Reducing values: dinitrosalicylate gives over-oxidation and invalid results whereas copper bicinchoninate gives no over-oxidation and valid results

Abstract

A comparative study was made between two carbohydrate reducing value methods, a relatively old, highly alkaline, 3,5-dinitrosalicylic acid (DNSA) method and a relatively newer, low alkaline (pH 10.5), copper bicinchoninate (CuBic) method. Reducing values for a series of equimolar amounts of maltose−maltohexaose, isomaltose−isomaltohexaose, and cellobiose−cellohexaose were compared by the two methods. The DNSA method gave over-oxidation for equimolar amounts of all three of the oligosaccharide series. The amount of oxidation increased as the sizes of the oligosaccharides increased, giving inflated, inaccurate reducing values. The CuBic method gave constant reducing values, for equimolar amounts of the oligosaccharides, indicating that there was no over-oxidation, as the sizes of the oligosaccharides were increased. The two methods were used to determine the number average molecular weights (MWn) for six polysaccharides. The DNSA method was not able to determine the MWn for any of the polysaccharides tested due to the low sensitivity of the method, compared with the CuBic method that did not give over-oxidation and gave reasonable MWn values for all six of the polysaccharides tested.