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Sedex 85LT Light Scattering Detector manual

1. 2 12 2 5 6 Connecting the Siphon Overflow 00 ceeceeeeeeeeeseeeesteeeennees 2 13 2 5 7 Connecting the Nebulization Gas to the Nebulizer 2 14 2 5 8 Connecting the Columitissac cciiccsssseasseasceve thedgonsscensecdsantenntees 2 14 2X9 Powerme Up theUpiti oa aiei e E ERR 2 14 3 Start up Procedure icwticai icnrastceatoetranisianieansinnttrinennmmatienteraientnesin 3 1 ess OVET eW n e ae Tar EE E Ea 3 1 3 2 The C ntro l Panel nei cs cine ecu d octet a eei ita a 3 1 3 2 1 The Digital Display jiisissscscduseacdasasepsaecaaviceeds inaia 3 1 3 2 2 Nhe User Intertates 2 0 ont Beye ei TAh 3 3 3 2 2 a The Status SCUC OM i i hie aio ses eats r Bleed aes 3 3 3 2 2 b The Offset Screen iicsdiecisistswe nana 3 3 3 2 2 c The Temperature Gadin Screen ccccscccceesseeeeeesteeees 3 4 3 2 2 d The Autozero Offset Screen ta ayscdi caidas 3 5 3 2 2 e The Noise Filter Pressure Unit Screen 3 5 3 2 2 f The Light Source Screen nn seccactiecntrienrcciesad en tneds 3 6 3 2 2 g The Light Source Normalization Screen 000 3 7 322b The Gas Valve Serei onirin 3 7 3 2 2 1 The Power Down Screen ccccccceccccccccesssssececcesseenes 3 8 3 2 2 k The Total Lifetime Elapsed Screen cccccceeseees 3 9 3 2 2 1 Phe Serial Number Screen 3 9 3 2 2 m The Firmware Screen sis pncasuaziees sack ntecceseawaeanke 3 9 3 2 2 n The Factory Menu Code Screen ccccccceeeseeeeenes 3 10 Seo
2. All electrical connections are made via the supply panel Figure 2 2 on rear panel a Connecting the Analog Signal Output If the Analog Signal output from the detector is to be employed plug the Signal cable that is supplied into the Signal socket on the detector Figure 2 2 and to the appropriate socket of the data collection device Make sure to use the correct polarity identified on the cable This is not required if a Chromatography Software Driver is used to control the detector In this case there is no need to use an A D convertor SEDEX Model 85LT 2 9 Evaporative Light Scattering Detector Chapter 2 b Connecting the External Autozero If the external autozero function is to be employed plug the cable that is supplied into the Ext Autozero socket on the detector Figure 2 2 and to the appropriate socket on the controlling device e g autosampler pump etc Refer to section 4 3 2 to operate external autozero signal If a TTL signal is used please make sure to use the correct polarity identified on the cable The external Autozero cable has to be connected to a Start information and not an AutoZero information on a controlling device e g autosampler only if a Chromatography Software Driver is used to control the detector In this mode the detector doesn t proceed to an Autozero it uses it as a signal synchronization for the driver Not using the Autozero connection for a Start
3. Gain 1 Figure 3 4 The Temp Gain Screen The Temp Gain screen is used to set the desired evaporation tube temperature and the photomultiplier Gain When the screen is accessed the cursor is on the Temp setting This evaporation tube temperature can be updated with the Q and buttons and validated with the button The evaporation tube temperature range is from 20 to 100 C ae Note When the detector is initially powered up or if you change the temperature the temperature may first overshoot the setpoint slightly and then stabilize at the desired point This initial overshoot is due to the regulation of the instrument and should not be a concern IE Note To maintain appropriate temperature control when the lowest temperature is required it should be set at least 5 C above ambient Temperature stabilization typical time is 30 minutes Please note that the stabilization time for temperature close to the ambient temperature may be longer When you press the v button the Gain setting field can be edited in the normal manner The gain range is from 1 to 12 and each increase in gain setting produces a twofold signal amplification e g if you change the gain from 1 to 4 the gain is increased by a factor of 8 and the full range of the gain from Gain 1 and Gain 12 is 1 to 2048 The Gain corresponds to the detector sensitivity achieved by controlling the high voltage applied to the photomultiplier to amplify response
4. Power up the detector Seal the solvent inlet connection 1 16 male fitting with a plug connection or a piece of Parafilm and fill the glass siphon with water Apply air or nitrogen pressure of 3 5bar 51 psi monitored by a regulator and checked with a pressure gauge The display should read 3 5bar 0 1 bar 51psi PRESSURE VALUE __ PASS FAIL Set the temperature to 50 C and wait until the temperature stabilizes 30 minutes The display should read 50 C 1 C DISPLAY TEMPERATURE PASS FAIL e Set the Gain to 1 Filter to 1s as described in section 3 2 2 e Temperature order to 50 and allow air or nitrogen gas flow through the instrument at a pressure of 3 5bar 51 psi Power on the light source Set Offset After Autozero to OmV as described in section 3 2 2 d and set Stray Light Adjustment to 100 as described in section 3 2 2 g Wait for temperature stabilisation Autozero the detector Raise the Gain to 12 monitor the signal for 5 seconds and enter the value below Observed Value mV The observed stray light readings should be in the range of 100 150mV PASS FAIL SEDEX Model 85LT A3 3 Evaporative Light Scattering Detector Appendix 3 f This test is performed in the same conditions as the Stray Light value determination Measure the noise over six 1 min segments with the ASTM method For some Chromatography Software the minimum method acquisition time must be at l
5. 2 4 5 Environmental Conditions This instrument has been designed for the following conditions Use inside buildings Altitude below 2000 meters Ambient temperature from 5 C to 40 C Maximum humidity of 80 for temperatures under 31 C with linear decrease down to 50 at 40 C e Maximum variations for main power voltage 10 from nominal voltage e Transitory overvoltage of class II Pollution degree 2 2 6 SEDEX Model 85LT Evaporative Light Scattering Detector Installation of the Detector 2 5 Installation of the Unit 2 5 1 Detector rear panel On the top of the detector rear panel are located the fan which provides cooling airflow through the detector and the black exhaust tube where gas and mobile phase vapour and solute particles exit the detector The exhaust tube installation is described in Section 2 5 3 On the back of the detector rear panel is located a supply panel Figure 2 2 with the gas inlet connection Section 2 5 2 the power module with main power switch and lines fuses and the electrical connection Section 2 5 4 Gas inlet 1V Output RS 232 Ext events Ext Autozero yi V Jo S A D A Figure 2 2 Supply Panel 2 5 2 Gas Supply The unit is connected to the gas supply via the 6 0mm plastic tubing supplied with the detector on the gas quick fitting inlet on the detector rear panel Figure 2 2 which requires no additional fitting The gas supply must be stable and
6. 14 Do not adjust any component inside the detector unless specifically authorized to do so by your dealer 15 If the instrument is used in a manner not specified by the manufacturer the protection ensured by the instrument can be ineffective 16 The user is responsible for decontamination if hazardous material is spilled on or inside the instrument 17 The user is responsible for detector end of life recycling You must not discard this electrical electronic product in domestic household waste This product is classed as a Monitoring and Control instrumentation product Detector internal parts present no danger for recycling Make certain than detector has been cleaned to ensure no solvent or solute can remain in detector drift tube b 18 The warning symbols on the instrument indicate the following Risk of burn Electric shock risk Warning The information in a warning statement relates to a condition or action that could lead to personal injury Table of Contents Warnings and Safety Precautions eececcceeeeeeeeseeeeeeeeeeeeeeeeseeeenees iii A Introduction inaasa aaa anana anana a anaE Saaana 1 2 1 1 The Low Temperature Evaporative Light Scattering Detector 1 2 1 2 Principle of Operation st aiesyce Sot i ee ies cet Rue a egg ame ation ats 1 3 I lt 4 ING DUI ZAR OM mn e e hg catacaiales crake E 1 4 1 2 2 Evaporation of the Solvent 2 4 4 1 2020 eee a ae 1 5 2 D gt ADGUCCH GN 2 6 isisis
7. O Q protocols demonstrate that the instrument performs according to its functional and operational specifications The procedure for the Performance Qualification is described on the worksheet presented in Section A3 4 P Q protocols demonstrate that the instrument performs according to sensitivity using Sulfanilamide as a test compound The overall Detector Performance report is presented in Section A3 5 IR Note The present document should be filled with blue ink except customer authorization for black ink SF Note Before starting S O P for new instruments or after storage the following operations must be performed Set gas pressure to 3 5bars 51 psi Set the temperature to 50 C and wait for stabilization Flush detector with 1mL min 25uL min for micro flow nebulizer of water at least 30 minutes The warm up period for the detector and light source is about 30 minutes S E D E R E recommends that maintenance visits and SOP certification should be performed once a year by a S E D E R E authorized representative SEDEX Model 85LT A3 1 Evaporative Light Scattering Detector Appendix 3 A3 2 Installation Qualification I Q Checklist Model Number SEDEX ELSD 85LT Instrument Serial Number Location of the Detector a b c d e g Has the instrument been delivered as ordered e g according to the U R S or purchase order YES NO Has the instrument been checked and ver
8. information will impair the synchronization of the Signal and may not generate the final report and or impair the retention time reproducibility c Connecting the External Events Cable If the external events functions are to be employed plug the cable that is supplied into the appropriate socket on the rear panel of the detector Figure 2 2 and to the appropriate socket on the controlling device e g autosampler pump etc The white cables are contact closure output cables that provide the ready non ready information to an external device The detector will be in the not ready mode the contact will be in closed position if any one of the following conditions is observed The light source is off The temperature is not at the indicated setpoint e The temperature is at the indicated setpoint but is not stable The pressure is below 3 0bar es Note The controlled device electrical consumption mustn t exceed 20mA under 12V DC The blue cables are contact closure TTL input cables that are used to power the unit down see Section 3 2 2 1 via a signal from an external device to the detector If the TTL signal is used please make sure to use the correct polarity identified on the cable 2 10 SEDEX Model 85LT Evaporative Light Scattering Detector Installation of the Detector d Connecting the RS232 Port If a computer is used to control the detector the detector should be connected to the
9. If the voltage is incorrect adjust with trimmer POT1 Do not plug in the light source connector now h Switch off the light source by pressing the button and plug in the white light source connector i Switch on the light source by pressing the button j Wait for 5 minutes and check the VLED voltage If necessary readjust the voltage with trimmer POT1 Eg Note If the potential is not set accurately the lifetime of the L E D will be adversely affected k In the LED menu Section 3 2 2 f access the Reset time elapsed line and press OK toreset the elapsed time counter for the L E D SEDEX Model 85LT 5 11 Evaporative Light Scattering Detector Appendix 1 Specifications Technical Specifications SEDEX 7 LSD COMPONENTS Light Source Selected High Efficiency Blue LED Elapsed time Counter Detection High Sensitivity Photomultiplier Sensitivity Mid Picogram Level LOD Stearic acid 500pg Dynamic Range Over rr sf 0 Linearity range R gt 0 99 Temperature Range Nebulizer 4PL mM in to 5mL min amp gh fora with 1 Nebulizer with DATA Analog Output 0 1 Volt Gain Setting 1 to 12 Factor 2 2048 Filter Moving Average 0 10 seconds Data Rate Analog 100Hz Digital 30Hz COMMUNICATION Selection amp Display d Keypad Event TTL for Ready Auto zero Power down Power down Method rce Heating and or PMT Computer Interface Soft
10. Screw the new light source cap Do not plug in the light source connector now e Switch on the detector and switch on the lamp by pressing the button Do not plug in the light source connector now 5 10 SEDEX Model 85LT Evaporative Light Scattering Detector Maintenance and Troubleshooting f Check the voltage at VLED test point on ALIMO004v2 Board Figure 5 4 Do not plug in the light source connector now The ground should be one of the fixing screws that attach the board to the detector a O6 ue I tt OEP Lp a l mi HO T oTa e OOk S HAFT E EO T T rer ie D Ko I E a ae HHR EO h hi hd hl Ga k P n NJE SEN Bt tof tah ig i tA h h UPT Ui y a ew tly ha m mh 3 Trimmer POT1 i act J POWER ACS ARS aA RO O Figure 5 4 The ALIMOO4V2 Board Lamp Setting g The voltage at VLED must be set to the value that is provided with the new L E D with a precision of 2 digits example 3 40V
11. collection e Chapter 4 Operating the Detector describes how to operate the Low Temperature Evaporative Light Scattering Detector It includes information about starting the unit on a routine basis collecting data and shutting the unit down e Chapter 5 Maintenance and Troubleshooting describes a series of activities that should be performed on a periodic basis to ensure maximum performance In addition this chapter includes a protocol that can be used to determine the cause of problems that are observed with the instrument e A series of appendices are provided which include product specifications a list of spare parts Standard Operating Procedures and Software Chromatography Drivers 1 8 SEDEX Model 85LT Evaporative Light Scattering Detector Introduction 1 4 For Further Information For additional information about Evaporative Light Scattering Detectors Applications Bibliography Sales or Maintenance Questions or Suggestions don t hesitate to contact S E D E R E and its worldwide network of authorized representatives to provide customers assistance Our Web site includes e mail addresses and phone numbers for representatives locations worldwide Please visit our website for additional information or assistance www sedere com 15 S E D E R E Location Information S E D E R E has two locations in France e Administration and head office are located in Paris e Production is located in Orl ans S E
12. 3 Cleaning Detector 5 8 Nebulizer 5 5 Column Connecting 2 14 Treatment 4 6 Configurations 2 1 Contents of Manual 1 8 Control Panel 3 1 D Decontaminating 5 8 Detection 1 6 Digital Display 3 1 E Electrical Connections 2 9 Electronic Noise Test 3 12 Environmental Conditions 2 6 Evaporation of the Solvent 1 5 External Autozero 2 10 4 2 External Events 2 10 Exhaust Requirements 2 5 2 9 F Factory Method Code Screen 3 10 Filter 3 5 4 6 Firmware version 3 9 G Gain Temperature Gain Screen 3 4 Gas Flow Problems 5 8 Flow Bypass Valve 5 8 Inlet Tube 2 7 Requirements 2 4 Supply 2 7 Gas Valve Screen 3 7 Glassware 1 5 Installing 2 12 I Initial Test Procedure 3 11 Installation 2 1 2 7 Nebulizer 2 12 Introduction 1 1 L Laboratory Requirements 2 4 Light source Screen 3 6 Lifting of Detector 2 3 Location of Detector in Laboratory 2 4 of Offices 1 9 M Maintenance 5 1 Manual Auto zeroing 4 2 Mobile Phase Optimizing 4 5 N Nebulization 1 4 Nebulizer Cleaning 5 7 Gas 2 4 Installation 2 12 Nebulizers 1 4 2 2 Noise Filter Pressure Unit Screen 3 5 Noise Tests 3 10 to 3 14 5 4 O Offset Screen 3 3 Operational Qualification Protocol 0 Q A3 3 Optimizing Mobile Phase 4 5 Performance 4 4 Temperature 4 4 P Power Cord 2 11 Requirements 2 4 Up the Detector 2 14 Power Down Screen 3 8 Powering Down 4 7 Preparing the Unit for Operation 4 1 Pressure Unit Noise Filter Pres
13. 4 6 AG SUMO WAL SEQUENCE ics cxcsesnscsiayeidaasucystes tdesa a e das t 4 7 5 Maintenance and Troubleshooting ccccccssseseseeeeeeeeeeeeeeeeeees 5 1 Dill OVERVIEW siiccsreehertnn itive didi Geet tb ecules aak beii ii 5 1 5 2 Mamnt in e 29 yee ie eee ieee ae Ses ey ie ae 5 1 5 37 Troubles hooting rires sat iiei a eles es eel ted eagle eae 5 3 5 3 1 General Troubleshooting Information 0 0 0 0 eeeeeeeeeeeeteeees 5 3 5 3 2 Initial Troubleshooting Activities eeeeeeseceeeseeeeteeeeeeeees 5 4 33 3 Pertotm the Noise TeStSa iei rn aaa E TERS 5 4 5 3 4 Specific Detector Troubleshooting se sssessesssesssesssseeessresseesse 5 4 5 3 5 Nebulizer Cleaning and Replacement Procedures 5 5 5 3 6 Gas Flow ProblemSs eeeeeeseeeeeseessiseresresseseresresseseresressessresees 5 8 5 4 Cleaning and Decontamination sesseeesseseeseseseeresseseresressessrsrressessresees 5 8 5 4 1 Instrument leanne nenea i aa 5 8 5 4 2 Instrument Decontamination ce eecceeeeeeeceeeeeeeteeeeneeeenaeeees 5 9 5 3 MESES TEP PACS MIST egos soc Ponsa iesit ete a Gaceaded oee gested eei 5 9 SG MIPICNG SOURCE AEE ESE NE AAE Ba Mian RG OR ha LR As 5 10 Appendix 1 Specifications cccccccceeeeeeeeeeeeeeeeeeeeeeeeeeeeeeneeeeeeeeeeeees A1 1 Appendix 2 Spare Parts List ccccccsssssseeeeseeeeeeeeeeeeeeeeeeeeeeeeeeeeeees A2 1 Appendix 3 Standard Operating Procedure and I Q 0 Q P Q A3 1 Appendix 4
14. 85 IT ELSD Gas Valve Open Prog Time O min Off Figure 3 9 The Gas Valve Screen The Gas Valve screen is used to open close the gas valve and to setup a program to close the gas valve after a user selected time period To use this feature move the cursor to the time field indicate the appropriate time then move the cursor to Off and use the Q or key to select On and press Q When you press the v button the External Shutdown screen Figure 3 10 will be displayed SEDEX Model 85LT 3 7 Evaporative Light Scattering Detector Chapter 3 3 2 2 1 The Power Down Screen The Power Down Mode screen Figure 3 10 is used to indicate which features should be shut down upon receipt of a power down signal from an external source or from the menu SEDERE SEDEX 85 Te IT ELSD Power down Mode General Activate Figure 3 10 The Power Down Screen The three options provided for external shutdown are summarized in Table 3 1 Mode Photomultiplier Lamp Heating Gas flow General Off Off Off Off Standby Off Off On Off Cleaning Off Off On On Table 3 1 Power Down Options To select the desired Power Down mode use the Q or Q key to access the desired mode and then press to validate the selection ee Note It will take a few minutes to attain operating status from General power down mode as the temperature must stabilize Once the Power Down mode has been chosen and validated
15. Chromatography Software Drivers sssse A4 1 vi 1 Introduction Thank you very much for purchasing a SEDERE LT ELSD Model 85LT detector which has been designed for analysis in a chromatography system As one of the originators of this detection mode SEDERE remains exclusively focused on this technology as a core competency and provides a complete and versatile product line dedicated to Low Temperature Evaporative Light Scattering Detector LT ELSD As the industry leader SEDERE leverages decades of experience and customer knowledge to continually raise the bar for High Sensitivity High Flexibility and High Fidelity detector performance for chromatography laboratories 1 1 The Low Temperature Evaporative Light Scattering Detector The SEDEX Model 85 Low Temperature Evaporative Light Scattering Detector Figure 1 1 is designed to detect compounds in the eluent from High Performance Liquid Chromatography HPLC Ultra High Performance Liquid Chromatography U HPLC Micro HPLC Gel Permeation Chromatography GPC Supercritical Fluid Chromatography SFC or Counter Current Chromatography CCC It is capable of monitoring eluent flow rates from 5uL min to 5mL min Evaporative Light ss Me Scattering Detector is a nearly universal ase technique which can detect any non volatile analyte Unlike other types of detection mode such as UV Detection it is not dependent on the absorption of radiation a
16. Formate NH4 Acetate C2Hs 3N can be used but they may increase the noise level at high gain settings In addition the solvent should not contain preservatives e g Tetrahydrofuran may contain BHT as a stabilizer The wetted parts of the detector are made from Teflon Stainless Steel and Glass Make sure that the solvents do not react with these materials ae Note Depending on the mobile phase nature and flow rate the suggested gas pressure 3 5bar 51psi may have to be adjusted in order to optimize the background noise and so Signal to Noise ratio SEDEX Model 85LT 4 5 Evaporative Light Scattering Detector Chapter 4 4 5 3 Sample Pretreatment If the sample contains any particulate matter it should be filtered through a 0 2um or 0 45um filter before injection 4 5 4 Column Treatment The chromatographic column typically contains microparticles which are used to separate the compounds of interest Under certain conditions the column packing will undergo chemical and or mechanical breakdown this may lead to the introduction of particulate matter into the detector which may lead to an increase in the noise The breakdown of the column packing is dependent on a variety of factors including the particle size type of column packing the manufacturer of the column and the nature of the mobile phase high pH may degrade silica based columns When you install a new column we suggest that you pump the mobile p
17. If you keep the key pressed the rate of change of the parameter will increase used to decrease the present value of a user settable parameter e g the offset by 1 unit If you keep the key pressed the rate of change of the parameter will increase validates the value of the parameter that you have edited Ae sets the present signal for the detector to zero by updated the offset value to set the present signal to OmV is used to power up the light source in the detector When the light g source is lit the keyboard orange L E D immediately above the button will be illuminated and blinks in case of light source time elapsed warning or trouble detected error A nep l changes the active line on the display to the next previous line or 7 the next previous screen lt gt moves the cursor on the display to the next previous field The keys on the control panel are deactivated to prevent unwanted changes during operation when the detector is controlled by a computer 3 2 SEDEX Model 85LT Evaporative Light Scattering Detector Start up Procedure 3 2 2 The User Interface The Status screen Figure 3 2 is the default screen and is presented after initialization of the detector In addition it will be automatically presented again if you have accessed another screen and have not made any keystroke within a few seconds Each parameter change must be validated with ox button or the change will not be ap
18. Starting kit refer to Table 2 2 Precise detector main Voltage csssscccssccsssscesesceeesceeeees Transparent Black front panel shield WINdOW c esccsseseseesesestesteseeseeneenees Spare Parts List Part Number 85650 85350 85050 85450 85950 85750 85135 85136 85159 REG 85159 CAR 85155 85154 4 85154 6 85156 6 85157 85158 85171 85172 85173 85174 85119 3 15T 85119 Kit 85138 13 85138 22 85145 85146 85137 85191 115 or 230 85144 A2 2 SEDEX Model 85LT Evaporative Light Scattering Detector Appendix 3 Standard Operating Procedure and 1 Q 0 Q P Q A3 1 Overview The Standard Operating Procedure S O P is provided to perform the Installation Qualification I Q Operational Qualification O Q and Performance Qualification P Q to confirm that the instrument is functioning in compliance with manufacturer specifications and therefore validate the instrument at your site The original Declaration of Conformity to manufacturing specifications is shipped with each instrument to certify that the instrument passed the final test An Installation Qualification checklist is presented in Section A3 2 and the installation of the unit is described in Chapter 2 I Q should be performed when the instrument is newly installed and each time it is moved to a different location The procedure for the Operational Qualification is described on the worksheet presented in Section A3 3
19. Union Column Bulkhead Bracket Gas one way Valve Siphon Overflow Figure 2 5 Installing the Nebulizer Glassware Chamber Assembly To install the assembly a Remove the Parafilm from all detector openings and from the nebulization cell these coatings are used to prevent dust particles from entering the detector during shipment b Position the glassware chamber as shown in Figure 2 5 and tighten the black nut at the bottom The glassware chamber should be placed on the backwall with the backwall as shown in Figure 2 6 2 12 SEDEX Model 85LT Evaporative Light Scattering Detector Installation of the Detector gt Figure 2 6 Fixing the Glass Chamber c Use the large black nut to position the nebulizer on the glass chamber d Screw the inlet fitting in the bulkhead on the left side of the compartment Special care must be taken when positioning this fitting The nebulizer is terminated with a small piece of Teflon tubing with an outer green sleeve For proper operation the Teflon tubing must extend less than 2mm past the end of the green sleeve Figure 2 7 r Figure 2 7 Nebulizer Inlet Fitting Fill the siphon overflow on the nebulizer glass chamber assembly with the mobile phase that will be used for the separation If you are using a very volatile solvent e g hexane or CH2Cl2 then use water to fill the overflow The liquid should fill the bent part of the siphon but should not pool in the bottom
20. also program a time table for gain filter temperature changes as a function of time and make LT ELSD detectors fully integrated in many HPLC equipments Drivers available are e Driver for ChemStation Part number 85210 Bundled with detector e Driver for EZChrom Elite Li Part number 85220 m meih e Driver for Xcalibur Part number 85230 e Driver for Clarity Part number 85240 The driver is included in Clarity software Q2EYECeBebe ans rig Administrator When using Drivers the AutoZero cable has to be connected to a Start information Blue wire to a Start and white wire to a GND Ground signal on a controlling device e g autosampler In this mode the detector doesn t proceed to an Autozero it uses it as a signal synchronization for the driver Not using the Autozero connection for a Start information will impair the synchronization of the Signal and may not generate the final report and or impair the retention time reproducibility For Installation and Operation with these drivers you can directly refer to their respective Operator s Manuals A4 1 SEDEX Model 85LT Evaporative Light Scattering Detector Appendix 4 This page intentionally left blank SEDEX Model 85LT A4 2 Evaporative Light Scattering Detector Index A Accessory Kit 2 2 Autozeroing the Detector 4 2 B Background Noise Test 3 11 C Carrying the Detector 2
21. by the following certified person Name Title Affiliation Date Signature A3 6 SEDEX Model 85LT Evaporative Light Scattering Detector Appendix 3 A3 5 Overall Detector Performance After the Installation Qualification I Q Operational Qualification O Q and Performance Qualification P Q procedures have been completed the Overall Detector Performance document should be completed to verify the completion of all tests Model Number SEDEX ELSD 85LT Instrument Serial Number Location of the Detector Part I Installation Qualification 1 Q Date PASS FAIL Part II Operational Qualification O Q Date _ PASS FAIL Part III Performance Qualification P Q Date PASS FAIL The above instrument was certified by the following certified manufacturer s representative Name Title Affiliation Date Signature The above instrument was certified with the presence of the following customer s representative Customer name Customer title Affiliation Customer Signature SEDEX Model 85LT A3 7 Evaporative Light Scattering Detector Appendix 3 This page intentionally left blank A3 8 SEDEX Model 85LT Evaporative Light Scattering Detector Appendix 4 Chromatography Software Drivers SEDEX LT ELSD detectors can be controlled by the keyboard however some Drivers can be proposed for a full control without need for an A D convertor Users can
22. detector is not ready ee Note Make certain that the pressure of gas supplied to the detector is less that 4 5bar 67psi If the pressure increases above 4 5bar the pressure sensor may be damaged This damage is not covered by the warranty SEDEX Model 85LT 3 11 Evaporative Light Scattering Detector Chapter 3 If you have an external gas gauge make sure that the external reading and the reading on the Status screen are in good agreement d Press the Q button The signal should be close to zero and remain constant e Set the noise filtering to 1s Refer to Section 3 2 2 e 3 3 2 Electronic Noise Test To determine the electronic noise a Do not switch the light source on Do not switch the HPLC pump on no solvent flow b Make sure that the siphon is filled and the bulkhead is blocked with Parafilm to avoid a Venturi effect c Set gas pressure to 3 5bar and temperature to 50 C Wait for stable temperature d Set gain 12 and monitor the signal for a period of 5 min The variation in the signal should be less than 2mV there may be some spiking of the signal e Record the level and autozero the detector again 3 3 3 Background Noise Stray Light Test To determine the background noise a Do not turn on the HPLC pump no solvent flow and make sure that the siphon is filled and the bulkhead is blocked with Parafilm to avoid a Venturi effect f Set gas pressure to 3 5bar and tempera
23. drain tubing Fix the drain tubing at the inlet of the waste container so that the end of the drain tubing never dives into the liquid in the container eS Note Ensure that the Parafilm is removed from the exhaust tube before installing the unit The drain tubing must be directed to an appropriate container regarding to the solvent nature The user is responsible for decontamination or recycling of any residue regarding to the local authorities environmental requirements Please check with your local regulatory authorities for health and safety requirements SEDEX Model 85LT 2 5 Evaporative Light Scattering Detector Chapter 2 2 4 4 Location of the Detector in the Laboratory All components of the system e g HPLC pumps detector should be located on a sturdy table The detector should be placed in an area that is free from drafts or significant temperature changes Do not place it near air conditioning vents windows ovens etc 25cm 10 When placing the detector in the laboratory access to the power to disconnect the device the appliance coupler or the mains plug must be kept accessible at all time ee Note The detector should be placed close to the outlet of the column to minimize extra column band broadening which would reduce the resolution of the chromatographic separation Re Note As a destructive detector the ELSD should be the last one in the flow path or alternatively be used with a splitter
24. it from the liquid chromatography flow path in order to avoid any clogging of the nebulizer or deposition of substances inside the detector SEDEX Model 85LT 5 1 Evaporative Light Scattering Detector Chapter 5 For an efficient Preventive Maintenance After each session and before shutting down the HPLC system the ELSD should be cleaned in order to ensure good performances Preventive maintenance consists in cleaning the detector before shutting down after the last analyses a Let the mobile phase or solvent flow to flush particles which could remain in the detector b Eventually increase temperature in order to dissolve possible deposit c Stop the mobile phase flowing but let the gas flow at least 30min to avoid particles deposit d Stop the gas flow if desired at the source or close the detector gas valve see section 3 2 2 h e Shut down the detector if desired The time required for each step depends on the application solvents type and concentration of the samples and should be determined accordingly Ise Note It is not necessary to access inside the instrument in routine operation If the suggestions provided in this chapter do not remedy the problem contact your local distributor ee Note The L E D used as the Light Source has a long but finite lifetime 5000 hours and should be replaced periodically by a skilled technician When this period has been reached a message indicating that the maximum
25. regulated by an external manometer The gas must be oil free dry and filtered by a 0 01um filter The typical gas pressure to operate the detector is 3 5bars Fire and explosion hazard Do not use gas that support combustion with combustible solvents Do not use air as a carrier gas when the mobile phase contains flammable components SEDEX Model 85LT 2 7 Evaporative Light Scattering Detector Chapter 2 eS Note Make certain that the pressure of gas supplied to the detector is less that 4 5bar 67psi If the pressure increases above 4 5bar the pressure sensor may be damaged This damage is not covered by the warranty The tubing should be cut and firmly inserted into the fitting as shown in Figure 2 3 after removing the Parafilm from the detector gas inlet Insert the tube into the Pull the tube to check a fitting until it bottoms engagement of the grab Figure 2 3 Inserting the Gas Inlet Tube Two pieces of tubing are provided If you are using the instrument with an external filter connect the gas source to the filter and then connect the filter to the back of the unit Make certain that no tube damage or inappropriate installation could allow a gas leak in laboratory To remove the gas inlet tube if necessary refer to Figure 2 4 To remove the tube disengage the grab ring teeth by a simple manual pressure on the push sleeve and withdraw the tube from the fitting Figure 2 4 Removal of the G
26. the black gas tubing has been removed make sure you are using the correct direction where the white one way valve is at the lower end near the gas arrival Avoid leaks at all connections and check for possible leakage when you turn the pump on again 5 3 6 Gas Flow Problems If the gas pressure is correct but the nebulizer does not produce any spray the liquid may be visible on the glassware make sure that the black gas tubing for the nebulizer is in the correct direction where the white one way valve is at the lower end near the gas arrival The gas pressure is indicated on the Status screen and is controlled by a valve inside the detector housing If the gas menu command does not work a manual bypass can be done on the gas valve to open it to set gas flow To bypass the gas valve a Switch off the detector and disconnect the detector from the main power supply b Remove the detector cover c The gas valve is located on the detector rear panel d Turn the red switch on the gas valve e Replace the cover and power up the instrument 5 4 Cleaning and Decontamination 5 4 1 Instrument Cleaning 1 Switch the instrument off 2 Disconnect all connection cables power cable signal cable autozero cable instrument gas input and nebulizer tubing 3 Allow the detector to cool down 4 Clean the outside of the detector with a non abrasive cloth If necessary a liquid such as soapy water or ethano
27. the detector can be powered down in two ways e With External event cable power down signal contact closure or TTL Signal at low level The detector will stay in the selected power down mode while the contact remains closed It comes back in normal mode when the contact closure is released e With Power down screen Press the v button to access the power down screen then press again the v button to place the cursor on the Power down activate line Validate with ox to put the detector in power down mode IE Note To leave the power down mode release the External Cable contact closure or TTL signal if power down has been activated by the external event cable or press any key if power down has been activated from the Power down screen When the cursor is on the Power down activate line pressing the v button will present the Total Lifetime Elapsed screen Figure 3 11 3 8 SEDEX Model 85LT Evaporative Light Scattering Detector Start up Procedure 3 2 2 k The Total Lifetime Elapsed Screen SEDERE SEDEX 85 WO ELSD Total Lifetime Elapsed HHHHHH hrs Figure 3 11 The Total Lifetime Elapsed Screen The Total Lifetime Elapsed information screen indicates the usage of the detector and cannot be edited by the user When you press the button the Serial Number screen Figure 3 12 will be displayed 3 2 2 1 The Serial Number Screen SEDERE SEDEX 85 Q LT ELSD Serial Number HHHHHHHHH Figure 3 1
28. 1 Set the conditions as follows and wait for equilibration time 30min Temperature 40 C Gain P12 Noise filtering Yes or 1S depending on detector version Gas pressure 3 5bar 51 psi air or nitrogen Stray light Adj 100 Offset after AZ Omv Siphon Filled with water Inlet tube 1 16 tubing loop 0 005 I D x 200cm between the pump and the autosampler to create backpressure Solute Caffeine Flow rate ImL min 20uL min for the micro flow nebulizer Mobile phase 100 fresh HPLC water 2 Autozero the detector 3 Make three 3 20uL injections of 5ppm S5ug mL caffeine standard for a full 20uL loop injection deliver 100uL of the standard each time to ensure that the loop is completely filled with the standard The average peak height mV should be At least 70mV under test conditions Peak height 1 mV Peak height 2 mV Peak height 3 mV Average Peak height mV PASS FAIL SEDEX Model 85LT A3 5 Evaporative Light Scattering Detector Appendix 3 4 Set gain 6 and then autozero the detector 5 Make six 6 20uL injections of a 250ppm 250ug mL caffeine standard at gain 6 The repeatability should be 3 0 RSD maximum Peak Area 1 Peak Area 2 Peak Area 3 Peak Area 4 Peak Area 5 Peak Area 6 Avg Peak Area Round Standard Deviation RSD PASS FAIL The manufacturer s procedure for the proper Performance Qualification of this instrument was completed
29. 2 The Serial Number Screen The Serial Number screen cannot be edited by the user The last character indicates the detector hardware revision When you press the v button the Firmware screen Figure 3 13 will be displayed 3 2 2 m The Firmware Screen SEDERE SEDEX 85 LT ELSD Firmware Version Firmware Date MM YY Figure 3 13 The Firmware Screen This information screen presents the firmware version and date release where MM is the month and YY the year The Firmware screen cannot be edited by the user When you press v the button the Factory Menu Code screen Figure 3 14 will be displayed SEDEX Model 85LT 3 9 Evaporative Light Scattering Detector Chapter 3 3 2 2 n The Factory Menu Code Screen z SEDERE SEDEX 85 Q LT ELSD Factory Menu Code Authorized Persons Onl Figure 3 14 The Factory Menu Code Screen The Factory Menu Code screen is reserved for qualified personnel only to access a variety of service features and is not designed to be employed by the user 3 10 SEDEX Model 85LT Evaporative Light Scattering Detector Start up Procedure 3 3 Initial Test Procedure 3 3 1 Preliminary Activities This section presents a protocol that can be used to ensure that the instrument is working in the proper way A detailed standard operating procedure 1 Q 0 Q P Q is presented in Appendix 3 ee Note When the instrument is set up the procedures indicated below should b
30. 3 4 provide results that are similar to that obtained when the unit was initially installed but the Column Noise test Section 3 3 5 provides a significantly different value than what was obtained at installation it is likely that the problem is in the column e g highly retained compounds are being eluted 5 3 4 Specific Detector Troubleshooting a The mist from the nebulizer should be homogeneous If it is not homogeneous the nebulizer the needle or the Teflon tube may be partially obstructed To remove the obstruction pump a solvent that can dissolve the foreign material As an alternative the nebulizer can be placed in an ultrasonic bath to dissolve the foreign material Instructions about cleaning of the nebulizer are presented in Section 5 3 5 Do not disassemble the nebulizer Disassembling the nebulizer will destroy it and this will void the warranty b If the sensitivity of the detector is low ensure that there are no leaks in the system Make sure you are using a fresh sample and consider running the test 5 4 SEDEX Model 85LT Evaporative Light Scattering Detector Maintenance and Troubleshooting using a backpressure loop instead of a column Alternatively the light source may need to be replaced or the nebulizer could be obstructed If the noise test did not show that the problem could be caused by the application or the system a decrease in the sensitivity is often caused by the nebulizer main cause Cle
31. 3 5bars make sure you are using the correct black gas tube direction fitting for the nebulizer where the white one way valve is at the lower end near the gas arrival on the front panel The installed Nebulizer Glass Chamber assembly is shown in Figure 2 5 To remove the nebulizer from the instrument a Switch off the pump and the ELSD detector b Remove the black front panel cover Pull its left side c Disconnect the nebulizer liquid inlet from the column and Disconnect the gas inlet from the nebulizer by pushing on the white inlet refer to Figure 2 4 for detail d Remove the nebulizer from the glass cell by unscrewing the black plastic nut with the right hand whilst maintaining the nebulizer with the left hand Take care not to pull or twist the nebulizer capillary e Remove the gas inlet quick fitting and the black plastic nut to avoid damaging the seals with the cleaning solvent ate 5 6 SEDEX Model 85LT Evaporative Light Scattering Detector Maintenance and Troubleshooting To clean the nebulizer a b c Fill an ultrasonic bath with water Fill a beaker 50 or 100mL with approximately 2cm of an appropriate solvent The solvent is dependent on the nature of the material that is present in the nebulizer In most cases ethanol is a satisfactory solvent Place the nebulizer vertically in the beaker 2cm solvent bath The nebulizer outlet should be placed at the bottom of the bath and the nebuli
32. After you have validated the desired gain setting press the v button again to display the Autozero Offset screen Figure 3 5 3 4 SEDEX Model 85LT Evaporative Light Scattering Detector Start up Procedure 3 2 2 4 The Autozero Offset Screen SEDERE SEDEX 5 o IT ELSD Output Signal Value After AZ O0OmV Figure 3 5 The Autozero Offset Screen This screen is used to allow the baseline level to reach the desired value after an AutoZero is performed with the keyboard or with the external AutoZero signal input on detector rear panel This function can be helpful when the user wishes to have a positive signal value instead of zero especially with some acquisition systems which have only positive signal acquisition capability This setting can be changed with the 0 and buttons and validated by the Q button After you have set the desired autozero offset press the v button to display the Noise Filter Pressure Unit screen Figure 3 6 3 2 2 e The Noise Filter Pressure Unit Screen SEDERE SEDEX 85 o LT ELSD Filter 1s Press Unit Bar Figure 3 6 The Noise Filter Pressure Unit Screen The Filter Pressure Unit screen is used to indicate if digital filtering is desired for the signal data improves signal to noise ratio and the desired units for the pressure display When the screen is presented the cursor is on the Filter field By pressing or keys you change the filtering strength time cons
33. D E R E S A S 9 Rue Parmentier Parc Volta BP 27 F 94141 Alfortville France Phone 33 0 1 45 18 05 18 Fax 33 0 1 45 18 05 25 e mail manager sedere com SEDEX Model 85LT 1 9 Evaporative Light Scattering Detector 2 Installation of the Detector 2 1 Overview This chapter describes how the laboratory should be prepared to optimize the performance of the SEDEX ELSD 85 Low Temperature Evaporative Light Scattering Detector and indicates how the unit is interfaced to other devices such as the column and the data recording device When you have successfully installed the unit refer to Chapter 3 for start up procedures The detector is provided in six different configurations as described in Table 2 1 Each configuration includes the indicated nebulizer to allow the user to employ the nebulizer to best match the application The first configuration Micro Flow nebulizer is dedicated to micro HPLC applications the second one Low Flow nebulizer is dedicated to small bore HPLC the third one is dedicated to standard HPLC the fourth one Combinatorial Chemistry is dedicated to high eluent flow rates the fifth one U HPLC is dedicated to Ultra fast Liquid Chromatography and the last one SFC is dedicated to SuperCritical Fluid Chromatography SEDEX ELSD 85LT Flow Rate Range 115V 230V Detector bundled with a U HPLC Nebulizer 200pL min 1 4mL min 85901 85900 Detector bundled with a f SFC Nebulizer 500pL min SmL mi
34. In liquid chromatography water and organic solvents with low boiling points are typically employed e g CHOH CHCl3 CH3CN A typical mobile phase for a reverse phase separation using Evaporative Light Scattering Detection might be CH30H H20 60 40 while a typical mobile phase for normal phase separation might be C6H14 CHCls 60 40 If acids bases and salts are used to modify mobile phase to provide the desired separation they should be able to be readily evaporated sublimed or decomposed into gases in the evaporation tube Mobile phase modifiers that are commonly used when an Evaporative Light Scattering Detector is employed include NH4OH C2Hs 3N NH Acetate NH Formate HCOOH CH3 COOH and CF3 COOH The exit of the heated tube leads directly into the detection chamber Figure 1 6 Evaporative tube SEDEX Model 85LT 1 5 Evaporative Light Scattering Detector Chapter 1 1 2 3 Detection Analyte particles pass through a detection chamber Figure 1 7 where they are hit by an incident light beam produced by a Blue Light Emitting Diode L E D The amount of light scattered by the particles is measured using a Photomultiplier which converts the scattered light to signal and which is positioned at an angle of 120 with respect to the light beam In SEDEX detectors a secondary gas inlet is used to concentrate the particles in the centre of the detection chamber to further increase sensitivity and to prevent it from contaminati
35. Initial Test Procedures onset nt oe ane wdc oe E ev 3 11 3 3 1 Preliminary ACUYIMES 53 4 725 as aactescusecadacocaschde coccteateemeadeasaaants 3 11 3 3 2 Electronic Noise Test ycicscaysicvstes ice sntistuesgiats a a 3 12 3 3 3 Background Noise Stray Light Test eee eeeeeeeeteeenees 3 13 3 34 Solvent N ise Test aeae a A e Re EA Ean 3 13 33 30 Golim Norse Testiran n ir t a E EERE 3 14 4 Operating the Detector ccccccccceseeeeeeeeeeeceeeeeesseeeeseeeeeeeeeeesssenees 4 1 AT SOWERVIG WE sii ce davetest de eands aster iatedeesteledeediedtieae nediaete a EES aE acdsee ee 4 4 2 Starp SECU TICE wads cca cyanea Sutaty dyansan sued seactubed ecedunedaday E T rs a Eest 4 4 3 Aut zeroing the Detect rse udeu ante ete Suan eases 4 2 4 3 1 Manual Auto zeroing of the Detector eeeeeeeeeesteeeenees 4 2 4 3 2 External Auto zeroing of the Detector eee eeeeceeeeteeeeteees 4 2 4 4 Routine Operation of the Detector 0 0 0 eee eeeeceessceeceneeeceeeeeeneeeeneeees 4 3 4 5 Optimizing Performance 25 2 c2ts 3c 293 sett hs se caads Santas eden taent tase vaaaia sce 4 4 4 5 1 Selecting the Optimum Temperature eeeeeeeeeeseeeeenees 4 4 4 5 2 Optimizing the Mobile Phase eee eeeseceeeseceesteeeesteeeenaees 4 5 453 Sample PICWEAUTIEIIL sspascgacastinastueaysey bets nitire e i 4 6 AO Column Treatment ense es i aa Oe ate 4 6 4 5 5 Optimizing the Noise Filter s 351cc2h2isssdeea5aecededecpccsenteeacdetaassessaars
36. SEDEX MODEL 85LT LOW TEMPERATURE EVAPORATIVE LIGHT SCATTERING DETECTOR OPERATOR S MANUAL SEDERE S A S PARC VOLTA BP 27 9 Rue Parmentier 94141 Alfortville Cedex FRANCE Telephone 33 1 45 18 05 18 Fax 33 1 45 18 05 25 Part Number 86000 Rev 9 Jan 2012 This product is covered by the following Patent Application Number Patent Application Number Patent Application Number Patent Application Number Patent Application Number Patent Application Number Patent Application Number Patent Application Number Patent Application Number Patent Application Number Patent Application Number Patent Application Number Patent Application Number 97 16240 date Dec 22 1997 FR 98 15518 date Dec 9 1998 US 6 148 661 date Nov 21 2000 FR 01 09 202 date Jul 11 2001 EP 02 290 580 6 date Mar 8 2002 FR 03 01652 date Feb 12 2003 US 2003 0086092 A1 date May 8 2003 FR 03 13706 date Nov 24 2003 EP 1 447 663 A1 date Aug 18 2004 US 6 936 174 B2 date Aug 30 2005 FR 06 50372 date Feb 2 2006 FR 07 52752 date Jan 19 2007 FR 2007 050669 date Jan 22 2007 SEDERE SEDEX and LT ELSD are trademarks of S E D E R E S A S Parafilm is a trademark of American National Can Co Teflon is a trademark of E I Dupont de Nemours Inc Tygon is a trademark of the Norton Corporation ChemStation and EZChrom are trademarks of Agilent Technologies Inc Xcalibur is a trademark of Thermo Fi
37. an the nebulizer as described in Section 5 3 5 If the sensitivity does not return to normal the nebulizer might need to be replaced Please note that the root cause might also be in different module i e volumes injected by the autosampler might be too low or dead volumes in capillary connections may cause peak broadening c If the detector signal is saturated or if there is a decrease in the dynamic range of the system it is possible that a residue is passing through the detector cell this will lead to an intense signal due to a significant amount of light scattering This residue may be a result of the elution of strongly retained materials from the column or may come from the solvent To determine the cause of the problem bypass the column and observe the signal intensity e If the signal returns to normal strongly retained materials are eluting from the column Flush the column with a strong solvent to elute all material e If the signal does not return to normal the solvent contains a too high residue material after evaporation and is not suitable for use with the detector d If the noise of the detector without solvent is high or if ghost peaks occur it is possible that foreign material is present in the drift tube In this situation increase the temperature to 100 C and pump appropriate solvent at the rate of 2mL min 40uL min with micro flow nebulizer using a gas pressure of 3 5bar 51psi The solvent will be determined
38. anel Figure 3 1 includes a digital display and a number of buttons that are used to enter data To increase value To decrease value To navigate into menu To validate To power on off light source To AutoZero the signal Figure 3 1 The Control Panel 3 2 1 The Digital Display The digital display presents information about the present status of the detector and is used to enter a variety of parameters When the detector is powered up the display will present a greetings message that includes the version number and date that the version was created for a few seconds SEDEX Model 85LT 3 1 Evaporative Light Scattering Detector Chapter 3 After the detector has completed the initialization procedures it loads the last conditions used before the power off and the Status screen Figure 3 2 will be presented SEDERE SEDEX 85 Te IT ELSD Gain 1 Temp Press 26 C 3 5b Signal 001 Figure 3 2 The Status Screen The user interface is provided via a series of screens that are described in Section 3 2 2 Some screens present information about the instrument status and cannot be edited by the user e g the Status screen while other screens e g the Temperature Gain screen Figure 3 4 are used to enter the desired set points The keys on the control panel are used to provide the following functions used to increase the present value of a user settable parameter e g r the offset by 1 unit
39. as Inlet Tube 2 8 SEDEX Model 85LT Evaporative Light Scattering Detector Installation of the Detector 2 5 3 Vent the Exhaust Line to a Fume Hood The black exhaust line on the back of the unit containing the volatilized mobile phase and sample particules and should be vented to a fume hood Make certain that the fume hood withdraws gas from the detector i e there should be a positive pressure between the detector and the fume hood Verify that no tube damage or inappropriate installation could allow a gas leak in laboratory The vacuum must be moderate to avoid turbulence in the glass cell siphon or liquid spilled into the evaporation tube excessive noise or loss of sample Avoid loops or bends in the black exhaust tubing which could create condensation traps resulting in bad measurement results If gas from the fume hood enters the detector i e a negative pressure exists between the detector and the fume hood it is possible that foreign material from the fume hood could contaminate the detector Install the vent tube so that it cannot become blocked or bent or restrict the gas flow from the detector to the fume hood in any way Avoid long tube installations in upward direction creating condensation dropping back into the detector If an extension tube is required i e the supplied tube is not long enough a suitable length of 34 ID of PVC tubing can be fitted over the exhaust tubing 2 5 4 Electrical Connections
40. ated once a year by a SEDERE authorized representative There are no components inside the detector that need to be serviced by the user 5 1 Overview This chapter describes e The maintenance procedures that should be performed by the operator on a routine basis Section 5 2 e Troubleshooting activities that should be useful in determining the cause of erratic or erroneous results Section 5 3 e Cleaning and decontamination procedure that should be performed to maintain instrument performance Section 5 4 e The replacement of main fuses Section 5 5 5 2 Maintenance The SEDEX ELSD 85 Low Temperature Evaporative Light Scattering detector is designed to require a minimum of maintenance activities If preventive maintenance activities are followed the detector should provide high sensitivity measurements without any further intervention by the operator The following general recommendations are proposed e Maintain the detector in a clean laboratory environment e If the instrument is not going to be used for a period of time flush out any mobile phase that contains acids bases or salts to prevent the deposition of foreign matter on components or corrosion of the instrument e Only use clean gas particle free and oil residue free Closing gas valve while the pump is still running may result in serious nebulizer damage If ELSD is used as a second detector and is not being used for some time it is recommended to remove
41. by the nature of the samples that were previously analyzed with the detector If you do not know the nature of the sample ethanol is a good choice Do not use solvents that can potentially corrode the instrument Maintain the flow and temperature during 3 hours at least 5 3 5 Nebulizer Cleaning and Replacement Procedures With time the nebulizer can get clogged by sample and mobile phase materials A dirty or clogged nebulizer can cause increased baseline noise and decreased sensitivity The following procedure can be used to clean the nebulizer If the mist of the nebulizer is not homogeneous the nebulizer the needle or the Teflon tube may be obstructed To remove the obstruction pump a solvent that can dissolve the foreign material As an alternative instructions about nebulizer cleaning are presented in this section SEDEX Model 85LT 5 5 Evaporative Light Scattering Detector Chapter 5 Handle the nebulizer carefully and do not disassemble the rear part of the nebulizer which is protected by the white thermal seal Improper handling of the nebulizer will destroy it and this will void the warranty The nebulizer rear part results from a very tricky setting which mustn t be dismounted for any reason In case it has been removed or unscrewed the only solution is to proceed to a nebulizer replacement In case of the nebulizer doesn t produce a spray and the liquid drawn directly to the siphon even if the pressure display is
42. ccesssssssssssssscssssssesssssseseseens Gas Gas Regulator with Filter and Manometer cccccssssssssssessessesesesesesseseenenes Cartridge O O1ym for Gas REQulator cceccsecssesesssceessseseessesesesesteseesteneenss Black Pneumatic tube for Nebulizer with fitting one way valve 3 Gas Wall Mounting Fitting 4 mm Diameter nebuliser c cece 6 mm Diameter rear panel cesses Gas Tube 6 mm diameter Meterii cccccccccccusessesssscescscssssesessscesssessseeeeeees Drain Exhaust Drain Tube Assembly includes stainless steel fitting and PTFE seal 4 Black exhaust tube 2M50 ccc ccc cscscscecsssseescsssssescscsssessscscssenssessseeses Cables Signal CADE shania eaten chess ees tue assess ATOZ O10 Call AEAEE EREE E ExtEvents Cables ssh cen ahi ei hee wih ih E hee ede RS232 Cablenk cit cu fie cele ds ck i tetas 8d ET oo A Bch Na he AG AA Be BA Fuses Main Power Fuses set of 2 FUSES cess csesceceescessescecsssssesesesssessseess Fuse Kitui mi maai a ts UR A eh eels Genii baer oes Miscellaneous Black Plastic Nut for Nebulization Chamber 13 mm Diameter Es aaa 30 mm Diameter cece sees 6 Union Bulkhead Fitting cccccccsessessssescsseesescsscscssesssscssescssseeeseesseees 7 Peek Nut 1 16 with Ferrule ccccccccccsssccsessssssscscssssesssssessescssseseens 8 Kit of 3 seals for GlaSSWAP Ee eee csecesssssscsesessescecscstsscstseseeseeeees 9
43. computer via a free RS232 port using supplied cable In this mode this avoids also to use an A D convertor to convert the Analog Signal from the detector The software drivers for Chromatography System i e ChemStation EZChrom Elite available are detailed in Appendix 4 e Connecting the Power Cord Place the ON OFF switch to the OFF position and plug the power cord into the socket on the rear panel of the detector The power cord of this instrument contains three wires which must be connected to a grounded line All components of the chromatographic system should be connected to a common ground If a two wire outlet is used make certain that an adapter is used to connect the third wire to ground Do not turn on the power at this time SEDEX Model 85LT 2 11 Evaporative Light Scattering Detector Chapter 2 2 5 5 Installing the Nebulizer Glass Chamber Assembly The transparent black front door can be removed to access the nebulizer and glassware chamber When removing this transparent black front door gently pull only its left side towards you To install the door first fix its right side and then gently push its left side Parafilm is used to cover various openings inside the compartment nebulizer and glassware to prevent dust particles from entering the detector during shipment The installed Nebulizer Glassware Chamber assembly is shown in Figure 2 5 Nebulizer Nut Glassware chamber Gas inlet Fitting
44. d for 1ISVAC or 230VAC with 50 60Hz input power depending on the country to which it is shipped Ensure that the voltage value indicated on the power connector on the rear panel corresponds to the line voltage in your facility The detector requires 115V 1 8A or 230V 1 7A Check that the power lines can provide sufficient current The detector must be connected to a properly grounded three prong plug to ensure proper operation of the instrument If a two prong outlet is used make certain that the ground wire is used to ground the instrument It is recommended that all components of the HPLC system are connected to a common ground The detector should not be connected to an electrical line which also serves units with a large power drain or which may be subject to power surges Such units include refrigerators ovens centrifuges and fume hoods 2 4 2 Gas Requirements A supply of filtered oil free clean gas e g N2 or air is required to operate the detector Pure gas is not required as gas is only a carrier vector for the solid particles e g air from an air compressor is acceptable if un reactive with analysis conditions Fire and explosion hazard Do not use gas that support combustion with combustible solvents Do not use air as a carrier gas when the mobile phase contains flammable components The gas supply should include a pressure gauge A manometer with a 0 01um filter part number 85159 REG is available as an o
45. e performed to determine the specific characteristics of your unit We suggest that you save the results in a permanent location as they can be very useful when you are performing troubleshooting activities Ine Note Before starting the tests for a new instrument or after storage flush the detector with water at a flow of ImL min 30uL min with a Micro Flow Nebulizer at least 30 minutes with a temperature set to 50 C and gas pressure to 3 5bar The warm up period for the detector and light source is about 60 minutes The following activities should be performed a Power up the instrument When the detector is shipped from factory the gain is set to 1 and the offset to OmV The Signal screen should indicate 000 or a very small signal b Access the Temperature Gain screen set the temperature to 50 C and press View the Status screen and verify that the temperature is rising to the setpoint on the Status screen Temperature regulation is stable when the Temperature value stops blinking es Note When the detector is initially powered up or if you change the temperature the temperature may first overshoot the setpoint slightly and then stabilize at the desired point This initial overshoot is due to the regulation of the instrument and should not be a concern c Provide gas to the detector and adjust the pressure to 3 5bar 51 psi If the pressure is less than 3 0bar 44psi the pressure value blinks indicating that the
46. e replacement fuses blow up again power off the detector and unplug the power cord from main power and contact your local distributor For safety protection replace fuses with only the same type of fuses 5x20mm and rated T3 15AL 250V fuses To avoid electric shock power off and unplug the power cord from the main power before examining fuses SEDEX Model 85LT 5 9 Evaporative Light Scattering Detector Chapter 5 5 6 Light source The L E D used as Light Source has a long but finite lifetime 5000 hours and should be replaced periodically A decreasing L E D light intensity will cause decreasing signal heights over time When this period of 5000 hours has been reached a message indicating that the maximum usage of the lamp has been exceeded will be displayed when the unit is powered up and the orange L E D on keyboard blinks when Light Source is power on The Light Source exchange part number is e 85120 for Sedex 85 Hardware from A to C 85121 for Sedex 85 Hardware from D Hardware revision can be checked as the last letter of the detector Serial Number refer to 3 2 2 k a Switch the detector on and switch off the Light Source by pressing the button b Switch the detector off and remove the detector cover c Disconnect the light source connector step 1 and unscrew the light source cap step 2 Note For reinstallation the white connector cannot be placed in the wrong way mechanical protection d
47. ease should be less than 200mV For mixed aqueous organic solvents the expected signal is approximately linear with respect to the concentration of organic phase in the solvent e g a water organic solvent 50 50 mixture should provide a signal increase of approximately less than 100mV Note The purity of the solvent is critical for a low background noise The sensitivity signal to noise ratio is inversely proportional to the solvent noise Note In most cases distilled water and HPLC grade solvents are satisfactory When you are comparing solvents from different sources the most critical parameter is the Residue After Evaporation this parameter should be less than 1ppm to maximize the sensitivity of the detector If the instrument fails the Solvent Noise test it is most likely due to an impurity in the solvent rather than a fault with the instrument If changing the solvent source does not solve the problem it may be necessary to decontaminate the instrument as described in Section 5 4 2 or clean the nebulizer as described in Section 5 3 5 SEDEX Model 85LT 3 13 Evaporative Light Scattering Detector Chapter 3 When filtering the solvent verify that it does not extract any contaminant from the filter The mobile phase should not contain non volatile solvent modifiers Volatile solvent modifiers e g CHOOH CH3 3COOH CF3 COOH NH Formate NH4 Acetate C2Hs 3N can be used but they may increase the noise lev
48. east 1 10min for the ASTM noise determination Segment Noise mV Segment Noise mV Segment Noise mV 1 3 5 See eC Minimum Value Maximum Value Mean Value The mean ASTM noise value should be less than 2mV PASS FAIL g This test is performed in the same conditions as the Stray Light value determination Collect data for 15 minutes Initial Signal Level mV Final Signal Level mV 15Min Drift __mV The baseline drift should be less than 2 0mV PASS FAIL IR Note If any part of the diagnostics fails the detector may need more time to equilibrate In any doubt please refer to your local distributor for service The manufacturer s procedure for the proper Operational Qualification of this instrument was completed by the following certified person Name Title Affiliation Date Signature A3 4 SEDEX Model 85LT Evaporative Light Scattering Detector Appendix 3 A3 4 The Performance Qualification P Q Protocol Model Number SEDEX ELSD 85LT Instrument Serial Number Location of the Detector Re Note Before this procedure is performed it is necessary to complete the Installation Qualification Part I and the Operational Qualification Part II The test report presented below or your own appropriate reference test should be used Remove the plug connector or Parafilm from the solvent inlet connector before connecting the capillary to the nebulizer liquid inlet union
49. el at high gain settings In addition the solvent should not contain preservatives e g Tetrahydrofuran may contain BHT as a stabilizer 3 3 5 Column Noise Test IR Note When strongly retained compounds are slowly eluted from the column excessive noise will be observed To determine the column noise a Turn off the pump and connect the column b Restart the pump and allow the mobile phase to flow through the system It is suggested that you flush the column with a strong solvent for a few minutes before attaching it to the detector The flow rate to be used is dependent on the column ID and is indicated in the following table Column ID Flow Rate mm uL min 1000 Table 3 3 Flow Rate versus Column Diameter Indication c Set the gain to 12 and monitor the baseline for a few minutes A suitable column will provide a baseline that is no more than 20 50mV above the solvent baseline ee Note If the mobile phase contains acidic modifiers e g CF COOH disconnect the detector and wash the HPLC system for 12h before starting to analyze unknown samples This wash should be performed after the column noise test is completed but need not be performed after each analysis 3 14 SEDEX Model 85LT Evaporative Light Scattering Detector 4 Operating the Detector 4 1 Overview This chapter describes the operations that should be performed on a routine basis when you want to collect chromatographic data using
50. er will destroy it and this will void the warranty Ise Note The control panel and instrument electronics do not contain any replaceable components If the suggestions provided in this chapter do not remedy the problem contact your local distributor SEDEX Model 85LT 5 3 Evaporative Light Scattering Detector Chapter 5 5 3 2 Initial Troubleshooting Activities a Make sure that the instrument and all components of the detector are grounded to a true ground b Check to ensure that the liquid level in the siphon is appropriate and there is no liquid pooling close to the evaporation tube inlet c Check that the gas pressure is sufficient and stable The selected pressure for most applications is 3 5bar Slpsi and gas consumption is 3L min for all nebulizers except for the CC nebulizer 4L min Pressure above 4 5bar 67psi can damage the pressure sensor The gas filter should be clean and in place Only use gas free of oil residue d Ensure that the flow rate of the pump is constant and check that there are no leaks in the chromatography system 5 3 3 Perform the Noise Tests Repeat the tests described in Section 3 3 and compare the observed data to the results that were obtained when the unit was initially installed These tests can be very valuable to isolate the problem As an example of this point if the Electronic Noise test Section 3 3 2 Background Noise test Section 3 3 3 and Solvent Noise test Section 3
51. gnificantly better sensitivity for volatile and thermally 40 ng injection sensitive compounds The minimum temperature that S can be used is dependent on the ARUN flow rate and the nature of the ks aM e mobile phase Figure 4 1 Chromatogram of Caffeine at Various Temperatures 4 4 SEDEX Model 85LT Evaporative Light Scattering Detector Operating the Detector 4 5 2 Optimizing the Mobile Phase Particulate matter in the mobile phase will increase the background noise The purity of the solvent is a critical issue in the noise When filtering the solvent verify that it does not extract any contaminant from the filter The purity of the solvent is critical for a low background noise The sensitivity is inversely proportional to the solvent noise In most cases distilled water and HPLC grade solvents are satisfactory When you are comparing solvents the most critical parameter is the Residue After Evaporation this parameter should be less than 1ppm to maximize the sensitivity of the detector As an example consider the analysis of a sample in a pure water mobile phase and a polluted water mobile phase It is clear that the use of an insufficient quality solvent can dramatically decrease your S N ratio Sensitivity Figure 4 2 Chromatogram with Various Solvent Quality The mobile phase should not contain non volatile solvent modifiers Volatile solvent modifiers e g CHOOH CHCOOH CF3 3COOH NH
52. hase through it for few minutes before connecting it to the detector This will flush out the microparticles that remained in the column after its manufacture After installing a new column we also suggest that you perform the Column Noise test Section 3 3 5 to obtain the baseline signal value corresponding to this column 4 5 5 Optimizing the Noise Filter The Digital Filter see section 3 2 2 e allows maximizing Signal to Noise ratio by filtering the noise The filter strength should be optimized according to the peak shape and more specifically to the peak width The following table proposes some Filter settings depending on peak width Peak Width Proposed Filter At 50 Second Second Table 4 1 Digital Filter versus Peak Width Indication These suggested values can be optimized depending on your specific chromatography by decreasing Filter if peaks are poorly resolved e g when Rs lt 1 5 or increasing Filter when optimizing Signal to Noise ratio 4 6 SEDEX Model 85LT Evaporative Light Scattering Detector Operating the Detector Example Comparison of digital filters with a signal with Peak width at half height of 2 5second Filter Os Filter 1s Filter 2s Signal Height 124 mV 122mV 110mV Noise ASTM Peak Width Table 4 2 Sensitivity improvement depending on Filter Signal to Noise ratio is multiplied by 2 when selecting Filter 1s without any peak broadening effect If Signal to Noise ratio is more impor
53. i eie s E ois S EE 1 6 1 3 Content of this Manual iiss 23i c cscceseglecceloeess ead ices aden deedtasanede eed 1 8 1 4 For Further Inrorma nn sce csc pstta ccsaccnaseycieacealiassasainceenaeaneusan 1 9 1 5 S E D E R E Location Informations sssesisiaverdedetceaeteti easiedeets Hie aaset 1 9 2 Installation of the DeteCtor ccssseeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeees 2 1 2M OM ORVICW isch S actions Geetha nied tba E E 2 1 2 2 Lifting and Carrying the Detector 3 22 2 3iscc2es ee aces deca hcs ear ject lence aera 2 3 2 3 Unpacking the Detectors 2 2 kt Be ee Oe BAe eae 2 3 2 4 Laboratory Requirements 925 55 ccnss2s seeds cies sdavasovqess bs desea Saso consadeemsaetasensieaaes 2 4 2 4 1 Power Requirements t 5 205002 f eres siiesesgsueissnesads ceebaandanesttas anes sibesesaee 2 4 2 4 2 Gas Requirements wsissivcckessscveaasesacdasspeacdaneaedanaideoandvassaseetebedeacs 2 4 2 4 3 Exhaust Venting and Drain Requirements sseececeeeeeeeeeeeee 2 5 2 4 4 Location of the Detector in the Laboratory seceecseseeeeeees 2 6 2 4 5 Environmental Conditions 0 ee eee eeseeeseceeeeeeeecneecnseesseeeenees 2 6 2 5 Installation Of the Unitasienres e a a anes e 2 7 201 Detector rear panel eenen a R 2 7 PAS A EIES E 8 D h AAE AEE A E E E E oleh gs 2 7 2 5 3 Vent the Exhaust Line to a Fume Hood seeeeceeereseeren 2 9 2 5 4 Electrical Connections 2 203 lnd ikaw is 2 9 2 5 5 Installing the Nebulizer Glass Chamber Assembly
54. ientation where the white one way valve is at the lower end near the gas arrival 2 5 8 Connecting the Column Attach the fitting from the union bulkhead to the outlet of the column 2 5 9 Powering Up the Instrument Place the ON OFF switch to the OFF position and plug the instrument into the wall socket Turn on the unit via the ON OFF switch The display will present the version number and date it was created for a few seconds the version number should be recorded as it may be required for service or troubleshooting and will then present the signal which should be OmV or very close to it the temperature which should be the ambient temperature the pressure which should be zero or very close to it and the gain Avoid leaks at all connections and check for leakages when you switch the pump on eS Note The liquid level in the siphon must be stable and should be equal at both sides If the vacuum is too strong liquid is drawn into the evaporation tube or generate air bubbles from the drain tube and both resulting in bad measurement results Refer to Chapter 3 to prepare the unit for routine operation 2 14 SEDEX Model 85LT Evaporative Light Scattering Detector 3 Start up Procedure 3 1 Overview This chapter describes e the role of the controls and the digital display on the control panel e the start up test procedure e how to prepare the instrument for operation 3 2 The Control Panel The Control P
55. ified as undamaged YES NO Has the required documentation been supplied Is it of correct issue and appropriately identified by Model Number Serial Number and Date YES NO Have details of all services and utilities required to operate the instrument been provided YES NO Have methods and instructions for user maintenance been provided along with contact points for service and spare parts YES NO Is the selected environment suitable for the instrument i e is adequate room provided for installation operation and servicing Have appropriate solvent services and utilities electricity nitrogen gas ventilation solvent waste recovery etc been provided YES NO Has health safety and environmental information relating to the operation of this instrument been provided YES NO The manufacturer s procedure for the proper Installation Qualification of this instrument was completed by the following certified person Name Title Affiliation Date Signature A3 2 SEDEX Model 85LT Evaporative Light Scattering Detector Appendix 3 A3 3 Operational Qualification O Q Protocol This procedure checks the proper operation of the detector with respect to stability of the electronic boards the energy of the light source and the sensitivity of the photomultiplier tube PMT Model Number SEDEX ELSD 85LT Instrument Serial Number Location of the Detector a b c d
56. imum performance 4 5 1 Selecting the Optimum Temperature There are two factors that should be taken into account when selecting the optimum temperature for the detector e Increasing temperature will optimize the evaporation of the mobile phase e Decreasing temperature will minimize the decomposition of thermally labile compounds and the volatilization of semi volatile compounds A very reasonable start is to set the temperature to 60 C if an aqueous mobile phase is used and 40 C if an organic mobile phase is used these temperatures are suggested for a flow rate of ImL min At higher flow rates more elevated temperatures may be required to minimize the noise Ine Note If the mobile phase used is not easily volatile such as DMSO or DMF temperature should be increased to allow correct evaporation process The temperature can be readily adjusted during the method optimization process If you suspect that the compound of interest is thermally labile or semi volatile a lower temperature could be used to improve the sensitivity by reducing the thermal decomposition or evaporation For a given flow rate and solvent there is however a point at which the noise in the chromatogram is dramatically increased because not all of the mobile phase is vaporized As an example consider the analysis of caffeine with evaporation temperatures of 30 C and 60 C Figure 4 1 It is clear that the use of a low temperature provides si
57. l can be used to remove stains or foreign material 5 8 SEDEX Model 85LT Evaporative Light Scattering Detector Maintenance and Troubleshooting 5 4 2 Instrument Decontamination Set the evaporation temperature to 100 C and the gas pressure to 3 5bar 51 psi Pump the appropriate solvent through the system at the rate of 2mL min 40uL min for micro flow nebulizer The solvent will be determined by the nature of the samples that were previously analyzed with the detector If you do not know the nature of the sample ethanol is a good choice Do not use solvents that can potentially corrode the instrument Maintain the flow and temperature during 3 hours at least 5 5 Fuses replacement If the digital display does not switch on when the detector is powered up power the unit off and inspect the main fuses The fuses are located inside the main power module on the rear panel Figure 2 2 A set of replacement fuses is delivered in the starting kit 1 Power off the instrument and unplug the power cord from the main power lines and from the power module on detector rear panel 2 Locate the fuse compartment on detector rear panel and withdraw the fuses holder 3 Remove and discard fuses Insert new fuses into the holder and the holder into the power module The fuse holder only fits in one way into the power module 4 Reconnect the power cord to the power module and to main power lines If the fuses are not blown or if th
58. lizer 5uL min 40uL min 15bar 40pL min 85650 Low Flow Nebulizer 40puL min 1 2mL min 44bar 1mL min 85350 HPLC Nebulizer 200uL min 2 5mL min 4bar 1mL min 85050 Nebulizer Model Flow Rate Range Combinatahial Chemistry ionann 4bar 1mL min 85450 Nebulizer U HPLC Nebulizer 200uL min 1 4mL min 8bar mL min 85950 The Micro Flow Nebulizer needs a specific glassware chamber Glassware for Micro HPLC Part Number 85136 Table 1 1 Nebulizers for the SEDEX Model 85LT Low Temperature Evaporative Light Scattering Detector Nebulizer Information f Part Number and Serial Number Liquid inlet Gas inlet Quick tubing fitting Figure 1 4 Nebulizer 1 4 SEDEX Model 85LT Evaporative Light Scattering Detector Introduction At the outlet of the nebulizer the aerosol travels through a glassware chamber Figure 1 5 With SEDEX detectors glassware chambers are designed to eliminate the biggest droplets of the solvent Large droplets in the aerosol are drawn to a siphon while the fine mist goes to the evaporation tube Siphon Figure 1 5 Glassware Chambers Standard on left and dedicated to Micro Flow Nebulizer on right 1 2 2 Evaporation of the Solvent This second step begins when the droplets are carried by the gas flow into the evaporation tube to evaporate the mobile phase SEDEX detector drift tube has been designed to optimize the efficiency of the required evaporation with the lowest temperatures
59. n 85701 85700 Table 2 1 SEDEX Model 85LT Low Temperature Evaporative Light Scattering Detector Versions Detector bundled with a Combinatorial Chemistry 1 0mL min 4 0mL min 85401 Nebulizer SEDEX Model 85LT 2 1 Evaporative Light Scattering Detector Chapter 2 Some accessories included with the detector are described in Table 2 2 Description Starting kit 1x Power cable 1x RS232 cable 1x Autozero cable 1x Signal cable 1x External Event cable 2 meters 1 meter of 6mm O D gas tubing 1x set of replacement fuses Table 2 2 Components Shipped with the SEDEX ELSD 85 Low Temperature Evaporative Light Scattering Detector The user can employ new nebulizers to best match his applications The complete range of nebulizers for SEDEX ELSD 85 is indicated in Table 2 3 Back Pressure Part with water Number Micro Flow Nebulizer 5uL min 40uL min 15bar 40pL min 85650 Low Flow Nebulizer 40puL min 1 2mL min 44bar 1mL min 85350 HPLC Nebulizer 200uL min 2 5mL min 4bar 1mL min 85050 Nebulizer Model Flow Rate Range Combinatorial Ghemistey I a Gay tain ADmin 4bar 1mL min 85450 Nebulizer U HPLC Nebulizer 200uL min 1 4mL min 8bar 1mL min 85950 The Micro Flow Nebulizer needs a specific glassware chamber Glassware for Micro HPLC Part Number 85136 Table 2 3 Nebulizers for the SEDEX ELSD 85 Low Temperature Evaporative Light Scattering Detector SEDERE provides a wide range of accessories e g Ga
60. nd is not affected by the absorption characteristics of the solvent Thus solvents which absorb UV radiation can be used As the solvent is completely evaporated a gradient can be performed to optimize the separation LT ELSD is a nearly Universal and powerful detector LT ELSD is used as a valuable alternative to Refractive Index and UV Vis detection for the latter particularly when analytes do not possess any chromophore and is also considered as Figure 1 1 The SEDERE relevant complementary equipment to LC MS LT ELSD 85 Detector 1 2 SEDEX Model 85LT Evaporative Light Scattering Detector Introduction The detector can be controlled via the keypad and digital display on the front panel The detector can be also controlled and monitored by an external computer using the RS 232 ports As an alternative the analog signal output can be sent to a recorder an integrator or a data station 1 2 Principle of Operation There are three discrete steps in the operation of the detector nebulization of the eluent evaporation of the solvent and detection of the compound s of interest Figure 1 2 NEBULIZATION gt EVAPORATION DETECTION Figure 1 2 Schematic Diagram of an Evaporative Light Scattering Detector Nebulization involves the conversion of the eluent into a fine aerosol This aerosol is directed to an evaporator to vaporize the solvent then the mist is irradiated by a light source and the scattered light is mea
61. of the evaporation tube Make sure that no liquid leak could affect the detector performance or create laboratory pollution 2 5 6 Connecting the Siphon Overflow Attach the drain tube assembly to the end of the siphon tube using the tapered hose connector and lead the tube to waste and drain Locate the tube in such a way that the discarded part of the solvent can flow freely from the siphon and ensure that the end of the tube never dives in the collected liquid Make sure that the liquid waste container complies with the solvent nature es Note A drain tube with a bend or diving in the liquid will create pressure fluctuations in the detector and will result in bad measurement results SEDEX Model 85LT 2 13 Evaporative Light Scattering Detector Chapter 2 Ensure that no siphon liquid leak could affect the detector performance or create laboratory pollution If the solvent that you are using is not compatible with Tygon e g THF use a piece of Teflon tubing or any material you know compatible with your solvent in its place Please check your local regulatory authorities for recycling solvents and health and safety requirements 2 5 7 Connecting the Nebulization Gas to the Nebulizer Attach the nebulization gas tube coming out from the front panel to the nebulizer gas inlet fitting located on the nebulizer side See Figure 2 5 In case the black gas tubing has been removed make sure you are using the correct or
62. on LED Light beam Photomultiplier Scattered light From particules Figure 1 7 Detection Chamber sieteltetetetete hi a Exhaust tube Optical head Detection chamber T ee 1 6 SEDEX Model 85LT Evaporative Light Scattering Detector Introduction The intensity of the scattered light is a function of the mass of the scattering particles and generally follows an exponential relationship which is shown in equation 1 1 I k m 1 1 where I is the intensity of light m is the mass of the scattering particles k and b are constants A plot of log I versus log m provides a linear response The values of the constants k and b are dependent on a variety of experimental conditions e g the temperature and the nature of the mobile phase SEDEX Model 85LT 1 7 Evaporative Light Scattering Detector Chapter 1 1 3 Content of this Manual This manual is designed to describe the installation operation maintenance and troubleshooting of the SEDEX ELSD 85 Low Temperature Evaporative Light Scattering Detector It includes e Chapter 2 Installation of the Detector describes suitable laboratory conditions for the detector and includes information about interfacing the detector to other devices e Chapter 3 Start up Procedure describes the role of the various controls and displays on the detector In addition this chapter discusses a number of activities to prepare the unit for routine data
63. or In general operation of an HPLC system with Evaporative Light Scattering Detection is similar to operation of the system with other detectors During operation of the detector the following points should be considered a Make certain that the exhaust from the detector is led into a fume hood or other device and make sure that there is a continuous flow of gas through the detector i e no constriction If a vacuum is used ensure that the vacuum effect will not disturb the detector Section 2 5 2 The exhaust gas should not be allowed to enter the laboratory to avoid any injury or laboratory pollution b Ensure that the siphon is filled with liquid at all times The overflow from the siphon should be collected in a suitable container c Make sure that all flow connections are properly tight In case of any leak switch off the pump immediately and remove the liquid Leakage of hazardous solvents may cause personal injury or laboratory pollution d Never exceed a gas pressure of greater than 4 5bar 67psi e Avoid the use of solvent or compounds that could corrode the detector The mobile phase is in contact with Glass and Teflon tubing and the evaporation tube is made of Stainless Steel SEDEX Model 85LT 4 3 Evaporative Light Scattering Detector Chapter 4 4 5 Optimizing Performance If you transfer a method from another manufacturer s ELS detector operating conditions will need to be optimized for max
64. or must be plugged into a grounded power line Make certain that all parts of the instrument are properly connected to a common ground If the mobile phase includes an organic solvent use an inert gas i e nitrogen to nebulize the mobile phase The exhaust from the detector should be vented into a fume hood or similar system Make certain that the output gas does not escape into the laboratory Take into consideration any solvent filter that could be required by your local environmental laws The gas pressure should not exceed 4 5bar 67psi Make certain that the gas flow is maintained while the mobile phase flows through the detector If the gas flow is interrupted for extended periods of time organic solvents could possibly damage the pressure sensor and or the photomultiplier Do not use corrosive materials that could damage the inner metal surfaces stainless steel of the detector Do not use any liquid or gas that support combustion under temperatures reached by the detector 10 Access inside the instrument is restricted to a suitably skilled technician 11 Do not remove the optical head or the photomultiplier tube while the instrument is powered up This can destroy the photomultiplier 12 The siphon overflow tube must contain liquid at all times 13 Do not disassemble the nebulizer or touch any component inside the nebulization chamber This can lead to the deposition of contaminants that could affect the signal
65. plied When the v button is pressed it displays the next line or the next screen 3 2 2 a The Status Screen The Status screen Figure 3 2 shows the present conditions of the detector This screen cannot be edited but the desired offset can be set via the Offset screen Figure 3 3 the temperature and gain can be set via the Temp Gain screen Figure 3 4 and the pressure units can be selected via the Noise Filter Pressure Unit screen Figure 3 5 Temperature value blinks if desired temperature is not reached or not stable Pressure value blinks if gas pressure is lower than 3 0bar When the v button is pressed the Signal Offset screen Figure 3 3 which is used to select the desired offset is displayed 3 2 2 b The Offset Screen d SEDERE SEDEX 85 Te IT ELSD Signal Offset mV 000 Figure 3 3 The Offset Screen To increase the offset value click on the Q button If you press the button quickly the offset will increase by 1 if you press and hold the button the value will increase at the rate of 20mV sec Once you have set the desired offset press the Q button to validate the new value When the instrument is Autozeroed the Autozero operation updates the Offset value to set the Signal to OmV Press the v button to access the Temp Gain screen Figure 3 4 SEDEX Model 85LT 3 3 Evaporative Light Scattering Detector Chapter 3 3 2 2 c The Temperature Gain Screen re SEDERE SEDEX 85 Q IT ELSD
66. ption Replacement filter cartridges are available as part number 85159 CAR 2 4 SEDEX Model 85LT Evaporative Light Scattering Detector Installation of the Detector 2 4 3 Exhaust Venting and Drain Requirements The carrier gas containing volatilized mobile phase and sample components exit the detector through the black exhaust tube located on the top detector rear panel The black exhaust tube from the detector can be cut and should be directed into a fume hood or exhaust vent If a vacuum is used it should be moderate so as to avoid turbulence in the glass cell siphon The exhaust should not be allowed to enter the laboratory atmosphere and any appropriate accessory e g solvent filter should be disposed of in a manner that meets the local regulatory authorities for health and safety requirements The vacuum must be moderate to avoid turbulence in the glass cell siphon or liquid spilled into the evaporation tube Avoid loops or bends in the black exhaust tubing which could create condensation traps resulting in bad measurement results If gas from the fume hood enters the detector i e a negative pressure exists between the detector and the fume hood it is possible that foreign material from the fume hood could contaminate the detector Install the drain tubing it can be cut in a way to the siphon outlet aligns straight to the waste container without loops or bends so that the waste liquid flows smoothly through the
67. s Regulator with Filter and Manometer to support the operation of the detector A complete listing of all spare parts and accessories is included in Appendix 2 2 2 SEDEX Model 85LT Evaporative Light Scattering Detector Installation of the Detector 2 2 Lifting and Carrying the Detector Once the instrument is unpacked ensure that no cables or tubing are connected when you carry the instrument The detector should be lifted by the bottom e g place your hands under the instrument Two persons are needed to ensure an easy transport and avoid physical injury Figure 2 1 Do NOT Lift Here Do NOT Lift Here Carry Here Carry Here Figure 2 1 Carrying the Detector 2 3 Unpacking the Detector Unpack components carefully making sure all items in the Table 2 2 have been included The glassware chamber and nebulizer are packed in separate boxes in the shipping box If there is any damage to a carton or its contents or if any component appears to be missing report to your local distributor immediately If there is any evidence that the main unit has been damaged do not plug the unit into the power line Contact your local distributor immediately It is strongly recommended that the shipping box be retained for future use to transport the detector or return to repair SEDEX Model 85LT 2 3 Evaporative Light Scattering Detector Chapter 2 2 4 Laboratory Requirements 2 4 1 Power Requirements The detector is configure
68. sher Scientific Inc Clarity is a trademark of DataApex Ltd Microsoft is a trademark of Microsoft Corporation All other trade names used in this document may be trademarks or names of the corresponding companies or their product All information in this manual is subject to change without notice and does not represent a commitment on the part of S E D E R E S A S Please consult S E D E R E S A S if you suspect any error or omission S E D E R E S A S believes that the information in its user manuals is accurate at the date of publication Copyright 2012 S E D E R E S A S All rights reserved No part of this manual may be reproduced adapted or transmitted in any materiel or electronic form without the prior written consent of S E D E R E S A S Printed in France Warnings and Safety Precautions The following precautions should be followed to minimize the possibility of personal injury and or damage to property while using the instrument 1 2 9 Read the Operator s Manual this manual thoroughly before you use the detector and keep this manual for future reference Maintain a well ventilated laboratory If the mobile phase contains a volatile organic solvent ensure that the laboratory is well ventilated so that a build up of vaporized solvent cannot occur Avoid open flames and sparks Do not use an open flame and do not use any equipment that can cause sparks in the same room as the instrument The detect
69. sure Unit Screen 3 5 Principle of Operation 1 3 R Recorder Connecting 2 9 Routine Operation 4 3 RS 232 Port 2 11 S Safety Precautions iii Sample Pretreatment 4 6 Screens External Power Down 3 8 Factory Method Code 3 10 Firmware version 3 9 Gas Valve 3 7 Light Source 3 6 Light S Normalization 3 7 Noise Filter Temperature 3 5 Offset 3 3 Serial Number 3 9 Status 3 3 Temperature Gain 3 4 Total Lifetime Elapsed 3 9 Shutting Down 4 7 Siphon Overflow Installing 2 12 Solvent Noise Test 3 13 Start Up Kit 2 2 Procedure 4 1 Stray Light Test 3 12 T Temperature Gain Screen 3 4 Temperature Optimizing 4 4 Tests Background Stray Light Noise 3 12 Column Noise 3 14 Electronic Noise 3 12 Solvent Noise 3 13 Total Lifetime Elapsed Screen 3 9 Troubleshooting 5 3 U Unpacking 2 3 User Interface 3 1 Vv Venting Exhaust Lines 2 9 Requirements 2 5 W Warnings iii Warning Symbols iv Z Zeroing the Detector 2 10 3 1 4 2
70. sured by a photomultiplier which is related to the concentration of the compound of interest in the sample A cross sectional view of the instrument is presented in Figure 1 3 Step 3 DETECTION Step 2 EVAPORATION Figure 1 3 Cross sectional View of the Detector SEDEX Model 85LT 1 3 Evaporative Light Scattering Detector Chapter 1 1 2 1 Nebulization This first step transforms the eluent from the chromatography system into fine droplets The smaller the droplet size the lower the temperature needed to evaporate the liquid phase With SEDEX detectors the nebulizer Figure 1 4 and the nebulization glassware chamber Figure 1 5 are designed to eliminate the biggest droplets of the solvent The transformation of the eluent into fine droplets is made by a nebulizer which uses typically nitrogen Six different nebulizers are available to optimize the performance of the detector at different HPLC flow rates and for U HPLC and SFC applications Table 1 1 The user should select the nebulizer to best match the flow rate that will be used with the application when the detector is ordered the optimal flow rate range for each nebulizer is indicated in Table 1 1 Nebulizer can be easily identified with the part number labelled on it Replacement or additional nebulizers are available from your local distributor and can be easily installed as described in Section 5 3 5 Back Pressure Part with water Number Micro Flow Nebu
71. tant within the following range e NO no filtering row data e 0 5s 0 5 second moving average filtering e 1s 10s 1 to 10 seconds moving average filtering SEDEX Model 85LT 3 5 Evaporative Light Scattering Detector Chapter 3 ee Note For better results the digital Filter should be used unless the peak s of interest are very poorly resolved e g when Rs lt 1 5 Default value is 1s corresponding to a peak width of approximately 2 seconds at half height User manual section 4 5 5 details Filter optimization If you have changed the value press fox to validate it before you press the v button to access the Press Unit line The pressure unit line allows for the selection of KPa bar or psi for pressure units the desired selection is made via the Q r key and validated by the Q key When you press the button the Light Source screen Figure 3 7 will be displayed 3 2 2 f The Light Source Screen z SEDERE SEDEX 85 Te IT ELSD LED ON H HH HHHH H Reset Time Elapsed Figure 3 7 Light Source Screen The Light Source screen is used to power the light source On Off and is equivalent to the Light Source button on the control panel Use the Q button followed by the fox button to power the light source on and the button followed by ox the button to turn it off The hours field indicates the number of hours that the Light Source has been in use and cannot be edited by the user When the life
72. tant than resolution a Filter 2s or higher can be set to improve sensitivity even better 4 6 Shutdown Sequence If desired some or all functions of the instrument can be powered down at the end of an automated series of analyses These power down features are described in detail in Section 3 2 2 1 To shut down the instrument a Turn off the mobile phase flow b Allow detector gas only to flow for few minutes 30minutes is recommended to drain the evaporation tube and detection chamber c Turn off the gas supply if desired at the source or close the detector gas valve see section 3 2 2 h d Power off the detector if desired If you are using a mobile phase which contains salts acids or bases pump few mL of water or ethanol through the system before switching off the detector to prevent any deposition of substances and possible corrosion of the instrument If the detector will not be used for some time it is recommended to remove it from the liquid chromatography flow path in order to avoid any clogging of the nebulizer or deposition of substances inside the detector Closing gas valve while the pump is still running may result in serious nebulizer damage SEDEX Model 85LT 4 7 Evaporative Light Scattering Detector Chapter 4 This page intentionally left blank 4 8 SEDEX Model 85LT Evaporative Light Scattering Detector 5 Maintenance and Troubleshooting It is recommended that the detector is calibr
73. tector 4 3 1 Manual Auto zeroing of the Detector To auto zero the detector a Set the Gain to the desired value The gain is set on the Temp Gain screen which is accessed by pressing the v button two times when the Signal screen is displayed b Press the button The detector will be automatically auto zeroed at this point c Ifthe signal is to be offset set the offset at this time The Offset screen is accessed by pressing the v button when the Status screen is displayed Re Note The offset must be selected after the detector is auto zeroed as the Auto zero operation sets the signal to 0 by modifying the offset es Note If you change the gain selection you may need to make an auto zeroed again before taking any measurement 4 3 2 External Auto zeroing of the Detector If desired the auto zero command can be initiated by an external device such as the HPLC system controller To employ this feature a cable from the external device is plugged into the Ext Autozero socket on the rear panel Section 2 5 3 To auto zero the detector a contact closure signal or a TTL signal is used to short circuit the contacts The signal should be at least 0 5sec long with a maximum current of 20mA at 5V If a TTL signal is used please make sure to use the correct polarity identified on the cable 4 2 SEDEX Model 85LT Evaporative Light Scattering Detector Operating the Detector 4 4 Routine Operation of the Detect
74. the SEDEX ELSD 85LT Low Temperature Evaporative Light Scattering Detector In this discussion we assume that you have demonstrated that the instrument is operating in an acceptable manner see Chapter 3 and that the general chromatographic conditions for the separation have been determined 4 2 Startup sequence To prepare the detector for operation a Power up the detector by pressing the switch on the rear panel b Open the gas distribution valve and set the pressure to 3 5bar 51psi The pressure is indicated on the Status screen c Ensure that the overflow siphon for the nebulization chamber contains sufficient liquid If necessary pump few mL of solvent through the instrument to fill the siphon d Select the desired temperature The temperature is set on the Temp Gain screen which is accessed by pressing the v button two times when the Status screen is displayed e Start the mobile phase flow through the instrument and allow the overall system to operate for at least 30 minutes to ensure that all components are equilibrated and a stable baseline is obtained es Note The Solvent Noise test Section 3 3 4 and the Column Noise test Section 3 3 5 should be performed to verify that the detector is functioning in a proper manner Ine Note The liquid level in the siphon must be stable and should be equal at both sides SEDEX Model 85LT 4 1 Evaporative Light Scattering Detector Chapter 4 4 3 Auto zeroing the De
75. time light source period has been reached approximately 5000h a message indicating that the maximum usage of the light source has been exceeded will be presented when the unit is powered up and the orange LED on the keyboard blinks when the light source is power on 1 Note The Reset Time Elapsed field should be validated with only when you change the lamp When you press the v button the Light Source Normalization screen Figure 3 8 will be presented 3 6 SEDEX Model 85LT Evaporative Light Scattering Detector Start up Procedure 3 2 2 2 The Light Source Normalization Screen SEDERE SEDEX 85 O LT ELSD Stray Light percent Value 100 Figure 3 8 The Light Source Normalization Screen The Light Source Normalization has been designed for QC and R amp D laboratories involved in regulated environment Indeed periodical controls are sometimes required using a specific standard with a same concentration and same chromatography conditions to ensure that the obtained response values are always identical However with time the light source intensity tends to decrease slightly Therefore a correction becomes necessary So the Light Source Normalisation screen is used to normalize the Stray Light value according to reference value used for detector Operation Qualification When you press the Q button the Gas Valve screen Figure 3 9 will be displayed 3 2 2 h The Gas Valve Screen SEDERE SEDEX
76. ture to 50 C Wait for a stable temperature b Switch on the light source c Change the Gain to 1 d Set the offset after Autozero to OmV Refer to Section 3 2 2 d and set Stray Light Value adjustment to 100 refer to Section 3 2 2 g e Autozero the detector f Change the Gain to 12 g Wait 15 minutes for stabilization and record the signal level The expected level is typically 100mV to 150mV The exact value will vary slightly and small deviations should not be a cause for concern 3 12 SEDEX Model 85LT Evaporative Light Scattering Detector Start up Procedure 3 3 4 Solvent Noise Test To determine the solvent noise a Make sure the siphon is filled and ensure that the gas is flowing at 3 5bar 51 psi the temperature is set to 50 C and stable and the pump is switched off b Switch on the light source wait 15 minutes and set the gain to 12 and monitor the signal Do not autozero the detector The signal may be negative c Bypass the column and connect the detector to the mobile phase delivery system and pump the solvent that you expect to use for your analyses through it at a flow rate of ImL min 30uL min for micro flow nebulizer d Monitor the baseline for a few minutes If water is used as the solvent the signal increase should be less than 10mV Higher values could be observed if non HPLC grade water with a higher non volatile residue is used If an organic solvent is used the signal incr
77. usage of the lamp has been exceeded will be presented when the unit is powered up and the orange L E D on keyboard blinks when light source is power on 5 2 SEDEX Model 85LT Evaporative Light Scattering Detector Maintenance and Troubleshooting 5 3 Troubleshooting 5 3 1 General Troubleshooting Information The SEDEX ELSD 85LT Low Temperature Evaporative Light Scattering Detector is designed to be incorporated into a Liquid Chromatography system It is important to note that the detector response reflects the overall performance of the system and a problem that is seen on the detector output may not necessarily be a detector problem In almost all cases there is one and only one cause for a problem As an example of this point if the user observes a noisy baseline it is possible that the problem is due to e The pump e g a defective check valve The mobile phase e g improper degassing or high residue after evaporation e The column e g elution of strongly retained components The nebulizer e g lack of maintenance The detector e g an electronic problem It is very unlikely that two problems occur at the same time The role of the troubleshooting activities is to determine the cause of the problem In the following we assume that the operator has already determined that other components of the system are operating in an appropriate way Do not disassemble the nebulizer Disassembling the nebuliz
78. ware ZChrom Elite ChemStation Xcalibur and f ELSD parameters EXTERNAL REQUIRE Power 60Hz 1 8A and patented auxiliary gas flow than 4L min for CC Nebulizer Gas Pressure Nitrogen Dimensions 10 W X 480mm 18 H Weight 16Kg 35lbs SEDEX Model 85LT A1 1 Evaporative Light Scattering Detector Appendix 1 Specifications This page intentionally left blank A1 2 SEDEX Model 85LT Evaporative Light Scattering Detector Appendix 2 Spare Parts List Nebuliser 1 Black Plastic Nut diam 22 6 Peek Nut 1 16 with Ferrule 8 Glassware 2 Union bulkhead fitting 7 Kit of 3 Seals 9 Drain Assembly 4 Black Pneumatic tube 3 tube fitting seal with stainless steel fittings one way valve Figure A2 1 Identification of Spare Parts SEDEX Model 85LT A2 1 Evaporative Light Scattering Detector Appendix 2 Part Item Nebulizer 1 Micro Flow Nebulizer requires a specific glassware CRAMBEP eceecescsscscseeveseseees Low Flow Neb liZenisoninnnrinan anen a A R A N RRR HPLC Nebulizer sen nth td te ete EER Combinatorial Chemistry NebuliZer ecescccsessesesecsssesesseesessessesestestesteseenes VU HPLC Nebulizer narnninninnianakneni n a e a ian SEC Neb lizen inen ke lt tae le Bel din S Glassware Nebulization Chamber 2 Standard Glass Nebulization Chamber ccccccccsessecsscscsscssessesessssssssssssseseeees Micro HPLC Glass Nebulization Chamber u ccccc
79. zer inlet liquid tubing should be pointing up Take care to ensure that the rear part of the nebulizer is not in contact with the solvent Clean the nebulizer for approximately 30 minutes with the solvent and then replace the solvent with water and clean for an additional 30 minutes If the nebulizer cannot be repaired by cleaning by pumping solvent through it or with an ultrasonic bath it requires a replacement To re install the nebulizer or replace it by a new one or another nebulizer model a After a nebulizer cleaning and for re installing the nebulizer re install the gas inlet quick fitting and the black plastic nut with its seal b Reverse the order of previous steps nebulizer removing In case the black gas tubing has been removed make sure you are using the correct direction where the white one way valve is at the lower end near the gas arrival c Make sure there is no liquid or gas leak at all connections and check for possible leakage that could affect the detector performance or create laboratory pollution when you turn on the pump d Install the black front panel cover first fix its right side and then push its left side e Test the nebulizer to ensure that it is working properly SEDEX Model 85LT 5 7 Evaporative Light Scattering Detector Chapter 5 eS Note If the Nebulizer cleaning procedure does not solve the problem contact your local distributor for a nebulizer replacement In case

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