16ANK4034

Kimia Farmasi Analisis Pustaka Pelajar

 Original Chromatography Experiment
Start: A glass End: A series of
column is filled colored bands is
with powdered seen to form,
limestone corresponding to
(CaCO3). the different
An EtOH extract Later pigments in the
of leaf pigments original plant
is applied to the extract. These
top of the column. bands were later
determined to be
EtOH is used to
chlorophylls,
flush the pigments
xanthophylls and
down the column.
carotenoids.
Solid Support Film of the liqiud
stationary Phase
 COO - H+ NH4+ OH-
Resin Resin
COO - H+
NH4+ OH-

Counter counter
ions ions

Anionic exchanger Cationic exchanger

very large molecules eluted first without separation

large molecules can enter some pores

very small molecules enter all pores and eluted at last

 Sample application

Bands of postivey Bands of negativey

charged compounds charged compounds
How Does Chromatography Work?

In all chromatographic separations, the sample is transported in a mobile phase. The

mobile phase can be a gas, a liquid, or a supercritical fluid.

The mobile phase is then forced through a stationary phase held in a column or on a
solid surface. The stationary phase needs to be something that does not react with the
mobile phase or the sample.

The sample then has the opportunity to interact with the stationary phase as it moves
past it. Samples that interact greatly, then appear to move more slowly. Samples that
interact weakly, then appear to move more quickly. Because of this difference in rates,
the samples can then be separated into their components.
Chromatography is based on a physical equilibrium
that results when a solute is transferred between the
mobile and a stationary phase.
K = distribution
A coefficient or
partition ratio
A A
A A
CS
A A
K =
A
CM
A Where CS is the molar
A concentration of the solute in the
A
A stationary phase and CM is the
molar concentration in the
mobile phase.
Cross Section of Equilibrium in a column.
“A” are adsorbed to the stationary phase.
“A” are traveling in the mobile phase.
Konsep dan teori kromatografi

Chromatogram
Konsep dan teori kromatografi
 Grafik Kromatografi K=1

0 2 4 6 Jumlah
8 Tahap
10 12 14 16 18

Grafik Kromatografi K=0,4 Grafik Kromatografi K=0,4

16 16
14 14
12 12
10 10
Axis Title

8 8
6 6
4 4
2 2
0 0
0 2 4 6 8 10 12 14 16 180 2 4 6 8 10 12 14 16 18
-2
Jumlah Tahap -2