Title: Paper Chromatography

Aim: To determine the different coloured components of methyl orange using paper chromatography

Apparatus and materials: methyl orange , equal 10cm x 3cm stips of chromatography paper, beaker of
water, wooden splint,ruler,capillary tube, white tile

Diagram:

DIAGRAM SHOWING HOW APPARATUS WAS SET UP TO DETERMINE THE DIFFERENT

COLOURED COMPONENTS OF METHYL ORANGE USING PAPER CHROMATOGRAPHY

Procedure:

1. Cut a 10cm long and 3cm wide strip of chromatography paper.

2. Draw a baseline and top line 2cm away from the edge of the chromatography paper.
3. Place the strip of chromatography paper on a white tile.
4. Using a fine capillary tube place a dot of methyl orange on the middle of the baseline.
5. Place the chromatography paper in a beaker of water keeping it in place with a wooden splint with
only the baseline being submerged in the ethanol.
6. Remove the strip of chromatography paper when the water has travelled to the top line and label
the different coloured components
7. Allow the chromatography paper to dry
8. Measure the distance travelled by the solute and solvent.
9. Observe the different colour components of the methyl orange.
 Variables:

Manipulated/Independent Variables:

Responding/Dependant Variable:

 Retention factor
 Different coloured spots which appears

Controlled Variable:

 Volume of solvent
 pH of water
 Dimensions and type of chromatography paper
 Size of dot (solute) placed on the base line of the chromatography paper

Results:

TABLE SHOWING THE RESULTS OBTAINED FROM DETERMINING THE DIFFERENT COLOURED
COMPONENTS OF METHYL ORANGE

DISTANCE MOVED DISTANCE NUMBER AND POSITION OF SPOTS COLOUR OF SPOTS

BY SOLUTE/CM MOVED BY OBSERVED
SOLVENT/CM

A (blue
pigment)

B(yellow
pigment)
Calculations:

Calculation for Retention Factor

The amount that each component of a mixture travels can be quantified using retention factors (Rf). The
retention factor of a particular material is the ratio of the distance the solute travelled to the distance the
solvent travelled. It can be calculated using the formula:

Retention factor (Rf) of solute A= distance travelled by solute A

Distance travelled by solvent

Retention factor (Rf) of solute B= distance travelled by solute B

Distance travelled by solvent

Discussion

Paper chromatography is used to separate mixtures of coloured compounds by

making use of differences in the solubility of the compounds. Paper
chromatography was carried out on chromatography paper. A spot of the methyl
orange was placed at the baseline on a strip of chromatography paper and the
paper was then placed upright in water. As the solvent soaked up the paper, it
carried the methyl orange with it. Different components of the mixture moved at
different rates. This therefore separated the mixture out.

. In the experiment, the methyl orange was distributed between a stationary phase
and a mobile phase. The stationary phase was the ethanol trapped between the
cellulose fibres of the paper. The mobile phase was the developing solution that
travelled up the stationary phase, carrying the samples with it. Components of the
sample separated readily according to how strongly they adsorbed onto the
stationary phase versus how readily they dissolve in the mobile phase.

When the methyl orange was placed on the paper, the colours separated from the
sample by placing one end of the paper in water. The solvent diffused up the
paper, dissolving the various molecules in the sample according to the polarities of
the molecules and the solvent. The sample of methyl orange contained more than
one colour, that means it must have more than one kind of molecule. Because of
the different chemical structures of each kind of molecule each molecule had at
least a slightly different polarity, giving each molecule a different solubility in the
solvent. The unequal solubility causes the various colour molecules to leave
solution at different places as the solvent continues to move up the paper. The
more soluble a molecule is, the higher it will migrate up the paper.From the
experiment it was found that the blue pigment was more soluble than the yellow
pigment.

Precautions:

1.) It was ensured that the strips of chromatography paper was cut precisely
2.) It was ensured that only the baseline was submerged in the solvent
Source of error: Parallax error occurred when measuring the distance travelled by the solute
and solvent

Limitations :The pigments did not completely separate before the solvent reached the topline

Conclusion: It is concluded that methyl orange separates into two coloured pigments; a blue
pigment and yellow pigment. It is also concluded that the blue pigment is more soluble than the
yellow pigment