Introduction
Chromatography is used to separate substances and provide information to help identify them
The components have different solubilities in a given solvent
E.g. Different coloured inks that have been mixed to make black ink
A pencil line is drawn on chromatography paper and spots of the sample are placed on it
A pencil is used for this as ink would run into the chromatogram along with the samples
The paper is then lowered into the solvent container, making sure that the pencil line sits above the level of the solvent so the samples don’t wash into the solvent container
The solvent used is usually water but it can be other substances such as ethanol
The solvent travels up the paper by capillary action, taking some of the coloured substances with it
Different substances have different solubilities so they will travel at different rates, causing the substances to spread apart
Those substances with higher solubility will travel further than the others
Interpret simple chromatograms
We can use a chromatogram to compare the substances present in a mixture to known substances and make assumptions
Pure substances will produce only one spot on the chromatogram
Impure substances will produce more than one spot on the chromatogram
If two or more substances are the same, they will produce identical chromatograms
If the substance is a mixture, it will separate on the paper to show all the different components as separate spots
It is common practice to include a known compound as a reference spot
This can help match up to an unknown spot or set of spots in order to identify it
We can draw several conclusions from this chromatogram:
The brown ink is a mixture as there are three dots
Red, yellow and blue are pure as there is only one dot for each
The brown ink contains red, blue and yellow as the dots are in line with one another horizontally



















How does the polarity of solvents and solutes affect separation in paper chromatography?
Why do some substances travel farther on the chromatogram than others?
What roles do capillary action and adsorption play in the effectiveness of paper chromatography?
How does changing the type of paper (e.g., porosity, thickness) impact chromatographic results?
How does the pH of the solvent affect the separation of amino acids or other biomolecules?
Can two compounds with similar Rf values be reliably distinguished using paper chromatography alone?
keywords
Paper chromatography
Rf value (Retention factor)
Mobile phase
Stationary phase
Solvent front
Capillary action
Adsorption and absorption
Chromatographic separation
Two-dimensional chromatography
Chromatogram