CHAPTER III

EXPERIMENTAL METHOD
A. Tools and Material
1. Proving the elements that exist in protein

Tools : Materials:
Test Tube
Concentrated NaOH
Drop pipette
White egg
Rack test tube
Soybean bar solution
Clamp test tube
Potato solution
Measuring cup
Potato solution
Spiritus burner
pH universal indicator
2. Albumin 2% Solubility in various solvents
Tools : Materials :
Test Tube
Solution of albumin 2%
Drop pipette
NaOH solution 0,2%
Rack test tube
NaCO3 solution 0,2%
Clamp test tube
HCl solution 0,2%
Measuring cup
Aquades
Vortexs

3. Proving the peptide bond constructed protein through biuret test

Tools :
Test Tube
Drop pipette
Rack test tube
Clamp test tube
Measuring cup
Vortex

Materials :
Solution of protein
NaOH solution 10%
CuSO4 solution 0,01 M
Soybean bar extract
Potato extractsolution
 4. Proving the amino acid through Ninhydrin test
Tools :
Test Tube
Drop pipette
Rack test tube
Clamp test tube
Measuring cup
Spiritus burner
Materials:
Arginin
Ninhydrin solution 0,1 %, protein reactant (3 kinds)
Soybean bar solution
Potato solution
B. Procedure
Proving the elements that exist in protein
1. Putting 5 drops albumin from white egg into dry test tube, then heating up on
spiritus burner directly. Observing the evidence happening (smell, colour, and
water vapor).
2. Putting 5 drops of albumin (white egg) into dy test tube, added with 10 drops
concentrated NaOH, then burning carefully on spiritus burner, and observing the
smell changed.
3. Putting 1 ml potato solution into dry test tube, then heating up on spiritus burner
directly. Observing the evidence happening (smell, colour, and water vapor).
4. Putting 1 ml potato solution into dy test tube, added with 2 ml concentrated
NaOH, then burning carefully on spiritus burner, and observing the smell
changed.
5. Putting 1 ml soybean bar solution into dry test tube, then heating up on spiritus
burner directly. Observing the evidence happening (smell, colour, and water
vapor).
6. Putting 1 ml soybean bar solution into dy test tube, added with 2 ml concentrated
NaOH, then burning carefully on spiritus burner, and observing the smell
changed.
Albumin 2% Solubility in various solvents
1. Prepare 4 test tubes, insert 1 ml of 2 % albumin solution in each test tube.
2. Then add in a test tube :
- First test tube : 1 ml of aquades
- Second test tube : 1 ml of 0,2% NaOH solution
- Third test tube : 1 ml of 0,2% HCl solution
- Fourth test tube : 1 ml of 0,2% NaCO3 solution
 3. Each test tube vortexed for one and a half minutes , leave for a moment
and observe what happens.
Proving the Peptide Bond Constructed Protein through Biuret Test
1. Preparing all the 3 types of protein solution (albumin, extract soybean bar,
and extract potato) and 6 reacts tube
2. Putting each protein solution into each react tube, each solution is 3 ml. (2
reacts tube for albumin, 2 reacts tube for extract soybean bar, and 2 reacts
tube for extract potatoes)
3. Adding 1 ml of 10% NaOH solution into every reacts tube.
4. Homogeny it by using vortex (each solution vortexted in 1.15 minutes)
5. Putting 3 drops 0.01 M CuSO4 into the first three reacts tube (albumin,
extract soybean bar, and extract potato)
6. Stirring the solution in few minutes. If the color do not changes, adding
the solution 1-2 drops again 0.01 M CuSO4
7. Observing the color changes
8. Putting 3 drops Biuret reagent into the second three reacts tube (albumin,
extract soybean bar, and extract potato)
9. Stirring the solution in few minutes. If the color do not changes, adding
the solution 1-2 drops again Biuret reagent
10. Observing the color changes

1. Proving the Amino Acid through Ninhydrin Test

2. Prepare 1 mL of protein solutions (albumin, potato, soybean bar) and arginin.
3. Putting 1 mL of albumin, 1 mL extract potato, 1 Ml extract soybean bar and 5
drops of arginin in the test tube, add each solution by 5 drops of 0,1%
ninhydrin solution.
4. Heat until boiling , then wait until the each solution cool and observe the color
change.