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Comparison of oligosaccharide labeling employing reductive amination and hydrazone formation chemistries
Author(s) -
Krenkova Jana,
Dusa Filip,
Cmelik Richard
Publication year - 2020
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201900475
Subject(s) - chemistry , reductive amination , hydrazone , sodium cyanoborohydride , yield (engineering) , derivatization , analyte , glycan , fluorescence , chromatography , hydrazide , propionitrile , pyrene , mass spectrometry , combinatorial chemistry , organic chemistry , biochemistry , materials science , physics , quantum mechanics , acetonitrile , glycoprotein , metallurgy , catalysis
In this work, we compare labeling by two negatively charged fluorescent labels, 8‐aminopyrene‐1,3,6‐trisulfonic acid (APTS) and 8‐(2‐hydrazino‐2‐oxoethoxy)pyrene‐1,3,6‐trisulfonic acid (Cascade Blue hydrazide [CBH]). Effectiveness of the labeling chemistries were investigated by 4‐hydroxybenzaldehyde and maltoheptaose followed by LC/UV‐MS and CE/LIF analysis, respectively. The reaction yield of APTS labeling was determined to be only ∼10%. This is due to reduction of almost 90% of the analyte by sodium cyanoborohydride to alcohol, which cannot be further labeled via reductive amination. However, the CBH labeling provides ∼90% reaction yield based on the LC/UV‐MS measurements. The significantly higher labeling yield was also confirmed by CE/LIF measurements. Finally, the more effective hydrazone formation technique of CBH was characterized and applied for N ‐linked glycan analysis by CE/LIF.