1.

Tittles:

Paper Chromatography

2. Objectives

- To separate the pigments into bands on the filter paper.

- To determine the distance travelled by the pigments in the filter paper.

3. Introduction

Basically speaking, pigment is a substance that absorbs light and change the color into the one
within the spectrum. The basic example is the rainbow where it absorbs light and produce color
which is have different wavelength. The pigment also has the same concept where it absorbs
light and emitted the color that based on the wavelength that the pigment have.

The simple example of the pigment in biology is the pigment in plant especially the leaf
parts. Although we see it only have one color, but it actually consist several other pigment that
have different color. The color of green in the leaf is the primary pigment. There are basically
two common pigment that can be found in a leaf that is chlorophyll a and chlorophyll b. These
chlorophylls are green pigment molecules the various colored lights such as red, blue, orange
except for one, which is green light as it is reflected back instead of being absorbed.

In this experiment we will use paper chromatography technique. This technique basically
depends on the solubility of the pigment in a solvent. For example if a pigment has the ability
dissolve in water (hydrophilic) will likely has less solubility in the hydrophobic solvent. This
technique take this advantage to make it visual able which pigment has less or more solubility.
It’s also helping the scientist to identify the type of pigment in the plant.
4. Apparatus and materials

Microburette, filter paper, cork stopper, pin, ruler, pencil, red spinach, green spinach, blue
marker, test tube

5. Procedure

1. The microburette was used as the dropper to drop the chloroplast extract on the prepared
chromatography paper.
2. The extract was dropped at about 1.0 cm from the pointing end of the paper. The drop was
then dried using a hair dryer and the process was repeated for 3-4 times until one small dot of
thick pigment was available.
3. The paper strip was attached at the cork stopper using a pin. The strip was vertically placed
straight into the test tube containing the solvent.
4. The solvent was left to move on its own and the paper was removed before the solvent front
would reach the top of the chromatography paper.
5. The last range of the solvent was marked with a pencil

.6. Results

Figure 6.0: Results of the experiment.

 Distance Distance
Pigment Colour traveled by traveled by Rf value Type of
compound(cm) solvent(cm) pigment

2.7 8.4 0.32 Chlorophyll b

1 Yellow -
green

orange 8.1 8.4 0.96 carotene

2

Table 6.0: The result for the red spinach

Distance Distance
Pigment Colour traveled by traveled by Rf value Type of
compound(cm) solvent(cm) pigment

5.5 7.0 0.78 Chlorophyll a

1 blue –
green

orange 6.4 7.0 0.91 carotene

2

Table 6.1: Result for the green spinach

Distance Distance
Pigment Colour traveled by traveled by Rf value Type of
compound(cm) solvent(cm) pigment

5.1 7.3 0.70 -

1 blue

Table 6.2: Result for the blue marker

 The formula for Rf

7. Discussion

In this experiment there is not much step so it’s hard to make an error or commit a mistake. The
crucial step is when you want to drop the sample onto the chromatography paper. You must
make sure the drop is not too big and it is important to make the drop to be a multiple layer.
Don’t forget to dry it first using a dryer each time you apply the drop.

From the result we can see that carotene has the highest Rf in each of the plant. Its also
prove that that carotene is more soluble than the other pigment. So, how do we decide the type of
the pigment? To decide the type of pigment you must observe the color of the pigment first. If
the color of the pigment is orange, the pigment is carotene. If the pigment has a yellow green
color, it is Chlorophyll b. If it is blue–green the pigment is Chlorophyll a. So, we can see that red
spinach and green spinach has different type of pigment. For the Rf values, it is indicate how
soluble it is in the solvent. Carotene is the most soluble than the other pigment.

As for blue marker experiment, we want to see just how soluble it is in the solvent.

8. Conclusion

As for the conclusions, the objectives has be achieve by using paper chromatography technique.