Paper Chromatography: Identifying The Components of A Mixture

Paper Chromatography
Identifying the components of a

mixture
Chromatography
Chromatography is a method for separating
mixtures into their components based on physical
and/or chemical properties of the components.
Developed around 1903

by Russian Mikhail
Semenovich Tswett in
which he separated
plant pigments on
“diatomaceous earth”
with alcohol
Uses for Chromatography
Chromatography can be used to:
• Qualitatively analyze the components of a mixture
• Qualitatively identify the components of a mixture
using known compounds
• Quantitatively determine the amount of a
component in a mixture using standard samples
• Purify individual components by separating them
from the other compounds in a mixture
The Basics
• Mixture is placed on stationary phase
• Mobile phase passes over the stationary phase
• Mobile phase dissolves the components
• Mobile phase carries the individual components a
certain distance through the stationary phase,
depending on their attraction to both of the
phases
Solvent Front
Chromatographed
Spot
Rf ratio of spot
distance to
Putsolvent front
the spotted paper
distance tank
in a developing
Origin line
Solvent
Principles of Paper
Chromatography
 Capillary Action – the movement of liquid within the

spaces of a porous material due to the forces of
adhesion, cohesion, and surface tension. The liquid is
able to move up the filter paper because its attraction to
itself is stronger than the force of gravity.
 Solubility – the degree to which a material (solute)

dissolves into a solvent. Solutes dissolve into solvents
that have similar properties. This allows different solutes
to be separated by different combinations of solvents.
Separation of components depends on both their

solubility in the mobile phase and their differential
affinity to the mobile phase and the stationary phase.
Investigation: What Dyes are in
the M&Ms and Food Colors?
Preparing the
Chromatography Paper
 Obtain an 7 x 8 cm of
chromatography paper
 With a pencil (why?), draw

an origin line 1 cm above the
bottom edge of the strip. Near
the top, ID the chromatogram
 Mark the origin line with a

lane mark each cm. No lane
can be closer than about 1 cm
from the edge
Preparing the
Chromatography Paper
 ID each lane and record what
each lane is spotted with
 Place a spot from each

solution on the origin
 More than one strip may be

required to analyze all of the
solutions
Developing the
Chromatograms
 Add chromatographic solvent to
a sufficiently large beaker to a
depth of less than 1 cm
 Place the spotted paper in the

beaker carefully
 Ensure the solution does not go

above the origin line
 Develop the chromatogram until

the solvent front is about 2 cm
from the top of the paper
Developing the
Chromatograms
 Remove the chromatogram and
place on a paper towel, trace
the solvent front with pencil,
and let the chromatogram dry
in the oven for a few minutes.
 After the chromatogram is dry,

calculate the retention factors,
Rf, and tabulate the values
 Determine the dye content of

each of the “unknowns”
Considerations
 Small spots are generally
better but harder to see after
developing
 UV lamps, if available can

(sun)burn your eyes but UV is
stopped by the plastic of your
goggles
 UV lamps are low power but

extended exposure can cause a
mild (sun)burn
 Don’t eat the M&Ms – they’ve

been in the lab awhile
Writing Procedure Proposals
 Clearly state your experimental methods and how
those procedures will yield solutions to the problems
addressed in the investigation.
 State what data you plan to collect and how you will
analyze the data.
 Include proposed data tables.
 State all the materials you plan to use including, if
possible, concentrations and quantities.
 Describe the safety hazards associated with your
analysis and appropriate precautions you will take to
avoid personal injury.
 Each team member must sign and date the procedure
proposal before turning it in.

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Paper Chromatography

Identifying the components of a

Developed around 1903

 Capillary Action – the movement of liquid within the

 Solubility – the degree to which a material (solute)

Separation of components depends on both their

 With a pencil (why?), draw

 Mark the origin line with a

 Place a spot from each

 More than one strip may be

 Place the spotted paper in the

 Ensure the solution does not go

 Develop the chromatogram until

 After the chromatogram is dry,

 Determine the dye content of

 UV lamps, if available can

 UV lamps are low power but

 Don’t eat the M&Ms – they’ve

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