CHROMATOGRAPHIC

SEPARATIONS

CHEM 314
7:30-8:30 TTH, P200
2 ND S E M E S T E R A Y 2 0 1 3 - 2 0 1 4
ENGR. RONALD RINGOR, CHE
 CHROMATOGRAPHIC SEPARATIONS

There are very few, if any, methods for chemical analysis that
are specific for a single chemical species. At best, analytical
methods are selective for a few species or a class of species.
Consequently, the separation from potential interferences is
quite often a vital step in analytical procedures. Until the
middle of the twentieth century, analytical separations were
largely carried out by such classical methods as
precipitation, distillation, and extraction.
Today, however, analytical separations are most commonly
carried out by chromatography and electrophoresis,
particularly with samples that are multicomponent and
complex.
 CHROMATOGRAPHY

 Chromatography was invented and named by the

Russian botanist Mikhail Tswett
 He employed the technique to separate various plant
pigments such as chlorophylls and xanthophylls by
passing solutions of these compounds through a
glass column packed with finely divided calcium
carbonate,
 The separated species appeared as colored bands on
the column, which accounts for the name he chose
for the method (Greek chroma meaning "color“ and
graphein meaning "writing")
 CHROMATOGRAPHY
 In all chromatographic separations the sample is dissolved in a
mobile phase, which may be a gas, a liquid, or a supercritical
fluid.
 This mobile phase is then forced through an immiscible
stationary phase, which is fixed in place in a column or on a
solid surface. The two phases are chosen so that the components
of the sample distribute themselves between the mobile and
stationary phases to varying degrees.
 Those components strongly retained by the stationary phase
move only slowly with the flow of mobile phase. In contrast,
components that are weakly held by the stationary phase travel
rapidly.
 As a consequence of these differences in migration rates, sample
components separate into discrete bands, or zones, that can be
analyzed qualitatively and quantitatively.
 CLASSIFICATION OF
CHROMATOGRAPHIC METHODS

Chromatographic methods can be categorized in two ways:

1)based on the physical means by which the stationary and
mobile phases are brought into contact
a)column chromatography – the stationary phase is held
in a narrow tube through which the mobile phase is
forced under pressure.

b) planar chromatography - the stationary phase is

supported on a flat plate or in the interstices of paper;
here, the mobile phase moves through the stationary
phase by capillary action or under the influence of gravity
 CLASSIFICATION OF
CHROMATOGRAPHIC METHODS

2) based on the types of mobile and stationary phases

a) Gas chromatography (GC),
b) liquid chromatography (LC),
c) and supercritical fluid chromatography (SFC).

 As the names imply, the mobile phases in the three

techniques are gases, liquids, and supercritical fluids,
respectively.
 Note that only LC can be performed either in columns or
on planar surfaces; GC and SFC, on the other hand, are
restricted to column procedures so that the column walls
contain the mobile phase,
 General Specific Method Stationary Phase Type of
Classification Equilibrium
Gas a.Gas-liquid Liquid absorbed or Partition between
Chromatography chromatography bonded to solid gas and liquid
surface

b.Gas-solid Solid Adsorption

Liquid a.Liquid-liquid, or Liquid absorbed or Partition between
Chromatography parition to solid surface immiscibleliquids

b.Liquid-solid, or Solid Adsorption

adsorption

c.Ion Exchange Ion-exchange resin ion exchange

d.Size exclusion Liquid in interstices Partition/sieving

of a Polymeric solid

Group specific liquid

e.Affinity bonded to a solid Partition between
surface surface liquid and
mobile liquid
Supercritical Fluid Organic species Partition between
Chromatography bonded to a solid Supercritical fluid