Direct Oligosaccharide Profiling Using Thin-Layer
Chromatography Coupled with Ionic Liquid-Stabilized Nanomatrix-Assisted
Laser Desorption-Ionization Mass Spectrometry
Posted on 2019-09-05 - 11:33
The
in-depth characterization of glycan structures is crucial to
understanding their structure–function relationships and their
effects on health and various diseases. Despite advances in rapid
analysis, the utility of matrix-assisted laser desorption/ionization
mass spectrometry (MALDI MS) is limited for complex mixtures of carbohydrates
due to their low ionization efficiency and the difficulty in separating
oligosaccharides because of their high structural similarity. In this
study, we developed an ionic liquid (IL)-stabilized, nanomatrix-decorated,
thin-layer chromatography (TLC)-MALDI MS method for simultaneous and
rapid separation, detection, and identification of oligosaccharides
to achieve efficient profiling. The IL demonstrated good dispersion
and stabilization for the spin coating of dihydroxybenzoic acid-functionalized
magnetic nanoparticles (DHB@MNPs) on the TLC plate with spot homogeneity,
which contributed to the observed high reproducibility (<20% CV)
and 12- and 28-fold signal enhancement. Although the TLC was not able
to separate isomeric glycans, the DHB@MNPs generate diagnostic glycosidic
and cross-ring cleavage ions, enabling on-spot structural elucidation
of composition, sequence, branching, and linkage of glycans in each
separated spot. Without chemical derivatization of glycan samples,
glycan visualization by TLC and tandem MS, our integrated platform,
allowed the identification of 25 oligosaccharides from human milk,
and heatmap analysis revealed the variability in the oligosaccharide
abundance in samples from individual donors at different lactation
times, which may provide insight into the microbiota and immunity
of infants. With the demonstrated simplicity of our sample preparation
method along with the achieved separation and in-depth structural
characterization, our approach can be used for the rapid screening
of other oligosaccharide-rich samples.
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Mernie, Elias
Gizaw; Tolesa, Leta Deressa; Lee, Ming-Jer; Tseng, Mei-Chun; Chen, Yu-Ju (2019). Direct Oligosaccharide Profiling Using Thin-Layer
Chromatography Coupled with Ionic Liquid-Stabilized Nanomatrix-Assisted
Laser Desorption-Ionization Mass Spectrometry. ACS Publications. Collection. https://doi.org/10.1021/acs.analchem.9b01241
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AUTHORS (5)
EM
Elias
Gizaw Mernie
LT
Leta Deressa Tolesa
ML
Ming-Jer Lee
MT
Mei-Chun Tseng
YC
Yu-Ju Chen
KEYWORDS
dihydroxybenzoic acid-functionalizedthin-layer chromatographysample preparation methodThin-Layer Chromatographyglycan visualizationchemical derivatizationcross-ring cleavage ionsisomeric glycansoligosaccharide abundanceheatmap analysisidentificationsignal enhancementIonic Liquid-Stabilized Nanomatrix-Assisted Laser Desorption-Ionization Mass Spectrometryseparationlactation timesTLC platecharacterizationionization efficiencyoligosaccharide-rich samplesspot homogeneityglycan structuresCV25 oligosaccharidesMALDI MStandem MSDirect Oligosaccharide ProfilingDHBILglycan samples