cytiva.com Good ÄKTA system practice Cue Cards 1. Good ÄKTA system practice 2 2. Run-Essentials 3 3. Fraction collector F9 and F9-C operation 4 4. Comprehensive System Wash and cleaning 5 5. UNICORN system notifications 6 6. Troubleshooting 7 6.1 Conductivity signal – a useful troubleshooting tool 7 6.2 Pressure alarms 8 6.3 Backpressure contributions 8 6.4 Fluctuating pressure 9 6.5 Fluctuating UV and conductivity signal, examples and solutions 10 6.6 Cleaning the pH electrode and UV flow cell 11 6.7 Air bubble in a flow cell 12 6.8 Broken connector tip stuck in valve 13 6.9 Tubing guide 13 7. Abbreviations and References 14
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cytiva.com
Good ÄKTA system practiceCue Cards
1. Good ÄKTA system practice 22. Run-Essentials 33. Fraction collector F9 and F9-C operation 44. Comprehensive System Wash and cleaning 55. UNICORNsystemnotifications 66. Troubleshooting 7
NOTE: TheFractioncollectorcontentdoesnotresetafterthemethodhasendedunlessthedoorhasbeenopened and closed.
The Fraction collector content is updated andallcassettesarereadytobeused.ToviewthecontentoftheFractioncollector,in the System control module select View:Fraction Collector Content.
Take out fractions during a run or in the middle of scouting or method queue runs
NOTE: Thereisapossibilitytotryoutotherdeep-wellplatesthentheonesapprovedbyCytiva,providedthatthedeep-wellplatespecificationsarefulfilled.Insomecases,theautomaticcassettescanningmayneedtobedisabledinSystem Control/System Settings/Fraction collector/Cassette configurationtomakethemwork.Makesuretomanually update the Fraction collector content in System settingseverytimethecontentischanged.ForfurtherdetailsseeÄKTAavantUserManualandÄKTApureUserManual(“References”onpage14).
Good ÄKTA system practice Cue Cards 29109616 AC 5
For further details see ÄKTA avant User Manual and ÄKTA pure User Manual.
4. Comprehensive System Wash and clean-ing
Cleanthe syringe port
of the injection valvewith a syringe
ComprehensiveSystem Washand cleaning
Clean thepH electrode
(if present)
Place allinlets in the firstcleaning solution
Start run of:
System PreparationOR
System CIP
method
Follow theinstructions in
UNICORN and moveinlets to the nextcleaning solution
1. In the System Controlmodule,selectSystem:Maintenance Manager to open the Maintenance Manager dialog.
2. In the Maintenance Managerdialog,clicktheNew System Notification.Result:TheNewNotificationfieldappears.
3. In the NewNotificationfield:a. Enteranameforthenotification,e.g.“Changepumprinsing
solution.”
b. Selectatimeintervalafterwhichthenotificationwillbeissued, e.g.oneweek.
c. Writeamessageinthemessageinputfieldthatwillbeshownforthemaintenancenotification,e.g.“Pleasechangethepumprinsingsolutionwithafreshlymade20%ethanolsolution.”
d. ClickApplytosavethechangesandapplythenotificationsettings.
conductivity signal is stablebeforesampleinjection
Note: The conductivity signal becomes distorted at higher salt concentration since the ion activity decreases when the concentration increases, i.e. a different curvature than the theoretical one is therefore normal at high salt concentrations.
Good ÄKTA system practice Cue Cards 29109616 AC8
For further details see ÄKTA avant User Manual and ÄKTA pure User Manual.
Dilute viscous samples or decreasetheflowrateduringsample application. Remove thesampleinletfilterandifus-ing the system pump to apply sample,bypassthemixer.
Toavoidover-pressure,ÄKTApure and ÄKA avant systems have optional pressure-con-trolledsampleapplication,wheretheflowrateisdecreased as the pressure increases and vice versa.
Column
Clean the column. Seecolumninstructionsforcleaning.
Donotusesmallerbeadsorcolumn diameter than the applicationrequires.
No air bubbles Small air bubles passing UV and cond flow cell
Small air bubles passing UV and cond flow cell
Small air bubles passing UV and cond flow cell
Small air bubles passing UV and cond flow cell
No air bubbles Air bubble flushed out with increased flow rate
Air bubble stuck in cond
UV
Conductivity
System flow
Airin buffers
De-gasbuffers
Temperate or de-gas
buffers
Coldbuffers
run at RT
Make sure the flow restrictor
is inline
Air inUV flow cell
Flow restrictoris offline
Set flowrestrictor
inline
Clean theUV flow cell
Dirt in theUV flow cell
Coldsample
run at RT
Reducethe flow rate
Effectsdue to air
Air spikes in UV signal
Large air segments passing the flow cells
Good ÄKTA system practice Cue Cards 29109616 AC 11
For further details see ÄKTA avant User Manual and ÄKTA pure User Manual.
6.6 CleaningthepHelectrodeandUVflowcell
Component Contaminant Cleaning solution
pHelectrode Salt deposits 0.1MNaOHand0.1MHCl
Lipid deposits Detergent or organic solvent
Proteindeposits 1%pepsinin0.1MHCl
UVflowcell Salt/lipid/proteindeposits
Detergent,suchasDeconex™
Good ÄKTA system practice Cue Cards 29109616 AC12
For further details see ÄKTA avant User Manual and ÄKTA pure User Manual.
6.7 Airbubbleinaflowcell
Large air segments assing UV and conductiv-ity flow cell
Air bubble stuck in cond flow cell
Air compressed int liquid and flushed out due to increased back pressure
UV
Conductivity
System flow
System pressure
YES
YES
YES
NO
NO
Is the flow restrictor
offline?
Set the flow restrictor inline and continue with the run.
Bypass the column and increase the flow rate to 5 ml/min.
Continuewith the run.
Connect an extra small i.d. tubing (see page 13) to the used outlet to increase the backpressure.
Remove the extra tubing and continue with the run.
Does the detector signal
go back to original level?
Does the detector signal
go back to original level?
Remove the extra tubing and flush the flow cells with 100% MeOH. (Make sure that the MeOH is not coming in contact with high salt concentration buffers.)
Good ÄKTA system practice Cue Cards 29109616 AC 13