Introduction:

Carbohydrate is a molecule consisting of carbon, hydrogen, and oxygen atoms.

Carbohydrates are divided into four groups: monosaccharides, disaccharides,
oligosaccharides, and polysaccharides. Monosaccharides are the most basic units of
carbohydrates. Examples of monosaccharides include glucose, fructose and galactose.
Monosaccharides are the building blocks of disaccharides and polysaccharides. A
disaccharide is the carbohydrate formed when two monosaccharides undergo a
condensation reaction while polysaccharides are produced when monosaccharides bind
together by glycosidic bonds.

Protein consists of carbon, hydrogen, oxygen and nitrogen atoms. It is made up of

one or more chains of amino acids, by joining the amino group of one amino acid with
the carboxyl group of another with a peptide bond. An amino acid join with another
amino acid to form peptide, two or more peptide that joined together will then produce
polypeptide. Different protein is made up of different sequence of amino acids. Also,
cells can build up thousands of protein from only 20 different amino acids.

Lipid is an organic compound that consists of carbon, hydrogen and oxygen. It is

one type of ester results from chemical reaction of fatty acids and alcohol.

Hypothesis:

Exercise 2.1: Carbohydrate

Glucose is a monosaccharide and reducing sugar.

Exercise 2.2: Protein

Albumin, egg yolk, egg white and milk contain protein.

Exercise 2.3: Lipid

Palm oil, corn oil and sunflower oil contain fats.

Exercise 2.1: Carbohydrate

Methods:

1) Molisch Test
a. Five drops of alfa naftol solution 5% is added into 2ml of each sucrose 2%,
glucose 2%, maltose 2% and starch 5% in 4 different test tubes. The test tubes
are shaken gently to mix the mixture.
b. 1ml of concentrated sulphuric acid is placed slowly into each of those test
tubes.
c. The colour changes are observed and the results are recorded.

2) Barfoed Test
a. 5ml of Barfoed tester is placed into 4 different test tubes.
b. 2ml of sucrose2%, glucose 2%, maltose 2% and starch 5 % are added into
each test tube separately.
c. The test tubes are soaked in boil water for 30 seconds.
d. The results are recorded.

3) Benedict Test
a. 1ml of sucrose 2%, glucose 2%, maltose 2% and starch 5% is placed into each
test tube separately.
b. 2ml of Benedicts solution is added into each test tube and mixed by gently
shaking.
c. Then, the test tubes are heat briefly in boiling water for 2 minutes.
d. The test tubes are cooled down. Any changes are observed and the results are
recorded.

4) Polysaccharides
a. 5 drops of sucrose 2%, glucose 2%, maltose 2% and starch 5% are placed onto
the tile patches
b. One or two drops of Iodine-potassium iodide solution are added and any
changes are observed.

Results:
 Sucrose 2% Glucose 2% Maltose 2% Starch 5%
Positive result Positive result
Positive result Positive result
Cloudy Cloudy
Cloudy Cloudy
solution with solution with
solution with solution with
Molisch Test large amount purple and
purple layer light purple
of light purple white layer
between is layer between
layer between between is
formed is formed
is formed formed
Positive result
Barfoed Test Negative result Red precipitate Negative result Negative result
is formed
Positive result
Negative result Positive result Negative result
Brown
Benedict Test Green solution Peach solution Blue solution
solution is
is formed is formed is formed
formed
Polysaccharid
Negative result Negative result Negative result Positive result
e

Exercise 2.2: Protein

Methods:

1) Biuret test
a. 3ml of albumin solution 5%, egg yolk, egg white and milk are added into 3
different test tubes.
b. 3ml sodium hydroxide 2% is added in and mixed gently.
c. Four drops of cuprum sulphate 0.5% is added and the colour changes are
observed.
d. The result is recorded.

2) Millon test
a. 3ml of albumin solution 5%, egg yolk, egg white and milk are added into 3
different test tubes.
b. Three drops of Millon solution is added into the solution.
c. The changes are observed and the result is recorded.

Result:
 Albumin
Egg yolk Egg white Milk
solution 5%

Two layered Two layered Two layered Two layered

Biuret solution is solution is solution is solution is
test formed with formed with formed with formed with
purple coloured purple coloured purple coloured purple coloured
solution on top solution on top solution on top solution on top
Millon White White White No observable
test precipitate precipitate precipitate change

Exercise 2.3: Lipid

Methods:

1) Acrolein test
a. Five drops of corn oil, palm oil and sunflower oil is placed into three different
test tubes.
b. 1g sodium bisulphate is added in and heated slowly using Bunsen burner.
c. The smell presence is indicated and the result is recorded.

2) Colouring test
a. Three drops of corn oil, palm oil and sunflower oil is placed into three
different test tubes.
b. 5ml of distil water is added and mixed by gently shaking.
c. A few drops of Sudan IV are added. The mixture is carefully mixed by hard
shaking. The mixture is then left for 1 minute and the changes are observed.

Result:

Palm oil Corn oil Sunflower oil

Acrolein test weaker smell strong smell strongest smell
Colouring No observable No observable
Red layer formed
test change change

Discussion:
 During Molisch test, all carbohydrates contained solution shows purple layer but
glucose 2% shows a more rapid result as compared to sucrose 2%, maltose 2% and starch
5%. This showed that glucose is a monosaccharide as monosaccharide will react rapidly
with the reagent. Sucrose, maltose and starch show a negative result in Barfoed test while
only glucose showing a positive test. This is because Barfoed test will only shows
positive result with the presence of reducing monosaccharides. Benedict test will shows
positive result when reducing sugar is present. Therefore, only glucose and maltose show
positive result for Benedict test. However, for polysaccharide test, only starch shows
positive result as starch is the only polysaccharides between sucrose, maltose and
glucose.

Biuret test and Millon test are held up to test for the presence of protein. All
experiments show positive result for Biuret test where two layered solution is formed
with purple coloured solution on top. This proved that albumin, egg yolk, egg white and
milk contained protein. However, for Millon test, only albumin, egg yolk and egg white
show white precipitate. The chemical change of milk is unable to be detected due to the
white coloration of milk.

For lipid test, Acrolein test and colouring test are held up. For Acrolein test, the
smell of burnt grease is detected with sunflower oil that has a stronger smell, followed by
corn oil then palm oil. However, for colouring test, only sunflower oil shows a positive
result by the red layer formed.

Conclusion:

These experiments confirmed the hypotheses. For carbohydrate tests, Molisch test is a
test for presence of carbohydrates and Barfoed test is a test for the presence of
monosaccharides. Benedict test is a test for the presence of reducing monosaccharides
sugar while polysaccharide test is a test for the presence of polysaccharides. For protein
tests, Biuret test and Millon test are tests for proteins. Acrolein test tests for the presence
of fats or glycerin while Colouring test is used to test the presence of lipids.