Chromatography

Chromatographic separation techniques are multi-

stage separation methods in which the components
of a sample are distributed between two phases, one
of which is stationary, while the other is mobile. The
stationary phase may be a solid or a liquid supported
on a solid or a gel. The stationary phase may be
packed in a column, spread as a layer, or distributed
as a film, etc. The mobile phase may be gaseous or
liquid. The separation may be based on adsorption,
mass distribution (partition), ion exchange, etc., or
may be based on differences in the physico-chemical
properties of the molecules such as size, mass,
volume, etc.
 Chromatography, in chemistry, analytical technique used for
the chemical separation of mixtures and substances. The
technique depends on the principle of selective adsorption (not
to be confused with absorption), a type of adhesion.
Chromatography was discovered in 1906 by the Italian-born
Russian botanist Mikhail Tswett, but was not widely used until
the 1930s. Tswett separated plant pigments (chlorophylls) by
pouring petroleum-ether extract of green leaves over a column
of powdered calcium carbonate in a vertical glass tube. As the
solution percolated through the column the individual
components of the mixture migrated downward at different
rates of speed, so that the column became marked with
horizontal bands of colors, called a chromatogram. Each band
corresponded to a different pigment.
Clinical In medicine, such as blood
cholesterol
Forensic Evidence in the study of crime
Geochemical Rocks and minerals
Inorganic Compounds in which carbon
combined with hydrogen or
oxygen is not the main
ingredient
Oceanographic Seawater and the ocean floor

Organic Compounds containing the

elements carbon and
hydrogen and certain element
groupings-functional groups-in
organic molecules
Petroleum Oil and petroleum products
Pharmaceutical Drugs

Pollution Toxic substances

Polymer Very large molecules, such as polyvinyl
chloride

Spatial Three-dimensional distribution of

elements or molecules in a sample

Surface Analysis of a sample's "skin" as

opposed to its interior

Trace Very small quantities or concentrations

Principles of separation,
apparatus and methods are given
in the following general methods
 Paper chromatography (Appendix III E;
Ph. Eur method 2.2.26))
 Thin-layer chromatography (Appendix III
A; Ph Eur method 2.2.27)
 Gas chromatography (Appendix III B;
Ph. Eur. method 2.2.28)
 Liquid chromatography (Appendix III D;
2.2.29)
Liquid Chromatography
Thin Layer Chromatography
Thank You