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2C - Chromatographic Techniques
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Coursework 2C for BTEC Foundation National Diploma Level 3 in Apllied Science. This has been given a distinction.
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Paper Chromatography of Chlorophyll Experiment ResultsLindsay Torres
The aim of this experiment is to test for pigments in spinach leaves.
The Rf value is calculated as follows:
Distance moved by the pigment
Rf= ------------------------------------------------------------------------------
Distance between the line of origin and solvent front
, Analysis and conclusion
Rf stands for retention factor or retardation factor. This value is most commonly used in
chromatography.
- The Rf value indicate how soluble the specific pigment is in the solvent by how high the pigment
moves on paper. Two pigments with the same Rf value are likely to be identical molecules. Small
Rf indicate larger, less soluble pigments while high soluble pigments have an Rf value near to
one.
- A particular compound will travel along the same distance along the stationary phase by a
specific solvent given that other experimental conditions kept constant. In short, every kind of
compound has a specific Rf Value
- There are 4 pigments carotene, chlorophyll a, xanophyll and chlorophyll b. Distance of solutes
go up in differently. As the solvent touches the paper the solutes will mix with it making it rise
up changing into different pigment colours. As we can see the higher the distance the lighter the
pigment.
- The solvent carries the dissolved pigments as it moves up the paper.
- The pigments are carried at different rates because they are not equally soluble. A pigment that
is the most soluble will travel the greatest distance and the pigment that is the less soluble will
travel at a shorter distance
An experiment to investigate the movement of chlorophyll pigments during paper chromatography. We
will use paper, a pencil, the selected pigments for the experiment, a measuring cup/jug and water. In
general, chromatography is a method used to separate chemical mixtures for analysis. In paper
chromatography, substances are separated as they travel in a mobile phase which passes over a
stationary phase. A mobile phase is the liquid that transports the substance through the absorbing
material which travels along the stationary phase and carried the substance components with it. A
stationary phase is the solid material that absorbs the mixing flowing through it. In this case, the mobile
phase is water and the stationary phase is the paper being used.
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