GE akta start - Quick Start Method Details; Affinity Chromatography (AC) Quick Start; Desalting;Buffer Exchange (DS) Quick Start

ÄKTA start Maintenance Cue Card, 29-0240-43 AC 9
System performance test
System performance test is performed to make sure
the system is performing within acceptable limits. It is
recommended to run this method during installation of
the instrument or after replacement of modules such
as Pump, UV, Conductivity or Valves. For a detailed
description of the test, refer to ÄKTA start Operating
Instructions, section 5.4 System performance.
Required solutions
Buer A: DM water
Buer B: 1.0% Acetone, 1.0 M NaCl
Sample: 1.0% Acetone, 1.0 M NaCl (Buer B)
Checklist
Calibration of all the modules: Pressure sensor, Pump,
UV and Conductivity.
Set the conductivity reference temperature to 20 °C.
No column should be present in the ow path.
Immerse Buer port A inlet in Buer A (DM water).
Immerse Buer port B inlet in Buer B (1.0% Acetone,
1.0 M NaCl).
Sample valve inlet immersed in sample (1.0% Acetone,
1.0 M NaCl).
Make sure that 1 ml Sample loop is lled with sample
(1.0% Acetone, 1.0 M NaCl).
Make sure that 2 m of 0.5 mm ID tubing is connected to
the Outlet valve at Waste position.
When performing System performance without Fraction
collector, make sure the outlet valve fractionation tubing
is inserted into a pre-weighed beaker.
When performing System performance with Fraction
collector, make sure the outlet valve fractionation tubing
is connected to Fraction collector with at least 5 pre-
weighed tubes.
Make sure that the system is prelled with DM water.
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Make sure to take note of all the required observations
by recording parameters while the performance run
is in progress. Enter the observed values in the ÄKTA
start report template available in ÄKTA start Operating
Instructions, section 10 Appendix.
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Operation from the Instrument Display
Method run
Prepare system
System performance
method
EndRun
Exit
ÄKTA start
Note the current temperature of the calibration
solution in the conductivity ow cell as displayed in the
Cell temperature eld.
In the Calibration screen, enter the conductivity value
at current temperature in the Set conductivity eld
and then tap Calibrate to carry out the calibration.
Note:
If a certied conductivity standard solution is used, use
the supplied theoretical conductivity value corresponding
to the temperature in question.
If a manually prepared 1.0 M NaCl calibration solution
is used, get the conductivity value at the current
temperature from the graph for conductivity of 1.0 M
NaCl as a function of temperature presented in the ÄKTA
start Maintenance Manual.
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Acceptance criteria
Time
(min)
Activity Check Aproved
interval
0 Pump wash Wash valve
position
Mobile phase
out through
Waste
1 1 ml/min, 0 %B, ow
through Outlet valve,
Waste position
Back pressure =< 0.05 MPa
2 Repeat UV Auto Zero
3 5 ml/min Back pressure 0.06 - 0.2 MPa
UV level ± 10 mAU
Cond. level ± 1 mS/cm
4 1 ml/min, Sample
valve, Sample position
Max UV level 300-380 mAU
Max Cond. level 65 - 95 mS/cm
7 1 ml/min, Sample
valve, Buer position
10 Request switch Injec-
tion valve to Inject
position
Max UV level 300-380 mAU
Max Cond. level 65 - 95 mS/cm
13 Request switch Injec-
tion valve back to
Load position
15 Start gradient,
0-100 %B in 10 min,
start fractionation/
collection
19 End fractionation
1
Weigh fraction no
2, 3 and 4
0.8 - 1.2 g
Max di. between
fractions
0.1 g
20
End collection
2
Weigh beaker 4.2 - 5.8 g
25
End gradient, stay at
100 %B
Gradient Straight, no
negative dips.
28
50% B Gradient level
3
45 - 55 %B
36 0% B (re-equilibrate)
41 End Check all connec-
tions for leakage
No leakage
1
with Fraction collector
2
without Fraction collector
3
UV 50% B / UV 100% B