Discussion: There are four major classes of macro-molecules; these are proteins,

carbohydrates, lipids and nucleic acids. As it pertains to food, nucleic acids are not
relevant. Proteins, carbohydrates and lipids are found in various types of food,
separately or in combination with each other. The presence of each of these
macro-molecules can be determined by various procedures grouped under a set of
procedures known as ‘food tests’. A food test may be defined as a method by which
certain components of a food mixture can be detected. Each macro-molecule has its
own procedure(s) for detection. For example the Biuret test is used to detect proteins
and the Benedict’s test is used to detect sugars. The food test is an important analytic
procedure as it is able to detect whether these key nutrients are present in a food
sample or not. So a manufacturer can’t truthfully proclaim that a particular food item
has in one or more of these nutrients, as these procedures can be used to determine the
presence of the nutrients. It can also be used to test foods for possible nutrients whose
nutrient content is unknown. In this experiment various food samples were tested for
the presence of these macromolecules. The food tests carried out were the Biuret test,
Benedict’s test (reducing and non-reducing sugar test), emulsion test and iodine test.
These test were carried out on five unknown food samples.

The results obtained showed that at least one of the unknown food samples tested
positive for each of the food tests carried out. In the iodine test, only unknown sample
A gave a positive result, which was indicated by the blue-black color seen when
iodine was added. This indicated that starch was present. The blue-black color change
observed is due to the starch iodide complex that was formed. The starch-iodide
complex is formed as electrons are transferred between the starch and iodide ions,
forming tri-iodide or penta-iodide ions.The transfer of charge between the starch and
the iodide ion changes the spacing between the energy levels. This change results in
the starch-iodide complex absorbing light at a different wavelength and hence the
sample gaining a different color. As a result a blue black color was observed.

For the Benedict’s test for reducing sugars, a positive result was obtained in test tube
A and a slight positive result in test tube D. This was indicated by the orange color
observed in test tube A and light green color in test tube B. the light green color
observed meant that the concentration of sugar was very low the solution. The main
component of Benedict’s solution is copper sulphate which contains copper (II) ions
(Cu2+). The reducing sugar possesses an aldehyde group that can reduce the Cu2+ ions
to Cu+ ions (copper I ions). The copper (II) ions are blue and the copper (I) ions are
red in color. As a result when Benedict’s solution is added to a solution containing a
reducing sugar, the copper (II) ions are reduced to insoluble copper (I) ions in the
form of compound copper (I) oxide. This accounts for the brick-red color and
precipitate seen.

A positive result was also obtained for the Benedict’s test for non-reducing sugar test.
This was indicated by the brick-red color observed in unknown solution X. Unlike
reducing sugars that have a free aldehyde group non-reducing sugars don’t and as a
result does not give a positive Benedict’s test unless modifications are made. Hence
the reason for the addition of hydrochloric acid and sodium bicarbonate. reducing
sugars (glucose) are monosaccharides and disaccharides (e.g. sucrose) are
non-reducing sugars. When non-reducing sugars are boiled in hydrochloric acid, the
bond (glycosidic bond) that joins the two monomers are broken producing back the
two constituent monosaccharides (reducing sugars). hence, when Benedict’s solution
is added the aldehyde group in present in the sugar will reduce the copper (II) ions of
the Benedict’s solution to copper (I) ions. Thus causing a brick-red color to be
obtained. The sodium bicarbonate was added to neutralize the solution after
hydrochloric acid was added. This was done because the reduction of the copper(II)
ions will not take place in acidic conditions.

A purple color was observed in unknown food sample B for the Biuret test. This
indicated the presence of proteins. The key components of the Biuret reagent are
sodium hydroxide and hydrated copper (II) sulphate. The sodium hydroxide is added
to raise the pH of the solution to alkaline levels. The copper (II) ions plays an integral
role in this process. If peptide bonds are present in this alkaline solution, the copper
(II) ions will form a coordination complex with four nitrogen atoms from peptide
bonds (bonds in the protein structure). The coordination complex causes the solution
to have a violet/purple color. This is because it absorbs light at 540nm.

A positive result was also obtained for the emulsion test. This was due to the
milky/cloudy white color observed in unknown food sample E. Lipids are non-polar
organic compounds. As a result they will dissolve in organic solvents such as ethanol
(alcohol) which is also non-polar. When ethanol is added to a sample containing lipids,
it extracts the lipids from the sample. The lipid molecules scatter light as it passes
through the sample causing it to appear white and cloudy.

In the experiment a possible limitation might be the amount of food sample and
reagents used. Certain foods contain more of a particular nutrient than other foods. As
a result if the quantity of nutrient is very the small then more of that sample as well as
more reagent should be used and maybe the results would have been different. During
the experiment it was ensured that the hydrochloric acid was handled with caution as
it is corrosive. A test tube holder was used to hold the test tubes, which were also held
away from the body and the other students near the water bath.

Conclusion: The nutrient protein was present in unknown food sample B, whereas,
starch and reducing sugar were present in unknown food sample A; reducing sugar
was also present in unknown food sample D; unknown food sample E contained lipids
and unknown food sample X contained non-reducing sugar.