The objective of our research was to modify the current
indirect casein method for bovine milk to enable it
to be applied to bovine milk, bovine milk concentrates,
and milks of other species that contain a protein concentration
up to 9% (wt/wt). Our work used a series of
bovine milk concentrates from about 3 to 9% protein
with the same casein as a percentage of true protein to
determine the amount of buffer required and pH of the
noncasein nitrogen (NCN) filtrate to achieve consistent
estimates of casein and casein as percent of true protein.
As the concentration of protein in milk increased
(either in bovine milk concentrates or in milks of other
species), the amount of buffer needed for the NCN sample
preparation method to achieve a filtrate pH of 4.6
increased. In the first part of the study using a series
of bovine milk concentrates, it was demonstrated that
the method gave more consistent predictions of casein
as a percentage of true protein when the final NCN
filtrate pH was between 4.5 and 4.6 at 38°C. When
the amount of buffer added to the sample was not sufficient
(i.e., the filtrate pH was too high), the filtrates
were not clear. A polynomial equation was developed
for prediction of the amount of acetic acid or sodium
acetate buffer required to achieve pH 4.5 to 4.6 for
milk protein concentrations from 3 to 9% protein using
bovine milk and milk concentrates. When the equation
developed using cow milk was applied to goat, sheep,
and water buffalo milks, it correctly predicted the volume
of reagents needed to achieve a final NCN filtrate
pH of 4.6 at 38°C. We also verified as part of this work
that the ability to measure NPN content of milk was
not influenced by protein content of milk in the range from 3 to 9% protein. The results of this study will be
used as the basis for proposed changes in the official
methods for measurement of the casein content of milk
to expand the scope of the method so it can be used to
achieve accurate results for milk concentrates and milks
of other species.
Key words: casein, Kjeldahl, noncasein nitrogen,
nonprotein nitrogen