BENEDICT’S TEST.

Objectives
To perform test to detect the presence of reducing sugar.

Introduction
Benedict’s test allows us to detect the presence of reducing sugars (sugars with
a free aldehyde or ketone group). All monosaccharides are reducing sugars
since all of them have active carbonyl group. Some disaccharides that exposed
carbonyl group are also reducing sugars but less reactive than monosaccharides.
By mixing the sugar solution with Benedict’s solution and heating them, a redox
reaction will occur. In this case, the reducing sugars as the reducing agent. The
copper (II) sulphate present in Benedict’s solution reacts with electrons from
aldehyde or ketose group of reducing sugars to form cuprous oxide, a red brown
precipitatate.

Hypothesis
If the reducing sugar is present in the food sample, the food sample will change
the original colour sample to brown when Benedict’s solution is added and heat
is apply.

Apparatus and Material

Test tubes, dropper, test tube rack, beaker, thermometer, Bunsen burner, tripod
stand, wire gauze, honey, onion juice, potato juice, sucrose solution, glucose
solution, starch solution, reducing-sugar solution, distilled water and Benedict’s
solution.

Procedures
1. 2mL of each of the sample is added into 8 clean test tubes. All the test
tube are labeled.
2. 2 drops of benedict’s solution is added into each test tube. The mixtures
are shaken gently.
3. Any colour change is observed.
4. Meanwhile, a beaker is half-filled with water. The water bath is heated until
60˚
5. All the test tubes are inserted into the water bath.
6. Any colour change is observed after about 5 minutes.
7. All the observation are recorded.
Results
Tube Solution Benedict’s colour reaction.
1 Honey Blue to light brown
2 Onion juice No colour change
3 Potato juice Blue to black with white precipitate
4 Sucrose solution No colour change
5 Glucose solution Blue to brown
6 Starch solution No colour change.
7 Reducing sugar solution Blue to brown
8 Distilled water No colour change

Discussion
Onion juice, sucrose solution, starch solution and distilled water did not show any
colour change. This show that they do not contain reducing sugar. Meanwhile,
honey, potato juice, glucose solution and reducing sugar solution show colour
changes. Honey, glucose solution and reducing sugar solution turn brown
because the copper(II) sulphate presence in the Benedict’s solution reacts with
electrons from aldehyde and ketone group to form cuprous oxide, a red brown
precipitate.

Conclusion
Reducing sugar is present in honey, potato juice, glucose solution and reducing
sugar solution.

Reference
www.biosci.ohiou.edu/introbioslab/Bios170/.../benedict.htm
IODINE TEST.

Objective
To detect the presence of carbohydrate in food sample.

Hypothesis
If starch is present n food sample, the food sample will turn to dark blue when
iodine solution is added to it.

Introduction
Iodine test is used to test for the presence of carbohydrates. Iodine solution
(iodine dissolve in an aqueous solution of potassium iodide) reacts with starch
producing a deep purple colour. The iodine test is used to distinguish starch from
monosaccharides, disaccharide and other polysaccharide. Due to its unique
coiled geometric configuration, it reacts with iodine to produce a blue black
colour and tests positive. A yellowish brown colour indicates that the test is
negative.

Apparatus and materials

Test tubes, droppers, test tubes rack, onion juice, potato juice, sucrose solution,
glucose solution, starch solution, reducing-sugar solution, distilled water and
iodine solution.

Procedures
1. 2mL of each samples are added into clean test tubes. The test tubes are
recorded.
2. 2 drops of iodine solution are added into each test tube. The mixtures are
shaken gently.
3. Any colour change are observed.
4. All the observations are recorded.

Result
Tube Solution Iodine colour change
1 Onion juice No colour change
2 Potato juice Yellow to blue
3 Sucrose solution No colour change
4 Glucose solution No colour change
5 Starch solution Yellow to blue
6 Reducing sugar solution No colour change
7 Distilled water No colour change
Discussion
Onion juice, sucrose solution, glucose solution, reducing sugar solution and
distilled water do not show any colour change. This indicate that they do not
contain starch. Meanwhile, the sample that change from yellow to blue show that
the starch is present in them since when iodine solution reacts with starch it will
produce blue black colour.

Conclusion
Starch present in potato juice and in starch solution.

Reference
en.wikipedia.org/wiki/Iodine_test
BIURET TEST.

Objective
To detect the present of of protein.

Hypothesis
If protein is present in food sample, the food sample colour will turn to violet
when Biuret’s reagent is added into it.

Introduction
Biuret test is a chemical test used to detect the presence of peptide bond.
Biuret’s reagent is made of sodium hydroxide and copper (II) sulphate. In a
positive test, copper (II) ion is reduced to copper (I) ion which form a complex
with the nitrogen and carbon of the peptide bond in an alkaline solution. The blue
reagent turns violet in the presence of protein and turn to pink when combined
with short-chain polypeptides. In this test for protein, there is a reaction between
the copper (II) ions and the amino group in the peptide bond.

Apparatus and materials

Test tubes, dropper, test tube rack, egg white, egg yolk, honey, corn oil, starch
solution, sugar solution, distilled water and Biuret’s reagent.

Procedure
1. 2mL of each sample is added into eight clean test tubes. All the test tubes
are labeled.
2. 4 drops of Biuret’s reagent are added into each of the test tubes. The
mixtures are shaken gently.
3. Any colour change is observed.
4. All the observations are recorded.

Result
Tube Solution Biuret colour reaction
1 Egg white Turn to purple
2 Egg yolk Turn to greenish yellow
3 Honey No colour change
4 Corn oil Turn to light yellow precipitate
5 Starch solution No colour change
6 Protein solution Turn to purple
7 Distilled water. No colour change
Discussion
Egg white, and protein solution did show change in colour: from their original
colour, they turn to purple. Egg yolk also contain protein but its colour did not turn
to purple when Biuret’s reagent is added. This is due to lack of peptide bond in
egg yolk. Meanwhile honey, starch solution and distilled water did not show any
change because they do not contain protein.

Conclusion
Protein present in egg white, egg yolk and in protein solution.

Reference
www.biosci.ohiou.edu/introbioslab/Bios170/170.../biuret.htm
SUDAN (III) TEST AND EMULSION TEST.

Objective
To detect the present of lipid.

Hypothesis.
If lipid is presence in food sample, the food sample will turn to red when Sudan
(III) solution is added into it.

Introduction.
Sudan III test.
Sudan III test detects the hydrocarbon groups that are remaining in the molecule.
Due to the fact that the hydrocarbon group are non polar and stick tightly
together with their polar surrounding, it is called a hydrophobic interaction and
this is the basis for Sudan III test. Since lipid molecules are insoluble in water,
consist mainly of carbon and hydrogen, with few oxygen containing functional
groups, they are considered as hydrophobic. Hence, if the result is visible orange
appear, it tested positive for lipid.
Emulsion test.
Lipids are heterogeneous group of compound that are categorized by the fact
that they are soluble in non-polar solvents such as alcohol and are relatively
insoluble in water. This characteristic of lipid is used in the emulsion test.

Apparatus and materials.

Test tubes, dropper, test tube racks, corn oil, honey, starch solution, egg yolk,
distilled water, lipid solution, tap water and Sudan (III) solution.

Procedures.
1. 2mL of each sample is added into eight clean test tubes. All the test tubes
are labeled.
2. 4 to 5 drops of Sudan (III) reagent are added into each of the test tube.
The mixtures are shaken gently.
3. Any colour change is observed.
4. All the observation are recorded.
Results.
Tube Solution Sudan III colour reaction.
Sudan III test
1 Corn oil+water No colour change
2 Corn oil+water+Sudan III Corn oil layer turns red
3 Honey+water+Sudan III No colour change
4 Starch solution+water+Sudan III No colour change
5 Egg yolk+water+Sudan III No colour change
6 Distilled water+water+Sudan III No colour change
7 Lipid solution+water+Sudan III Lipid solution turns red
Emulsion test
8 Corn oil+dilute alcohol solution Three layers formed
(80% ethanol solution.)

Discussion
There is no change in test tube 1and 6. In test tube 2, the corn oil layer turns red
which shown that lipid is present. In test tube 3, two layer form which is light
orange on the top and dark orange on the bottom. It is because the density of
honey is higher than water where the light orange layer is water and dark orange
is honey. In test tube 4, the solution just turn cloudy orange. In test tube 5, the is
no change in colour but egg yolk do has lipid. The situation occur because. In
test tube 7, two layer formed whish are lipid layer and water layer. The lipid layer
turn to dark orange. It suppose to turn to red but due to some error the result is
not accurate. This is might happen because the lipid solution is too dilute. In test
tube 8, three layers formed which is ethanol layer at the top, corn oil at the
middle and water at the bottom of the test tube. Thus, lipid is present.

Conclusion
Lipid present in corn oil, egg yolk and lipid solution.

Reference
www.scienceteacherprogram.org/biology/Lillien02.html