Monitor UPC-900 User Manual 18-1125-55
Important user information All users must read this entire manual to fully understand the safe use of ÄKTA™ monitor UPC-900 Workstation. Safety symbols The following Warning symbols highlights instructions that must be strictly followed in order to avoid personal injury. Be sure not to proceed until the instructions are clearly understood and all stated conditions are met. WARNING! Read the instruction to avoid hazardous conditions.
Contents Contents 1 Introduction 1.1 1.2 2 Installation 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 2.10 3 Unpacking..............................................................................................................3 General precautions ...............................................................................................3 Installing the optical unit .......................................................................................4 Installing the conductivity flow cell ..............
Contents 4.8 4.9 5 Cleaning the pH electrode ...................................................................................25 Changing the pH electrode ..................................................................................26 Trouble-shooting 5.1 5.2 5.3 General ................................................................................................................27 Faults and actions ................................................................................................
Contents Short instructions on back page 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 operating the instrument.
Contents iv Monitor UPC-900 User Manual 18-1125-55 Edition AE
Introduction 1 1 Introduction 1.1 General Monitor UPC-900 is a high precision on-line monitor for the combined measurement of UV absorption, pH and conductivity in liquid chromatography. The UPC-900 features: • Fixed wavelengths of 254, 280 nm (Hg lamp), or 214 nm (Zn lamp). • Other wavelengths of 313, 365, 405, 436 and 546 nm through filter change. • Two alternative flow cells with pathlengths of 2 mm and 5 mm.
1 Introduction 1.2 Safety • The module is designed for indoor use only. • Do not use in a dusty atmosphere or close to spraying water. • Operate in accordance with local safety instructions. WARNING! The module must be connected to a grounded mains socket. WARNING! Always disconnect the power supply before attempting to replace any item on the module during maintenance. WARNING! The module must not be opened by the user. It contains high voltage circuits that can deliver a lethal electric shock.
Installation 2 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. Note: It is important that the filters, flow cells and lamps are not handled during unpacking. For protection of these items, they should remain in their packing materials until required for use. We recommend that all packing materials be retained if onward transport of the module is expected.
2 Installation 2.3 Installing the optical unit UPC-900 Holder Installing the holder 1 Conductivity cell Hook the holder into the slot on the right hand side of the module. Secure it by pushing up the slide clamp. Optical unit Flow cell installation There is one analytical (5 mm) and one preparative (2 mm) flow cell available (stand-alone units only include the preparative flow cell).
Installation 3 Connect the lamp cable to the socket the module. 4 Connect the signal cable to the socket panel of the module. Lamp 2 on the rear panel of Optical unit on the rear Changing lamp assembly WARNING! The module uses high intensity ultra-violet light. Do not remove the UV lamp while the monitor is running. Before changing a defective lamp, ensure that the lamp cable is disconnected from the monitor to prevent injury to eyes.
2 Installation Connection to the column 1 Fix the optical unit directly under the column if possible. Note: Always position the optical unit with the filter wheel cover facing upwards. 2a Connect the column outlet tubing directly onto the top (a) of the optical unit for the 5 mm flow cell using a fingertight connector. 2b Connect the column outlet tubing onto the bottom (b) of the optical unit for the 2 mm flow cell using a fingertight connector.
Installation 2 2.5 Installing the pH flow cell and electrode Mounting the flow cell holder In ÄKTAFPLC systems, the pH electrode is optional. 1 Hook the flow cell holder on the right hand side of the housing. Secure it with the slide clamp. If the flow cell holder is not used, the flow cell must still be installed at an angle of 30° from the vertical with the outlet placed higher than the inlet to prevent air bubbles being trapped in the cell.
2 Installation 2.6 Connecting electrical signal cables The sockets for electrical signals are located on the rear panel. Optical Unit Conductivity Flow Cell Analogue out 0-1 V UniNet 1 pH-Ground pH-Probe Mains Lamp Voltage Frequency Power, max 100-200 V- 50-60 Hz 30 VA Connecting to chart recorder (if used) The external chart recorder outputs for UV, pH and conductivity from the monitor are 0–1 V. 1 Connect the chart recorder to the mini-DIN-socket Analogue out 0-1 V using the cable supplied.
Installation 2 Connect two UniNet 1 cables to the UniNet 1 connectors. The module can be connected in series anywhere between the PC and a termination plug. The UniNet 1 link connects, in series, the PC with Pump P-920, Monitor UPC-900 and the Frac-900. The termination plug is connected to the last module in the chain. 2.8 Connecting to supply voltage WARNING! The module must be connected to a grounded mains socket. 1 Make sure the on/off switch is in the OFF-position (0).
2 Installation 2.10 Installing optical filters (optional) The Hg optics with 254 and 280 nm filters and the Zn optics with the 214 nm filter are delivered with filters installed. If other filters are to be used, install them as follows: 1 If the Zn lamp is attached, remove the lamp housing as described in section Changing the UV lamp. 2 Remove the four screws in the filter housing. Separate the filter housing from the detector housing. 3 Carefully remove the filter wheel from the filter housing.
Operation 3 3 Operation 3.1 On/off Selftest Please wait... UPC-900 Calibrating Switch on the module at the mains switch on the rear 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. All parameters are set to factory default values. Calibration ok AU Cond% pH The selftest takes approx. 1 minute.
3 Operation Main menus Main menu 1 3 click CW with the dial Main menu 2 Sub menus ESC or 3 click CCW with the dial Submenu 2.1 OK Sub menu 2.1.1 OK ESC ESC Sub menu 2.2 OK Sub menu 2.3 Sub menu 2.3.1 ESC Sub menu 2.3.2 Return to main menu Pressing ESC repeatedly always returns to the main menu 1, which is the main operating menu. Main menu 1 ESC Main menu 2 ESC ESC ESC ESC Select value A cursor below a text or numerical value shows what is affected by the dial.
Operation 3 3.3 Main menu overview AU Cond%Tc pH Main menu 1. This menu is accessed from all positions by pressing ESC. Lamp control. This menu is accessed by turning the dial one step counter-clockwise from main menu 1. It is used to manually switch the lamp on/off. Lamp (on) Autozero UV Autozero. Used to zero the UV signal from the monitor. Eventmark Eventmark. Used to set eventmarks on the UV curve. pH 12.50 22.4 Main menu 2.
3 Operation 3.5 Reading the UV absorbance value AU Cond%Tc pH Absorbance value The main menu 1 shows the absorbance value with four digits for the chosen wavelength. This menu is reached from any other menu by pressing ESC. If the lamp is off, Lamp Off is displayed instead. 3.6 Autozeroing UV The autozero function in the UV part of the module sets the detected absorbance to zero when OK is pressed. Using autozero is recommended at the start before the sample is injected.
Operation 3 Set Cond Zero Level 2 Press OK to access the settings menu. Press OK. Set Cond Zero Level 3 Set the desired zero value. The range is 0.000 - 999.9 mS/cm. Press OK to acknowledge. Set Cond Full Scale 4 Turn the dial clockwise to access the settings menu. Press OK. Set Cond Full Scale 5 Set the desired full scale value. The range is 0.000 - 999.9 mS/cm. Press OK to acknowledge. Press ESC two twice to return to menu Set Parameters.
3 Operation Set UV Range 2 Press OK to access the settings menu. The current setting is displayed. Press OK. Set UV Range 3 Set the desired full range value. Press OK to acknowledge. Set UV Zero Level 4 Turn the dial clockwise to access the settings menu. The current setting is displayed. Press OK. Set UV Zero Level 5 Set the desired zero level value. Press OK to acknowledge. Press ESC twice to return to menu Set Parameters. The module has an automatic over-range function.
Operation 3 Calibrating with the electrode outside the flow cell When calibrating the electrode out of the flow cell and changing from one buffer to another, rinse the electrode tip with distilled water and dab it carefully with a soft tissue to absorb the remaining water. Do NOT wipe the electrode as this may charge it and give unstable readings. The steps below describe the procedure used with the electrode removed from the flow cell.
3 Operation Calibr pH Buffer 2 9 This text disappears when the reading is stable and the following text is then shown: Calibr pH Buffer 2 10 Adjust the pH value in the display using the dial so that it corresponds to the known pH value of the second buffer solution and press OK.
Operation 3 3.11 Filtering noise in the UV signal To filter the noise in the UV-signal, a moving average filter is used. The averaging time is the time interval used for calculating the moving average of the absorbance signal. A long averaging time will smooth out noise efficiently, but it will also distort the peaks. Peaks narrower than the minimum peak width value according to the table below may be distorted. Because of this, the averaging time should be as short as possible.
3 Operation 3.13 Reading pH, temperature and conductivity values AU Cond%Tc pH* 0.00002 015.0 4.02 The main operating menu 1 shows UV absorption, conductivity in percentage of full range and pH. If the pH value is not stable or is changing, an asterisk is displayed, i.e. pH*. This menu is reached from any other menu by pressing ESC. If temperature compensation is switched on, the display will show Cond%Tc instead of Cond%, see section B.2.
Operation 3 Storage of the pH electrode CAUTION! Never leave the pH electrode in the flow cell for any period of time when the system is not used, since this may 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 1:1 mixture of pH 4 buffer and 2 M KNO3. Do NOT store in water only. The pH electrode should always be stored in a 1:1 mixture of pH 4 buffer and 2 M KNO3 when not in use.
4 Maintenance 4 Maintenance WARNING! Always disconnect the power supply before attempting to replace any item on the module during maintenance. CAUTION! Only spare parts approved or supplied by Amersham Biosciences may be used for maintaining and servicing the module. 4.
Maintenance 4 When necessary, replace the lamp according to Section 4.10 Changing the UV lamp, or contact Amersham Biosciences for lamp replacement. Autozero The module internal absorbance value for autozero can be checked to test the consistency of buffers. Check Autozero and press OK. 1 Select menu 2 Select menu Check Autozero. The autozero absorbance value for the wavelength used is shown. Check 4.3 Cleaning the UV flow cell in-place WARNING! NaOH is injurious to health. Avoid spillage.
4 Maintenance 4.5 Cleaning the conductivity flow cell in-place WARNING! NaOH is injurious to health. Avoid spillage. Remove the pH electrode and install the dummy electrode in the pH flow cell. Pump a cleaning or sanitising agent through the flow cell. The standard recommendation is to pump 1 M NaOH for 30 minutes and then wash out with buffer. 4.6 Cleaning the conductivity flow cell off-line WARNING! NaOH is injurious to health. Avoid spillage.
Maintenance 4 4.8 Cleaning the pH electrode Note: The pH electrode has a limited lifetime and should be replaced every six months, or when the response time is slow or the slope is out of range (<80%). WARNING! HCl and NaOH are injurious to health. Avoid spillage. Use one of the following procedures to clean the electrode to improve the response: • Salt deposits: Dissolve the deposit by immersing the electrode first in 0.1 M HCl, then in 0.1 M NaOH, and again in 0.1 M HCl.
4 Maintenance 4.9 Changing the pH electrode See section 2.4 Installing the pH flow cell and electrode.
Trouble-shooting 5 5 Trouble-shooting 5.1 General UPC-900 Version 1.00 When contacting Amersham Biosciences for support, state the program version of the module, which is shown for 2 seconds during start-up. WARNING! The module must not be opened by the user. It contains high voltage circuits that can deliver a lethal electric shock. 5.2 Faults and actions If the suggested actions do not correct the fault, call Amersham Biosciences.
5 Trouble-shooting Fault Low sensitivity Action 1 Aging lamp. Check the lamp and replace if necessary 2 Wrong lamp position. Check that the lamp position and the wavelength used (filter position) fit together. 3 Dirty on-line filter. Clean or replace the filter. Error in external chart recorder 1 2 Check the chart recorder in accordance with its manual. Test the recorder function by selecting recorder test according to section B.1 in Reference Information.
Trouble-shooting 5 Fault Action 10 There may be interference from static fields. Connect the pH flow cell and the rear panel of the monitor using a standard laboratory 4 mm “banana plug” cable. 11 Check that the pH electrode has been calibrated at the correct temperature. 12 In organic solvents such as ethanol, methanol and acetonitrile, stable pH measurements are not possible since dehydration of the membrane will occur.
5 Trouble-shooting Fault Ghost peaks appear in the gradient profile Action 1 2 A charged sample has been detected (e.g. protein). Air bubbles are passing through the flow cell. Check for loose tubing connections. If necessary, use a flow restrictor after the conductivity flow cell. 1 2 Check the chart recorder in accordance with its manual. Test the recorder function and input voltage, which should be 1 V full scale. Check the conductivity scaling and pH scaling, see section 3.
Trouble-shooting 5 5.3 Error messages If the suggested actions do not correct the fault, call Amersham Biosciences. Message 34 Start up failed Retry/Call service 35 WARNING wrong averaging time set 50 Electrical error Call for service Action 1 Perform a new start-up. The preceding message may tell more about the cause. 2 If not, call service. 1 Wrong value for averaging time set. See section 3.11 on how to set the averaging time relative to peak width. 1 Call for Service.
5 Trouble-shooting 83 WARNING temp_cal will be changed 1 2 Press OK to accept the change. Press ESC to skip the change. 1 The difference between 0 and 100% must be at least 0.1 mS/cm. Increase the span between zero and full scale setting. See section 3.8. Check that the conductivity cell is connected. Recalibrate temperature. If problem remains, replace the conductivity cell. Check the pH electrode connection. Clean the pH electrode. If problem remains, change the pH electrode.
Trouble-shooting 98 Cal failed. Cell constant not 0.1-300 1 2 4 5 1 2 3 Conductivity cell constant is out of range. Wrong solution used during calibration. Use 1.00 M NaCl and recalibrate. Air in conductivity cell during calibration. Flush the flow cell with calibration solution and recalibrate. Dirty conductivity cell. Clean the flow cell and recalibrate. If the problem remains, change the conductivity cell. The key was pressed during self-test, or is faulty. Switch off the module. Switch on the module.
A Reference information Reference information A Description A.1 Module Monitor UPC-900 is an on-line monitor for measurement of UVabsorption, pH (optional in ÄKTAFPLC) and conductivity. The monitor can work with standard glass pH electrodes with a built in liquid-filled reference electrode and a BNC connector.
Reference information A A.2 UV optical unit The UV optical unit houses the lamp (Zn or Hg), the wavelength filter and the UV flow cell. The light beam is directed through a double conical or straight flow-through cuvette (6 ml or 2 ml illuminated volume) to a photodetector. The photodetector current is fed to the signal processing circuitry in the module.
A Reference information A.3 pH electrode The pH electrode is of the sealed combination double junction type. It contains a sealed Ag/AgCl reference which cannot be refilled, an internal electrolyte bridge of 4 M KCl saturated with Ag/AgCl, an outer electrolyte bridge of 1 M KNO3, an annular ceramic reference junction and a low profile pH membrane. The pH electrode is delivered with a transparent cover. The flow cell should not be used with any other pH electrode.
Reference information B B Menus B.1 Check menus B.1.1 Checking autozero level The module internal absorbance value for autozero can be checked to test the consistency of buffers. Check Autozero AZ 0.00006 AU 1 Select menu Check and press OK. 2 Select sub menu Check Autozero. The autozero absorbance value for the used wavelength is shown. B.1.2 Checking lamp run time The lamp run time can be checked to determine the need for lamp replacement. Run times for both Hg and Zn lamps are monitored.
B Reference information Telephone Service: 012345678901 Contract Number: 012345678901 Serial Number: 01234567 YM 012345 UPC-900 V1.00 Date of Maintenance: ? Buzzer Test 3 The service telephone number is displayed, press OK. 4 The service contract number is displayed, press OK. 5 The module serial number is displayed, press OK. 6 The module name and software version are displayed, press OK. 7 The date of the last service is displayed, press OK.
Reference information B B.2.3 Setup show temperature The display of the temperature in the conductivity flow cell, shown in the main operating menu 2, can be enabled or disabled. 1 Select sub menu Setup Temp and press OK. Setup Temp Cond pH UV Setup Show Temp (on) 2 Select sub menu Setup Show Temp and press OK. The current status for showing temperature is displayed. If on is shown, current temperature is displayed in main menu 1 and 2. If off is shown, no temperature is displayed in the main menus.
B Reference information before adjusting the cell constant (see section B.2.2). 1 Prepare a calibration solution of 1.00 M NaCl, 58.44 g/l. Let the solution stand until it is at room temperature. This is important for exact measurements. 2 Fill the flow cell completely with the calibration solution by pumping at least 15 ml through the cell with a syringe. 3 Stop the flow and wait 15 minutes until the temperature is constant in the range 20–30 °C.
Reference information B B.2.7 Setup adjust cell constant After replacing the flow cell, the cell constant has to be set. (The cell constant is shown on the packaging). Setup Temp Cond pH UV Setup Adj Cell Const (83.56cm-1) Warning! This will change cell calib Setup Adj Cell Const (83.56-1) 86.6 1 Select sub menu Setup Cond and press OK. 2 Select sub menu Setup Adjust Cell Const and press OK. The current cell constant is shown. 3 A warning message is shown until confirmed by pressing OK.
B Reference information Setup Show Cond (on) on off 3 Change the setting if desired and press OK. B.2.9 Setup pH temperature compensation The relationship between pH and the output signal from the pH electrode is temperature dependent. For accurate measurements during temperature changes, the pH measurement can be temperature compensated. In normal applications, when the temperatures of the buffers and calibration buffers are identical, temperature compensation is not necessary.
Reference information Set Averaging (1.3S) 1.3 Set Averaging (1.3S) 0.64 B 2 Select sub menu Set Averaging and press OK. The current filter constant is shown. Valid values are 0.02, 0.04, 0.08, 0.16, 0.32, 0.64, 1.3, 2.6, 5 and 10 seconds. 3 Change the setting if desired and press OK. B.2.12 Setup lamp run time When the UV lamp is replaced, the Lamp Run Time counter should be reset.
B Reference information Setup Language (GB) GB D E F I 2 Select sub menu 3 Select the desired language. GB = English D = German F = French E = Spanish I = Italian Setup Language and press OK. B.2.13 Setup unit number The unit number is the identification the Monitor UPC-900 has on the UniNet-bus. It should correspond to the number set in UNICORN for the Monitor UPC-900. The number should be set to 0 if one UPC-900 is used.
Reference information B B.2.14 Setup display angle Sets the display angle to compensate for different viewing heights. Set Display Angle and press OK. 1 Select main menu 2 Select sub menu 3 Select viewing angle (->\ Up, Setup Set Display Angle ->| and press OK. Mid or ->/ Down). B.3 Setting and using the alarm timer You can set the alarm function to either a fixed alarm time or use a count-down timer. You cannot set both an alarm time and the count-down timer.
B Reference information Lamp (on) Lamp AU Cond%Tc pH 0.00002 015.0 12.50 Autozero UV Calibrated Electrode Eventmark Calibr pH Buffer 2 (10.00) Set Cond Full Scale pH 12.50 22.4 735.8mS/cm Tc 78.8% Calibr pH Buffer 2 (10.00) Set Cond Full Scale Calibr pH Buffer 2 Set Cond Zero Level Set parameters Calibr pH Buffer 1 (7.00) Set Cond Zero Level Calibr pH Buffer 1 (7.00) Set Cond Analog Out Set Cond Analog.
Reference information B Continued from previous page Setup Setup Adjust Temp Temp sub menus Setup Adjust Temp (22.3°C) Setup Show Temp (on) Warning! This will Setup Show Temp (on) on off Setup Cond Temp Comp (0.0%) 0.0 Setup Cond Ref Temp (25.0%) 25.0 Cond sub menus Setup Cond Temp Comp (0.0%) Setup Cond Ref Temp (25.0%) Setup Adjust Cond (83.53mS/cm) Setup Adj Cell Const (83.56mS/cm) Setup Show Cond (on) Warning! This will change cell calib Setup Adjust Cond (83.53mS/cm) 86.
C 48 Reference information Monitor UPC-900 User Manual 18-1125-55 Edition AE
Reference information C B.5 Menu overview B.5 Menu overview (continued) C Technical specifications The full specifications apply only after at least 1 hour warm-up. C.
C Reference information Temperature sensor accuracy drift Flow rate sensitivity ±2.0 °C ±0.5 °C per 10 h ±1% within 0–100 ml/min Conductivity flow cell Flow rate Max. pressure Generated backpressure 0 - 100 ml/min 5 MPa (50 bar, 725 psi) Max. 0.01 MPa at 100 ml/min pH measurement pH range 0 to 14 (spec. valid between 2 and 12) Accuracy temperature compensated ±0.
Reference information compensated Response time Long term stability Flow rate sensitivity pH flow cell Flow rate Max. pressure Generated back-pressure D ±0.2 pH within +15 to +25 °C ±0.5 pH within +4 to +15 °C and +25 to +40 °C <10 s (0 - 95% of step) Dev. max. 0.02 pH/h (measured at pH 4.0) Dev. max. 0.1 pH units 0.1 - 100 ml/min 0.5 MPa (5 bar, 72 psi) Max. 0.02 MPa (0.2 bar, 2.9 psi) at 100 ml/min C.
December 2002
Short instructions The following short instructions are intended as a guide to users who are fully familiar with the safety precautions and operating instructions described in this manual. These short instructions assume that the module is installed according to the installation instructions. OK ESC 1 Switch on the module with the mains switch on the rear panel. Calibrating 2 Switch the lamp on/off Lamp 3 The main operating menu displays measured parameters. AU Cond%Tc pH 0.00002 015.0 12.
