PARTITION CHROMATOGRAPHY
By: Claudia V. Payumo
Lordelle D. Ragas
Introduction
Chromatography is a technique for separating or
analyzing a mixture of gases, liquids, or dissolved
substances. This is brought about by means of two
immiscible substances, one of which (the mobile
phase) transports the sample mixture through the
other (the stationary phase).
The mobile phase may be a gas or a liquid; the
stationary phase may be a liquid or a solid, and may
be in a
 column,
 on paper,
 or in a thin layer
on a glass or plastic support.
Introduction
The components of the mixture are adsorbed or
impeded by the stationary phase to different
extents and therefore become separated.
The technique is used for both qualitative and
quantitive analyses in biology and chemistry.
Column chromatography
In column chromatography, the stationary
phase, a solid adsorbent, is placed in a
vertical glass (usually) column and the
mobile phase, a liquid, is added to the top
and flows down through the column (by
either gravity or external pressure).
Column chromatography is generally used
as a purification technique: it isolates
desired compounds from a mixture.
Typical Column
Chromotography
Column chromatography on a large scale in
the 1950s. The chemist
uses a ladder to refill
eluent. He operates not
one but 11 columns.
Barely
visible
are
Erlenmeyer receptacles
on the floor.
Paper Chromatography
Paper chromatography is an analytical
technique for separating and identifying
mixtures that are or can be colored,
especially
pigments. This can also be
used in secondary or primary schools in
ink experiments. This method has been
largely
replaced
by
thin
layer
chromatography, however it is still a
powerful teaching tool
Thin Layer Chromatography (TLC)
Thin layer chromatography (TLC) is a
technique used to separate mixtures. It
involves a stationary phase consisting of a
thin
layer
of
adsorbent
material,
immobilized onto a flat, inert carrier sheet.
A liquid phase consisting of the solution to be
separated is then dissolved in an
appropriate solvent and is drawn up the
plate via capillary action, separating the
experimental solution based on the polarity
of the components of the compound in
question.
Partition Chromatography

Separation of similar substances by repeated
extraction by two miscible liquids.

Partition chromatography is referred to as liquidliquid chromatography (LLC).

Separation is based upon Nernst’s Law:
Nernt’s Law
When the ratio of concentrations
expresses a distribution value for a single
chemical species, the constant is designated
a partition coefficient or distribution
coefficient K and may be expressed
mathematically as:
K = Cl
--Cu
Where Cl represent concentration in the
lower phase; while Cu represent concentration
in the upper phase.
In order to explain the mechanics involved
in transferring solute molecules through a
column during a chromatographic process, the
plate theory is illustrated as follows:
Plate Theory
In the plate theory the entire length of the
chromatographic column is considered to be composed of
many small, identical cells called theoretical plates. Each cell
contains a pair of immiscible solvents (ether & water). The
lower phase is called the stationary phase, and the upper
phase, the mobile phase. A mixture of solutes is introduced
in the the first cell (0). Equilibrium is established, and the
upper phase of each cell is then transferred to the next cell
down to the next cell down the column. Each volume of
upper phase eluting from the column is called a plate volume.
The process of equilibration between transfers and the
transfer of the upper phase of each cell to the next cell is
continued until negligible amounts of solute remain in the
stationary phase.
Container #
0
1
2
3
4
ether
Row A
water
Row B
50
50
Row C
Row D
Row E
Row F
25
25
25
25
12.5
25
12.5
12.5
25
12.5
6.25
18.75
18.75
6.25
6.25
18.75
18.75
6.25
3.125
12.5
18.75
12.5
3.125
3.125
12.5
18.75
12.5
3.125
Detector
Exit-Outlet
Theoretical Distribution of a single solute after
varrying numbers of transfers.
Distribution of Solute in a Five-Plate Partition Chromatography System
Plate Volume No.
Fraction of Solute
0
0.00
1
0.00
2
0.00
3
0.00
4
0.00
5
0.031
6
0.078
7
0.117
8
0.137
9
0.137
10
0.123
11
0.103
12
0.081
13
0.061
14
0.044
15
0.031
16
0.020
17
0.014
18
0.009
Distribution of solute in partition-elution chromatography
0.16
Fraction of Solute
0.14
0.12
0.10
0.08
0.06
0.04
0.02
0.00
1
2
3
4
5
6
7
8
9
10 11 12 13 14 15 16 17 18 19
Plate Volume No.
Thank you!