Enzymatic spectrophotometric reaction rate determination of aspartame

Aspartame is an artificial sweetener of low caloric value (approximately 200 times sweeter than sucrose). Aspartame is currently permitted for use in food and beverage production in more than 90 countries. The application of aspartame in food products requires development of rapid, inexpensive and accurate method for its determination. The new assay for determination of aspartame was based on set of reactions that are catalyzed by three different enzymes: ?-chymotrypsin, alcohol oxidase and horseradish peroxidase. Optimization of the proposed method was carried out for: (i) ?-chymotrypsin activity; (ii) time allowed for ?-chymotrypsin action, (iii) temperature. Evaluation of the developed method was done by determining aspartame content in ?diet? drinks, as well as in artificial sweetener pills.