Comparison of HPLC Methods for the Determination of Amino Sugars in Soil Hydrolysates

Abstract

A study on the suitability of chromatographic techniques such as high performance anion exchange chromatography (HPAEC) with fluorescence detection (FL) and pulsed amperometric detection (PAD) and reversed phase (RP) chromatography for the determination of galactosamine, glucosamine, mannosamine, and muramic acid in soil hydrolysates was carried out. The reversed phase fluorescence method was rapid, provided good validation parameters, and employed relatively inexpensive instrumentation. The HPAEC methods had slightly higher limits of quantification, 0.6-5.0 lmol L À1 (HPAEC-FL) and 1.0-10.0 lmol L À1 (HPAEC-PAD), compared to the reversed phase fluorescence method (0.5-5.0 lmol L À1 ). Various sample pretreatment methods and chromatographic methods were investigated and the advantages and disadvantages of the HPLC methods are discussed.

Selective and Accurate Quantification of N-Acetylglucosamine in Biotechnological Cell Samples via GC–MS/MS and GC–TOFMS

“…Generally, the analysis of monosaccharide derivatives is achievable using gas (GC) and liquid chromatography (LC) based separation. While several separation approaches are described in the literature for hexosamine stereoisomers, − to our knowledge, no analytical method separating all three biologically relevant N-acetylated hexosamines is available for absolute quantification. To increase the (steric) selectivity and realize separation, derivatization often comes into play .…”

Metabolic tracing unravels pathways of fungal and bacterial amino sugar formation in soil

“…The cation exchange column (AG 50W‐X8 Resin, H + form, mesh size 100–200, Biorad, Munich, Germany) was adapted from Indorf et al . (): a glass wool layer was installed at the bottom of the glass columns (inner diameter, 0.8 cm), then 4 cm of cation exchange resin was placed into the column and preconditioned with ∼10 ml of 0.1 m HCl solution to ensure the H + ‐form of the sorbent. Afterwards, 5 ml of water was added to reach the neutral pH of the mobile phase.…”

Improved δ13 C analysis of amino sugars in soil by ion chromatography-oxidation-isotope ratio mass spectrometry

“…Liquid chromatography requires column purification to remove hydrolysable non‐cationic compounds such as monosaccharides and carboxylic acids from the extract. A cation‐exchange column (AG 50 W‐X8 resin, H + form, mesh size 100–200; Biorad, Munich, Germany) was used as suggested by Indorf et al ., and a thin layer of clean glass wool was installed under 4 cm of cation‐exchange resin in the glass column (inner diameter: 0.8 cm). The resin was filled in by rinsing with ~10 mL of 0.1 M HCl solution to ensure the H + form of the sorbent, covered with a thin layer of glass wool and preconditioned with 5 mL of water.…”