Amersham Pharmacia Biotech AKTA P-920 User manual

User Manual
18-1125-54
Pump P-920
18-1125-54_ab.qxd 1998-10-22 12:49 Sida 1 (1,1)

Trademarks
ÄKTA, FPLC and UNICORN are trademarks of
Amersham Pharmacia Biotech Limited or its
subsidiaries.
Amersham is a trademark of Nycomed Amersham plc.
Pharmacia and Drop Design are trademarks of
Pharmacia & Upjohn Inc.
Terms and Conditions of Sale
All goods and services are sold subject to the terms
and conditions of sale of the company within the
Amersham Pharmacia Biotech group which supplies
them. A copy of these terms and conditions of sale is
available on request.
Adresses
Amersham Pharmacia Biotech UK Limited
Amersham Place
Little Chalfont
Buckinghamshire
England HP7 9NA
Amersham Pharmacia Biotech AB
SE-751 84 Uppsala
Sweden
Amersham Pharmacia Biotech Inc
800 Centennial Avenue
PO Box 1327
Piscataway
NJ 08855 USA
© Amersham Pharmacia Biotech AB 1998
– All rights reserved
Important user information
Meaning: Consult the instruction manual to avoid
personal injury or damage to the product or other
equipment.
WARNING!
The Warning sign is used to call attention to the
necessity to follow an instruction in detail to avoid
personal injury. Be sure not to proceed until the
instructions are clearly understood and all stated
conditions are met.
CAUTION!
The Caution sign is used to call attention to
instructions or conditions that shall be followed to
avoid damage to the product or other equipment. Be
sure not to proceed until the instructions are clearly
understood and all stated conditions are met.
Note
The Note sign is used to indicate information
important for trouble-free or optimal use of the
product.
Should you have any comments on this instruction, we
will be pleased to receive them at:
Amersham Pharmacia Biotech
SE–751 84 Uppsala
Sweden
!
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Contents
1 Introduction
1.1 General ..................................................................................................................1
1.2 Accessories.............................................................................................................2
1.3 Safety.....................................................................................................................3
2 Installation
2.1 Unpacking..............................................................................................................4
2.2 General precautions ...............................................................................................4
2.3 Connecting electrical signal cables .........................................................................4
2.4 Connecting to UniNet-1 communication link ........................................................5
2.5 Connecting to supply voltage.................................................................................5
2.6 Rinsing tubing .......................................................................................................6
2.7 Connecting the inlet and outlet tubing...................................................................6
2.8 Running-in the new pump .....................................................................................7
2.9 Installation of accessories.......................................................................................8
3 Operation
3.1 On/off....................................................................................................................9
3.2 Menu selection and settings ...................................................................................9
3.3 Menu overview ....................................................................................................10
3.4 Starting and stopping the pump...........................................................................11
3.5 Setting the flow rate and starting the pump.........................................................12
3.6 Setting concentration B........................................................................................12
3.7 Running a simple gradient ...................................................................................13
3.8 Ending the run and storage..................................................................................13
3.9 Changing eluent ..................................................................................................14
3.10 Restart after power failure...................................................................................15
4 Maintenance
4.1 Periodic maintenance ...........................................................................................16
4.2 Pump cleaning .....................................................................................................16
4.3 General care.........................................................................................................17
4.4 Changing rinsing solution....................................................................................17
4.5 Testing leakage ....................................................................................................18
4.6 Replacing the piston seal .....................................................................................19
4.7 Replacing a damaged piston ................................................................................22
4.8 Cleaning the 6-port pump valve...........................................................................22
Pump P-920
User Manual 18-1125-54 Edition AB i
Contents
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5 Trouble-shooting
5.1 General ................................................................................................................23
5.2 Faults and actions................................................................................................23
5.3 Error messages.....................................................................................................25
5.4 Checking the pump pressure................................................................................27
A-D Reference information
A Description ..........................................................................................................28
A.1 Module .....................................................................................................28
A.2 Rear panel.................................................................................................28
A.3 Fluid delivery ............................................................................................29
A.4 6-port pump valve.....................................................................................29
A.5 Pressure monitoring ..................................................................................30
A.6 Protective covers .......................................................................................30
A.7 Using an external chart recorder ...............................................................30
B Menus..................................................................................................................31
B.1 Check menu ..............................................................................................31
B.1.1 Checking piston status..............................................................31
B.1.2 Checking the number of piston strokes.....................................31
B.1.3 Checking mixer run time ..........................................................31
B.1.4 Checking service mode..............................................................31
B.2 Setup menu ...............................................................................................32
B.2.1 Set pressure limit.......................................................................32
B.2.2 Set wash pressure limit .............................................................32
B.2.3 Set pressure unit........................................................................32
B.2.4 Set gradient base.......................................................................32
B.2.5 Set mixer operation...................................................................32
B.2.6 Set pulse compensation mode ...................................................33
B.2.7 Set pressure offset .....................................................................33
B.2.8 Setup language..........................................................................33
B.2.9 Setup unit number ....................................................................33
B.2.10 Setup display angle ...................................................................33
B.3 Alarm timer...............................................................................................34
B.4 Service displays .........................................................................................34
B.5 Menu and text overview ...........................................................................35
C Technical specifications .......................................................................................36
C.1 Operating data..........................................................................................36
C.2 Physical data .............................................................................................36
D Accessories and consumables...............................................................................38
Short instructions on back page
Pump P-920
User Manual 18-1125-54 Edition AB
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Contents
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About this manual
This manual comprises two parts; a practical part (sections 1 – 5)
and a reference part (sections A – D). Sections 1 – 5 contain the
necessary information for installing, operating and maintaining
the instrument.
Contents
Pump P-920
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Contents
Pump P-920
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1 Introduction
1.1 General
Pump P-920 is a high precision laboratory pump for use in liquid
chromatography and other applications where constant flow is
required. The performance of Pump P-920 is accurate and
reproducible from low flow rates over the whole pressure range. The
chemical resistance of the pump allows its use with corrosive liquids
and organic solvents as well as aqueous solutions with high salt
concentrations.
Pump P-920 facilitates routine chromatography work.
It is utilised as system pump in ÄKTA™ FPLC™chromatography
systems, or is used as a stand-alone unit. The flow rate is set locally
with a front dial, or from a PC running UNICORN™ 3.0 or higher.
Pump P-920 works with a wide range of columns and media
supplied by Amersham Pharmacia Biotech.
The pump can be used to form accurate and reproducible gradients.
In stand-alone configurations, setup parameters are stated in a Setup
menu, see Reference information B.2.
Pump P-920 features:
• A pressure sensor connected to pump module A and B.
• Pulse compensation
P-920
0
1
Pump P-920
User Manual 18-1125-54 Edition AB 1
Introduction 1
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Pump P-920
User Manual 18-1125-54 Edition AB
2
1.2 Accessories
Mixer M-925 can be connected to the pump. When P-920 is
connected to UNICORN, the mixer can be controlled on/off/auto
from UNICORN. When P-920 is run as a stand-alone unit, the
mixer is operated via the local user interface.
M-925
1 Introduction
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1.3 Safety
•The module is designed for indoor use only.
• Do not use in a dusty atmosphere or close to spraying water.
• Do not block the air inlet and outlet of the unit.
WARNING! Hands and fingers risk being squeezed between the
piston driving arm and the pump housing when running the pump
without the protective covers fitted.
WARNING! During normal operation, all protective covers over
internal capillaries, piston rods and glass cylinders must be in
place when running the pump.
WARNING! Incorrectly fitted tubing may loosen, causing a jet of
liquid to spray out. This is especially dangerous if hazardous
chemicals are being used. Connect the tubing by first inserting the
tubing fully, then tightening the connector fingertight.
WARNING! NaOH is injurious to health. Avoid spillage.
WARNING! When using hazardous chemicals, all suitable
protective measures, such as protective glasses, must be taken.
WARNING! The module must not be opened by the user. It
contains high voltage circuits that can give a lethal electric shock.
WARNING! Always disconnect the power supply before
attempting to replace any item on the module.
WARNING! The module must be connected to a grounded mains
socket.
Pump P-920
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Introduction 1
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Pump P-920
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4
2 Installation
2 Installation
2.1 Unpacking
Unpack the module and check the items against the supplied packing
list. Inspect the items for obvious damage that may have occurred
during transportation.
We recommend that all packing materials be retained if onward
transport of the module is expected.
2.2 General precautions
The module should be installed in a non-corrosive atmosphere.
The module should be located in a place of low temperature
variations, away from heat sources, draughts and direct sunlight.
The module may be operated at normal ambient temperatures in the
range +4 to +40 °C.
The module should be installed on a stable laboratory bench or in
an ÄKTAdesign chromatography system. To ensure correct
ventilation, a free space of 0.1 m is required behind and in front of
the module. Place the module directly on the bench. To ensure that
the ventilation inlet below the front is not blocked, do not place soft
material under the module.
2.3 Connecting electrical signal cables
The sockets for electrical signals are located on the rear panel.
Voltage
WARNING! For continued protection against
risk of fire, replace only woth fuse
of the specified type and current ratings
Frequency Power, max Fuse
100-200 V- 50-60 Hz 600VA T 6,3 AL
Leakage current, max
3,5 mA
Mains
Mains output
UniNet 1
UniNet 2
Remote
Pressure
Analogue out 0-1 V
CAUTION! Read the following information carefully to ensure
that the module is installed correctly.
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Pump P-920
User Manual 18-1125-54 Edition AB 5
Installation 2
Connecting to chart recorder (if used)
1 Connect the chart recorder to the Mini-DIN-socket Pressure
Analogue out 0-1 V using the cable supplied.
Pin Signal
1 Pressure signal
2 Signal ground
3–6 Not used (reserved for factory testing)
Note: The signal cable is delivered with protective covers on each
wire. Do not remove the protective covers from unused connections
as a short circuit may disturb the measurement.
2 Set the recorder to 0–0.5 V input, full scale. 0.1 V corresponds to
1 MPa.
2.4 Connecting to UniNet-1 communication link
When used in ÄKTAdesign chromatography system, the pump is
controlled from a PC running UNICORN version 3.0 or higher via
UniNet-1 cables.
1 Connect two UniNet-1 cables to the UniNet-1connectors. The
module can be connected in series anywhere in the chain between
the PC and the termination plug. The UniNet-1 link connects, in
series, the PC with Pump P-920 and other modules. The
termination plug is connected to the last module in the chain.
2.5 Connecting to supply voltage
1Make sure the on/off switch is in the OFF-position marked O.
2 Connect a mains cable between the module and a grounded
mains socket. Any voltage from 100–240 V AC, 50–60 Hz can be
used.
The module contains one user-replaceable fuse. See Technical
Specifications for fuse ratings.
WARNING! The module must be connected to a grounded
mains socket.
CAUTION! The mains power to the ÄKTAdesign
chromatography system must be switched OFF before connecting
the module to the UniNet-1 link.
1
2
3
4
5
6
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2.6 Rinsing tubing
The rinsing tubing for the piston seals is installed at the factory and
filled with 20% ethanol. Check that the rinsing solution is present
behind the pistons and that the rinsing tubing is connected and
undamaged. Always use 20% ethanol as rinsing solution. Change
rinsing solution at least every month.
2.7 Connecting the inlet and outlet tubing
In ÄKTAFPLC,the inlet and outlet tubing are installed at the factory.
When using P-920 in stand-alone applications, the inlet and outlet
tubing must be installed.
The outlet tubing (PEEK tubing, i.d. 0.50 mm, o.d. 1/16”) has
tubing connectors attached at both ends.
The inlet tubings (A1, B1, teflon tubing i.d. 1.6 mm, o.d. 1/8”) have
tubing connectors attached at one end and inlet filters at the other
end. The inlet filters have replacable filter inserts.
The inlet tubings are connected to the inlets marked A IN and B IN
on the pump valves and the outlet tubing is connected to the upper
connection on the pressure sensor marked OUT. The other end of the
outlet tubing is connected to a mixer inlet.
Pump P-920
User Manual 18-1125-54 Edition AB
6
2 Installation
Pump rinsing tubing
Pump rinsing tubing
Inlet pump
module A Inlet pump
module B
Outlet to
mixer
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2.8 Running-in the new pump
General
To be able to follow this section, you must know how to operate the
pump. Read through section 3 Operation before continuing.
Note: Running with air is not harmfull to the pump sealings.
However, the sealing ring lifetime might be shortened if the pump
pumps air for long periods.
The pump is tested at the factory prior to delivery and washed with
20% ethanol. Remains of this is removed by running-in the pump
with a liquid such as distilled water that is miscible with ethanol.
After running-in, the pump is primed for the chromatographic run.
Running-in
1 Submerge the inlet tubings in a suitable reservoir filled with
degassed distilled water. Place the outlet in a waste container.
2 Switch the mains power switch on the front panel to ON. A
selftest is carried out.
Note: When the pump is run for the first time after replacing a
sealing, friction between the glass cylinder and the dry sealing may
give rise to some noise. This will disappear when the sealing has
been wetted.
3 Leakage is tested by checking the pump pressure against a
blocked outlet using distilled water and a 5/16” stop plug. The
test is carried out from the pump dial and display. A chart
recorder connected to the analogue pressure output can be useful
to record the pressure reading during this test.
See section 4.5 Testing leakage for details.
4 Remove the stop plug and reconnect the outlet tubing. The pump
is then ready for operation. Before changing solvent, see section
3.9 Changing eluent.
Selftest
Please wait...
Pump P-920
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Installation 2
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2.9 Installation of accessories
Install Mixer M-925 as follows:
0
1
M-925
UniNet 1
UniNet 2
Remote
Pressure
Analogue out 0-1 V
CAUTION! Before connecting the mixer M-925, make sure the
power switch is in OFF position.
-Loosen the
two screws
-Mount the attached
mounting bracket with
the two screws Use one of the
mounting rails
Pump P-920
User Manual 18-1125-54 Edition AB
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2 Installation
The mixer is
connected to
the UniNet 2
connector
Stop plug
Mixer outlet
Inlet tubing A1
Inlet tubing B1
Outlet tubing
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3 Operation
3.1 On/off
Switch on the module at the mains switch on the front panel. At
switch on, the module performs a selftest. Several beeps are heard
during this process. If an error is detected, an error message is
shown.
Name and software version number is shown for 2 seconds.
The selftest takes approximately 1/2 minute. When start-up is
completed with no errors, the display shows the main menu with the
pump in End mode. All parameters are factory set to default values.
3.2 Menu selection and settings
Menu selection
A specific menu is selected by turning the front selection dial
clockwise or counter-clockwise. When the required menu is visible,
the menu or selection is accepted by pressing the OK-button.
Menu selection OK-button ESC-button
Go down one menu Return one menu
level, or accept setting level
Select sub-menu Return one menu level
If a menu has sub levels, the sub menu is displayed by pressing OK.
Pressing ESC moves back one menu level.
Return to higher menu levels
Pressing ESC repeatedly returns to the main operating menu. Turn
the dial one click counter-clockwise to return to the work mode
changing menu.
Work mode menu
Main operating menu
End 0.00ml/min
0.00MPa 0.0%B
Pump P-920
<version no.>
Selftest
Please wait...
Pump P-920
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Operation 3
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Selecting a value
A cursor below a text or numerical value shows what is affected by
the dial. To increase the value, turn the dial clockwise. To decrease
the value, turn the dial counter-clockwise. The value can be reset by
turning the dial several clicks counter-clockwise.
To simplify entering large numerical values, the cursor moves up to
the next digit if the dial is turned quickly in one direction. The
cursor moves back one place to the right every two seconds if the
dial is not turned. The text or numerical value displayed is accepted
by pressing OK. To cancel, press ESC.
3.3 Menu overview
Work mode changing menu. From here the pump is started,
stopped, held, paused and continued. This menu is accessed from all
positions by turning the dial one click counter-clockwise. The
appearance of this menu depends on the current mode.
Main operating menu. The menu is accessed from all positions by
pressing ESC repeatedly.
Setting flow rate in ml/min.
Setting concentration and gradient values.
Wash program selection. Individual pump cylinder assemblies or
a complete pump wash can be selected.
Pump synchronisation selection. This means that the pump
cylinder(s) are run to one stop position. Individual pump cylinder
assemblies or a complete pump synchronisation can be selected.
Change piston direction selection. Individual piston direction or
both piston directions can be selected.
Check internal operating values. See Reference information section
B.1.
Setup language, pressure limits and unit, etc. See Reference
information section B.2.
Set different timer options. The pump can be started or stopped at
set times. See Reference information section B.3.
Alarm/Timer 12:30:52
Setup
Check
Change Direction
Pump Sync 20ml/min
Pump Wash 20ml/min
Set Conc./Gradient
(0.0%B)
Set Flow Rate
(0.00ml/min)
Run 13.40ml/min
2.00MPa 45.5%B
End 1.00ml/min
Run
Pump P-920
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3 Operation
Setup Hi Press Limit
(5.00MPa) 5.00
Parameter
Current value New value to be set
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3.4 Starting and stopping the pump
Main operating menu
The main operating menu shows the current flow rate together with
work mode indication, pressure and %B, if used. The available work
modes are:
Run The pump is running with set flow rate.
End The pump is not running. Flow rate and gradient are reset.
Pause The pump is stopped but the set flow rate and the
gradient are retained.
Hold The gradient is held at the value displayed and the
pump continues to run.
Continue Not a work mode. Used to continue from Hold or Pause.
Work mode changing menu
Work mode changes are made in the work mode changing menu
above the main operating menu (turn dial counter-clockwise). The
current work mode is shown in the upper left corner of the display.
Available actions are shown at the lower right. There are four
different displays for this menu and the menu displayed will depend
on the current mode (see below). When a new mode is selected, the
appearance of the menu will change.
Pressing OK in a work mode changing display selects the underlined
work mode. Different work modes are selected by turning the dial.
• To start the pump, select Run and press OK.
• To stop the pump, select End and press OK.
Hold 1.00ml/min
End Continue Pause
Pause 1.00ml/min
End Hold Continue
End 1.00ml/min
Run
Run 1.00ml/min
End Hold Pause
Run 13.40ml/min
2.00MPa 45.5%B
WARNING! All protective covers over internal capillaries, piston
rods and glass cylinders must be in place when running the pump.
WARNING! Incorrectly fitted tubing may loosen, causing a jet
of liquid to spray out. This is especially dangerous if hazardous
chemicals are being used. Connect the tubing by first inserting
the tubing fully, then tightening the connector fingertight.
Pump P-920
User Manual 18-1125-54 Edition AB 11
Operation 3
Current work
mode Available actions for
the current
work mode
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Preparation before starting
1 Check that there is sufficient solvent present for the run, and that
the solvent filter is fully immersed. If the eluent is to be changed,
see section 3.9 Changing eluent.
Note: The pump may not work if the buffer vessels are sealed, or if
they are placed too far below the pump inlets. Do not close the
vessels off completely. Place the buffer vessels on the workbench or
on the buffer tray on top of ÄKTAFPLC.
2 Check that there is rinsing liquid (20% ethanol) in the rinsing
system (behind the piston heads).
3 Set the pressure limit. Refer to section B.2.1 in Reference
information. If the pressure limit is exceeded, the pump is
stopped.
Emergency stop
Use the front power switch, or disconnect the mains supply from the
pump.
3.5 Setting the flow rate and starting the pump
1Select operating menu Set Flow Rate and press OK.
2 Set the desired value and press OK.
3 Go to the work mode menu, select Run and press OK.
4 To change the flow rate while the pump is running, repeat steps 1
and 2.
5 To stop the pump, go to the work mode menu, select End and
press OK.
In UNICORN, select instruction Flow in
System Control:Manual:Pump. Click on Execute and the pump will
start.
3.6 Setting concentration B
A percentage of eluent B can be set.
1 Select operating menu Set Conc./Gradient and press OK.
2 When the display shows Set Concentration B, press OK.
3 Set the desired %B value and press OK.
In UNICORN, select instruction Gradient in
System Control:Manual:Pump. Set the desired %B value.
Set Concentration B
(0.0%B) 5.0
Set Conc./Gradient
(0.0%B)
Set Flow Rate
(0.00ml/min) 2.50
CAUTION! Before the start of each run, always ensure that there
is an adequate supply of eluent in the reservoirs. Do not allow the
pump to run dry, since this will affect the lifetime of the sealings.
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3 Operation
!
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Pump P-920
User Manual 18-1125-54 Edition AB 13
Operation 3
3.7 Running a simple gradient
Gradients can be run in time or volume base. The default is time
base. To change base, see B.2.4 in section Reference information.
The gradient is run from current
concentration to target concentration,
over a set time or volume. The
gradient can be set in any operating
mode.
1 Select operating menu Set Conc./Gradient and press OK.
2 When the display shows Set Concentration B, turn the dial to
select operating menu Set Gradient Length and press OK.
3 Set the desired value and press OK.
4 Turn the dial to select operating menu Set Gradient Target and
press OK. Set the desired gradient target value in %B and press
OK. If the pump is in work mode Run, the gradient starts
immediately.
5 At the end of the set gradient time, the pump continues to run at
the target concentration.
In UNICORN, select instruction Gradient in
System Control:Manual:Pump. In the Parameters window set Target
and Length. Click on Execute.
3.8 Ending the run and storage
Overnight storage: The pump can be left filled with a buffer.
Note: If buffers or water are stored at room temperature, bacterial
growth may occur.
If no further runs are planned, the pump should be washed
immediately with pure eluent. If aqueous buffers have been used,
washing with pure distilled water is particularly important to
prevent salt precipitation.
Weekend and long term storage: Wash the pump with water and
then fill it with 20% ethanol.
Set Gradient Target
(0.0%B) 50.0
Set Gradient Length
(0.0min) 8.0
Set Conc./Gradient
(0.0%B)
0%
100% Target
concentration
Current
concentration
Start of
gradient
Gradient length
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Pump wash operation
1 Immerse the inlet filters in a wash solution. Select operating menu
Pump Wash and press OK.
2 When the display shows Pump Wash A, Band B&A, select B&A
and press OK.
3 First pump B and then pump A is washed in sequence. The pump
wash is performed with 20 ml/min for seven strokes each.
In UNICORN, select instruction PumpWash in
System Control:Manual:Pump. In the Parameters window set
PumpA and PumpB. Click on Execute.
3.9 Changing eluent
Note! When changing from one eluent to another, it is extremely
important that the two eluents are totally miscible with one another.
If the two eluents are immiscible, the pump should be washed first
with an intermediate liquid that is miscible with both eluents. Failure
to do this will disrupt the flow from the pump.
When changing from a salt-containing buffer to an organic solvent,
use water as the intermediate liquid to prevent precipitation.
1 Stop the pump by setting it in Pause mode.
2 If not carried out from UNICORN, use a bypass tubing to
replace the column.
3 Transfer the inlet tubing into the new eluent or into the
intermediate liquid.
4 Perform a pump wash.
5 If an intermediate liquid is being used, transfer the inlet tubing
into the final eluent and repeat step 4 with the new eluent.
In UNICORN, select instruction PumpWash in
System Control:Manual:Pump. In the Parameters window set
PumpA and PumpB. Click on Execute.
Note: When high buffer purity is important, use an intermediate
step. This avoids remains of the previous buffer in the inlet filter
diffusing out in the new buffer.
CAUTION! To prevent precipitation of crystals when changing
from a salt-containing buffer to organic solvent, always wash the
system with water as the intermediate liquid.
Washing
Please wait......
Pump Wash 20 ml/min
A B B&A
Pump Wash 20 ml/min
Pump P-920
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3 Operation
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