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Stationary phase
The phase that stays in place inside the column.
Adsorption Chromatography
Has a solid stationary phase and a liquid or gaseous
mobile phase
Solute is adsorbed on the surface of the of the solid
particles .
The more strongly a solute adsorb the slower it
travels through the column
Solute adsorbed on
the surface of
stationary phase
Partition chromatography
Most common in gas chromatography. Stationary
phase is a liquid bonded on a solid surface
Solute equilibrates between the stationary liquid and
mobile phase
Small molecules
penetrate pores
of particles
Affinity chromatography
Selective kind of chromatography.
Involves specific interaction between solute
molecules and stationary phase i.e antibody antigen
reactions
One kind of molecule in
complex mixture
becomes attached to
the molecule that is
Covalently Bound to
stationary phase
or
N=16() 2
N=16 ()2
Or alternatively
N=5.55()2
N=5.55()2
The larger the value of N is for a column,
the better the column will be able to separate two
compounds that have small differences in
retention time
H = A + B/m + Cm
where:
m = linear velocity (flow-rate x V m/L)
H = total plate height of the column
A = constant representing eddy diffusion &
mobile phase mass transfer
B = constant representing longitudinal diffusion
C = constant representing stagnant mobile phase
& stationary phase mass transfer
RS = 2
Example
α ==
Example
In a chromatographic analysis of low-molecular-
weight acids, butyric acid elutes with a retention
time of 7.63 min. The column’s void time is 0.31
min. Calculate the capacity factor for butyric acid.
SOLUTION
In the same chromatographic analysis for low-molecular-
weight acids considered in the Example above, the retention
time for isobutyric acid is 5.98 min. What is the selectivity
factor for isobutyric acid and butyric acid?
SOLUTION
First we must calculate the capacity factor for isobutyric
acid. Using the void time from the example above, this is
Nonretained 3.1 -
A 5.4 0.41
B 13.3 1.07
C 14.1 1.16
D 21.6 1.72