Determination of Mono- and Oligosaccharides Derivatized with p-Aminobenzoic Ethyl Ester by Reverse Phase HPLC

  • Kwon, Hyokjoon (Analytical Sciences Group, Specialty Chemicals Research, LG Chem Research Park) ;
  • Kim, Joon (Analytical Sciences Group, Specialty Chemicals Research, LG Chem Research Park)
  • Published : 1995.12.25

Abstract

Mono- and oligosaccharides are derivatized with p-aminobenzoic ethyl ester (ABEE), strongly absorbs UV light at 254 nm, in the presence of sodium cyanoborohydride. C18-bonded silica column is used for the separation of sugar-ABEE derivatives in an isocratic mode. RP-HPLC conditions are optimized by using ternary mixture as mobile phase and $45^{\circ}C$ as a column temperature. Sugar-ABEE derivatives are separated well within a short run time (ca. 25 min) by reverse-phase partition chromatographic mode. The ($1{\rightarrow}6$) linkage type of dihexose-ABEE derivatives has shorter retention time than ($1{\rightarrow}4$)-linkage type. After acid hydrolysis of glycoproteins with 2M trifluoroacetic acid, monosaccharide composition and contents are determined. This procedure is very useful for the simultaneous analysis of neutral and amino sugars in a single chromatographic step using RP-HPLC without reacetylation of deacetylated amino sugars, which are produced by acid hydrolysis.

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