The results for the Biuret’s Tests (Figure 1) shows that the solution didn’t

react at all. The solution should have turned violet because of the presence
of peptide bonds -- the chemical bonds that hold amino acids together. The
proteins detected must have at least three amino acids, which means that
the protein must have at least two peptide bonds. The reagent’s copper
ions, with a charge of +2, are reduced to a charge of +1 in the presence of
peptide bonds, causing the color change.

Figure 1. Solution for Biuret’s

Test

For Iodine Test, a blue violet coloration also is expected for a positive result. This indicates that the mixture
has a presence of starch. The figure below shows the data obtained.

Figure 2. Spot test for saliva, iodo-starch complex

It can be seen that test tube A showed negative results while test tube B and C acquired positive results.
The starch-iodide complex is formed as charge - recall electrons are charged particles – is transferred
between the starch and iodide ions - tri-iodide or pentaiodide. The transfer of charge between the starch and
the iodide ion changes the spacing between the energy levels/ orbitals. This change results in the starch-
iodide complex absorbing light at a different wavelength - than any other species aforementioned - resulting
in an intense purple colour; biologists call this colour blue-black.

Standard Protein 0.4

Concentration
Concentration Absorbance 0.3

X Y 0.2
0 0.076 0.1
2.5% 0.136
0
5% 0.175
0 5 10 15 20 25
10% 0.152
Absorbance
20% 0.368
Table 3. Result of absorbance of enzyme using UV- Graph 1. Relationship between the concentration and
visible spectrophotometer absorbance data

The trend above shows the scatterplot of the concentration and absorbance of saliva solution analyzed using
UV-visible spectrophotometer.