Name: Bradlee Blair

Form: 6B-2

Date:

Subject: Biology

Aim: To determine the macronutrients in unknown samples, A, B, and C.

Materials and Apparatus: test tubes, water bath, samples A, B and C, Benedict’s solution,

potassium iodide solution, 5% potassium hydroxide solution, 10% copper sulphate solution,

ethanol, three test tubes, droppers, tongs, beakers, hydrochloric acid, sodium hydrogen

carbonate.

Method:

Biuret Test

1. 2cm of the samples A, B and C were added to their respective test tubes.

2. Equal volumes of 5% potassium hydroxide solution was added to each solution and

mixed.

3. Two drops of 1% copper sulphate solution were then added to the solutions and were

mixed.

4. The colour change of each solution was recorded.

Emulsion Test

1. 2cm of the samples, A, B and C were added to their respective test tubes.

2. 2cm ethanol solution was added to each test tube and then shaken vigorously.

3. An equal amount of cold water was then added and the colour changes were recorded.

KI/I Test

1. Ten drops of the samples B and C were added to their respective test tubes.
 2. Five drops of KI solution were then added to each solution.

3. The colour changes were then recorded.

Reducing Sugar Test

1. Ten drops of the samples B and C were added to their respective test tubes.

2. An equal amount of Benedict’s solution was then added to each solution.

3. The solutions were shaken and then put to boil in an electric water, for 1 minute.

4. The colour changes were recorded.

Non-Reducing Sugar Test

1. 2 cm of Sample C was added to its respective test tube.

2. 1 cm of dilute HCL was added and the solution was boiled for 1 minute.

3. The solution was then neutralized with sodium hydrogen carbonate.

4. The Benedict’s test was then carried out on the solution.

5. The colour change was recorded.

Observation:

Table Showing the results of Tests of the Various Samples

 Sample Test

Buiret Emulsion Benedicts N.R.S Starch

A yes yes - - no

B no no yes - no

C no no no yes yes

Discussion:

Sample A:

Upon the completion of the biuret test (a test that is used to determine the presence of peptide

bonds) and emulsion test (a test to determine the presence of lipids), it was deduced that

sample A contained both proteins and lipids. In the biuret test, the nitrogen atoms in the

peptide chain formed a purple complex with copper (II) ions (Cu²ᐩ ) because of the presence

of dilute copper sulphate in an alkaline solution. In the emulsion test, water was added to the

sample in ethanol where a cloudy white suspension was produced. Adding water to a

solution of the lipid in ethanol resulted in emulsion of lipid droplets in the water. This

reflected light giving it a cloudy white appearance.

Sample B:
Sample B was found to contain reducing sugar as upon addition of Benedict’s solution to the

sample, a red precipitate evolve since the the reducing sugars reduced the soluble blue copper

sulphate, with copper (II) ions to insoluble red-brown copper oxide containing copper (I).

Sample C;

Sample C was found to contain starch and non-reducing sugar. When starch is mixed with

iodine in water, an intensely blue colored starch/iodine complex is formed. This is because

iodine gets stuck in the coils of amylose molecules. Sucrose is called a non-reducing sugar as

it does not reduce copper sulphate, however, if it is first hydrolysed to glucose and fructose, it

will then give a positive Benedict’s Test.

Precautions:

1. All measurements that were taken was read at eye level to ensure that accurate

volumes of samples and solutions was added to get accurate results.

2. Each test tube that was placed in the water bath was placed for the correct amount of

time to allow for accurate results.

Conclusion:

In conclusion, sample A contained both proteins and lipids, sample B contained reducing

sugars and sample C contained starch and non-reducing sugars.