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Analysis of mono-, di- and oligosaccharides by CE using a two-stage derivatization method and LIF detection.

Electrophoresis (2009-03-26)
Hua-Ming Tseng, Stefano Gattolin, Jeremy Pritchard, H John Newbury, David A Barrett
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
4-Fluoro-7-nitrobenzofurazan, ≥98% (elemental analysis)
Sigma-Aldrich
4-Fluoro-7-nitrobenzofurazan, BioReagent, suitable for fluorescence, ≥98.0% (HPLC)

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