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  • Combination of Microwave-Assisted Girard Derivatization with Ionic Liquid Matrix for Sensitive MALDI-TOF MS Analysis of Human Serum N-Glycans.

Combination of Microwave-Assisted Girard Derivatization with Ionic Liquid Matrix for Sensitive MALDI-TOF MS Analysis of Human Serum N-Glycans.

Journal of analytical methods in chemistry (2018-11-14)
Hoa Thi Le, Kyu H Park, Woong Jung, Hyung Soon Park, Tae Woo Kim
ABSTRACT

We developed a new method for MALDI-TOF MS detection of N-glycans derived from human serum. The synergistic combination of microwave-assisted Girard T derivatization, solid-phase extraction desalting, and an ionic liquid matrix (2, 5-dihydroxybenzoic acid/aniline) (GT-SPE-DHB/An) allowed of more sensitive N-glycans detection than a conventional ionic liquid matrix in MALDI-TOF MS. The superior sensitivity of our method was confirmed by the number of assigned N-glycans in 900-2,000 m/z range. Using our GT-SPE-DHB/An method, we were successfully able to assign 31 glycans. However, with the established method, i.e., DHB/An method, only 15 glycans were assigned. To the best of our knowledge, this GT-SPE-DHB/An method is the first to combine cationic derivatization of N-glycan and ionic liquid matrix for N-glycan analysis in MALDI-TOF MS.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
α-Cyano-4-hydroxycinnamic acid, ≥98% (TLC), powder
Sigma-Aldrich
Maltohexaose, ≥65% (HPLC)
Sigma-Aldrich
Human Serum, from human male AB plasma, USA origin, sterile-filtered
Sigma-Aldrich
2,5-Dihydroxybenzoic acid, 98%
Sigma-Aldrich
Trifluoroacetic acid, ReagentPlus®, 99%
Sigma-Aldrich
Ammonium bicarbonate, ReagentPlus®, ≥99.0%
Sigma-Aldrich
DL-Dithiothreitol, ≥98% (HPLC), ≥99.0% (titration)
Sigma-Aldrich
p-Coumaric acid, ≥98.0% (HPLC)
Sigma-Aldrich
Peroxidase from horseradish, lyophilized, powder, ~150 U/mg
Supelco
Vials, screw top with polypropylene open-top cap, pre-assembled, pkg of 100, volume 4 mL, amber glass vial, O.D. × H 15 mm × 45 mm, tan PTFE/silicone septum