Models 500, 504, 506 - Scientific Systems
Contents
1. REAR PANEL See Figure 2 2 1 Earth Ground This terminal is continuous with the earth ground 2 Recorder Outputs Two terminals supply an analog output to be used with a strip chart recorder or integrator The full scale outputs of these terminals are selectable from 1Omv 1 00mv and 1 0 V This output is dependent upon the position of the range and rise time controls 3 Integrator Output Two terminals supply a fixed 1 V AU analog output to be used with an integrator This output is independent of the range control rise time control short and event switches but dependent upon the auto zero control 4 Remote Auto Zero Two terminals provide a remote auto zero function A momentary contact closure or TTL low activates the auto zero circuit 5 Remote Event Two terminals provide an event mark with a momentary contact closure or TTL low 6 Remote Lamp Shut Off Two terminals provide a remote means of turning the detector s lamp off Lamps are shut off by providing a continuous contact closure or TTL low Lamps are reignited by the interruption of a contact closure or TTL low 7 Earth Ground This terminal is continuous with the earth ground Recorder Full Scale Voltage Selection Switches Three two position rocker switches control the full scale voltage of the recorder output These switches do not effect the integrator output 8 Pressing the top portion of this switch so that it rocks upwards to the on po2. 3 3b 2 Fasten the W lamp assembly using the same two screws and aluminum standoffs that are used to fasten the D2 lamp assembly to the detector 3 Plug the W lamp power cord into the lower of the two receptacles located on the detector 4 Replace the lamp housing and its retaining screws TO REMOVE THE W LAMP 1 2 3 9 Make sure that the power cord is disconnected from the rear panel of the detector Unscrew the screw and remove the lamp housing the rear housing on the left side of the detector to expose the W lamp WARNING The W lamp gets quite hot Allow sufficient time for it to cool before attempting Its removal Disconnect the W lamp lead from the detector by gently pulling straight back toward you Do not twist the connector while pulling Unscrew the thumbscrews holding the lamp assembly in place and pull the lamp assembly straight back toward you Be careful not to lose the aluminum standoffs or the thumbscrews Be careful not to get fingerprints on the lamp SERVICE SECTION 4 TROUBLESHOOTING TROUBLESHOOTING Most problems with HPILC detectors are in fact caused by other parts of the system Noisy and drifting baselines poor reproducibility in quantitative analysis and similar problems are more often the result of dissolved air bubbles contaminated eluants dirty samples or damaged columns rather than of actual problems with detector hardware In order to focus mor
3. and optional Tungsten lamp 380 800 nm light sources with concave holographic grating monochromator with double beam optics provisions for purging the monochromator 2 0 1 0 0 5 0 2 0 1 05 02 01 005 002 001 and 0005 AUFS Single output with 10 mV 100 or 1 0 V full scale capability with a switch providing 10 full scale offset at each of the aforementioned settings 1 0 V AU analog output independent of range control but dependent upon auto zero function A switch supplying an additional 10 0mV is provided on the rear of the unit Rear Panel input for Auto Aero Event Mark and Remote Lamp Shut Off 2 x 10 AU 30 sec from 220 280 nm with 1 0 sec rise time static dry flow cell lt 2 10 4 AU hour after 1 hour warm up Auto zero circuit capable of offsetting greater than 1 5 AU with standard flow cell Chart Recorder Filter Display Flow Cells Dimensions Line Voltage Second Order Bessel filter with four user selected rise times 0 1 0 3 1 0 and 3 0 seconds Rise time in seconds is 2 x Time constant in seconds A filter bypass switch located on the rear panel provides an equivalent rise time of 0 1 seconds digit LED displays absorbance and relative sample and reference light intensities Pathlengths from 10 mm to 0 0 mm cell volumes from 0 to 15 microliters stainless steel titanium or Ke F contact materials sapphire windows 1000 psi pressure r
4. 6 The cables should have a 14 or so of bare wire or more ideally a spade connector Connect the positive input of the recorder to the screw labeled positive for the recorder output Connect the input of the recorder to the screw labeled negative for the recorder output INTEGRATOR CONNECTIONS The detector also provides a fixed span output I V AU for use with an integrator or data acquisition system This is independent of the rise time control full scale range and voltage settings of the unit However its output will reflect the zeroing of the Auto zero circuit Thus a changing baseline can be corrected by pushing the Auto zero button Connect the input line of the integrator in an identical manner as was outlined for the strip chart recorder OTHER CONNECTIONS The detector provides for a number of other connections for accessories You do not need to connect these in order to finish the initial installation and check out If you do not wish to connect the Remote Auto Zero Event Switch and Lamp Shutoff at this time you may proceed directly to the FLUID CONNECTIONS sections AUTO ZERO The Auto zero connection on the Rear Panel See Figure 2 6 duplicates the function for the Zero Button on the Front Panel whenever it is connected to a momentary contact closure or TTL low OPERATION Connect the triggering device so that the positive line is connected to the positive pole and the negative line is
5. Kel F HPLC Flow Cell Variable Pathlength Preparative Scale HPLC Flow Cell Stainless Steel Body Variable Pathlength Preparative Scale HPLC Flow Cell inert Titanium Body Capillary Flow Cell for off column detection suitable for Micro LC amp Super Critical Fluid Chromatography 250 gm Pathlength and 35 nL volume Capillary Flow Cell for on column detection suitable for Micro LC amp Capillary Zone Electrophoresis High Pressure Microbore Cell suitable for pack column Super Critical Fluid Chromatography 2mm Pathlength 250 nL volume Remote Auto Zero and Event Marker Cable Variable Regulated Back Pressure Device
6. capabilities and specifications of the unit Section 2 This section describes the procedure for unpacking and installing the instrument and for routine operation as an LC detector Section 3 This section describes routine maintenance and service Section 4 This section is a troubleshooting guide Section 5 This section describes the theory of operation Section 6 The appendices describe the warranty accessories and replacement parts INTRODUCTION CAPABILITIES SECTION 1 CAPABILITIES AND SPECIFICATIONS CAPABILITIES This unit provides a unique combination of application flexibility and analytical power in an easy to use low cost package The instrument is capable of functioning as 1 A variety Wavelength Detector from 190 380 mn with the standard deuterium lamp 2 A variety Wavelength detector from 390 800 mn with the optional tungsten lamp 3 An Analytical High Performance Liquid Chromatography HPLC Detector with the standard 6 mm pathlength 9uL volume stainless steel flow cell Model 9550 0100 4 An Analytical HPLC Detector with an optional 10 mm pathlength 15uL volume stainless steel flow cell Model 9550 0122 5 An Analytical HPLC Detector with an optional 6mm pathlength 9uL volume inert surface flow cell Model 9550 0103 6 Microbore HPLC Detector with an optional 3mm pathlength 1 2uL volume stainless steel flow cell Model 9550 0053 7 Asemi preparative HPLC Detector
7. lamp Do not loosen this screw Do not loosen these screws FIGURE 3 3b Tungsten Lamp Loosen these thumbscrews to remove tungsten lamp Lamp alignment pin 3 7 3 8 THE TUNGSTEN LAMP W The lifetime of the tungsten lamp is approximately 2 500 hours To check the W lamp intensity 1 Power up the unit if it is not already approximately 10 minutes 1 Adjust the wavelength drive to read 550 nm from a position at least 10 0 nm below 3 Push the Reference Light Intensity Switch 4 If the displayed value is equal to or greater than 100 the lamp is good If the displayed value is between 99 and 50 the lamp is marginal If the displayed value is less than 50 the lamp should be replaced In general a value less than 15 for the 380 450 nm range and less than 50 for 450 800 nm range is indicative of a bad W lamp The W lamp should be used for all wavelengths above 380 nm The user may notice higher relative light intensities for the D2 than the W lamp from 380 450 nm However this light represents the second order diffraction spectra of the D2 lamp and should not be used In order to install the Tungsten Lamp Assembly the D2 lamp assembly must first be removed To remove the D2 lamp see the instruction entitled To Remove the D2 Lamp just prior to this section NSTALL THE W LAMP 1 Slide the W Lamp Assembly along the same alignment dowel used for the D2 lamp see Figure
8. the lamp ignition period adjust the chart recorder pen to your desired position After lamp ignition the appropriate lamp indicator UV for D2 VIS for W will light an absorbance will be displayed on the LED display and the Waft Light will still be ft During this interval the recorder output will be fixed at zero volts while the integrator output will transmit a voltage related to the present flow cell absorbance and last zero value stored in the auto zero circuits memory To activate the recorder output press the Auto zero button This feature protects the chart recorder from rapidly moving and slamming into its margin if the detector is inadvertently set at a sensitive range and a large discrepancy exists between flow cell absorbance and the last stored zero value After the lamp has ignited and the Auto zero button has been pushed ft will be necessary to push the SHORT Switch in order to set the recorder output to zero volts to allow adjustment of the chart recorder pen The SHORT Switch does not effect the integrator output Set the detector to a more sensitive range such as 0 01 AUFS and monitor the baseline until a straight non drifting baseline is noted SETTING DETECTOR QONTROLS After the column has equilibrated and the detector has warmed up prepare a sample to be injected Set the detector parameters according to the following guidelines WAVELENGTH Turn the wavelength selector until the wavelength indicator coincides
9. 00 hours merely that its intensity will be reduced to 50 of the original output Each D2 lamp assembly is equipped with a chronometer indicating the total hours of operation The chronometer is read by noting the position of the gap in the mercury tube against the graduated background To check the lamp intensity 1 Power up the unit if it is not already on Waft for a period of approximately 10 minutes 2 Adjust the wavelength drive to read 254 nm from a position at least 10 nm below 3 Push the Reference Light Intensity Switch 4 If the displayed value is equal to or greater than 100 the lamp is good If the displayed value is between 99 and 50 the lamp is marginal If the displayed value is less than 50 the lamp should be replaced As a general rule the D2 lamp should produce reference light intensities greater than 50 from 190 380 nm TO REMOVE THE D2 LAMP 1 Make sure that the power cord is disconnected from the rear panel of the detector 2 Unscrew the screw and remove the lamp housing the rear housing on the left side of the detector to expose the lamp see Figure 2 3 SERVICE 3 5 NOTE No harm will be done if the lamp is plugged into the wrong connector but the lamp will fail to light 36 2 Unscrew the screw remove the lamp housing the rear housing on the left side of the detector to expose the lamp see Figure 2 3 CAUTION UV light can damage eyes and skin Always disco
10. FORMING A TEST RUN After setting the detector parameters the instrument should be zeroed As a general rule it is a good idea to auto zero the detector prior to each injection Zero the detector inject your sample and activate the event mark You should note an approximate 20 deflection on the recorder Note the peaks as they appear on the strip chart recorder Readjust the parameters of wavelength range and rise time to optimize the chromatography OPERATION NOTE It is possible to turn this NOTE It is possible to turn this control beyond 190 or 800 nm These wavelengths are outside the instruments specified range NOTE For maximal wavelength reproducibility a given wavelength value should always be approached from the same direction For example if initial work at 254 nm was performed moving the wavelength control from 210 nm to 254 nm all subsequent work at 254 nm should be performed by arriving at that wavelength from a value about ten nanometers less NOTE Although the Second Order Bessel filter may distort peak shape at long rise times peak area is always conserved Thus integration can be safely performed for the purpose of quantitative analysis from the recorder output NOTE Typically rise time in seconds is equivalent to 2 times a time constant in seconds 2 15 NOTE After the lamp is remotely reignited the strip chart recorder output will transmit a voltage related to the current flow c
11. Models 500 504 506 Detector equivalent to Linear Instruments Models 200 204 206 User Manual SOIN LabAlliance Kel F is a registered trademark of the 3M Corporation PS 2 is a registered trademark of the International Business Machines Corporation CONTENTS TABLE OF CONTENTS Introduction i SECTION 1 CAPABILITIES AND SPECIFICATIONS Capabilities 1 1 Specifications 1 2 SECTION 2 INITIAL OPERATION Unpacking 2 1 What You Will Need 2 1 Controls and Conneciions 2 2 Front Panel 2 2 Display 2 2 Rise Time Selection Switch 2 2 Range Selection Switch 2 2 Wavelength Selector 2 2 Wavelength Indicator 2 2 Ultra Violet UV Lamp Indicator 2 2 Visible VIS Lamp Indicator 2 4 Sample Light Intensity Switch 2 4 Reference Light Intensity Switch 2 4 Short Switch 2 4 Even Switch 2 4 Wait Indicator Light 2 4 Over Range Light 2 4 Auto Zero Switch 2 4 Rear Panel 2 5 Earth Ground 2 5 Recorder Outputs 2 5 Integrator Output 2 5 Remote Auto Zero 2 5 Remote Event 2 5 Remote Lamp Shut Off 2 5 Earth Ground 2 5 Recorder Full Scale Voliage Selection Switches Filter By Pass Switch 2 7 Integrator Offset Switch 2 7 Recorder Offset Switch 2 7 Power Switch 2 7 Power Connector 2 7 Fuse Block 2 7 Voltage Control Selector 2 7 Left Bulkhead 2 8 Lamp Housing Cover 2 8 Flow Cell Housing Cover 2 8 Flow Cell Exit Tubing 2 8 Flow Cell Inlet Line 2 8 Setting Up 2 10 Location 2 10 Electrical Connections Setting Voltage Power Cor
12. aft Indicator Light A green LED indicates a fixed zero volt recorder output This occurs a upon powering up the unit After the lamp has ignited it is necessary to push Switch Number 14 the Auto Zero control The Waft Indicator will then shut off and the recorder outputs will become active b upon pushing the Auto Zero button After the new zero value has been stored the light turns off and the recorder outputs will become active 13 Over Range Light A green LED indicates a total absorbance in the flow cell which exceeds the auto zero circuits capacity The unit will still output voltages proportional to absorbance upon both recorder and integrator outputs There will simply be an offset proportional to difference of the new baseline to that of a true zero baseline For instance if the auto zero circuit functions to set the new zero value to 002 AU all output absorbance values will contain a 0 002 AU offset The auto zero circuit is capable of zeroing greater than 1 5 AU with the standard flow cell 14 Auto Zero Switch A momentary switch activates an auto zero circuit capable of zeroing greater than 1 5 AU After pushing this button the Waft Light will turn on and the recorder output will be shunted to zero volts Within three seconds a new zero value will be calculated and the Light will turn off The Over Range Light will illuminate when the total absorbance within the flow cell exceeds this circuits capacity
13. ating for stainless steel cells 500 psi pressure rating for Kel F cells 2 000 psi pressure rating for variable pathlength preparative cells 7 000 psi pressure rating for high pressure microbore cell 6 1 4 inches high 13 4 inches deep 9 34 inches wide Weight 20 Ibs 100 120 220 240 10 Cx 50 OE 60 Hz Kel F is trademark of the 3M Corporation SPECIFICATIONS 1 3 SECTION 2 INITIAL OPERATION UNPACKING Carefully unpack the detector from the shipping container and inspect both the unit and packing for any signs of damage If any damage is noted contact the shipping company immediately In addition to this manual the shipping container contains a power cord and any options which you ordered Carefully check the packing list against the contents of the container If anything is missing check the packing materials carefully for the overlooked items If items are missing contact the factory or your dealer Place the detector on the bench where it will be used and familiarize yourself with the location and function of the controls and connections WHAT YOU WILL NEED In addition to the detector itself you will need the following items for setup and initial operation 1 Strip chart recorder or integrator and connecting cables 2 Liquid Chromatograph 3 Column 4 Standard test mix 5 Appropriate solvents reagents etc 6 Nuts ferrules appropriate to the column end fittings be
14. connected to the negative pole of the remote Auto Zero terminal The remote auto zero may be triggered by shunting across its two input terminals EVENT The Event connection on the Rear Panel see Figure 2 6 duplicates the function of the Event Button on the front panel whenever it is connected to a momentary contact closure or TTL low Connect the triggering device so that the positive line is connected to the positive pole and the negative line is connected to the negative pole of the remote Event terminal The remote Event may be triggered by shunting across its two input terminals REMOTE LAMP SHUT OFF The Remote Lamp Shut Off connection on the Rear Panel see Figure 2 6 turns the lamp off when supplied with a constant contact closure or TTL low Connect the triggering device so that the positive line is connected to the positive pole and the negative line is connected to the negative pole of the Remote Lamp Shut off terminal The Remote Lamp Shut off may be triggered by shunting across its two input terminals Interruption of this shunt reignites the lamp FLUID CONNECTIONS The Detector fluid inlet is the lower tube which protrudes from the rear wall of the cell compartment see Figure 2 3 As a general rule the less tubing between the column outlet and the flow cell the better Ideally the column outlet should be connected directly to the detector inlet line If this is not possible you should use a minimu
15. d Setting the Recorder Full Scale Voltage Recorder Connections Integrator Connections Other Connections Auto Zero Event Remote Lamp Shut Off Fluid Connections Operation Setting Detector Parameters Wavelength Range Rise Time Performing a Test Run Shut Down 2 10 2 10 2 10 2 12 2 12 2 12 2 12 2 12 2 13 2 13 2 13 2 14 2 14 2 14 2 15 2 15 2 15 2 16 SECTION 3 ROUTINE MAINTENANCE AND SERVICE Changing the Flow Cells Cleaning and Disassembly of Flow Cells Changing Lamps The Deuterium Lamp To Remove the D2 Lamp To Install the D2 Lamp The Tungsten Lamp W To Install the W Lamp To Remove the W Lamp SECTION 4 TROUBLESHOOTING Light Intensity Diagnostics Proper Full Scale Voltage Output Filter Bypass Switch General Problems SECTION 5 THEORY OF OPERATION SECTION 6 APPENDICES A Accessories and Replacement Parts B Warranty C Flow Cell Instructions 3 1 3 4 3 5 3 5 3 5 3 6 3 8 3 8 3 9 4 1 4 1 4 2 4 3 5 1 INTRODUCTION The Model 500 is a manual variable wavelength ultraviolet visible absorbance monitor for liquid chromatography The unit is capable of performing applications from capillary to preparative scale making it unmatched by any other detector in its class This manual describes the installation operation troubleshooting maintenance and service for the MODEL 500 detector The manual is organized in the following manner Section 1 This section briefly describes the
16. d not be used 13 Recorder Offset Switch A single two position rocker switch provides a 100 fixed offset to the recorder output This offset is independent of full scale range or full scale voltage This recorder offset does not effect the integrator output 14 Power Switch A single two position rocker switch turns the instrument on and off Pressing the top portion of this switch so that it rocks upwards powers up the unit while pressing the bottom portion of the switch so that it rocks downwards shuts the unit off 15 Power Connector A three pin receptacle is provided that accepts a standard modular power line cord 16 Fuse Block Pry out this block to allow access to the fuses and voltage control It contains one 1 0 amp slo blow fuse for 100 120 VAC operation and two 0 5 amp slo blow fuses 220 240 VAG operation 17 Voltage Control Selector A four position voltage selector allows the instrument to be operated at 100 120 220 or 240 VAC 50 Hz 60 Hz OPERATION NOTE If any of these three switches are simultaneously turned on the recorder output will reflect an uncalibrated full scale Insure that only one switch is on at a time 2 7 NOTE If a second detector is to be used in series with this detector be sure to minimize the total tubing length from this outlet tube to the inlet tube of the second detector This prevents excessive band broadening NOTE The outlet and inlet tubes O D I D and p
17. e effectively on troubleshooting detector problems we will first discuss on board diagnostic tips and later present a troubleshooting table organized by symptom cause remedy LIGHT INTENSITY DIAGNOSTICS The detector provides the capability of monitoring relative light intensities at both the sample and reference photodiodes If an unusually noisy baseline is noted relative intensities of reference and sample light should be assessed Acceptable values are dependent upon the flow cell used wavelength of operation and background absorbance The basic guidelines are as follows 1 A clean flow cell and good lamp will yield a reference to sample light ratio of approximately 2 1 2 An unusually high reference to sample light ratio may indicate a dirty flow cell b excessive absorbance by solvent 2 Anacceptable ratio of reference to sample light accompanied by a reference light level less than 50 indicates a bad lamp PROPER FULL SCALE VOLTAGE OUTPUT An exceptionally noisy baseline may also be due to an inappropriate full scale voltage output setting for the strip chart recorder To test if the detector s full scale output voltage is properly configured 1 Press the short switch and move the recorder pen to a good reference point 2 Release the Short Switch and press the Event Switch 3 The event mark should be approximately 20 full scale if the output voltage is properly configured 4 If the event mark is too
18. ell absorbance and the last stored zero value in the auto zero circuit 2 16 SHUT DOWN As a general rule it is recommended that the flow cell be flushed with several volumes of clean non ionic eluant This is especially important if ionic buffer solutions have been used After flushing simply turn the the power switch on the back panel to the off downwards position The lamp may be shut off remotely with the rear panel located Lamp Shut off terminal This will prolong lamp life while the detector is not in use The lamp is shut off by providing a continuous contact closure or TTL low The lamp is reignited by interrupting the contact closure or TTL low SECTION 3 ROUTINE MAINTENANCE AND SERVICE CHANGING THE FLOW CELLS The detector accommodates a variety of flow cells with different pathlengths illuminated volumes and wetted materials for different applications A complete listing of the available cells is provided in Appendix A All flow cells are provided premounted in a holder assembly to minimize alignment problems Detailed instructions specific to the various flow cells are included with the flow cells themselves This section of the manual provides general guidelines for flow cell service and maintenance The f low cell is located in the forward housing on the left hand side of the detector Figure 3 1 To change the flow cell 1 Make sure the power cord is disconnected from the rear panel of the detec
19. g wavelength CAUTION Do not force the knob below 180 nm or exceed 820 nm Damage to the wavelength drive may result 5 Wavelength Indicator A mechanical three digit indicator displays the wavelength of operation 6 Ultra Violet UV Lamp Indicator A green LED indicates that a Deuterium D2 lamp is present in the instrument and is fit FIGURE 2 1 DETECTOR FRONT PANEL RISE TIME 20 ES R 0005 RANGE AUFS WAVE LENGTH LAMPS EFR LIGHT LEVEL SHORT EVENT 5 2 3 7 Visible VIS Lamp Indicator A green LED indicates that a Tungsten W lamp is present in the instrument and is fit 8 Sample Light Intensity Switch A momentary switch functions to display a value upon the LED display which is proportional to the light intensity at the sample cell photodiode 9 Reference Light Intensity Switch A momentary switch functions to display a value upon the LED display which is proportional to the light intensity at the reference photodiode 10 Short Switch A momentary switch functions to short the recorder output terminals to zero volts This switch must remain depressed to function This allows the user to set the pen position of a chart recorder This switch does not effect the integrator output 11 Event Switch A momentary switch functions to send an event mark of approximately 20 deflection to the recorder output This switch does not affect the integrator output 12 W
20. ing used 7 Wrenches appropriate to column end fittings 8 Connecting tubing and union if column cannot be connected directly to the cell OPERATION 2 1 N N CONTROLS AND CONNECTIONS FRONT PANEL See Figure 2 1 The following controls and indicators are located on the front panel 1 Display A 3 1 2 digit LED display provides absorbance values up to 1 999 AU This display also provides relative sample and reference light intensities when Switches 8 Sample Light Intensity and 9 Reference Light Intensity are depressed respectively 2 Rise Time Selection Switch A four position rotary switch controls the degree of filtering performed by a Second Order Bessel filter Rise times of 0 1 0 3 1 0 and 3 0 seconds can be chosen Typically Rise Time in seconds is equivalent to 2X the Time Constant in seconds 3 Range Selection Switch A twelve position rotary switch controls the full scale output range for the rear panelpositioned Recorder Output Full scale ranges of 2 0 1 0 0 5 0 2 0 1 05_02_01_005_002 001 and 0 0005 AUFS are provided This switch does not effect the fixed 1 VAU output of the rear panel located integrator output 4 Wavelength Selector A mechanical continuous turn control selects wave lengths from 190 to 800 nm Rotating this control clockwise decreases wavelength while counterclockwise rotation will increase wavelength The arrow indicates the direction of rotation for increasin
21. large the output voltage needs to be reduced NOTE This reference light value of 50 is only applicable for wavelengths within the acceptable range for a given lamp NOTE The user may notice higher relative light intensities for the deuterium lamp than the tungsten lamp from 380 450 nm However this light represents the second order diffraction spectra of the deuterium lamp and should not be used 4 1 FILTER BYPASS INT OFFSET RECORDER PS REC OFFSET Figure 4 1 NOTE If any of these three switches are simultaneously set to the on position the recorder will reflect an uncalibrated full scale output Insure that only one switch is closed ata time 5 If the event mark is too small the output voltage needs to be increased The instrument is factory configured to 10 mV full scale To change the full scale voltage 1 Press the bottom portion of Switch 3 so that it rocks downward to the off position see Figure 4 1 2 100 mV scale output press the top part of Switch 2 so that it rocks to the on position upwards 3 1 0 V full scale output press the top portion of Switch 1 so that it rocks to the on position upwards FILTER BYPASS SWITCH An exceptionally noisy baseline not responsive to the Rise Time Selector on the front panel may be the result of an activated Filter Bypass Switch When the Filter Bypass Switch is on the front panel rise time control is bypa
22. les trapped in the sample cell Output span of detector does not match input span of recorder or integrator Large temperature fluctuations are occurring Flow cell photodiode assembly or flow cell cover is loose Check lines for leakage Increase flow rate and or back pressure Press Event Mark to see if a spike of 20 of full scale occurs Remove detector from the source of drafts of hot and cold air Tighten thumbscrews fastening flow cell holder and flow cell cover TROUBLESHOOTING SECTION 5 THEORY OF OPERATION THEORY OF OPERATION Figure 5 1 illustrates the optical system for the detector Only one lamp deuterium or tungsten can be mounted ad a time Both lamps are continuum lamps that jointly provide consistent intensity across the entire spectrum from 190 to 800 nm Two sets of balf les are used to minimize stray light Wavelength selection is provided by a concave holographic grating actuated by a mechanical wavelength drive True double beam operation is provided by a fiber optic beam splitter A reference photodiode continuously monitors the light from one leg of the beam splitter The other leg is imaged by a lens through a sample cell onto the sample photodiode The photodiodes are connected to individual preamplifiers The preamp output voltages are directly proportional to the light intensity at the photodiode active surfaces see Figure 5 2 The outpu
23. m length of narrow bore 0 010 inch I D connecting tubing and a zero dead volume union Because different columns use different fittings the detector is supplied with a bare tube end to allow connection to any column accepting 1 16 inch O D tubing You should use nuts and ferrules suitable to your column Connect the cell outlet the upper of the two tubes which protrude from the rear wall of the cell compartment to a line leading to an appropriate waste reservoir If bubble formation in the detector cell causes problems you may wish to connect the cell outlet to a restrictor or back pressure device providing 20 60 psi back pressure 2 13 OUTPUT REMOTE RECOADER Zeno EVENT LAMP EKISEEEEEE Fig 2 6 NOTE If excessive noise is noted after interfacing the detector with a remote triggering device a ground loop may have been established It may be necessary to insure that the remote device has a negative output which is isolated from the earth ground NOTE If the lamp is turned off by the Remote Lamp Shutoff and then reignited the fixed zero volt recorder output function indicated by the front panel located Light will not be maintained The recorder output will return to a level representative of the current absorbance and last stored zero value The Auto zero circuit should be activated NOTE Detailed descriptions of the flow cells you specified are contained in the appendices of this ma
24. mn to the input of the sample cell for laake c Increase flow rate and or back pressure on cell d Check recorder with voltmeter to see if either of the signal inputs is grounded to case or earth ground 4 3 4 4 3 Recorder base line drifts excessively e Photodiode window is dirty or not held down properly to the cell holder f Sample cell is not screwed down to the main unit Output span of the detector does not match input range of recorder or integrator External trig gering device is causing a ground loop problem a Contamination sample cell windows has occurred b Solvent from column is chang ing absorption e Remove and clean photodiode window f Check sample cell mounts and cell holder assamhlv g Press event mark to see if the spike is approximately 20 of scale h Use only triggering device with ground isolated from earth ground a Clean cell by flush ing with solvents methanol acetone water nitric 5N acid water Inspect cell and photodiode for fingerprints and smudges and clean if necessary b Column is filled with UV absorbers that are bleeding replace column impure sol vent is equilibrating with the column replace solvent with more pure grade switch to a longer wavelength so that background absorb tion does not fluctu ata as mich Leakage in the lines from column to flow cell Tiny bubb
25. mp Cover Lamp Cover Screw FIGURE 3 2 FLOW CELL ASSEMBLY _ 2 MOY 8814 SM dwe joes MSJOSTUNUJ siy 95007 0 SMAJOS WNU assu 95007 Sul Ajqwesse SM8IOSQUNU 9594 U6S00 HNO 3 3 NOTE Use only spectroscopic grade solvents 3 4 CAUTION Avoid scratching or putting fingerprints on the flow cell windows or the monochromator lens which Is revealed upon cell removal These Items should be cleaned with spectroscopic grade methanol and lint free paper 8 Slide the cell holder assembly onto the alignment dowels The inlet line should enter the bottom of the cell in order to provide efficient bubble flushing Securely fasten the cell holder assembly with the two thumbscrews 9 Slip the inlet and outlet tubes into the slots in the strain relief clamp and tighten the thumbscrews holding the clamp in place 10 Replace the photodiode mount and fasten it securely with the two thumbscrews 11 Replace the cell cover be careful not to pinch the cable or the tubing and fasten it securely with the thumbscrew 12 Reconnect the inlet line to the column or connecting tubing and reconnect the outlet tubing to the fraction collector back pressure device or appropriate waste reservoir 13 Reconnect the power cord
26. nect the negative input of the recorder to either of the two earth ground terminals located on the rear of the instrument This could establish a ground loop resulting in increased noise see the Troubleshooting Section NOTE A fixed 10 0 mV offset can be set for the integrator output by pushing the top portion of the Integrator Offset Switch until it rocks upwards to the on position Switch 5 Figure 2 5 This integrator offset is supplied to aid integrators incapable of zeroing for a negative drifting baseline In most instances it need not be used OUTPUT REMOTE INT RECORDER ZERO EVENT LAMP SE SS Se eee ep Fig 2 6 2 12 SETTING THE FULL SCALE VOLTAGE The detector provides a single strip chart recorder channel The full scale voltage for this channel maybe set at 10 mv 100 or 1 using a bank of switches located on the Rear Panel See Figure 2 5 The instrument is factory configured to 10 mv full scale To change the full scale voltage 1 Press the bottom portion of Switch 3 so that it rocks downward to the off position 2 Fora 100 mv full scale output press the top part of Switch 2 so that it rocks to the on position 3 For a 1 0 V full scale output press the top portion of Switch 1 so that it rocks to the on position RECORDER CONNECTIONS The recorder cables are connected at the VO terminal on the Rear Panel of the detector see Figure 2
27. nnect the power cord before working In the vicinity of the lamp The D2 lamp gets quite hot Care must be taken while handling It to prevent burns Always allow the lamp to cool before removing It 3 Disconnect the UV lamp lead from the detector by gently pulling it straight back toward you DO NOT twist the connector while pulling see Figure 3 3a 4 Unscrew the two thumbscrews holding the lamp mount in place and pull the lamp mount straight back towards you Be careful not to lose the two aluminum standoffs or the thumbscrews Be careful not to get fingerprints on the lamp TO INSTALL THE Q2 LAMP 1 Slide the lamp mount onto the alignment dowel located to the left of the monochromator s aperture the mount has a pre drilled hole to accommodate the dowel see Figure 3 3a The lamp leads should emerge from the top of the lamp 2 Use the thumbscrews and aluminum standoffs to attach the lamp assembly to the detector Connect the lamp lead to the upper of the two terminals in the lamp compartment 4 Replace the lamp housing and its retaining screw CAUTION NEVER loosen the screw holding the lamp to the mount and never attempt to rotate or move the lamp up or down In the mount Doing so will degrade the system performance The lamp is provided as a pre aligned assembly FIGURE 3 3 LAMP ASSEMBLIES FIGURE 3 3a Deuterium Lamp Do not loosen these screws Loosen these thumbscrews to remove deuterium
28. nual NOTE Tubing size and position is different for the adjustable pathlength preparative flow cells high pressure microbore flow cell off column capillary flow cell and on column capillary flow cell See their owner s manuals for details NOTE Before connecting any new tube or column to the detector flow several mL of clean solvent through the new tube to a waste reservoir This will clean any particulates or oil that may be residing in the tube that could clog the heat exchanger or contaminate the sample cell of the detector NOTE When the unit is first turned on the LED indicator will indicate random absorbances Typically a 1 or 1 will be indicated with no significant figures to the right of the decimal point The Light will be lit The deuterium lamp will require approximately 20 seconds for ignition while the tungsten lamp lights almost instantaneously NOTE Allow approximately 1 0 hour warm up period for the detector to reach its optimal performing state Po R OPERATION Turn on the HPLC System and allow the column to equilibrate with the flowing eluant the time required will depend on your particular column and eluant If you have not already done so turn ON the power to the detector using the switch located on the lower rear panel Turn the Range selector knob to 2 0AUFS and Rise Time selector to 1 0 seconds Adjust the wavelength drive to the appropriate wavelength for your test mix During
29. or the voltage appropriate to your country 100 120 220 or 240 VAC 50 or 60 Hz If the voltage is incorrectly set you should reset it to the proper value before proceeding further Insert the blade of a small screwdriver into the slot next to the connector and pry open the fuse block Pull the fuse block straight out Using tweezers or a pair of longnose pliers pull the voltage selector card straight out Orient the plastic indicator for the proper voltage as indicated in Figure 2 4 then press the selector card back into place Insure that the fuse block is properly oriented for the selected voltage This is done by rotating the block along its longitudinal axis until a The long single fuse faces outward for 100 and 120 VAC 1 amp slo blow b The two short fuses face outwards for 200 and 220 VAC 0 5 amp slo blow Snap the fuse block cover back into place POWER CORD Power to the detector is provided by a standard modular power cord assembly Connect the power cord to the receptacle next to the fuse block 2 10 R VOLTA ELE E E 2 4 Voltage Selector Card Indicator Pin Fuse Block 90 2 11 Fig 2 5 NOTE If any of these switches are simultaneously set to the on position rocked upwards the recorder output will reflect an uncalibrated full scale Insure that only one switch is closed at a time NOTE These switches do not effect integrator output NOTE DO NOT con
30. ositions may vary depending upon the flow cell specified See the flow cell owner s manual for more details 2 8 LEFT BULKHEAD See Figure 2 3 1 Lamp Housing Cover Remove the screw fastening this cover to gain access to the lamp WARNING NEVER remove the lamp housing cover when the detector power line Is connected UV radiation from the deuterium lamp can damage skin and eyes Both the deuterium and tungsten lamps get quite hot Care must be taken while handling them to avoid burns 2 Flow Cell Housing Cover Remove the thumbscrew fastening this cover to gain access to the flow cell and sample photodiode 3 Flow Cell Exit Tubing Connect this tube to a fraction collector back pressure device or appropriate waste receptacle 4 Flow Cell Inlet Line Connect this tube directly to the column outlet N possible FIGURE 2 3 DETECTOR LEFT BULKHEAD 2 9 SETTING UP LOCATION Place the detector on a Laboratory benchtop in close proximity to the LC column outlet Allow at least 5 inches of clear space between the rear panel of the unit and any wall or obstruction This provides both access to the rear panel connections and a free f low of air ELECTRICAL CONNECTIONS See Fiqure 2 4 SETTING VOLTAGE Check the voltage selector block located next to the power cord connector on the rear panel The white plastic tab indicates the voltage for which the instrument has been configured This has been factory set f
31. sition sets the full scale recorder output to 1 0V when Switches 9 and 10 are rocked downwards to the off position 9 Pressing the top portion of this switch to the on position sets the full scale recorder output to 100 mV when Switches 8 and 9 are in the off position OPERATION NOTE DO NOT use this terminal as a negative ground for any output or input function It may create ground loops resulting in excessive noise see the Troubleshooting Section FIGURE 2 2 DETECTOR REAR PANEL OUTPUT REMOTE iN 4UTO RECORDER tawy ZERO EVENT LAMP ZAS pee ES Ce FILTER BYPASS PE pO EA oom INT OFFSET REC OFFSET 10 Pressing the top portion of this switch to the on position sets the full scale recorder output to 10 mV when Switches 8 and 9 are in the off position 11 Filter By Pass Switch A single two position rocker switch controls a bypass circuit for the Second Order Bessel filter Pressing the top portion of this switch so that it is on renders the front panel Rise Time Control inert and results in effective rise time of 0 1 seconds 12 Integrator Offset Switch A single two position rocker switch provides an additional 10 mV offset to the fixed 1 VAU signal of the integrator output when the switch is in the on position This switch does not effect the recorder output This integrator offset is supplied to aid integrators incapable of zeroing for a negative drifting baseline In most cases it nee
32. ssed and a resultant rise time of 0 1 seconds is created To deactivate the Filter Bypass Circuit 1 Push the bottom portion of Switch 6 see Figure 4 1 until it rocks downward to the off position 2 Rise Time Selector should now function TROUBLESHOOTING GUIDE GENERAL PROBLEMS Remaining noise and drift problems can be due to various sources within and external to the system The following table provides a checklist of symptoms possible causes and some TROUBLESHOOTING SYMPTOM CAUSE 1 Spikes on a Bubbles recorder base passing thru line cell b External trig gering device is creating elec trical noise c Extremely large supply voltage transients on the AC line 2 Noisy baseline a Sample cell on recorder windows are random contaminated b Sample input line has a leak c Bubble trapped in the sample cell d Recorder or integrator is grounded an causing a ground loop problem REMED a Degas solvent and or supply back pressure to the sample cell also check all high pressure fittings for leaks both liquid and gasses b Check electrical lines for good con nection and or inter ference from broad cast radiation Check for ground loops c Remove systems that consume high power from the AC line a Flush cell with sol vents methanol acetone water nitric 6N acid water and check for leaks b Check all lines from the output of the colu
33. to the rear panel of the detector CLEANING OF FLOW CELLS If at all possible we discourage the disassembly of flow cells for routine cleaning purposes Most cells can be adequately cleaned by flushing with several milliliters of appropriate solvent We recommend the following solvents for this purpose 1 Methanol 2 Tetrahydrofuran 3 Methylene Chloride 4 HPLC Grade Water 5 6N Nitric Acid followed by flushing with HPLC Grade Water If flushing proves to be inadequate consult the owner s manual for the cell model with which your detector is equipped for flow cell disassembly directions CHANGING LAMPS The detector has provisions for accepting two light sources A deuterium D2 lamp assembly 1 90 380nm and tungsten W lamp assembly 380 800nm If your dete ctor was ordered ready for work in the visible spectrum it contains the W assembly If you ordered your detector in the standard configuration it is equipped with the D2 assembly Only one lamp can be mounted In the Instrument at a time Consequently moving from the UV to the visible requires the removal of the D2 assembly and installation of the W assembly All lamp assemblies are supplied pre aligned and no further alignment adjustments are needed when changing from one lamp to another THE DEUTERIUM LAMP The Deuterium Lamp 132 is rated for 1 000 hours of life to 112 the original intensity this does not mean that the lamp will burn out after 1 0
34. tor 2 Disconnect the cell inlet tube from the column or connecting tube and free the cell outlet tubing 3 Remove the cell cover by unscrewing the thumbscrew and pulling the cover straight back to expose the photodiode mount 4 Unscrew the two thumbscrews on the photodiode mount and pull the photodiode mount straight back see Figure 3 2 The connecting cable is long enough to allow the photodiode mount to rest on the bench top CAUTION Be careful to avoid scratching or putting fingerprints on the photodlode The photodiode surface should be cleaned with spectroscopic grade methanol and lint free lons paper 5 Loosen the thumbscrew holding the tubing strain relief clamp in place and gently pull the clamp towards you far enough to disengage the tubing from the clamp 6 Unscrew the two thumbscrews securing the cell holder assembly in place and pull the cell holder assembly straight back toward you to remove it 7 Replacement cells are mounted pre aligned in a cell holder assembly Installation reverses the removal process 3 1 3 2 OO FIGURE 3 1 FLOW and LAMP ASSEMBLIES Cell Cover Photodiode Retaining Screw Photodiode Mount Tubing Strain Relief Thumbscrew Cell Retaining Screws Tubing Strain Relief Clamp Cell Body Assembly Cell Holder Cell Holder Thumbscrews Lamp Light Shield Screw Lamp Light Shield Deuterium Lamp Assembly and Plug Lamp Thumbscrew Standoff Lamp Thumbscrew La
35. ts of the preamplifiers are sent to an analog ratiometer The output of this circuit is a voltage that is directly proportional to the negative logarithm of the ratio of the sample signal to the reference signal This voltage is supplied to the rear panel as the 1 V per absorbance unit 1 VAU integrator output and as the adjusted absorbance full scale recorder output THEORY 5 1 FIGURE 5 1 DETECTOR OPTICAL SYSTEM HOUSING MONOCH SAMPLE PHOTODIODE 5 2 I 5 2 DETECTOR ELECTR FI 1 1 0 NV AL LndiLno HOLVUDILNI U S Bol 0 232 1 0832 OLNV UJIdITdWYV d Lb SHOLOJLI 5 5 5 3 APPENDIX A ACCESSORIES AND REPLACEMENT PARTS PRODUCT NO 201305 TS 9051 0205 TS 9551 0022 TS 9550 0100 TS 9550 0053 TS 9550 0101 TS 9550 0103 TS 9551 0070 TS 9550 0147 TS 9550 0149 TS 9550 0155 TS 9550 0150 TS 0203 0780 TS 9051 0143 DESCRIPTION Pre aligned Deuterium Lamp Assembly Mounting plate to be used with Pre aligned Tungsten Lamp Assembly You must purchase this to be used with TS 0203 7078 Pre aligned Tungsten Lamp Assembly replacement lamp 9 uL 6mm Stainless Steel HPLC Flow Cell 0 8 UL 2mm Stainless Steel Microbore HPLC Flow Cell 3 UL 2mm Stainless Steel Semi Preparative HPLC Flow Cell 9 uL 6mm
36. with an optional 3mm pathlength 4 5 uL volume stainless steel flow cell Model 9550 0101 8 Preparative Scale HPLC Detector with a variable pathlength 0 3mm variable volume 0 4 6 uL with stainless steel body Model 9551 0070 and inert titanium body Model 9550 0147 9 UV Visible detector for use in pack column Supercritical Fluid Chromatography or other high pressure applications with an optional 2mm pathlength 250 nL volume high pressure 7 000 psi microbore cell Model 9550 0150 NOTE Consul your sales representative for other cells 10 An UV Visible detector for psuedo on column capillary detection applications such as Supercritical Fluid Chromatography or Micro LC with an optional 250 um pathlength 35 nL volume capillary flow cell Model 9550 0149 11 An UV Visible detector for on column capillary detection applications such as Micro LC and Capillary Zone Electrophoresis with an optional on column detection flow cell Model 9550 0155 1 1 1 2 SPECIFICATIONS Wavelength Wavelength Drive Band Width Wavelength Accuracy Wavelength Precision Optical Methodology Range Selections Recorder Output Integrator Output Remote Controls Noise Drift Zero Adjust 190 380 nm standard Deuterium lamp 380 800 nm with optional Tungsten lamp Manual drive with mechanical wavelength indicator 6nm 1nm 0 1 nm Standard Deuterium lamp 190 380 nm
37. with the wavelength of maximal absorbance for your sample The best wavelength ranges are a 190 380 nm for the standard deuterium lamp b 380 800 nm for the optional tungsten lamp Rotating this control clockwise decreases wavelength while a counter clockwise rotation will increase wavelength The arrow indicates the direction of rotation for increasing wavelength CAUTION Do not try to go below 180 nm or exceed 820 nm Damage to the wavelength drive may result RANGE Rotate the Range Selector Switch to an appropriate full scale absorbance for your sample Full scale ranges of 2 0 1 0 0 5 0 2 0 1 0 05 0 02 0 01 0 005 0 002 0 001 and 0 005AUFS are provided This switch does not effect the fixed I V AU output of the rear panel located integrator output RISE TIME As a general rule a rise time equivalent to 1 10 of the fastest peak base width should be used Too short a rise time results in an unnecessarily noisy baseline while too long a rise time may distort the shape of the peak For most LC applications a rise time value of 1 0 second is sufficient The Second Order Bessell filer provides user selectable rise times of 0 1 0 3 1 0 and 3 0 seconds For extremely fast peaks a filter by pass switch is provided on the rear panel Figure 2 5 Switch 6 Pressing the top portion of this switch turns the circuit on and renders the rise time selector inert and results in an effective rise time of 0 1 seconds PER
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