DATE: 2018

PRACTICAL #:

TITLE: FOOD TEST

PROBLEM STATEMENT:
Evaluate the protein content of protein shakes versus local foods high in protein,

HYPOTHESIS:
The protein shakes have a higher concentration of protein than the foods high in protein.
Because all the concentrations of the protein shake will produce a purple or mauve colour to
indicate it has the highest protein content.

AIM
To investigate how much protein is in protein shakes compared to regular foods with high
protein content.

MATERIALS/APPARATUS:
Test tubes, test tube holders, test tube racks, measuring cylinder, mortar and pestle, dropper,
food samples, Biuret solution, distilled water.

PROCEDURE:

1. If the substance to be tested is a solid, it was grinded and made a solution with water.
2. If liquid 1cm3 of liquid was placed in a test tube.
3. The same volume of Biuret solution was added to the test tube.
4. This was mixed by shaking the test tube from side to side.
5. 50 and 25% solutions of the food sample were produced and steps 1 to 4 were
repeated on each solution.
6. Step 1 and 5 were repeated, using the other food sample.
7. The colour changes where observed in each concentration of the different food sample
and the results tabulated accordingly.
VARIABLES:
MANIPULATED –The food sample used in the experiment.
RESPONDING –The colour changes observed when each sample reacts with the biuret
solution.
CONTROL – The biuret reagent used in the experiment.

RESULTS:
TABLE SHOWING THW BURIET TESTS DONE ON AT DIFFERENT
CONCENTARTIONS FOR VARIOUS FOOD SAMPLES
FOOD CONC. OF INITIAL(COLOUR) FINAL(COLOUR)
SAMPLE SAMPLE (%)
Protein shake 100 blue Purple
50 blue Purple
25 blue Purple
cheese 100 Blue Purple
50 Blue Purple
25 blue Pink
milk 100 Blue Purple
50 Blue Purple
25 Blue Pink
peanuts 100 Blue Purple
50 Blue Pink
25 Blue Pink
eggs 100 Blue Purple
50 Blue Purple
25 Blue pink

DISCUSSION
The Biuret test is based on the ability of Cu (II) ions to form a violet-colored chelate
complex with peptide bonds (-CONH- groups) in alkaline conditions.
Lone electron pairs from 4 nitrogen atoms in the peptide bond coordinate a copper (II) ion to
form the chelate complex.
The chelate complex absorbs light at 540 nm so appears violet. Hence a color change from
blue to violet indicates that proteins are present. The greater the concentration of peptide
bonds, the greater the color intensity. If the concentration of peptide bonds is low – such as
when short-chain peptides are present - the color change is from blue to pink.

With this information an experiment was done to determine if protein shakes have a higher
concentration of protein compared to regular foods with high protein content , the results
were as expected. To carry out this experiment each food sample (protein shake, peanut,
cheese, milk, egg) had to undergo the biuret test at different concentrations (100%, 50%, and
25%) to determine which would produce the expected purple or mauve colour at each
concentration as a opposed to a pink colour which would indicate the protein contents are
weak. The protein shake had a colour change from blue to purple at all three concentrations
therefore; it had the highest concentration of peptide bonds. The cheese, milk and produced a
colour change from blue to purple at concentrations of 100% and 50%, and at 25% there was
a colour change of blue to pink, this determined these had the second highest protein
concentrations relative to the protein shake and had similar concentrations of protein. The
peanut had a colour change of blue to purple at only a concentration of a 100% and turned
pink at 50% and 25% proving it had the lowest concentration of protein and peptide bonds.
Based on these finds it can then be said that the protein shake contains a higher concentration
of protein compared to regular foods with high protein content.

REFLECTION
The experiment was a success and most of the results where as expected and some were just
as bit off. Initially when deciding which method to use a few were taken into consideration
such as such as the Bradford assay, BCA assay and Absorbance. However these were not
chosen because it required a colorimetric or spectrometer, and/or other reagents which might
not be in the school’s lab. These methods were also more complicated even though they
might have produced more quantitative results. The method used was based on the Modified
Lowry test which has the ideology that based on the frequency of the colour or how close it is
to the expected purple or mauve the protein contents can be determined. This was done by
using different concentration of the same food products to see which one will not produce the
blue colour as the concentration gets lower, hence less protein. After the experiment was
carried out the hypothesis was proven to be correct as the protein shake produced the purple
colour at all there concentrations. However a few predictions were made previously, as I said
I expected the cheese and the peanut to have the second highest protein concentration relative
to the protein shake and the milk and egg to have the least protein as it contents more water
and dilution would have a more profound effect on its protein contents, however I was proven
wrong by the experiment as the cheese, milk and egg both produced a purple colour at 100%
and 50% while the peanut only produced a purple colour at 25% . This proved the protein
shake had the highest protein concentration, followed by the cheese, egg and milk then the
protein with the least. In essence the hypothesis that reads “The protein shakes have a higher
concentration of protein than the foods high in protein. Because all the concentrations of the
protein shake will produce a purple or mauve colour to indicate it has the highest protein
content” was proven to be correct. However the determinations made about the other food
samples were wrong. The most important thing I leant from this experiment and my research
is that the biuret test has a wide range of quantitative test and that protein shakes are a
essential source of protein.

SOURCES OF ERROR/LIMITATIONS:

1. The mortar pestle might have not been cleaned after each used which may cause
cross- contamination and affect the results.
2. The food sample might have been spoiled or affected by the environment which alter
the protein contains.

PRECAUTIONS:

1. Ensure the food samples used were not processed or alter with to gain the most
accurate results.
2. Clean the mortar and pestle, and test tubes after each biuret test.

CONCLUSION
Based on the results obtained from carrying out the experiment, the hypothesis was proven to
be true that, “The protein shakes have a higher concentration of protein than the foods high in
protein. “