Development of a new high-throughput method to determine the composition of ten monosaccharides including 4-O-methyl glucuronic acid from plant cell walls using ultra-performance liquid chromatography

Abstract

Plant cell walls are an important dietary source for livestock, and could be an enormous resource for production of next-generation bioethanol and more valuable materials. Because polysaccharides are major components of plant cell walls, analysis of their composition is important. In this report, we established a high-throughput method to determine the composition of ten monosaccharides from plant cell walls simultaneously using ultra-performance liquid chromatography with p-aminobenzoic ethyl ester-labeling technology. Complete separation of a mixture of internal standards, 2-deoxyglucose and 3-O-methyl glucose, and ten monosaccharides, consisting of seven neutral and three acidic sugars including 4-O-methyl-D-glucuronic acid, which are frequently found in plant cell wall polysaccharides, can be obtained within 7 min using this system. Relative standard deviations of retention time and peak area value are lower than 1%. Linearity for broader dynamic ranges (0.02–2000 mg l −1), faster analysis and higher sensitivity than other traditional methods, including one that employs widely used high-performance anion exchange chromatography, are achieved. We evaluated this new method by analyzing the composition of cell walls from three model plants (Arabidopsis thaliana, rice and hybrid aspen) and confirmed that the obtained results for most monosaccharides are consistent with those in previous studies. These data suggest that our newly developed system could greatly contribute to the study of plant cell walls, especially research requiring highthroughput analysis.