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

Abstract

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

Capillary electrophoretic analysis of neutral carbohydrates using ionic liquids as background electrolytes

“…Many suitable co-ions have been reported, such as 2,6-pyridinecarboxylic acid [16], sorbic acid [17], 1-naphthylacetic acid [18] and peptides [19]. Several derivatization techniques have been devised [20][21][22].…”

Method development for mono‐ and disaccharides monitoring in cell culture medium by capillary and microchip electrophoresis

“…Lastly, the incubation time and temperature were evaluated. Fructose, as a ketose sugar, is more difficult to derivatize [39,43] and was not detected at any of the previous reaction conditions. It was reported that fructose could be derivatized with APTS when using a longer incubation time [23].…”

Capillary Electrophoresis

“…Carbohydrates. Tseng et al described a CE-LIF assay for small molecule carbohydrates, a class of molecules traditionally difficult to detect (132). Carbohydrates first underwent reductive amination with ammonia followed by fluorogenic labeling with 4-fluoro-7-nitrobenzofurazane, achieving limits of detection of 50-250 nM.…”