An o-toluidine method for detection of carbohydrates in protein hydrolysates.
The o-toluidine high-performance thin-layer chromatography (HPTLC) method for detection of reducing sugars has been demonstrated to be a facile method for composition analysis of protein hydrolysates with a maximum sensitivity range of 50-100 pmol. The solution phase reaction of o-toluidine with reducing sugars has been previously used for spectrophotometric detection of glucose at 480-630 nm. In contrast, the heterogeneous reaction of o-toluidine with reducing sugars resolved by thin-layer chromatography produces chromophoric derivatives which have a broad absorbance at 295 nm. Detection of these chromophoric derivatives is achieved by uv diffuse reflectance scanning densitometry. It is demonstrated that detection limits of less than 10 ng can be achieved by using HPTLC plates and is therefore equal or more sensitive for some sugars than recently reported high-pressure liquid chromatography methods using amperometric or fluorescence detection.
Morcol
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Department of Chemical Engineering, Virginia Polytechnic Institute and State University, Blacksburg 24061.
Velander
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WH
D000143:Acids; D002241:Carbohydrates / Q000032:analysis*; D002855:Chromatography, Thin Layer / Q000379:methods; D006801:Humans; D006868:Hydrolysis; D007074:Immunoglobulin G / Q000737:chemistry*; D007700:Kinetics; D015394:Molecular Structure; D011486:Protein C / Q000737:chemistry*; D011516:Prothrombin / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014052:Toluidines
D000143:Acids; D002241:Carbohydrates; D007074:Immunoglobulin G; D011486:Protein C; D014052:Toluidines; D011516:Prothrombin; C023622:2-toluidine