Improved RP-HPLC and anion-exchange chromatography methods for the determination of amino acids and carbohydrates in soil solutions

Abstract

In spite of their low concentrations in soil solutions, low-molecular weight organic substances (LMWOS) such as amino acids, sugars, and uronic acids play a major role in the cycles of C and N in soil. With respect to their low concentrations and to possible matrix interferences, their analysis in soil leachates is a challenging task. We established two HPLC (high-performance liquid chromatography) methods for the parallel determination of amino acids and carbohydrates in soil leachates. The pre-column derivatization of amino acids with an o-phthaldialdehyde (OPA) mercaptoethanol solution yields quantitation limits between 0.03 and 0.44 μmol L-1 and SD values of <8.3% (n = 9). High-performance anion-exchange chromatography (HPAEC) on a Dionex CarboPac PA 20 column with a NaOH acetate gradient combined with pulsed amperometric detection (PAD) was used for the determination of carbohydrates. The calibration curves obtained for 11 carbohydrates showed excellent linearity over the concentration range from 0.02 to 50.0 mg L-1. Recovery studies revealed good results for all analytes (89%-108%). Interferences from Hg(II) salts and chloroform used for stabilization of the leachates did not occur with both chromatographic methods. The optimized method was successfully used for quantitative determinations of amino acids and carbohydrates in soil leachates. © 2008 Wiley-VCH Verlag GmbH & Co. KGaA.