CHROMOTOGRAPHY -VANI PRIYADHARSHINI K

1. Chromatography.
K.VANI PRIYADHARSHINI
ROLL NO: CB16S075914
II BIOTECHNOLOGY

2. Definition of chromatography
 a process in which a chemical mixture carried by a liquid or gas is separated
into components as a result of differential distribution of the solutes as they
flow around or over a stationary liquid or solid phase.
 The various constituents of the mixture travel at different speeds, causing them
to separate.

3. types of chromatography
 There are four main types of chromatography. These are
 Liquid Chromatography,
 Gas Chromatography,
 Thin-Layer Chromatography and
 Paper Chromatography

4. Principle of Chromatography
 Chromatography is based on the principlewhere molecules in mixture
applied onto the surface or into the solid, and fluid stationary phase
(stable phase) is separating from each other while moving with the aid
of a mobile phase.

5. applications of chromatography
 Chromatography has various applications.
 It is used for the separation of different colors of ink.
 It is also used to identify and separate the preservatives and additives
added in the food items.
 It is also used in DNA fingerprinting and bioinformatics.

6. Paper chromatography
 In paper chromatography, the stationary phase is a very uniform absorbentpaper.
 The mobile phase is a suitable liquid solvent or mixture of solvents.
 It is used in the sequencing of DNA and RNA.
 Paper chromatography is used as a qualitative analytical chemistry technique for
identifying and separating colored mixtures like pigments.
 It is used in scientific studies to identify unknown organic and inorganic compounds
from a mixture.

8. Thin-layer chromatography (TLC)
 Thin-layer chromatography (TLC) is achromatography technique used to
separate non-volatile mixtures. ...
 The mobile phase has different properties from the stationary phase.
 For example, with silica gel, a very polar substance, non-polar mobile phases
such as heptane are used.
 Chromatography works on the principle that different compounds will have
different solubilities and adsorption to the two phases between which they are
to be partitioned.
 Thin Layer Chromatography (TLC) is a solid-liquid technique in which the two
phases are a solid (stationary phase) and a liquid (moving phase).
 used separation technique used in life sciences and chemistry studies.
 It is easy, cheap and gives rapid results.
 It is used to isolate and analyse mixtures of compounds.

10. Gas chromatography
 Gas chromatography is a common type of chromatography used in analytical
chemistry for separating and analyzing compounds that can be vaporized without
decomposition.
 Typical uses of GC include testing the purity of a particular substance, or separating
the different components of a mixture
 Principle of gas chromatography:
 The sample solution injected into the instrument enters agas stream which transports
the sample into a separation tube known as the "column." (Helium or nitrogen is used
as the so-called carrier gas.)
 The various components are separated inside the column.
 Two types of gas chromatography are encountered:
 gas-solid chromatography (GSC) an
 d gas-liquid chromatography (GLC)

12. Liquid chromatography
 Liquid chromatography (LC) is a separation technique in which the mobile phase is a liquid, where sample ions or molecules are dissolved.
 It is carried out either in a column or a plane.
 used to separate proteins, nucleic acids, or small molecules in complex mixtures.
 Liquid chromatography (LC) separates molecules in a liquid mobile phase using a solid stationary phase.
 Liquid chromatography can be used for analytical or preparative applications.
 Types
 Partition chromatography.
 Normal–phase chromatography.
 Displacement chromatography.
 Reversed-phase chromatography (RPC)
 Size-exclusion chromatography.
 Ion-exchange chromatography.
 Bioaffinity chromatography.
 Aqueous normal-phase chromatography.