Pigments and Chromatography

Absorbance spectrum: A graph of absorbance of different light wavelengths by various compounds

(i.e. photosynthetic pigments).
Action spectrum: A graph showing the effect of different wavelengths on a process (i.e.
photosynthesis)

Pigment

Colour of Pigment

Colour Absorbed by
Pigment

Relative Rf Value of
Pigment

Chlorophyll a

yellow-green

violet-blue and orangered (400-500nm and

650-700nm)

in between chlorophyll b
and carotenoids

Chlorophyll b

blue-green

violet-blue and yellowred (400-500nm and

600-700nm)

much lower

Carotenoids

xantophylls are yellow

and carotenes are
orange

blue-green (450-500)

- Similarity between absorbance and action spectrums

The spectrums are similar in the location of their peaks. This is because when violet-blue and
orange-red light is shone on the pigments, more light energy is absorbed and converted to
chemical energy in the light dependent reactions. More light energy is absorbed with these
wavelengths because of the cumulation of the absorbance spectrums of the pigments present
in the photosystem.
The opposite goes for yellow-green light.
- Unravelling the mystery behind the difference between absorbance and action spectrums.
The peak nearing the red side of the spectrum is greater than the peak on the blue side in the
action spectrum even though the opposite is true for the absorption spectrum.
- This is because P700 and P680 are directly activated by wavelengths of red light. Light of
other spectrums must travel through accessory pigments and be funnelled to the reaction
centre. As this happens, energy is lost.
The
space between the peaks is higher in the action spectrum than in the absorption

spectrum.
- This is because photosynthesis (shown in the action spectrum) depends on the light
independent reactions as well as the light dependent reactions. In the presence of light, the
rate of photosynthesis can thus not drop below a certain level.
- Additionally, the carotenoids absorbance spectrum covers some of the 500-600nm range
and thus partially compensates for the drop in the absorption of chlorophyll a and b.
- Chlorophyll illuminated by UV light or a standard florescence tube
When this happens, chlorophyll glows red. This is because electrons are excited as the
pigment absorbs light energy, but since there is only extracted pigment the absorbed energy
cannot be usefully passed on to work and thus lose their energy to the surroundings in the
form of florescence (less energetic/ greater wavelength) and heat.