DISCUSSION

In this experiment, crude protein of sample which is the salted fish is determined by using
Kjeldahl method. This method only determines crude protein because this method measures the
total nitrogen content in protein which includes protein nitrogen and non-protein nitrogen in the
form of NH4+ . This total nitrogen that indicated by the free amino acid, amide, purine, alcohol
and others compound that contain nitrogen is accounted as crude protein. In the Kjeldahl method,
proteins and other organic food components in sample are digested with sulphuric acid in the
present of catalysts. The total organic nitrogen is converted to ammonium sulphate. The digest is
neutralized with alkali and distilled into a boric acid solution. The borate anions formed are
titrated with standardized acid, which is converted to nitrogen in the sample. The result of the
analysis represents the crude protein content of the food since nitrogen also comes from nonprotein components [Nielsen S.S., 2003].
During the experiment, the solution was digested in the digester in the fume area. The
digestion is completed when the black color solution become clear and has blue-green color.
This happen when all hydrocarbons are oxidized and nitrogen change to ammonium ion (NH4+)
form, which gives a clear color solution [Pomeranz, Y., 1987]. After cooling, sodium hydroxide
solution was added slowly to form layers. The top layer is acid while the bottom layer is alkali.
In the distillation machine, the nitrogen gas moves upward because the machine produces hot
vapor. Boric acid then capture the nitrogen gas and change the color from purple to green.
heat
(NH4) 2SO4 + 2NaOH --------- 2NH3 + Na 2SO4 + 2H2O
Ammonia gas

The ammonia was titrated with 0.05N sulphuric acid (H2SO4 ) solution. H2SO4 was used
because it is a strong acid and a good oxidizing agent. It oxidizes all the organic compounds and
itself is reduced to SO2. Compared to HCl, H2SO4 gives a higher oxidizing rate. In the presence
of sulfuric acid, all the nitrogen elements will be converted into ammonium sulfate after full
digestion [James, 1995]. The ammonium sulphate produced is non toxic substance.
From this experiment, the percentage of crude protein that we obtained from the sample
which is salted fish are 34.38% and 36.34%. By comparing the results obtained from another
group (Group 4) as they used the same sample, the range of percentage of crude protein they
have obtained is between 34% to 36%. This can conclude that the results obtained is in
acceptable range. By comparing the results we obtained with Group 1, the range of percentage of
crude protein they have obtained is around 28%. The results are different due to different sample
used. Group 1 used soya bean as their sample. 28% of crude protein indicates that soya bean has
less nitrogen compared to salted fish.
There are many advantages of Kjeldahl method. This method is applicable for most of the
food samples, easy to handle and low cost. It also gives accurate result for rough protein content
and used as standard method for comparison against all other methods. However, this method is
time consuming to carry-out [Pomeranz, Y., 1987]. It does not give a measure of the true protein,
but only determine crude protein content. Kjeldahl method has low precision compared to other
method such as Biuret method. The use of concentrated sulfuric acid at high temperatures poses
a considerable hazard. Different proteins need different correction factors due to their sequences
of amino acids.