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R.ARUNA VIGNESHWARI
96507214001
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Term Definition
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Ú Among all the chromatography methods paper
chromatography is an inexpensive and rapid method that
provides graphic and clear results.
Ú Used as a qualitative method for identifying the
components in a mixture.
Ú The separated spots on the finished and dried
chromatogram can be cut out and re-dissolved to obtain a
pure sample of component of the sample mixtures.
Ú Used in several scientific studies in identification of
unknown organic and inorganic compounds from a
mixture.
Ú In forensic studies paper chromatography is used in crime
scene investigation and DNA and RNA sequencing along
with other studies.
Ú Paper chromatography is used as an analytical chemistry
technique for identifying and separating colored mixtures
like pigments.
Ú Sugars, amino acids, lipids and nucleic acids and other
biomolecules can be easily identified by spraying with
appropriate reagents to detect these specific compounds.
Ú Paper chromatography can be reproduced easily as long
as the conditions are controlled and maintained
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This includes chromatographic methods in
which:
The stationary phase is packed into a
column.
The mobile phase is a moving liquid or gas.
Column
chromatography
Stationary phase is
held in a narrow
tube through
which the mobile
phase is forced
under pressure or
under the effect
of gravity
Factors affecting solutes separation in
( Factors affecting column efficiency)
Factor Effect
Particle size of solid
Decrease of size improves separation (but very
stationary phase (or of
small particles need high pressure).
support)
Efficiency increases as ratio length / width
olumn dimensions
increases.
Non uniform packing results in irregular
movement of solutes through column & less
Uniformity of packing
uniform zone formation, (i.e. band broadning or
tailing).
mncrease in column temperature results in
olumn temperature speed of elution but does not improve
separation (tailing).
Solvents should be of low viscosity (to give
Eluting solvent efficient resolution) & h igh volatility (to get
rapid recovery of the substances).
Solvent flow rate Uniform & low flow rate gives better resolution.
ontinuity of flow Discontinuous flow disturbs resolution
Deactivation of adsorbent decreases
ondition of adsorbent
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Adsorbents:
The most common are Alumina & Silica gel in which the
interactions with solute molecules is due to OH groups
present on their surface.
More polar molecules are adsorbed more strongly &
thus, will elute more slowly
Strength of adsorption of polar groups (solutes) on polar
support is in the following order:
- - < O-H3 < -OOR < > O < -HO < -NH2 < -
OH < -OOH
Olefins < Ethers < Esters < Lactones < Aldehydes <
Amines < Phenols < Acids.
Alumina: sterols, dyestuffs, vitamins,
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esters,
alkaloids & inorganic compounds.
Not used for compounds containing phenolic or
carboxylic groups