Reaction between albumen and 3,3',5,5'-tetramethylbenzidine as a method to evaluate egg freshness

The 3,3',5,5'-tetramethylbenzidine colorimetric test is proposed as a method for the evaluation of shell egg freshness. A simplification of the original method was tested on albumen, and the modified method was used to study the development of albumen-tetramethylbenzidine reaction in eggs stored at three temperatures for different times. The stability of the reagent solution and the repeatability of the analytical method were also studied. The reaction mechanism showed the predominant role of iron ions on the colorimetric response, and the secondary role of copper ions added to the reaction mix in order to enhance the response. The response coefficient of variation, due to the natural variability in eggs laid by hens of two breeds and different ages, was 10.3%, which is similar to the value obtained for furosine, but lower than those previously observed for other egg freshness indices such as Haugh Unit and air-cell height.