Direct and Indirect Determination of True Protein Content of Milk by Kjeldahl Analysis: Collaborative Study

Currently, the reference procedure for determination of the "protein" content of milk is based on measurement of the total nitrogen content of milk by the Kjeldahl method (AOAC method, 920.105). About 6% of the total nitrogen content of milk Is nonprotein nitrogen. Therefore, total nitrogen multiplied by the conversion factor 6.38 overestimates the true protein content of milk on average by about 6%. In the present study, new direct and Indirect methods were developed for measurement of the true protein content of whole milk by Kjeldahl nitrogen determination. Both new methods are sample preparation procedures used to fractionate the nitrogen-containing compounds In milk prior to measurement of the nitrogen content of these fractions by Kjeldahl analysis. The collaborative study consisted of 9 pairs of blind duplicate milk samples that were analyzed for total nitrogen, nonprotein nitrogen, and protein nitrogen by each of 10 laboratories. Both methods for true protein measurement (direct and Indirect) gave acceptable statistical performance characteristics and good agreement between methods. The new direct method requires about half the laboratory analysis work of the indirect method (i.e., total minus nonprotein nitrogen). The methods have been adopted official first action by AOAC as (1) a new method for nonprotein nitrogen determination in milk, (2) a new method (direct) for determination of protein nitrogen content of milk, and {3) an alternative method (indirect) for determination of protein nitrogen content of milk.