Chromatography

• the separation of the components of a

substance by passing it over or through a
substance that absorbs selectively
Chromatography
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• a modern method of continuous or batch
materials Research II | CN: 4, 14, 30

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chemical analysis and separation based on
the preferential adsorption of different
components of a gas, liquid, or solid mixture
or solution as it contacts an adsorbent
medium

• the objective of chromatography is to

determine the different substances that make
up the mixture, by dissolving and separating
it. 
 Chromatography
• In a chromatography, we can identify the
"mobile phase" and the "stationary phase".

Chromatography
• The "mobile phase" is the phase that moves
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in the process. (mostly
II |liquids
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materials Research 30

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types • On the other hand, the "stationary phase" is
the phase that doesn't move within the
process. (e.g. paper, thick liquid)

• As phases, they don't show time or the form

of an object. It is the "object" itself.
 Chromatography
• After a chromatography process, the
different pattern of spots within the
stationary phase is identified as a
Chromatography
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"chromatogram". 
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materials Research II | CN: 4, 14, 30

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types •  It is a column or strip of material
containing constituents of a mixture
separated by chromatography.
 Chromatography
In chromatography, the symbol Rf is
used to denote the position of a
Chromatography
dissolved component on a
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chromatogram relative to the distance
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the solvent moved.
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To calculate it, measure the distance of
the soluble from the line, and the
solvent distance, which is also from the
line.

This value, however, is just to measure

its distance from the starting line.
 Types of Chromatography
• Paper Chromatography
• Thin Layer Chromatography (TLC)
• Gel Chromatography
• Chromatography
Column Chromatography
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materials • ResearchChromatography
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• Gel Filtration Chromatography
• Gas Liquid Chromatography
• Affinity Chromatography
 Materials
• beaker (or any container)
• any soluble chemical
• any solvent
• Chromatography
filter paper (varies depending on the type of
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chromatography; in this case,4,we14,
are30
doing a

materials
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paper chromatography)
• any solid object 
• pencil

1. Make sure that the soluble is compatible with the solvent, in order for it to be
dissolved.
2. The solid object will be used to cover the beaker in the process.
3. The pencil will be used to draw a line that will be used in the process.
4. Any variable that would be changed (e.g. use a normal paper instead of a filter one),
will have different result than what is expected.
 Procedure
1. Get a piece of filter paper and use a pencil to draw a line near the bottom of a
sheet which we can label it as a "baseline".

2. Add your sample ink that you think contains a lot of different substances in the
pencil line.
Chromatography
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3. Get the beaker and add a shallow Research
or little amount
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solvent
4,in14,
it. (e.g.
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ethanol)

4. Place the filter paper inside the beaker that has a shallow amount of solvent.
MAKE SURE that the pencil line and the solvent does not submerge or meet up
to each other. Place a lid at the top of the beaker to stop the solvent from
evaporating.

5. Wait for the solvent to seep up the filter paper. Doing it so will result on the ink
dissolving and separating the different substances of the ink. Each substance have
different rates when separating, which tells that the ink MUST have different
substances. The travel of the different substances will go from bottom to top.
 Procedure
6. This will take about 15 minutes, but may take longer depending on the
solvent and paper type. Be sure to remove your chromatography paper from
the solvent once it has reached one to one half cm from the top. Do not

Chromatography
allow the solvent to reach all the way to the top of the filter paper!
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7. After the solvent has reached up nearly the top of the paper, remove it

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inside the beaker, and leave it to dry. The spots in the paper is the
chromatogram.

8. The chromatography is now done, and it is possible to calculate the Rf

value. And chromatography is done.

Reminder: The chemical, or ink that will be used should be soluble, in order
to be dissolved.
Having a non-soluble ink will not be separated, hence it will just stay in the
baseline.
 Advantages
• Chromatography is the simplest method for the separation of
components.
• Chromatography can be controlled by a single person.
Chromatography
• Components from a complex mixture can separate using
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chromatography method.

materials
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• The small amount of sample can be detected by this

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chromatography.
• It is a rapid and precise method of separation.
• Very few sample volume/quantity is required for analysis.
• It works on a broad range of samples.
• In some chromatography techniques, it is possible to separate
different components of a complex mixture.
• The separation of components can be achieved in different
methods.
 Disadvantages
• The chromatography equipment can only be operated by a trained
person.
• Chromatography instruments are expensive.
Chromatography
• An error occurs due to the overloading of the samples.
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• Chromatography equipment must Research
be handled
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these parts are expensive and sensitive.
• Some of the chromatography techniques require more solvent to
separate the analytes.
• Periodic maintenance and parts need to be changed
• Some of the chromatography methods require high power
consumption.
• High operational pressure may be required to achieve efficient
separation.