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chromatography

chromatography

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Published by: Alyaa on Mar 31, 2009
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06/15/2013

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Definition and Classification of Chromatography
Chromatography course
 
Dr.Ehab Aboueladab
 
(Assistance. Prof.of Biochemistry, Mansoura University)
 ةحفص
1
Aim Separation Techniques
1-Biological fluids are extremely complex in composition.2-Chemical analysis would be
impossible
if it were necessary tocompletely isolate each substance prior to its measurement.3- An optimal method tests for a specific substance in thepresence of all others, requiring no isolation of the substanceunder analysis.4- A test is specific when none of the other substances presentinterfere. However, virtually all chemical tests are subject to atleast some interference.5-This is one of the most important problems in clinicalchemistry. Therefore some type of separation procedure isrequired.7-Separation in clinical chemistry usually is
based ondifferences
in the
size, solubility or charge
of the substancesinvolved.
 
Definition and Classification of Chromatography
Chromatography course
 
Dr.Ehab Aboueladab
 
(Assistance. Prof.of Biochemistry, Mansoura University)
 ةحفص
2
INTRODUCTION
The Russian botanist
M. S. Tswett
is discovery of chromatography. He used a column of powdered
calciumcarbonate to separate green leaf pigments
into a series of colored bands by allowing a solvent to percolate through thecolumn bed. Since these experiments by Tswett many scientistshave made substantial contributions to the theory and practice of chromatography
.
Not least among these is
A. J. P. Martin
whoreceived the Nobel Prize in 1952 for the
invention of partitionchromatography
(with R. L. K. Synge) and in the same yearwith
A. T. James
he introduced the technique of 
gas-liquidchromatography
. Chromatography is now an important toolused in all branches of the chemical and life sciences.
1-Definition of Chromatography
Chromatography is essentially a physical method of separation in which the components to be separated aredistributed between
two phases
one of which is stationary
(stationary phase)
while the other
(the mobile phase)
throughit in a definite direction.
2- Classification of chromatographic methods
The common feature of all chromatographic methods istwo phases, one stationary and the other mobileA classification can be made depending upon whether the
stationary phase is
 solid or liquid 
. If it is solid, the method is
 
Definition and Classification of Chromatography
Chromatography course
 
Dr.Ehab Aboueladab
 
(Assistance. Prof.of Biochemistry, Mansoura University)
 ةحفص
3
termed
adsorption chromatography
; if it is
liquid 
the methodis
partition chromatography.
 One of the two phases is a
moving phase (the mobile phase)
,while the other does
not move (the stationary phase).
The
mobile phase
can be either a
gas or a liquid
, while the
stationary phase
can be either a
liquid or solid
.
 
3- Classification scheme
One classification scheme is based on the nature of the twophases. All techniques which utilize
a gas for the mobile phase
 come under the heading of 
gas chromatography (GC)
. Alltechniques that utilize
a liquid mobile phase
come under theheading of 
liquid chromatography (LC)
. Additionally, wehave gas
 – 
liquid chromatography
(GLC)
, gas
 – 
solidchromatography
(GSC)
, liquid
 – 
liquid chromatography
(LLC)
,and liquid
 – 
solid chromatography
(LSC)
,
4- Main Type of Chromatography
In general, there are four main types which can beclassified as follows:
4.1-Liquid-Solid
chromatography
 
 
Classical
adsorption chromatography
(Tswett column)
 
Ion-exchange chromatography
4.2
.
Gas-Solid
chromatography
4.3
.
Liquid-Liquid
chromatography
 
Classical
 partition chromatography
 
 
Paper chromatography

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equation (2) is sometimes employed to calculate Neff. show that this relation is equivalent to the more classical equation(1)
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