Background Considerations - GE AKTA pilot 600 User Manual

4.1.1

Background considerations

Chromatography techniques
Different chromatography techniques make different demands on sample application,
as summarized in the table below. These considerations can influence the choice of
sample application technique.
More information about chromatography techniques may be found in the Life Science
Handbooks, available for download from the Support pages on www.gelifesciences.com.
Technique
Size exclusion chro-
matography (SEC)
Adsorption chromatogra-
phy, (e.g., affinity or ion
exchange chromatogra-
phy)
Sample application options
Samples may be applied to the column from a system inlet (A inlet) or from a Superloop
150 SNAP mounted on the column valve.
Option
Apply sample from
a system inlet
ÄKTA pilot 600 User Manual 29274559 AA
Demands
A sharply defined sample band with constant sample
concentration throughout. Any broadening of the sample
band will be reflected in broadening of the elution peaks.
The sample volume applied to the column should ideally
not be more than 10% of the column volume. Precise
requirements will depend on the separation being per-
formed.
Sample is concentrated by adsorption on to the chro-
matography resin during sample application. The volume
and concentration of sample is not critical, provided that
the capacity of the column is not exceeded.
When?
Recommended for sam-
ple volumes larger than
50 mL and for column
performance tests where
sample is readily avail-
able.
4.1 Sample application
4.1.1 Background considerations
Description
The sample is pumped onto the
column from one of the A inlets
using system pump A. The sample
passes through the system pump,
flow restrictor and any other mod-
ules before the column valve in the
flow path.
The sample volume that can be
applied from a system inlet is in
principle unlimited.
4 Operation
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