Optimisation of amino sugar quantification by HPLC in soil and plant hydrolysates

Amino sugars are increasingly used as indicators for the accumulation of microbial residues in soil and plant material. A reverse-phase high-performance liquid chromatography method was improved for the simultaneous determination of muramic acid, mannosamine, glucosamine and galactosamine in soil and plant hydrolysates via ortho-phthaldialdehyde (OPA) pre-column derivatisation and fluorescence detection. The retention time was reduced, and the separation of muramic acid and mannosamine was optimised by modifying the mobile phase. The effects of excitation wavelength, OPA reaction time, tetrahydrofuran concentration and pH value of the mobile phase on the amino sugar separation were tested. Quantification limits were in the range of 0.13 to 0.90 μg ml−1. No interferences exist from amino acids or other primary amines, occurring in soil and plant hydrolysates.