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Analysis of mono‐, di‐ and oligosaccharides by CE using a two‐stage derivatization method and LIF detection
Author(s) -
Tseng HuaMing,
Gattolin Stefano,
Pritchard Jeremy,
Newbury H. John,
Barrett David A.
Publication year - 2009
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.200800517
Subject(s) - derivatization , chemistry , chromatography , reductive amination , ammonia , honeydew , reproducibility , detection limit , pentose , sugar , mass spectrometry , organic chemistry , botany , fermentation , biology , catalysis
A sensitive CE with LIF method has been developed for quantitative analysis of small carbohydrates. In this work, 17 carbohydrates including mono‐, di‐ and oligosaccharides were simultaneously derivatized with 4‐fluoro‐7‐nitrobenzofurazane (NBD‐F) via a two‐step reaction involving reductive amination with ammonia followed by condensation with NBD‐F. Under the optimized derivatization conditions all carbohydrates were successfully derivatized within 2.5 h and separated within 15 min using borate buffer (90 mmol/L, pH 9.2). For sugar standards LODs were in the range of 49.7 to 243.6 nmol/L. Migration time and peak area reproducibility were better than RSD 0.1 and 3%, respectively. The method was applied to measure sugars in nanoliter volume samples of phloem sap obtained by stylectomy from wheat and to honeydew samples obtained from aphids feeding from wheat and willow.