Development of a precolumn derivatization HPLC method with diode‐array detection for the determination of amino sugars in peat and soil humic acids

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EarlyView Article

  • Published: Jan 2, 2018
  • Author: Erik Beňo, Róbert Góra, Milan Hutta
  • Journal: Journal of Separation Science


The work is focused on the development of a high‐performance liquid chromatography method with diode‐array detection for the separation and quantitation of the three most abundant amino sugars; d‐glucosamine, d‐galactosamine, and d‐mannosamine. The high‐performance liquid chromatography separation was carried out by reversed‐phase chromatography on Chromolith Performance RP‐18e monolithic column after acid hydrolysis (5 M HCl) and precolumn derivatization of samples using diethyl ethoxymethylenemalonate. Gradient elution and a mobile phase composed of ammonium formate buffer solution (10 mmol/L, pH 3.60) and methanol with flow rate of 1.0 mL/min were used. The monitoring wavelength was set at 280 nm. The limits of detection and quantitation for analytes ranged from 0.017 to 0.122 mg/L and from 0.057 to 0.407 mg/L, respectively. The proposed method was successfully applied for the determination of amino sugars in samples of humic acids isolated from different soils and peat.

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