11/18/22, 1:29 PM Photosynthetic Pigments

Photosynthetic Pigments
There are various methods by which you can investigate aspects of photosynthe
research by biologists to shed light on the many different reactions that occur during p

One method of investigating photosynthesis involves  chromatography, which analy

pigments in leaves. Photosynthetic pigments are small molecules that are capable of

 Remember that photosynthesis occurs in the chloroplasts of a plant, and the chlorophyll in t
a plant their green colour. This is because chlorophyll, the most common pigment found in l
reflects green light. 

Chloroplasts  are an essential part of the light-dependent reaction during photosynth

chlorophyll contained within them, the plant wouldn't absorb light energy, and photoio
curious about how light energy is converted into chemical energy via photosynthes
dependent and light-independent reactions. 

What is chromatography? 

Chromatography is a laboratory technique used by biologists and biochemists. It allo

which is achieved by passing it in a solution or suspension in which the mixture's c
rates. It is often used to separate a product from any unreacted solution. 

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There are two primary types of chromatography: thin-layer chromatography (sometim

chromatography. 

The function of both methods are the same, but each uses different equipment to s
components. You are most likely to use thin layer chromatography during your studie
professional laboratory, you will make use of both. The differences between the two a

How do you investigate photosynthesis using chromatograp

For your A-level course, you will be asked to use chromatography to investigate the
of different plants. You might, for example, be asked to compare the leaves of shade-
or leaves that are different colours from the same plant. To do this, you will need to ca
below. 

Equipment:

Leaf samples
Filter paper
Distilled water
Pestle and mortar
Ruler
Capillary tube
Chromatography solvent
Acetones
Pipette

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11/18/22, 1:29 PM Photosynthetic Pigments

Pencil

Method:

1. Using the pencil, draw a straight line around 1 cm above the bottom of the fil
pencil for this step, not a pen, as the ink will get into your solution and mess up y
2. Take a section of the leaf, and place it into the mortar. Add 20 drops of acetone
acetone using the pestle. This will release the pigments in the leaf. 
3. Using the capillary tube, extract the pigment and drop it onto the centre of the pe
4. Suspend the filter paper with the pigment in the solvent; however, make sure
above the pencil line. 
5. Leave the paper until the solvent has risen close to the top. Make sure that the
paper.
6. Remove the paper from the solvent, and draw a pencil line marking where the s
notice that pigment has separated into different compounds, which should all be
the first pencil line. 

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Fig. 1 - Example of Thin Layer Chromatography

Although this won't look exactly like your experiment, it will be set up similarly. 

How do you calculate an Rf value?

Once you have collected your filter paper with the different pigments separated, you
spot.  Rf stands for  retention factor, and this formula is used to help identify
chromatography sample.

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The equation for the Rf value is as follows:

 When calculating the Rf value for each spot, always measure to the centre of each spot, not

When you have your set of Rf values, you can then compare them to the known Rf
identify which pigments are present in your leaf.  If you need to find your own da
specifically for paper chromatography. Use the same solvent as you since these
travelled by certain pigments!

Which photosynthetic pigments are in chloro

You are likely to find the following pigments in a leaf during chromatography:

Chlorophyll a and Chlorophyll b: These pigments are green in colour. Chlorophy

pigments, absorbs blue and red light and reflects green light.  Chlorophyll b
however it can absorb a wider wavelength of light energy than chlorophyll a. C
light and reflects green light, like chlorophyll a; however, the green light reflecte
a molecular level, chlorophyll has a  porphyrin ring, which helps absorb light ene
Carotenoids: These pigments reflect orange, yellow and red light waves. In the
cluster next to chlorophyll molecules to efficiently hand off any absorbed photon
Xanthophylls: like carotenoids, xanthophylls pass along light energy
antioxidants. Xanthophylls reflect yellow light waves.

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Anthocyanin: a pigment that absorbs blue and green light. Anthocyanin molecul

plant cell.  

Deep dive

Chlorophyll c is another important photosynthetic pigment; however, you will no
experiment.  This is because it is not found in plants but in  microorga
photosynthesis!

Why are photosynthetic pigments

photosynthesis? 

Photosynthetic pigments are crucial to photosynthesis as they absorb  photo

wavelengths. As you will see in the diagram below, different pigments can absorb an
light, which allows a plant to get as much energy as possible from a single light source

After absorbing photons, the electrons contained in the pigments become ex

level. Accessory pigments such as carotenoids, anthocyanins, and xanthophylls can th
pigments like chlorophyll a, which can oxidize and donate an electron to the ele
dependent reaction. 

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Photosynthetic Pigments - Key ta

We can investigate the different photosynthetic pigments present in leaves via c
isolating the pigments in a solution and then using filter paper to separate each
We can identify different pigments using an Rf value, which you can calculate fr
has travelled on the filter paper. You can work out the Rf value using the followin

Depending on the kind of leaf you experiment on, you will find a range of pigm
and Chlorophyll b, Carotenoids, Xanthophylls, and Anthocyanin. Chlorophyll a is
pigment. 
Photosynthetic pigments are crucial to photosynthesis as they absorb photo
wavelengths. 
Different pigments can absorb and reflect different wavelengths of light, which
energy as possible from a single light source. 

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