This work is a part of research on bioethanol technology from poplar wood. Analysis of products is an important question at each stage of the process. Enzymatic hydrolysis of pre- treated wood or delignified polysaccharide fraction yields mixture of soluble part of cellulose or hemicelluloses, oligosaccharides and simple sugars. HPLC is commonly applied for determination of sugars, using –NH2 column and acetonitrile-water as eluent. The main obstacle is in our case the presence of higher sugars in analysed samples. Due to extremely long retention times, they lead to column blocking, pressure increase and decrease of resolution. Aqueous size exclusion chromatography was applied to separate monosaccharides from higher fractions of saccharides. Two Phenomenex PolySep-GFC series columns were used – narrow pore P 1000 and mixed-bed P Linear. Resolution was found to low for separate glucose and xylose from each other even using narrow-pore column, though satisfactory in the case of glucose-cellobiose mixture. Buffer used for pH stabilization and enzymes were found to give peaks overlapping the sugar ones. Phosphate, citrate and acetate buffers were removed from samples using two Dowex ionites mixture – 50W x4 and 2×8, 50-100 mesh each. Enzyme adsorption on the same mixed ionite was found unsuccessful. The kinetic of adsorption was determined at room temperature and the amount of ionite needed to complete buffer removal was estimated.
Keywords: size exclusion chromatography, monosaccharides, ionite, buffer, enzyme
Authors
Lewandowska A.
Wood Technology, Warsaw University of Life Sciences, Warszawa
Radomski A.
Wood Technology, Warsaw University of Life Sciences, Warszawa
Szadkowski J.
Szadkowska D.
Marchwicka M.
Archanowicz E. I.
Lisiecka E.
Drożdżek M.
Login to download the PDF