Pump P works with a wide range of columns and media supplied by Amersham Biosciences. The pump can be used to form accurate and reproducible gradients. Pump P features:. Great functional condition. Tested switching valves and verified flow rates for both pumps. Pistons are in excellent, leak free condition. NOTE: There is some cosmetic flaking of the paint on the top of the unit. See pictures. The flow cell can be changed when required, for example from 2 mm to 5 mm to increase the sensitivity, or from 5 mm to 2 mm to decrease the sensitivity.
Change the flow cell as follows: 1. The inlet port of the 5 mm UV cell is above the optical unit. If the locking nut is not tightened sufficiently, the monitor will function poorly e. Place the protective cover around the flow cell to protect the electronics inside the optical unit from liquid spillage. The Hg lamp housing has two positions, one for nm, marked by on the filter housing, and the other one marked by for all other wavelengths. The Zn lamp housing has only one position, for nm.
There will be a faint click when the housing is correctly slide into either position. Set the wavelength to be used by selecting lamp position indicated by a dot on the lamp housing in combination with the appropriate filter, i. A click will indicate that the filter is in position. This must be entered by the user. When starting a method, a question appears in which the user is prompted to state the wavelength to be used in the run.
Fractionation is selected and specified in the fractionation dialogs in the Method Wizard. This fractionation method is available in all methods except gel filtration methods.
Fixed volume fractionation can be combined with peak fractionation, which means collecting peaks during elution. The peak size is set to a volume which is larger than the actual peak volumes. The peak size is set to a volume which is smaller than the actual peak volume. The basic steps required to create a method are: 1. Select the appropriate parameter values in the subsequent pages. Click OK to go to the next page. Check and fine-tune the values for the method variables, e.
Before starting the run, check that the system is in End mode, and that the valves are in their initial positions. Conductivity flow cell Cell constant Temperature Entering a new cell constant Sample pump optional. Must be done when changing the conductivity flow cell Temp. Whenever the running conditions are changed.
Check that the waste bottle is not full and will accept the volume diverted to it during the run. Check that the pump has been purged i. If not, purge the pump as described in the P User Manual. Check that the correct wavelength is set on the optical unit and that the correct UV flow cell is installed.
Click the Customize panes toolbar button or choose View:Panes from the menu to select which panes to display. Run Data. The run data pane displays the current values for selected run parameters. Right-click in the Run Data pane and select Properties. Select the run data items to be displayed and click OK.
The curves pane displays the monitor signal values graphically. Rightclick in the Curves pane and select Properties to select the curves to display. All curves are always stored in the result file. Flow scheme. The flow scheme is a graphical representation of the flow path in the chromatography system.
During a run, the flow scheme shows open flow paths and monitor signals with numerical displays. All actions and unexpected conditions such as warnings are logged for every run, with date, time and current username. The logbook provides a complete history of any given run. The log is saved in the result file. Front panel displays. In each case, the main operating menu display shows the most important parameters.
The available modes are: Run End Pause Hold. The pump is running with the set flow rate. The system is not running. The pump is stopped but the set flow rate and the gradient values are retained. The gradient is held at the value displayed and the pump continues to run. The main operating menu 1 of Monitor UPC shows the absorbancy value with 6 digits for the selected wavelength, the conductivity as a percentage of full scale and the pH value optional.
By turning the dial one click, an alternative display of the conductivity is shown main menu 2. These instructions can be used during a run to alter system conditions in response to the results observed. The Manual menu in System Control opens a dialog box similar to the text instruction box in the Method editor. Manual instructions are entered as follows: 1. Highlight the instructions list by clicking on a button on the left of the instruction panel and select the required instruction s from the list displayed.
Some instructions, for example, gradient or fraction instructions, may take time to complete. To print all instructions with explanations, click on Print in the Method Editor:File menu. This opens a window containing all that may be printed. Make sure that the Instruction Set box is checked and clear any unwanted items. Click OK to print the instructions.
From the modules. Manual changes are normally recorded in the log book. Perform a PumpWash with distilled water to wash P If you are not using the separation unit for a few days or longer, perform a PumpWash with distilled water.
Remove the column and the pH electrode optional. Replace the column by a bypass capillary and fit the pH dummy electrode if applicable. Make sure that all tubing and all flow paths used are rinsed. Never leave the pH electrode in the flow cell for any period of time when the system is not used, since this might cause the glass membrane of the electrode to dry out.
Dismount the pH electrode from the flow cell and fit the end cover filled with a mixture of pH 4 buffer and 2 M KNO3. Do NOT store in water only. When the pH electrode is removed from the flow cell, the dummy electrode supplied shall be inserted in the flow path. Place the computer outside the cold room. A 15 m UniNet cable is available as an accessory and should be used to connect the computer to the separation unit.
Any parameters can be scouted, provided that they can be defined as variables in the method used. Feedback tuning is useful in applications where high back-pressure can be expected, or when the back-pressure fluctuates, for example, when using samples with high viscosity.
The feedback tuning is set in the Method Wizard and it is activated only during sample application and wash-out of unbound sample. Feedback tuning is set up and used in slightly different ways when applied to the system pump and the sample pump, which is described in the following sections. There are two regulators involved in tuning the system pump.
The first one is active as long as the pressure is below the set point. This regulator tunes the actual pump flow rate to the set flow rate. If the pressure exceeds the set point, a second regulator decreases the flow rate in order to reduce the pressure.
When the pressure falls below the set point, the first regulator takes control again and tunes the actual flow rate to the set flow rate, and so on.
The regulators use so-called PID feedback tuning, where P, I and D are parameters that determine the tuning characteristics. However, the parameters can be further optimized to suit a specific application see section Optimizing the PID parameters. Feedback tuning of the system pump in a method 1 To include feedback tuning in the Method Wizard, select Flow Regulation of the system pump. Type the pressure control set point slightly below the column pressure limit and the minimum allowed flow. Note: If the flow rate falls below the MinFlow value, an Alarm is raised and the system set to Pause.
A suitable action is to continue to the next block. To prevent pressure peaks when continuing, use a lower flow rate in the block after the Watch instruction than used when the Watch instruction was activated. Feedback tuning can also be added manually to a method in the Method Editor module. Feedback tuning instructions. Feedback tuning can also be used when running the system pump manually. The instructions are found in the System Control module by selecting Manual:Pump and are explained in Table To activate feedback tuning, select PressFlowControl.
PressLevel is the pressure control set point MinFlow is the minimum allowed flow rate. Active below the PressLevel value. Active above the PressLevel value.
The two regulators for the system pump have separate PID parameters. However, the parameters can be further optimized to suit a specific application. The table below describes the three PID parameters. The P parameter reduces the effect of an error but does not completely eliminate it.
A simple P-regulator results in a stable stationary error between actual and requested flow. The I parameter eliminates the stationary error, but results in a slight instability leading to oscillations in the actual flow. The I parameter can have values between 0 and infinity. Smaller values have a greater effect and a value of infinity has no effect. In certain cases, the D parameter can reduce the oscillations introduced by a PI-regulator.
D can have values between 0 and infinity, where larger values have a greater effect and a value of 0 has no effect. To set the default values, select System:Settings in the System Control module. The parameters are found in Specials. The feedback tuning of the sample pump is simpler than the feedback tuning of the system pump.
When the pressure reaches the maximum allowed pressure, the flow is decreased. After a short while, the flow slowly increases towards the set flow rate, and so on. The tuning regulator is rather simple and does not use PID-parameters. The parameters that control the tuning characteristics can not be changed. Select Pressure Control for Sample pump.
The default value for the maximum allowed pressure is 2. Feedback tuning can also be applied in an existing method in the Method Editor module. To change the maximum allowed pressure: Alternative 1 1.
Follow the recommendations in this chapter to keep the system in good working order. Do not allow spilt liquid to dry on the instrument. Wipe the surface regularly with a damp cloth. Let the system dry completely before using it.
For details of how to perform the various actions, please refer to the individual User Manuals and Instructions. If there is a risk that large volumes of spilt liquid might penetrate the casing of the instruments and come into contact with the electrical components, immediately switch off the system and contact an authorised service technician.
Xenon lamps emit a high intensity continuous spectrum of light, your chosen wavelengths are selected using a monochromator. The lamp is triggered only when needed, this extends its lifespan, meaning the average lamp can withstand many years of normal use. Wavelengths are selected using a band pass filter.
When the system is operated at room temperature with a wavelength of nm, the average Hg lamp lifetime is hours. In the cold room the average lifetime is reduced to hours.
Therefore, the first thing that you should do if you get this message is give your system a good clean. There are two positions for aligning the Hg lamp with the filter housing, one for nm marked by a filled white circle and one for all other wavelengths marked by a white ring.
Having the lamp and filter housings incorrectly aligned can result in a low light intensity warning. How can I use my system in cold rooms? Purifying some samples at room temperature can lead to increased levels of degradation; performing your purifications at 4C can help. The computer systems are not however cold room compatible, and can be damaged by being placed at 4C. When moving a system to or from a cold room, time must be allowed for the system to adjust to its new temperature.
You may also find that you need to tighten the connectors on your system slightly to prevent leaks when you bring a system out from the cold, and loosen them slightly before you put a system into the cold to prevent pressure build up.
Changes in temperature can also affect the viscosity of your buffers so keep a close eye on your back pressure. We ship the most recent available manual and support documentation at the time we ship the software to you.
Retain this manual as it is the correct one for the version of software you purchased. Cleaning the system The procedures described below are for system cleaning. To bypass the column, use a piece of i. If the column is to be left in the flow path, make sure that the maximum allowed flow and pressure for the column are not exceeded. For column cleaning procedures and column storage instructions, please refer to the Instructions supplied with the column.
At the end of the day The system can be left filled with a buffer overnight. If the system will be used with other buffers next day, rinse the pump and the system with distilled water using the PumpWash instruction as follows:. Leaving the system for a few days Perform a PumpWash with distilled water. Clean the system every month or when problems such as ghost peaks occur. The system is cleaned as follows:.
Other cleaning considerations After repeated separation cycles, contaminating material may progressively build up in the system and on the columns. This material may not be removed by the cleaning step described above. The nature and degree of contamination depends on the sample and the chromatographic conditions employed. Planned maintenance can be part of a service agreement, scheduled to service your system before it is in need of attention.
We can help you design a schedule and routine to allow you to maintain your system, please contact your local Cytiva service representative. So what can you expect from a planned maintenance visit from Cytiva service representative?
A complete overhaul, once a year, ensures that your instrument is running at peak performance so you can be confident of your scientific results. In addition, wear and tear on systems under constant use by multiple end users is minimized, giving the system a longer life and better value for money. To find out more about service possibilities contact your local Cytiva service representative. How can I determine the delay volume?
Three different methods are described below: Method I - Determining the delay volume of your system by measuring different retention volumes. Chemical resistance guide and chemical compatibility The chemical resistance to some of the most commonly used chemicals in liquid chromatography is indicated in the table below.
The ratings are based on the following assumptions: 1. The synergistic effects of the chemical mixtures have not been taken into account. Room temperature and limited over-pressure is assumed. Note: Chemical influences are time and pressure dependent. Pressure limit for PEEK decreases. Hydrochloric acid, 0. Connect stop plugs to non-used ports in the sample valve.
For information regarding each component, place the cursor over the component of interest. The sample inlet tubing should be adjusted to a suitable length. Flushed loop injection With flushed loop injection means that the sample loop is completely filled with sample.
Partial loop injection With partial loop injection means that the loop is partially filled with sample. Seals are considered as consumables and should be replaced annually or when needed.
Please keep a set of each in your stock to prevent long downtime. For code numbers refer to the table below. Alternatively you can create a system report, which will list all components and the current Module Software version.
Find solutions to product related issues. For unlisted issues please contact local Cytiva service representation. There might be dirt in the check valves. Clean the valves according to instructions in the Pump P User Manual. Tap the flow cell carefully or tilt it to remove the air. Use suitable flow restrictor after the pH electrode.
FR 0. Clean the pH electrode according to instructions. In organic solvents such as ethanol, methanol and acetonitrile, stable pH measurements are not possible since dehydration of the membrane will occur. We recommend that the pH electrode is not used in applications using organic solvents.
The calibration curve shows the relation between pH and the output signal from the monitor in mV. Read more about the calibration curve in the instructions. The electrode may be restored by soaking in buffer over night. Check the connection to the pump. The valve should be connected to the UniNet 2 socket, not to the UniNet 1 socket. Remove the air in the eluent or buffers by degassing. Preventive action: It is recommended to use degassed buffers for best performance. Check the mixer function by placing a stirrer bar on top of the mixer housing.
The stirrer bar should rotate when the system is in Run mode. The mixer function can also be checked by running the installation test. Check that most suitable mixing chamber, with regards to the volume, is used. Check if the signal is still noisy in water. Preventive action: Make sure the buffers have been filtered. Check that the lamp is in a poper position and that the correct filter is used according to instructions in the Monitor UPC User Manual.
Normal and abnormal pressure traces together with comments are shown in the Troubleshooting section of the Pump P User Manual.
Preventive actions: Make sure the piston rinsing system is always used when working with aqueous buffers with high salt concentration. Check the mixer chamber. If liquid has penetrated the mixer chamber walls and sealings, replace the chamber according to instructions Mixer M Instructions.
Prepare a new calibration solution and recalibrate the conductivity cell according to instructions in the Monitor UPC User Manual.
The conductivity of the solution changes with temperature. If temperature compensation is being used, incorrect temperature compensation factor might be in use. Tighten the connectors. If necessary replace the connectors. Please refer to the order information. Check the motor operation. Place the hand on the mixer and start it by starting the pump at low flow rate. You should both hear and feel the mixer motor and stirrer when they are spinning. Replace the mixing chamber to a chamber with smaller volume according to instructions Mixer M Instructions.
Check that the pump is operating and is programmed correctly according to instructions in the Pump P User Manual. Replace the mixing chamber to a chamber with larger volume according to instructions in the Mixer M Instructions. Check the chart recorder in accordance with its instructions. Check that the lamp position and the wavelength used filter position fit together according to instructions in the Monitor UPC User Manual.
Regular maintenance is important for safe and trouble-free operation of your instrument. The user should perform daily and monthly maintenance. Preventive maintenance should be performed on a yearly basis by qualified service personnel. For maintenance of a specific component, carefully read the component manual and follow the instructions. Change the solvent to distilled water and flush out all salt before flushing the 6-port pump valve.
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 finger tight. Do not allow solutions that contain dissolved salts, proteins or other solid solutes to dry out in the flow cell. Do not allow particles to enter the flow cell as damage to the flow cell may occur.
Electrical shock hazard. All repairs should be done by service personnel authorized by Cytiva. Do not open any covers or replace parts unless specifically stated in the user documentation. Disconnect power. Always disconnect power from the instrument before replacing any component on the instrument, unless stated otherwise in the user documentation.
Hazardous chemicals during maintenance. When using hazardous chemicals for system or column cleaning, wash the system or columns with a neutral solution in the last phase or step. Do not perform any type of maintenance work while the system is powered electrically or when the piping system is pressurized.
Note that the piping system can be pressurized even when the system is closed down. Fire hazard. Follow instructions in the User Manual for correct installation of a new UV-lamp. If the lamp is not installed properly it may be overheated and cause a fire hazard. When using hazardous chemicals, take all suitable protective measures, such as wearing protective glasses and gloves resistant to the chemicals used.
Follow local regulations and instructions for safe operation and maintenance of the system. Keep the instrument dry and clean. Wipe regularly with a soft damp tissue and, if necessary, a mild cleaning agent. Let the instrument dry completely before use. Make sure that the piping system is completely leakage free before performing any CIP on the system. NaOH is corrosive and therefore dangerous to health. When using hazardous chemicals, avoid spillage and wear protective glasses and other suitable personal protective equipment.
After assembly, the piping system must be tested for leakage at maximum pressure for continued protection against injury risks due to fluid jets, burst pipes or explosive atmosphere.
Hands and fingers risk being squeezed between the piston driving arm and the pump housing when running the pump without the protective covers fitted. This symbol indicates that the waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected separately.
Please contact an authorized representative of the manufacturer for information concerning the decommissioning of equipment. Check for leakage. If there are signs of liquid leaking out from the cylinder assemblies, the on-line filter may require replacement more often. Change rinsing solution. An increase in the volume of rinsing solution behind the pistons indicated internal pump leakage. Replace the piston seals Refer to procedure below.
Put the open end in the waste container. Note the back-pressure on the pump display or in the Run Data window. Note the pump display or in the Run Data window. Replace it if it is not within limit. Check for external or internal leakage.
Run the Replace channel plate and distribution plate procedure yearly or when required. Replace the on-line filter. If Pump P is used for sample application, the on-line filter may require replacement more often. Loosen each one equally in turn so the distribution plate comes off parallel to the valve body. Using the Allen key, tighten the 4 screws in turn, a little at a time, until the distribution plate is fixed to the valve body.
Faulty operation of the 6-way pump valve is usually indicated by no flow and that the pressure sensor gives no indication that the back pressure has increased. The probable cause is that one or more channels are blocked. Identify the relevant pump cylinder assembly by observing which pump head is not delivering flow. First, try to clean the inlet channel in—place by disconnecting the inlet tubing and the tubing from the corresponding pump cylinder assembly.
Clean the channels in the valve by flushing nitrogen in reverse flow direction through the valve.
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