Separation and detection of cellooligosaccharides on cellulose thin-layer chromatography Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/1c18dj35g

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  • Linear oligosacchardies of 1,4 linked β-D-glucopyranose are commonly referred to as cellodextrins (CD) or cellooligosaccharides (CO). They are of interest to those working in disciplines involving cellulose chemistry because they are often used as model substrates for cellulose itself. They are of interest to food scientists and nutritionists because they are easily incorporated into foods as non-digestible oligosaccharides, a category of food ingredients that is thought to be beneficial lo human health. The intent of the research presented in this thesis was to evaluate the potential of using cellulose supports for the chromatographic separation of soluble CDs differing in their degree of polymerization (DP; a numerical value indicating the number of glucose substituents per molecule). Soluble CDs range in DP from 2 to 8. Thin layer chromatography (TLC), using cellulose-coated TLC plates, was used as a model chromatographic system. Mixed CD preparations, containing CDs ranging in DP from 2 to 8 were prepared by incomplete acid-catalyzed hydrolysis of cellulose. The DP profiles of the different CD preparations were qualitatively demonstrated by TLC using silica-coated plates, an organic solvent-based mobile phase, and a standard carbohydrate visualizing reagent (p-anisaldehye in sulfuric acid). CD-preparations were then chromatographed on cellulose-coated TLC plates. Visualization of the chromatographed CDs was accomplished using a silver nitrate-sodium hydroxide reagent system, a reducing-sugar visualizing reagent. The silver nitrate-sodium hydroxide system was found to be the most appropriate, based on detection limits, simplicity and safety, of the several visualization reagents tested. Eight different mobile phases, all aqueous-based, were tested as potential solvents for the resolution of CDs, differing in DP, on cellulose-coated tlc plates at room temperature. The optimum solvent was found to be 60% ethanol/40% water. This solvent clearly resolved CDs of DP 3, 4 and 5. CD preparations chromatographed with the same mobile phase, but with silica-coated TLC plates, were not resolved. These combined results suggest that the TLC system with the cellulose stationary phase behaves similar to an affinity system, since silica and cellulose are both relatively hydrophilic stationary phases (i.e. both systems are typically considered examples of normal phase adsorption chromatography). The results further illustrate that cellulose supports have potential for use in the preparation of CDs of defined DP.
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