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Chromatography Is the collective term for a set of laboratory techniques for the
separation of mixtures. Chromatography is a method by which a mixture is
separated by distributing its components between two phases. The stationary
phase remains fixed in place while the mobile phase carries the components of
the mixture through the medium being used The separation of materials is based
on differential partitioning [retardation]between the mobile and stationary phases.
The mobile phase (also known as solvent) may be either liquid or gas.
The stationary phase (also known as sorbent) can be either a solid or liquid,
a liquid stationary phase is held stationary by a solid(support or matrix).
The molecules in the mixture to be separated are the
Classification of chromatography according to mobile phase:
1. Liquid chromatography: mobile phase is a liquid. (LLC, LSC).
2. Gas chromatography : mobile phase is a gas. (GSC, GLC)
It is a method for identifying substances and testing the purity of compounds. TLC
is a useful technique because it is relatively quick and requires small quantities of
material.
The stationary phase:
is a thin layer of adsorbent (usually silica gel or alumina) coated on a plate.
[Rf] Value:
-If Rƒvalue of a solute is closer to zero, the solute has more attraction to stationary
phase.
-If Rƒvalue of a solute is closer to 1,then the solute has more affinity for the
mobile phase and travels further.
-The final chromatogram can be compared with other knownmixture
chromatograms to identify sample mixes, using the Rf value in an experiment
Principles
Applications
• Used to determine impurities in pharmaceutical raw materials and formulated
products.
• Often used as a basic identity check on pharmaceutical raw materials.
• Potentially useful in cleaning validation, which is part of the manufacture of
pharmaceuticals.
determine the number of components in a mixture
• verify a substance’s identity
• monitor the progress of a reaction
• determine appropriate conditions for column chromatography
• analyze the fractions obtained from column chromatogrpahy
Paper Chromatography
1. Descending
-In this method, the solvent is kept in a trough at the top of the chamber and is
allowed to flow down the paper.
-The liquid moves down by capillary actionas well as by the gravitational force.
-In this case, the flow is more rapid as compared to the ascending method.
2. Ascending
In this method, the solvent moves upward against gravitational force.
The only force that cause the motion[of solvent and the componds] is
capillary force. So the speed of the process is slow.
References:
1. owlcation.com/stem/tlc-thin-layer-chromatography-Principle-Procedure
2. [David_G_Watson]_Pharmaceutical_analysis__a_textb(BookZZ.org)
3. paper_and_thin_layer_chromatography_tlc
4. http://cdn.biologydiscussion.com/wp-
content/uploads/2015/10/clip_image028.jpg