Capillary electrophoresis as a simple and sensitive method to study polysaccharides of Sinorhizobium sp. NGR234

Abstract

We report the minimum amounts of polysaccharides (6.5 ng) that can be studied using capillary electrophoresis-laser-induced fluorescence (CE-LIF) and describe the separation of the various monosaccharides susceptible to occur in Sinorhizobium cell wall. On-gel hydrolysis of the polysaccharides is described. It allowed the easy and rapid determination of the monosaccharide composition of the excised lipopolysaccharides. The composition obtained fits with gas chromatography-mass spectrometry (GC-MS) results. This study underlines the simplicity of using CE-LIF, which remains an entirely "water medium" method, unlike GC-MS which is, in addition, less sensitive (at least 100-fold) than CE-LIF.

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

“…It is interesting to note that fructose resulted in two peaks after the two‐step derivatization and the other saccharides resulted in single peaks. This phenomenon, which may be due to the instability of the ketose sugar skeleton, is in agreement with previous results 7, 24 using UV‐detectable 4‐aminobenzonitrile as a labeling reagent. Unlike a previous report of APTS labeling of fructose 13 we observed a substantially lower labeling efficiency for fructose compared with glucose.…”

“…8‐aminopyrene‐1,3,6‐trisulfonate (APTS), one of the most popular labeling reagents for the derivatization, was first introduced by Fraysse et al . 7. Due to its extensive aromaticity, APTS–carbohydrate derivatives have a substantially higher molar absorptivity and quantum efficiency than most of the commonly used fluorophore carbohydrate derivatives 5.…”

Coupled Mass Spectrometric Strategies for the Determination of Carbohydrates at Very Low Concentrations: The Case of Polysaccharides Involved in the Molecular Dialogue Between Plants and Rhizobia

“…Most of the time 1-aminopyrene-3,6,8-trisulfonate (APTS) is used for monosaccharide, their migration time allow the identification of the different derivatized alditols, it is very convenient for small quantities of natural polysaccharides and specially polysaccharides extracted from DOC PAGE. An acidic compound is more easily prepared than with classical alditol strategies for GC analysis [81].…”

Separation of R‐form lipopolysaccharide and lipid A by CE–Fourier‐transform ion cyclotron resonance MS

“…CE-MS was successfully applied to the analysis of derivatized and underivatized polysaccharides from various origins including part structures of LPS [20][21][22][23][24]. Also phospholipids [25][26][27] and gangliosides [28,29] have been analyzed by CE-MS, in some cases using non-aqueous CE solvent systems.…”