2 - 20486252
AN - SCOPUS:77952907264
VL - 24
SP - 1565
EP - 1574
JO - Rapid Communications in Mass Spectrometry
JF - Rapid Communications in Mass Spectrometry
SN - 0951-4198
IS - 11
ER -