Sumehra PAG 6.

3 9/1/23

Investigation using thin layer chromatography to separate photosynthetic pigments

Solvent Front (distance travelled by solvent) = 7cm

Table 1:

Colours: Distance travelled from lower Rf Value calculated Pigment

pencil line (n/7cm)
1. Grey 0.75cm 0.11 N/A
2. Yellow 1.11cm 0.16 N/A
3. Yellow 1.65cm 0.24 N/A
4. Orange 2.3cm 0.33 N/A
5. Green 2.65cm 0.38 N/A
6. Dark Green 3cm 0.43 Chlorophyll B
7. Faint Yellow 5.15cm 0.74 Xanthophyll
Extension Questions:

 Why is a small spot of pigment needed on the TLC plate?

A small spot is needed to ensure it doesn’t spread to other spots and so it can separate
properly as it goes up the TLC plate.

 Why is propanone used as the extraction solvent?

It is slightly polar and so can dissolve polar substances, such as the pigment.

 Why is the chromatogram run in the dark?

Because the pigment is photosynthetic, the light can affect the result of our chromatogram
and also to make it easier to see the different colours shown on the chromatogram.

 How could you find out whether the separated pigment spots you have observed on
your chromatogram are single, pure pigments or a mixture of two or more?

We can compare the chromatogram to another chromatogram with single pigments to see
which pigments are shown on our chromatogram. We could also use our Rf values to
compare with the Rf values of pigments and see similarities between both datas. We can
also do the experiment again with a different solvent to separate pigments that were in a
mixture and then identify them.