BIOCHEMICAL TESTS FOR BIOMOLECULAR

Objectives :
1.
2.
3.
4.

Carry
Carry
Carry
Carry

out
out
out
out

benedict tests for reducing and non-reducing sugars,

the iodine in potassium iodide solution test for starch
the emulsion test for lipids
the biuret test for proteins

Theory :
All monosaccharides and some disaccharides including maltose and lactose are
reducing sugars. These can be tested for, by adding Benedict's reagent to the sugar and
heating in a water bath. If a reducing sugar is present, the solution turns green, then yellow
and finally produces a brick red precipitate. Non-reducing sugars can also be tested for using
Benedict's reagent but first require addition of an acid and heating to hydrolyse (break apart)
the sugar. The acid must then be neutralised using an alkali like sodium hydroxide before
carrying out the test as described above.
Iodine solution or potassium iodide solution can be used to test for the presence of
starch. A positive result changes the solution from an orange-brown to a blue-black colour. refer to gcse and title biochemical test for carboydrate.
The reagent used to test for proteins is called biuret reagent. It can be used as two
separate solutions of copper sulphate and potassium or sodium hydroxide or as a ready-made
biuret solution. In either case, a purple colour indicates a positive result.
Materials :
1. Test tube
2. Mortar and stamp
3. Spatula
4. Tube rack
5. Gas burner
6. Beaker glass
7. Drop pipette
8. Sample of food :
A. Mineral Water
B. Liquid fructose
C. Sugar
D. Potato
E. Rice
F. Bread

G.
H.
I.
J.
K.

Frying Oil
Egg Yolk
Tempe
Egg white
Mineral Water

Reagents :
1. Benedict solution
2. Dilute HCL
3. Dilute NaOH
4. Dilute NaHCO3
5. Iodine solution
6. Biuret solution
7. Ethanol
8. Aquades

Procedures :
A. Reducing Sugars Test
Add benedicts reagent to a sample (A AND B) and heat it. Make sure the solution
doesnt boil. Carry out tests to identify which unmarked solution is fructose solution
only, which is water only. Look at the result and describe the tests made and the results
obtained.
B. Non-Reducing Sugars Test
Boil the test solution (C AND K) with dilute HCL and then neutralize it with Sodium
hydrogen carbonate. Carry out the reducing sugars test. Look at the result and describe
the tests made and the results obtained.

C. Proteins Test
Add the sample of food (I and J) to keep the alkaline situation by adding NaOH solution.
Use the biuret test on a solution of egg white and tempe. Use the same volumes and
time for unknown solutions and compare the intensity of colour obtained with the
standards.
D. Fats Test
Add the food sample (G and H) with ethanol and shake it until they are mixed. Add the
water and see what is happened and describe the tests made and the results obtained.
E. Starch test
Add the sample of food (D, E, and F) with 3-4 drops of iodine solution. A dark blue
color should be produced