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KMID : 0380020040190040269
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Korean Journal of Biotechnology and Bioengineering
2004 Volume.19 No. 4 p.269 ~ p.273
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Simple and Quantitative Analysis Method for Total Carbohydrate Concentration in Oligosaccharides by using TLC
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Lee Jin-Ha
Lee Hyung-Woo
Lee Hyung-Ki
Cho Dong-Lyun
Sunwoo Chang-Shin
Park Ki-Deok
Choi Jung-Sik
Kim Do-Won
Kim Do-Man
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Abstract
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A simple, fast and reproducible quantitative analysis method for sugar concentration composed in oligosaccharide mixture was developed. Two glass TLC plates were prepared per sample. After dipping one plate into the copper bicinchoninate reagent and the other plate into 5% sulfuric acid solution, both plates were baked in microwave oven until sugar spots were developed or the surface temperature of TLC plate becomes 60 to 70 . The corrective factor values [F value =(the value of total sugar concentration converted as glucose unit/the value of reducing sugar concentration converted as glucose unit)/(polymerization degree of sugar)] of different molecular weight sugars were determined. Within the concentration of 0.25∼1.0 in each sample loaded, the fructose-F (corrective factor value of fructose) was 0.45, yet for the higher concentration (2.5∼7.5 ) fructose-F was 1.0. In case of glucose, in the range of 0.5∼7.5 , glucose-F was same as fructose-F, 1.0. However, as the molecular weight of sugar was increased, the F values were decreased in both maltodextrin and isomaltodextrin oligosaccharides in 0.5∼7.5 of each sample loaded. Interestingly, F values were equal for the same molecular weight sugars, although the structures were different from each other. Using F value of each sugar, we could determine and compare the exact total sugar concentration of different molecular weight maltooligosaccharide and isomaltooligosaccharide. We also could determine if the unknown sugar was a reducing or non-reducing compound by using optimized TLC with microwave oven method.
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KEYWORD
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Quantitative analysis, TLC, reducing value, total carbohydrate, microwave Oven
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