Carbohydrate analysis by a phenol–sulfuric acid method in microplate format

Among many colorimetric methods for carbohydrate analysis, the phenol–sulfuric acid method is the easiest and most reliable method. It has been used for measuring neutral sugars in oligosaccharides, proteoglycans, glycoproteins, and glycolipids. This method is used widely because of its sensitivity...

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Bibliographic Details
Published in:Analytical biochemistry 2005-04, Vol.339 (1), p.69-72
Main Authors: Masuko, Tatsuya, Minami, Akio, Iwasaki, Norimasa, Majima, Tokifumi, Nishimura, Shin-Ichiro, Lee, Yuan C.
Format: Article
Language:English
Subjects:
Analysis
Carbohydrate
Carbohydrates - analysis
Colorimetry
Microplate
Phenol
Phenols - chemistry
Phenols - metabolism
Sensitivity and Specificity
Sulfuric acid
Sulfuric Acids - chemistry
Sulfuric Acids - metabolism
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Summary:Among many colorimetric methods for carbohydrate analysis, the phenol–sulfuric acid method is the easiest and most reliable method. It has been used for measuring neutral sugars in oligosaccharides, proteoglycans, glycoproteins, and glycolipids. This method is used widely because of its sensitivity and simplicity. In its original form, it required 50–450 nmol of monosaccharides or equivalent for analysis and thus is inadequate for precious samples. A scaled-down version requiring only 10–80 nmol of sugars was reported previously. We have now modified and optimized this method to use 96-well microplates for high throughput, to gain greater sensitivity, and to economize the reagents. This modified and optimized method allows longer linear range (1–150 nmol for Man) and excellent sensitivity. Moreover, our method is more convenient, requiring neither shaking nor covering, and takes less than 15 min to complete. The speed and simplicity of this method would make it most suitable for analyses of large numbers of samples such as chromatographic fractions.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2004.12.001