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Paper chromatography

Paper is used to separate mixtures of substances. These are often coloured substances such as food colourings, inks, dyes or plant .

Image gallerySkip image gallerySlide1 of 3, A pencil line is drawn across a sheet of chromatography paper and spots of ink or plant dye are placed along it. The paper is held abovea basin containing solvent., Paper chromatography 1. Water and ethanol solution is heated

Phases

Chromatography relies on two different 'phases':

  • the , which in paper chromatography is absorbent paper
  • the is the that moves through the stationary phase - in this instance, the paper - carrying different substances with it

The different substances in a mixture are attracted to the two phases to different degrees. This causes them to move at different rates through the paper.

Interpreting a chromatogram

Separation by chromatography produces a .

A paper chromatogram can be used to distinguish between and impure substances:

  • a pure substance produces one spot on the chromatogram
  • an impure substance produces two or more spots

A paper chromatogram can also be used to identify substances by comparing them with known substances. Two substances are likely to be the same if:

  • they produce the same number of spots, and these match in colour
  • the spots travel the same distance up the paper (have the same Rf value)
Results of a chromatogram of three pure substances and brown ink
Figure caption,
Interpreting the chromatogram for a brown ink

In this chromatogram, the brown ink is made of a mixture of the red, blue and yellow inks. This is because the spots in the brown ink reach the same heights (and have the same Rf value) as the reference inks.

Rf values

Rf values can be used to identify unknown chemicals if they can be compared to a range of reference substances. The Rf value is always the same for a particular substance if run in the same solvent system in the same conditions.

The Rf value of a spot is calculated using:

Rf = \(\frac{\textup{distance~travelled~by~substance}}{\textup{distance~travelled~by~solvent}}\)

Rf values vary from 0 (the substance is not attracted at all to the mobile phase) to 1 (the substance is not attracted to the stationary phase).

Chromatography paper next to a measurement scale, showing the distance travelled by the solvent, and a coloured dot showing the distance travelled by the substance
Figure caption,
Measurements needed to calculate an Rf value

Different compounds have different Rf values in different solvents, which can be used to identify them. The compounds in a mixture may separate into different spots depending on the solvent, but a pure compound produces a single spot in all solvents.