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Is the collective term for a family of laboratory techniques
for the separation of mixtures
Involves passing a mixture which contains the analyte
through a stationary phase, which separates it from other
molecules in the mixture and allows it to be isolated

 

M
    
  
  
  
 
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R
, Mikhail Semyonovich Tsvet, who used columns of

calcium carbonate for separating plant pigments
During the R 
s and R 
s. They established the
principles and basic techniques of partition chromatography
Since then, the technology has advanced rapidly
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The analyte is the substance that is to be separated
during chromatography
Stationary phase: Phase that stays in place inside the
column. Can be a particular solid or gel-based packing (LC)
or a highly viscous liquid coated on the inside of the
column (GC)
Mobile phase: Solvent moving through the column,
either a liquid in LC or gas in GC
 Eluent: Fluid entering a column
Eluate: Fluid exiting the column
Elution: The process of passing the mobile phase
through the column.
Chromatogram: Graph showing detector response
as a function of a time.
Flow rate: How much mobile phase passed /
minute (ml/min).
Linear velocity: Distance passed by mobile phase
per Rmin in the column (cm/min)
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The stationary phase (column


packing) in the column is very polar

÷ 
One starts will a less polar solvent to ÷


 ÷÷÷
remove
the less polar compounds, and then
you slowly
Increase the polarity of the solvent to
remove the more polar compounds

Figure 1. column chrotography a

  
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Here the mobile phase is a liquid
Flowing past a thin layer of powder on a solid
support
Substances that are less attracted to the solid or are
more soluble in the liquid move faster
And so move further up the plate by the time that the
process has been stopped by taking the plate out of
the liquid. - larger Rf
Figure 2. principles in TLCa
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Figure 3. Paper chromatography jar

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For substances that are very soluble in the liquid
Rf will be close to ....
R
For substances that are rather insoluble in the liquid
Rf will be close to ....
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Gel-Permeation Chromatography is a mechanical sorting of

molecules based on the size of the molecules in solution.
Small molecules are able to permeate more pores and are,
therefore, retained longer than large molecules.
 
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Separation in Ion-exchange Chromatography is based on

the competition of different ionic compounds of the sample
for the active sites on the ion-exchange resin (column-
packing).
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Liquid chromatography (LC) is a separation technique in
which the mobile phase is a liquid. Liquid
chromatography can be carried out either in a column or
a plane

Present day, use high

performance liquid
chromatography (HPLC)

Figure 4 . HPLC 1525

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Principals of Separation
Column is selected, packed with Liquid Phase, and installed.
Sample injected with microliter syringe into the injection
port where it is vaporized and mixed into the Carrier Gas
stream (helium, nitrogen, argon).
Sample vapor becomes partitioned between Moving Gas
Phase and Stationary Liquid Phase
The time the different compounds in the sample spend in
the Vapor Phase is a function of their Vapor Pressure.
The more volatile (Low Boiling Point / Higher Vapor
Pressure) compounds arrive at the end of the column first
and pass into the detector
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Figure 5. chromatograph
Figure 6. chromatographic peak is fronting

Figure 7. chromatographic peak is tailing

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Figure 5. HPLC
Figure 6. Gas chromatography
Chromatography is used by scientists to:

Analyze ² examine a mixture, its components,

and their relations to one another

Identify ² determine the identity of a mixture or

components based on known components

Purify ² separate components in order to isolate

one of interest for further study

Quantify ² determine the amount of the a mixture

and/or the components present in the samplecxxc
 Pharmaceutical Company ² determine amount of
each chemical found in new product

Hospital ² detect blood or alcohol levels in a

patient·s blood stream

Law Enforcement ² to compare a sample found at

a crime scene to samples from suspects

Environmental Agency ² determine the level of

pollutants in the water supply

Manufacturing Plant ² to purify a chemical needed

to make a product

So on
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