ÄKTA start

Make sure that:

• The UV flow cell is assembled tightly.
• The inlet and the outlet tubing, and connectors are tightened to block any stray light from entering.
• The system is not exposed to direct sunlight.

Tap Diagnostics.

Result: The Diagnostics screen opens.

Tap Dark current test.

The result is displayed as Pass or Fail:

• Pass: the UV Monitor is working as it should.
• Fail: replace the UV Monitor.

Tap Stray light test.

The result is displayed as Pass or Fail:

Pass: the UV Monitor is working as it should.

Fail: clean, reassemble and secure the UV flow cell and the connected tubing. Make sure that the detector is not exposed to e.g., direct sunlight.

If the test fails again, replace the flow cell and then possibly the UV Monitor, or contact a Cytiva Service Engineer.

Tap Flow cell path length.

Result: The following screen opens.

Flush the UV flow cell thoroughly and leave it filled with demineralized water.

Tap Set baseline to capture a new reference value for the test.

Result: Abs should show a value close to 0 mAU.

Flush the UV flow cell with 1.0% acetone solution, and then leave it filled.

Note the new Abs value.

Calculate the actual Flow cell path length by using the following formula:

Cell (mm) = 2.00 * (new Abs value / 340 )

Update the calculated Cell length value by using up/down buttons or enter the value. Tap Save path length to save the value to permanent memory.

Note: The Abs value should be 340 ± 5% mAU, confirming that normalization has been done.

Note: For higher precision, instead of using acetone, use the Fe2(SO4)3 kit, product number: 18-1129-63.

Tap Configuration.

Result: The Configuration screen opens.

Tap Reset.

Result: A confirmation screen opens.

If the UV Monitor has been replaced, tap Yes.

Or,

If the UV Monitor has not been replaced, tap No to cancel the action.

In the Settings and service screen, tap Pump.

Result: The Pump screen opens.

Tap to select the intended option.

In the Pump screen, tap Diagnostics.

Result: The Diagnostics screen opens.

Use the up/down arrows to set the desired Flow Rate value in the range 0.5 to 5 ml/min.

• Prime the Pump and the flow path. Tap Start flow to run the Pump.
• Place the outlet tubing from the Wash valve (Waste port) in a pre-weighed collection tube.

Note: Before starting Diagnostics, prime the flow path with water and make sure that the outlet tubing where the pumped water is collected, is filled with demineralized water. This ensures that the volume of the collected water corresponds to the pumped volume.

• Tap Stop flow to turn the Pump OFF.
• Note the volume of the collected water (weigh the pre-weighed collection tube containing the collected water and estimate the volume of the pumped water).
• Calculate the flow rate and verify that it corresponds to the set flow rate.

Note: If the collected water does not correspond to the required volume (i.e., the Pump does not deliver water with the intended flow rate), inspect the condition of the pump tubing, then re-calibrate the Pump and re-diagnose.

If the collected water does not correspond to the required volume replace the Pump tubing.

After replacing the pump tubing, calibrate and diagnose the Pump again. If the collected water does not correspond to the required volume replace the Pump.

• Immerse the buffer inlet tubing A in demineralized water.
• Place the outlet tubing from the Wash valve in a pre-weighed collection tube.

Note: Before starting the calibration, prime the flow path with water and make sure that the outlet tubing where the pumped water is collected is filled with demineralized water. This ensures that the volume of the collected water corresponds to the pumped volume.

In the Pump screen, tap Calibration.

Result: The Calibration screen opens.

Use the up/down arrows to set the desired Flow rate.

• Tap Start flow to start the Pump.
• Collect water for at least one minute in the pre-weighed collection tube.
• Tap Stop flow to turn the Pump OFF.
• Note the volume of the collected water (weigh the pre-weighed collection tube containing the collected water and calculate the volume of the pumped water) and then set the Collected Volume value equal to this volume.

Tap Calibrate.

Result: The following screen opens.

Tap Pump tubing log.

Result: The Pump tubing log screen opens.

In the Pump tubing log screen, tap Reset.

Result: A confirmation screen opens.

Tap Yes if the pump tubing has been replaced.

Or,

If the pump tubing has not been replaced, tap No to cancel the action.

• Immerse the buffer inlet tubing A and B in demineralized water.
• Place the outlet tubing from the Wash valve in the waste bottle.

Start the Pump:

In the Settings and service screen, access Pump:Diagnostics screen.

Enter the desired flow rate and then start the Pump by tapping Start flow, and then tap Back:Back to return to the Settings and service screen.

In the Settings and service screen, tap Next to access the 2^nd screen.

Result: The following screen opens.

In the Settings and service screen, tap Buffer valve.

Result: The Buffer valve screen opens.

Check that the valve position Buffer A is selected.

Tap Turn valve to check that the Buffer valve switches between Buffer A and Buffer B on the Display. There is also a click sound when the valve switches.

Check that the flow is delivered from the Buffer A inlet or the Buffer B inlet, according to the selected valve position.

Note: The Buffer valve is by default in the Buffer A position (the liquid is delivered from the Buffer A inlet).

Visually inspect the flow at the outlet.

If the water is not flowing properly, check the following and fix accordingly:

• Is there a blockage in the tubing?
• Are the connectors tightly connected?

Stop the Pump:

• In the Settings and service screen, access Pump:Diagnostics screen.
• Stop the Pump by tapping Stop flow.

If the valve has been replaced, tap Reset to set the Valve switches counter to 0.

Result: A confirmation screen opens.

Tap Yes to confirm the reset if the valve has been replaced.

Or,

If the valve has not been replaced, tap No to cancel the action.

• Immerse the buffer inlet tubing A and the sample inlet tubing in demineralized water.
• Place the outlet tubing from the Wash valve in the waste bottle.
• Check that Buffer A is selected in the Buffer valve screen.

Start the Pump:

• In the Settings and service screen, access Pump:Diagnostics screen.
• Enter the desired flow rate and then start the Pump by tapping Start flow.

In the Settings and service screen, tap Next to access the 2nd screen.

Result: The following screen opens.

In the Settings and service screen, tap Sample valve.

Result: The Sample valve screen opens.

• Check that the Valve position Buffer is selected.
• Tap Turn valve to check that the Sample valve switches between Buffer and Sample on the Display. There is also a click sound when the valve switches.
• Check that the flow is delivered from the Buffer inlet or the Sample inlet according to the selected valve position.

Note: The Sample valve is by default in the Buffer position (the liquid is delivered from the Buffer inlet).

Visually inspect the flow at the outlet. If the water is not flowing properly, check the following and fix accordingly:

• Is there a blockage in the tubing?
• Are the connectors tightly connected?

Stop the Pump:

In the Settings and service screen, access Pump:Diagnostics screen.

Stop the Pump by tapping Stop flow.

If the valve has been replaced, tap Reset to set the Valve switches counter to 0.

Result: A confirmation screen opens.

Tap Yes to confirm the reset if the valve has been replaced.

Or,

If the valve has not been replaced, tap No to cancel the action.

• Immerse the buffer inlet tubing A in demineralized water.
• Place the outlet tubing from the Wash valve and the outlet tubing from the Outlet valve in the waste bottle.
• Check that:
  • Buffer A is selected in the Buffer valve screen
  • Buffer is selected in the Sample valve screen

Start the Pump:

In the Settings and service screen, access Pump:Diagnostics screen.

Enter the desired flow rate and then start the Pump by tapping Start flow.

In the Settings and service screen, tap Next to access the 2^nd screen.

Result: The following screen opens.

In the Settings and service screen, tap Wash valve.

Result: The Wash valve screen opens.

• Check that the valve position Waste is selected.
• Tap Turn valve to check/listen that the Wash valve switches between Waste and Column. There is a click sound when the valve switches.
• Check that the flow is diverted to the Waste outlet or the Column outlet according to the selected valve position.

Note: The Wash valve is by default in Waste position (the flow is diverted to waste).

Visually inspect the flow at the outlet. If the water is not flowing properly, check the following and fix accordingly:

• Is there a blockage in the tubing?
• Are the connectors tightly connected?

Stop the Pump:

• In the Settings and service screen, access Pump:Diagnostics screen.
• Stop the Pump by tapping Stop flow.

If the valve has been replaced, tap Reset to set the Valve switches counter to 0.

Result: A confirmation screen opens.

Tap Yes to confirm the reset if the valve has been replaced.

Or,

If the valve has not been replaced, tap No to cancel the action.

• Immerse the buffer inlet tubing A in demineralized water.
• Place the outlet tubing from the Outlet valve in the waste bottle.
• Check that:
  • Buffer A is selected in the Buffer valve screen
  • Buffer is selected in the Sample valve screen
  • Column is selected in the Wash valve screen

Start the Pump:

• In the Settings and service screen, access Pump:Diagnostics screen.
• Enter the desired flow rate and then start the Pump, tap Start flow.

In the Settings and service screen, tap Next to access the 2^nd screen.

Result: The following screen opens.

In the Settings and service screen, tap Outlet valve.

Result: The Outlet valve screen opens.

• Check that the valve position Waste is selected.
• Tap Turn valve to check that the Outlet valve switches between Waste and Collection. There is also a click sound when the valve switches.
• Check that the flow is diverted to the Waste outlet or the Fraction collector outlet according to the selected valve position.

Note: The Wash valve is by default in Waste valve position. Hence, the flow is diverted to Waste.

Visually inspect the flow at the outlet. If the water is not flowing properly, check the following and fix accordingly:

• Is there a blockage in the tubing?
• Are the connectors tightly connected?

Stop the Pump:

In the Settings and service screen, access Pump:Diagnostics screen.

Stop the Pump by tapping Stop flow.

If the valve has been replaced, tap Reset to set the Valve switches counter to 0.

Result: A confirmation screen opens.

Tap Yes to confirm the reset if the valve has been replaced.

Or,

If the valve has not been replaced, tap No to cancel the action.

In the Settings and service screen, tap Next:Next to access the 3^rd screen.

Result: The following screen opens.

Tap Conductivity.

Result: The Conductivity screen opens.

Tap Conductivity.

Result: The Configuration screen opens.

Set the Cell constant value (Set cell const) for the new Conductivity flow cell by using the up/down arrows.

Tap Save to save the new value.

Note:

• The Cell constant value is provided on the packaging of the Conductivity flow cell.
• If a Cell constant value is not available, calibrate the Conductivity flow cell as explained in Temperature sensor calibration and Conductivity flow cell calibration.

In the Conductivity:Configuration screen, set the reference temperature (Set ref temp) value in the range 4°C to 35°C.

In the Conductivity screen, tap Advanced calibration.

Result: The Advanced calibration screen opens.

Note the temperature and enter it into the Set actual temp text box, and then tap Calibrate to carry out the temperature calibration

Note: 

• Make sure that the temperature of the Conductivity flow cell has stabilized and measure the temperature of the calibration solution with a precision thermometer.

In the Conductivity screen, tap Calibration.

Result: The Calibration screen opens.

Note: Make sure that the checkbox for Enable Temperature Compensation is checked.

In the Calibration screen, enter the conductivity value at the current temperature in the Set Conductivity field and then tap Calibrate to carry out the Conductivity calibration.

Note: 

• If a certified conductivity standard solution is used, use the supplied theoretical conductivity value corresponding to the temperature in question.
• If a manually prepared 1.00 M NaCl calibration solution is used, get the conductivity value at the current temperature from the graph for conductivity of the 1.00 M NaCl as a function of temperature presented below.

Disconnect the inlet tubing from the Pressure sensor to expose the sensor to atmospheric pressure only.

Note: Make sure that the Pump is OFF before disconnecting the tubing.

In the Settings and service screen, tap Pressure sensor.

Result: The Pressure sensor screen opens.

In the Pressure sensor screen, tap Zero offset.

Result: A Message screen opens.

Make sure that there is no back pressure in the system, and then tap OK.

In the Settings and service screen, tap Fraction collector.

Result: If the Fraction collector is enabled, the following Fraction collector screen opens.

If the Fraction collector is not enabled, the following screen opens.

Tap Enable Frac to enable the Fraction collector.

Or,

To disable the Fraction collector, tap Disable Frac.

In the Fraction collector screen, tap Diagnostics.

Result: The Diagnostics screen opens.

Tap Home test and visually observe that Frac30 rotates to home position (tube no. 1) from current position.

Note: If Frac30 does not rotate the Bowl assembly, check if the Frac30 cable is connected properly to ÄKTA start. If the cable is not connected properly, connect the cable properly and secure it in position with a screw driver if required.

In the Fraction collector screen, tap Run log.

Result: The Run log screen opens.

If the drive sleeve has been replaced, tap Reset to set the Run time to 0.

Result: A confirmation screen opens.

Tap Yes to confirm the reset if the drive sleeve has been replaced.

Or,

If the drive sleeve has not been replaced, tap No to cancel the action.

In the Settings and service screen, tap Next:Next to access the 3^rd screen.

Result: The following screen opens.

Tap Display to access the Instrument Display options.

Result: The Display screen opens.

In the Display screen, tap Touch screen calibration.

Result: A warning screen opens.

Tap Yes to proceed with the calibration of the Touch screen.

Result: The following screen opens.

Tap precisely on the marked circle 1.

Result: The following screen opens.

Tap precisely on the marked circle 2.

Result: The following screen opens.

Tap precisely on the marked circle 3.

Result: The following screen opens.

Tap precisely on the marked circle 4.

Result: The following screen opens.

Tap Exit.

Note: If the calibration fails, repeat the test. If the calibration fails again, contact a Cytiva Service Engineer.

In the Display screen, tap Color test.

Result: The following screen opens.

Tap Exit.

Result: The Color test is completed.

Note: If the test fails, contact a Cytiva Service Engineer.

In the Display screen, tap Diagnostics.

Result: The following screen opens.

In the Select backlight brightness screen, tap a radio button to select the intended brightness (%).

Result: The rectangle shows the backlight brightness at the selected brightness level.

In the Display screen, tap Log book.

Result: The following screen opens.

Note the number of hours the Instrument Display has been used.

If the Instrument Display has been replaced, tap Reset to set the number of hours to 0.

Result: A confirmation screen opens.

Tap Yes to confirm the reset if the Instrument Display has been replaced.

Or,

If the Instrument Display has not been replaced, tap No to cancel the action.

In the Settings and service screen, tap System

Result: The following screen opens.

In the System screen, tap Firmware update.

Result: The following screen opens.

Tap Microprocessor firmware update.

Result: The following screen opens.

Tap Yes and then wait for approximately 3 minutes for the update to be done.

Note: When the firmware update is completed, the instrument automatically restarts and displays the version of the firmware.

Tap FPGA update.

Result: The following screen opens.

Tap Yes. The updating may take up to 5 minutes.

Result: The following screen opens.

Once the firmware is updated, perform the following calibration sequence:

• Display:Touch screen calibration
• Pressure sensor:Zero offset
• Pump : Calibration:Flow rate
• UV:UV LED calibration
• Conductivity:Advanced calibration (temperature sensor calibration)
• Conductivity:Calibration (flow cell calibration)

In the System screen, tap Export System Report to USB.

Result: The files INSTLOG.TXT and ERRORLOG.TXT are exported to the USB memory stick.

In the System screen, tap Delay volume setting.

Result: The following screen opens.

Enter the internal diameter (ID) and length of the tubing from the Outlet valve to Frac30 in the respective fields, and then tap Save.

Result: The total delay volume from the UV Monitor to Frac30 is displayed in the Total Delay volume field.

Note: The delay volume from the UV Monitor to the Outlet valve is constant (0.27 ml) in all ÄKTA start instruments.

In the Settings and service screen, tap System.

Result: The following screen opens.

In the System screen, tap Switch valve timing.

Result: The following screen opens.

Tap the radio button to select Switch valve timing B (switch time 5 sec.).

Tap Save to save the timing.

If wavy gradients still are obtained, or if fluctuations on step gradient levels are large, then select Advanced timing.

Result: The following screen opens.

Set switch valve time in the range of 3.0 to 5.0 sec (0.1 sec increments) by pressing the up/down arrows.

Tap Save to save the optimized timing.

Loosen the M3 screw from the valve using a T10 torx driver, supplied with the equipment at delivery.

Note: The screw can be located on different places on the top circumference of the holder.

Replace with a new Buffer valve, connect the cable, and reconnect the tubing to the valve ports.

Note: Before replacing the valve, make sure that the part number (P/N) on the new valve matches with that of the old valve (29003271, Buffer valve CTV-31-516U-1).

Disconnect the RJ45 connector from the UV Monitor side.

Note: Make sure that the RJ45 connector is not moved inside the cabinet. The connector needs to be retained outside the cabinet for re-assembling of a new UV module.

Place the protective cover around the flow cell to protect the electronics inside the optical unit from liquid spillage.

Note: The protective cover should be assembled after completing the assembly of the UV flow cell inside the UV Monitor by just press-fit.

In the Settings and Service screen, tap UV. In the UV screen, perform a UV LED calibration and a Flow cell path length test.

This is described further in the "Access the modules" section above.

Loosen the screw from the valve using a T10 torx driver, supplied with the equipment at delivery.

Note: The screw can be locate on different places on the top circumference of the holder.

Replace with a new Sample valve, connect the cable, and reconnect the tubing to the valve ports.

Note: Before replacing the valve, make sure that the part number (P/N) on the new valve matches with that of the old valve (29003272, Sample valve CTV-31-32U-3).

Loosen the two M4 screws from the Pump using a T20 screwdriver, supplied with the equipment at delivery.

Note: Make sure that the pump connector is not moved inside the cabinet. The connector needs to be retained outside the cabinet for re-assembling of a new Pump module.

Note: While loosening the screw, make sure to hold the screw in your hand before it is completely out of the Pump. There is a risk that the screw will fall into the pump head if proper care is not taken during the disassembly.

Remove the Pump.

Note: Make sure that the pump cable is placed away from the fan, on the left side of the instrument after removing the Pump. This safety precautions is to ensure that the cable does not get stuck in the ventilation fan.

Place the tubing between the rollers and the track, press against the pump head inner wall.

Note: Make sure that the pump tubing is not twisted or stretched against the rollers.

Lower the flip top until it clicks into its fully closed position.

The track closes automatically and the tubing is stretched correctly as the track closes.

Loosen the M3 screw from the Wash valve using a T10 torx driver, supplied with the equipment at delivery.

Note: The screw can be located on different places on the top circumference of the holder.

Loosen the M3 screw from the Outlet valve using a T10 torx driver, supplied with the equipment at delivery.

Note: The screw can be located on different places on the top circumference of the holder.

Replace with a new Outlet valve. Connect the cable, and then reconnect the tubing to the valve ports.

Note: Before replacing the valve, make sure that the part number on the new valve matches with that of the old valve (29003274 Outlet valve CTV-31-32U-1).

Remove the Conductivity Monitor and leave the connector as it is.

Note: Make sure that the connector is not moved inside the cabinet. The connector needs to be retained outside the cabinet for re-assembling of a new Conductivity module.

Hold the Bowl assembly and orient the Bowl with the aligning ribs oriented towards the aligning groove on the Bowl holder.

Note: Do not lift the fraction collector by holding the Dispenser arm.

Tip: If it is hard to locate the alignment feature in corresponding parts, hold the Bowl assembly near the top of the Bowl holder and rotate the Bowl holder unless the aligning ribs gets located.