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G4218-90002 - Agilent Technologies

1. Se e e s e e ee Notices Agilent Technologies Inc 2008 2009 2010 2011 2012 Microsoft is a U S registered trademark of Microsoft Corporation Parafilm is a trademark of American National Can Co Tygon is a trademark of the Norton Corpo ration No part of this manual may be reproduced in any form or by any means including elec tronic storage and retrieval or translation into a foreign language without prior agree ment and written consent from Agilent Technologies Inc as governed by United States and international copyright laws Manual Part Number 64218 90002 Rev B Edition 05 2012 Printed in Germany Agilent Technologies Inc Hewlett Packard Strasse 8 76337 Waldbronn This product may be used as a com ponent of an in vitro diagnostic sys tem if the system is registered with the appropriate authorities and com plies with the relevant regula tions Otherwise it is intended only for gen eral laboratory use Warranty The material contained in this docu ment is provided as is and is sub ject to being changed without notice in future editions Further to the max imum extent permitted by applicable law Agilent disclaims all warranties either express or implied with regard to this manual and any information contained herein including but not limited to the implied warranties of merchantability and fitness for a par ticular purpose Agilent shall not be li
2. V All tubing should be in good shape Any damaged tubing or tubing with kinks should be replaced V All cables should be in good shape Any electrical cables that are frayed or damaged should be replaced V Only use filtered oil free pressurized gas Agilent 1260 Infinity ELSD User Manual Maintenance 7 Cleaning and Decontamination Cleaning the Detector WARNING Shock hazard and damage of detector Liquid drip into the detector could cause shock hazard and damage the detector gt Do not let liquid drip into the detector Turn the instrument off 2 Unplug all connection cables the gas input and nebulizer tubing before cleaning 3 Ifthe unit has been powered up wait for it to cool down before proceeding 4 Clean the instrument with a clean non abrasive rag If necessary a solvent such as isopropanol can be used Decontaminating the detector WARNING Health risk by potentially harmful substances The toxicological properties of many samples separated by the HPLC system are usually not well known They could cause toxications and other health risks gt f you have any doubt about the cleanliness of a detector treat the detector as if it contained a potentially harmful substance and decontaminate it before working on it Agilent 1260 Infinity ELSD User Manual 83 7 84 Maintenance To decontaminate the detector 1 Power up the detector and allow nebulizer gas to flow through it in th
3. 3 Use the large black nut to position the nebulizer on the nebulization chamber 4 Screw the inlet fitting into 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 PTFE tubing with an outer green sleeve For proper operation the PTFE tubing must extend less than 2 mm past the end of the green sleeve Figure 13 Figure 13 Nebulizer Inlet Fitting 44 Agilent 1260 Infinity ELSD User Manual Installing the Detector 3 WARNING Health risk caused by liquid leaks Liquid leaks could cause personal injury or laboratory pollution or negatively affect the detector performances by pressed out liquid gt Make shure that all connections are tight and that there is no liquid leak 5 Fill the siphon overflow on the nebulizer glass tube assembly with the mobile phase that will be used for the separation If you are using a highly volatile solvent like hexane or CH2C12 use water to fill the overflow The liquid should fill the bent part of the siphon but should not pool in the bottom of the condenser tube Agilent 1260 Infinity ELSD User Manual 45 3 Installing the Detector Connecting the Siphon Overflow 1 Attach the Tygon drain tube assembly to the end of the siphon tube using the tapered hose connector and lead the tube to a waste container 2 Locate the tube in such a way that condensed solvent can flow freely from th
4. 50 To prepare the system for operation 1 2 Power up the detector by pressing the switch on the rear panel Open the gas distribution valve and set the pressure to 3 5 bar 51 psi The pressure is indicated on the Status screen Ensure that the overflow siphon for the nebulization chamber contains sufficient liquid If necessary pump a few ml of solvent through the system to fill the nebulization chamber Select the desired temperature using your control software or the instrument control see Appendix 9 The temperature is set on the Temp Gain screen which is accessed by pressing the button two times when the Status screen is presented Start the mobile phase flow through the system and allow the overall system to operate for at least 15 minutes to ensure that all components are equilibrated and a stable baseline is obtained Please refer to Chapter 5 for choosing temperature settings solvents and columns Agilent 1260 Infinity ELSD User Manual Using the System 4 Auto zeroing the Detector Manual Auto zeroing of the Detector To auto zero the detector 1 Set the Gain to the desired value The gain is set on the Temp Gain screen which is accessed by pressing the 7 button two times when the Signal screen is presented 2 Press the button The detector will be automatically auto zeroed at this point 3 Ifthe signal is to be offset set the offset at this time The Offset screen is accessed by pressing the
5. 7 button when the Status screen is presented The offset must be selected after the detector is auto zeroed as the auto zero operation sets the signal to 0 If you change the gain selection make sure that the system is auto zeroed again before taking any measurements 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 AUTO ZERO socket on the rear panel See Additional Options on page 40 Agilent 1260 Infinity ELSD User Manual 51 4 Using the System Routine Operation of the System 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 system the following points should be considered Health risk by exhaust gas Exhaust gases could cause personal injury or laboratory pollution when it is lead into the laboratory gt Make sure that the exhaust gas from the detector is led into a fume hood or other device V Make sure 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 system i e no constrictions or condensation traps see Chapter 3 Ifa vacuum is used ensure that the vacuum effect will not disturb the detector Health risk by potentia
6. OFF switch The display will present the firmware 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 0 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 Refer to Chapter 4 for preparing the unit for routine operation Agilent 1260 Infinity ELSD User Manual 47 3 Installing the Detector Installing the Nebulizer and Nebulization Chamber Assembly 48 Agilent 1260 Infinity ELSD User Manual Agilent 1260 Infinity Evaporative Light Scattering Detector A User Manual c00 ee 4 Using the System e a e g Preparing the System for Operation 50 Auto zeroing the Detector 51 Routine Operation of the System 52 Installation Test Procedure 54 Powering Down and Shutting Down the System 56 This chapter describes the operations that should be performed on a routine basis when you want to collect chromatographic data using the Agilent 1260 Infinity Evaporative Light Scattering Detector In this discussion we assume that you have demonstrated that the system is operating in an acceptable manner see Chapter 4 and that the general chromatographic conditions for the separation have been determined ig Agilent Technologies 49 4 Using the System Preparing the System for Operation
7. and Diagnostics Specific Detector Troubleshooting V The mist from the nebulizer should be homogeneous If it is not homogeneous the nebulizer the needle or the PTFE tube may be 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 for cleaning are presented in Cleaning the Nebulizer on page 86 Do not disassemble the nebulizer this will void the warranty V If the sensitivity of the system is low ensure that there are no leaks in the system In some cases a small increase in the gas pressure e g 0 1 or 0 2 bar may solve the problem Alternatively a new LED may be required or the nebulizer might be obstructed V 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 derived from the elution of strongly retained materials from the column or may be derived from the solvent For determining the cause of the problem bypass the column and observe the signal intensity 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 if the signal
8. and firmly inserted into the 2 Cut the tube square fitting as shown after removing Parafilm from detector gas inlet 3 Insert the tube into the fitting until it bottoms 4 Pull the tube to check engagement of the grab 34 Agilent 1260 Infinity ELSD User Manual Installing the Detector 3 WARNING Tube damage or inappropriate installation Health risk due to gas leakage gt Make sure that no tube damage or inappropriate installation could allow a gas leak in laboratory Two pieces of tubing are provided If you are using the system with an external filter connect the gas source to the filter and then connect the filter to the back of the unit Make sure that no tube damage or inappropriate installation could allow a gas leak in laboratory Agilent 1260 Infinity ELSD User Manual 35 3 Installing the Detector Removing the Gas Inlet Tube To remove the gas inlet tube if necessary 1 dissengage the grab ring teeth by a simple manual pressure on the push sleeve and withdraw the tube from the fitting 36 Agilent 1260 Infinity ELSD User Manual Installing the Detector 3 Vent the Exhaust Line to a Fume Hood 1 Vent the black exhaust tube on the back of the unit to a fume hood exhaust line or similar installation 2 Make sure that the fume hood withdraws gas from the detector i e there should be a positive pressure between the detector and the hood Danger of poison
9. angle of 120 with respect to the light beam Figure 6 When the carrier gas contains microparticles the light is scattered and is detected by the photomultiplier 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 I km 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 depend on a variety of experimental conditions e g the temperature and the nature of the mobile phase A gas inlet before the detector chamber provides a concentric shield for the carrier gas This serves to eliminate diffusion of the carrier gas and eliminates contamination of the detector cell Agilent 1260 Infinity ELSD User Manual Agilent 1260 Infinity Evaporative Light Scattering Detector User Manual 2 Site Requirements and Specifications Site Requirements 20 Specifications 24 ee Agilent Technologies 2 Site Requirements and Specifications Site Requirements A suitable environment is important to ensure optimum performance of the Agilent 1260 Infinity detector Power Considerations The detector is configured for either 100 V AC 50 60 Hz 115 V AC 60 Hz or 230 V AC 50 Hz input power depending on the country to which it is shipped Ensure that the voltage value indicated
10. batteries shall be disposed off locally according to national waste disposal regulations for batteries Lithiumbatteri Eksplosionsfare ved fejlagtig handtering Udskiftning ma kun ske med batteri af samme fabrikat og type Lever det brugte batteri tilbage til leverand ren Lithiumbatteri Eksplosionsfare Ved udskiftning benyttes kun batteri som anbefalt av apparatfabrikanten Brukt batteri returneres appararleverandoren 114 Bij dit apparaat zijn batterijen geleverd Wanneer deze leeg zijn moet u ze niet weggooien maar inleveren als KCA Agilent 1260 Infinity ELSD User Manual Appendix 10 Agilent Technologies on Internet For the latest information on products and services visit our worldwide web site on the Internet at http www agilent com Select Products gt Chemical Analysis It will provide also the latest firmware of the Agilent 1260 Infinity modules for download Agilent 1260 Infinity ELSD User Manual 115 10 Appendix Agilent Technologies on Internet 116 Agilent 1260 Infinity ELSD User Manual Index A accessory kit 30 32 Auto zeroing of the Detector 51 auto zeroing of the detector 51 background noise test 69 battery safety information 114 C carrying the system 28 cleaning detector 83 nebulizer 86 column connecting 47 treatment 63 column noise 77 configurations 27 control panel 96 D Date Time screen 105 decontamination 83 detection 17 digital di
11. dry to avoid particles deposit 4 Stop the gas flow 5 Shut down the detector The time required for each step depends on the application used solvents type and concentration of samples and should be determined practically Agilent 1260 Infinity ELSD User Manual 57 4 Using the System Powering Down and Shutting Down the System 58 Agilent 1260 Infinity ELSD User Manual Agilent 1260 Infinity Evaporative Light Scattering Detector User Manual 5 Optimizing Performance Selecting the Optimum Temperature 60 Optimizing the Mobile Phase 62 Optimizing the Filter 64 ee Agilent Technologies 59 5 Optimizing Performance 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 evaporation 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 1 ml min At higher flow rates higher temperatures may be required for minimizing the noise If a mobile phase such as DMSO or DMF has a relatively low volatility temperature should be increased to allow correct evaporation process The temperature can be adjuste
12. on the power connector on the rear panel corresponds to the line voltage in your facility The detector requires G4218 64000 230 V 1 7 A 400 W 50 Hz G4218 6410 115 V 1 8 A 210 W 60 Hz G4218 64020 100 V 2 1 A 210 W 50 60 Hz 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 system If a two prong outlet is used make sure 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 WARNING Incorrect line voltage at the instrument Shock hazard or damage of your instrumentation can result if the devices are connected to a line voltage higher than specified gt Connect your instrument to the specified line voltage 20 Agilent 1260 Infinity ELSD User Manual Site Requirements and Specifications 2 Gas Considerations A supply of clean filtered oil free inert clean gas typically nitrogen is required to operate the detector The gas supply needs to be free of particles as particles will create background noise in the chromatograms In case of such noise for example for newly installed gas lines flush the ga
13. polar solvents like hydrocarbons For details please refer to compatibility data of manufacturer Saint Gobain 112 Agilent 1260 Infinity ELSD User Manual Appendix 10 The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC Abstract The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC adopted by EU Commission on 13 February 2003 is introducing producer responsibility on all Electric and Electronic appliances from 13 August 2005 This product complies with the WEEE Directive 2002 96 EC marking requirements The affixed label indicates that you must not discard this electrical electronic product in domestic household waste Product Category With reference to the equipment types in the WEEE Directive Annex I this product is classed as a Monitoring and Control instrumentation product Do not dispose off in domestic household waste To return unwanted products contact your local Agilent office or see www agilent com for more information Agilent 1260 Infinity ELSD User Manual 113 10 Appendix Lithium Batteries Information Danger of explosion if battery is incorrectly replaced Replace only with the same or equivalent type recommended by the equipment manufacturer Lithium batteries may not be disposed off into the domestic waste Transportation of discharged Lithium batteries through carriers regulated by IATA ICAO ADR RID IMDG is not allowed Discharged Lithium
14. specified type normal blow time delay and so on are used for replacement The use of repaired fuses and the short circuiting of fuse holders must be avoided Do not install substitute parts or make any unauthorized modification to the instrument Do not disassemble the nebulizer or touch any components inside the nebulization chamber This can lead to the deposition of contaminants which could affect the signal Safety Symbols Table 11 shows safety symbols on the instrument Table 11 Safety Symbols Symbol Description Hot surface Risk of burn Electric shock risk The apparatus is marked with this symbol when the user should refer to the instruction manual in order to prevent risk of harm to the operator and to protect the apparatus against damage Agilent 1260 Infinity ELSD User Manual Appendix 10 WARNING A warning alerts you to situations that could cause physical injury or damage to the equipment gt Do not proceed beyond a warning until you have fully understood and met the indicated conditions CAUTION A caution alerts you to situations that could cause a possible loss of data gt Do not proceed beyond a caution until you have fully understood and met the indicated conditions Agilent 1260 Infinity ELSD User Manual 111 10 Appendix Solvent Information Observe the following recommendations on the use of solvents Solvents Brown glass ware can avoid growth of algae Always
15. the rear panel of the detector Figure 10 and to the appropriate socket on the controlling device 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 of the following conditions is observed The lamp is off The temperature is not at the indicated setpoint The temperature is at the indicated setpoint but is not stable The pressure is below 3 0 bar The controlled device electrical consumption mustn t exceed 20 mA under 12 V DC The blue cables are contact closure input cables that are used to power the unit down see The Gas Valve Screen on page 103 via a signal from an external device to the detector A contact closure signal must be used from the controlling device to short circuit the contacts If a TTL signal is used please make sure to use the correct polarity identified on the cable e RS 232 Port If a personal computer is used with the detector the detector should be connected to the computer via the RS 232 port using the supplied cable Agilent 1260 Infinity ELSD User Manual 41 42 3 Installing the Detector 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 Do not turn on the power at this time The power cord of this system contains three wires wh
16. 0 cm 0890 1727 Tubing 5m 1 5061 3378 Cable remote APG 1 optional G4218 60100 Gas regulator with filter 0 01 pm and manometer 1 optional see Table 9 Nebulizer Agilent Technologies provides a wide range of accessories e g Gas Regulator with Filter and Manometer part number G4218 60100 to support the operation of the detector A complete listing of all spare parts and accessories is included as Chapter 8 Damaged Packaging Upon receipt of your detector inspect the shipping containers for any signs of damage If the containers or cushioning material are damaged save them until the contents have been checked for completeness and the detector has been checked mechanically and electrically If the shipping container or cushioning material is damaged notify the carrier and save the shipping material for the carriers inspection Delivery Checklist Ensure all parts and materials have been delivered with the detector The instrument box contains the instrument and an Accessory kit A separate box contains the reference manual and the power cable Table 2 on page 30 lists the content of each accessory kit Please report missing or damaged parts to your local Agilent Technologies sales and service office Agilent 1260 Infinity ELSD User Manual 31 3 Installing the Detector Installing the Detector CAUTION Defective on arrival problems If there are signs of damage to the autosampler please do not attempt to instal
17. 01 Signal Cable G4218 81100 RS232 Cable Female Female Straight 3 m G4218 81103 External Event Cable G4218 81102 APG Remote Cable 5061 3378 Caffeine Standard 250 pg ml in water G4218 85000 Agilent 1260 Infinity ELSD User Manual 93 8 Parts for Repair Table9 Nebulizers for the G4218A Agilent Evaporative Light Scattering Detector Nebulizer Useable Flow Optimum _ Nebulizer Back Identifying Part Number Range Flow Range Pressure bar Marks with water Micro Flow 5 0 pl min 15 0 pl min 24 100 pl min Blue Seal G4218 20003 Nebulizer 40 0 pl min 25 0 pl min 2 Rings Semi Micro Flow 0 04ml min 0 1ml min 44 1 ml min Yellow Seal G4218 20001 Nebulizer 1 0 ml min 0 3 ml min 2 Rings Standard Flow 0 2ml min 0 5ml min 4 1 ml min Black Seal G4218 20000 Nebulizer 2 0 ml min 1 2 ml min 2 Rings Large Flow 1 0ml min 2 0 ml min 4 1 ml min Red Seal G4218 20002 Nebulizer 5 0 ml min 3 0 ml min 1 Ring Rapid Resolution 0 2ml min 0 4 ml min 14 White Seal G4218 20004 Nebulizer 1 4 ml min 1 0 ml min 1 Ring The optimum flow range provides highest sensitivity and repeatability t 14 bar at 1 4 ml min with water acetonitrile 50 50 94 Agilent 1260 Infinity ELSD User Manual Agilent 1260 Infinity Evaporative Light Scattering Detector o User Manual 209 The Control Panel e 7 j The Control Panel 96 This chapter describes the role of the controls and the digital display on the control panel ee Ag
18. 2 Noise Filter Pressure Unit 101 Offset 99 Serial Number 105 Status 98 Temperature Gain 99 Total Lifetime Elapsed 105 Serial Number screen 105 shutting down 56 siphon overflow installing 46 solvent noise 75 solvent noise test 75 start up kit 27 procedure 50 Status screen 98 stray light test 74 zeroing the detector 51 T temperature optimizing 60 Temperature Gain Screen 99 Temperature Gain screen 99 tests background stray light noise 74 Total Lifetime Elapsed screen 105 troubleshooting 65 U unpacking 30 user interface 98 V venting exhaust lines 37 requirements 22 www agilent com In This Book This manual is designed to describe the installation operation maintenance and basic troubleshooting of the G4218A Agilent Evaporative Light Scattering Detector It includes e Introduction Installation of the Detector e Start Up Procedure e Operating the System e Maintenance and Troubleshooting e Specifications e Spare Parts List e Electrical connections e Safety Information Agilent Technologies 2008 2009 2010 2011 2012 Printed in Germany 05 2012 2 Rev Agilent Technologies
19. 90 Agilent 1260 Infinity ELSD User Manual Agilent 1260 Infinity Evaporative Light Scattering Detector User Manual 8 Parts for Repair Evaporative Light Scattering Detector Parts 92 ee Agilent Technologies 91 8 Parts for Repair Evaporative Light Scattering Detector Parts Evaporative Light Scattering Detector Parts lt s N Figure 18 Evaporative Light Scattering Detector Parts 92 Agilent 1260 Infinity ELSD User Manual Parts for Repair 8 Table 8 Evaporative Light Scattering Detector Parts Part Part Number 1 Nebulizer Assembly 1 see list below 2 Nebulization Chamber glass 2 G4218 40000 3 Black Plastic Nut for Nebulization Chamber 13 mm Diameter 3 G4218 40010 9 Black Plastic Nut for Nebulization Chamber 30 mm Diameter 9 G4218 40011 Seal kit for nebulization chamber G4218 68010 Gas Regulator with Filter and Manometer G4218 60100 Cartridge for Gas Regulator G4218 40150 4 5 Drain Assembly includes fitting 5 4 G4218 40100 Drain Bottle Assy G1946 60111 inch PE Tubing for Drain Bottle 1 unit 1 cm 0890 1727 Tube Fitting 6 mm diameter for Gas Regulator G4218 40160 6 Pneumatic tube diameter 4 mm for Nebulizer G4218 40220 includes stainless steel fitting 6 Detector Gas Inlet Tube 6 mm 0 D 10 m G4218 40170 7 Wall Mounting Fitting 4 mm diameter 7 G4218 40140 8 Bulkhead Fitting 8 G4218 40130 11 Gas check valve G4218 40200 Front panel shield window G4218 40400 Autozero Cable G4218 811
20. Cleaning the Detector 83 Decontaminating the detector 83 Removing the Nebulization Chamber and Nebulizer 84 Cleaning the Nebulizer 86 Adjusting Stray Light Settings 89 8 PartsforRepair 91 Evaporative Light Scattering Detector Parts 92 9 The Control Panel 95 The Control Panel 96 The Digital Display 97 The User Interface 98 Agilent 1260 Infinity ELSD User Manual 7 Contents 10 Appendix 107 Safety Information 108 Solvent Information 112 The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC 113 Lithium Batteries Information 114 Agilent Technologies on Internet 115 Index 117 8 Agilent 1260 Infinity ELSD User Manual Agilent 1260 Infinity Evaporative Light Scattering Detector User Manual 1 Introduction The Low Temperature Evaporative Light Scattering Detector Principle of Operation 12 s j Agilent Technologies 10 1 Introduction The Low Temperature Evaporative Light Scattering Detector The Agilent 1260 Infinity Evaporative Light Scattering Detector Figure 1 is designed to detect compounds in the eluent from high performance liquid chromatography HPLC micro HPLC gel permeation chromatography GPC or counter current chromatography CCC It is capable of monitoring eluent flow rates from 5 ul min to 5 ml min Evaporative light scattering detection is a universal technique which can detect any non volatile analyte Detection does not depend on the absorbtion of radiation and is not a
21. Press Unit bar Figure 24 The Noise Filter Pressure Unit Screen The Filter Pressure Unit screen displays the settings for digital filtering of the signal data and the selected measurement unit bar kPa or psi for the pressure display When the screen is displayed the cursor is on the Filter field By pressing or keys you change the filtering strength within the following range e NO no filtering e 0 58 0 5 second moving average filtering e 15 10S 1 to 10 seconds moving average filtering 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 See Optimizing the Filter on page 64 for details on filter optimization If you have changed the value press to validate it before you press the button to access the Press Unit line The pressure unit line allows for the selection of bar kPa or psi for pressure units the desired selection is made via the or key and validated by the key When you press the 7 button the LED screen Figure 25 will be shown Agilent 1260 Infinity ELSD User Manual 101 9 102 The Control Panel The LED Screen LED ON H Reset Time Elapsed Figure 25 LED Screen The LED screen is used to turn the light source on off and is equivalent to the Light source button on the control pan
22. S 232 serial connection as a standard connection to a control PC with Agilent Data System for control and data evaluation Different options are available for connections to third party devices see Appendix 4 2 Connect the detector to your personal computer via the RS 232 port using the cable supplied with the instrument If you use a different cable make sure to use a straight RS 232 cable which directly connects pins of same numbers Most RS 232 cables are cross over or null modem cables and cannot be used for this connection 3 Connect the ELS detector to the APG remote control of your LC modules Use the supplied cable Cable remote 5061 3378 and plug it to the connectors labeled with REMOTE Not using the APG connection will impair the retention time reproducibility 4 Place the ON OFF switch to the OFF position and plug the power cord into the socket on the rear panel of the detector Do not turn on the power at this time 5 Connect the three wires of the power cord of this system 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 sure that an adapter is used to connect the third wire to ground 38 Agilent 1260 Infinity ELSD User Manual Installing the Detector 3 Electrical Connections 1V Output REMOTE MAIN POWER C oo Ext events Ext Autozero a Figure 9 Supply panel Agilent 1260 Infinity ELSD User Manual 39 3 I
23. Unit screen Figure 24 The temperature value blinks if desired temperature is not reached and stable The pressure value blinks if the gas pressure is lower than 3 0 bar Agilent 1260 Infinity ELSD User Manual The Control Panel 9 When the button is pressed the Offset screen Figure 21 which is used to select the desired offset is presented The Offset Screen Signal Offset mV 000 Figure 21 The Offset Screen To increase the offset value click on the key 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 20 mV s Once you have set the desired offset press the button to accept the new value Please note that for user convenience a fast offset setting can be done in the Status screen Figure 20 simply by pressing the or key This will change the offset value immediately Press the 7 button to access the Temp Gain screen Figure 22 The Temperature Gain Screen Temp 50 C Gain 1 Figure 22 The Temp Gain Screen The Temp Gain screen is used to set the desired Temperature and Gain When the screen is accessed the cursor is on the Temp setting This setting can be changed with the and buttons and validated by the button The temperature range is 20 to 100 C To maintain appropriate temperature control the temperature should be set at least 5 C above ambient A wait time of 15 to 30 mi
24. able for errors or for incidental or consequential damages in connec tion with the furnishing use or per formance of this document or of any information contained herein Should Agilent and the user have a separate written agreement with warranty terms covering the material in this document that conflict with these terms the warranty terms in the sep arate agreement shall control Technology Licenses The hardware and or software described in this document are furnished under a license and may be used or copied only in accor dance with the terms of such license Restricted Rights Legend If software is for use in the performance of a U S Government prime contract or subcon tract Software is delivered and licensed as Commercial computer software as defined in DFAR 252 227 7014 June 1995 or asa commercial item as defined in FAR 2 101 a or as Restricted computer soft ware as defined in FAR 52 227 19 June 1987 or any equivalent agency regulation or contract clause Use duplication or disclo sure of Software is subject to Agilent Tech nologies standard commercial license terms and non DOD Departments and Agencies of the U S Government will receive no greater than Restricted Rights as defined in FAR 52 227 19 c 1 2 June 1987 U S Government users will receive no greater than Limited Rights as defined in FAR 52 227 14 June 1987 or DFAR 252 227 7015 b 2 November 1995 as applicable in
25. ail in The Gas Valve Screen on page 103 To shut down the system 1 Turn off the pump 2 Allow the nebulization gas to flow through the detector for a few minutes to drain the evaporation tube and detection chamber 3 Turn off the power to the detector if desired If you are using a mobile phase which contains salts acids or bases pump a few ml of water or methanol through the system before turning off the detector to prevent the deposition of substances and corrosion of the system If the ELSD is used as a second detector and is not used for some time it is recommended to remove it from the liquid chromatography flow path in order to avoid blockage of the nebulizer or deposition of substances in the detector If the LC system shall be used without the ELSD it must be switched off and removed from the software configuration for avoiding problems due to a missing instrument power failure 56 Agilent 1260 Infinity ELSD User Manual Using the System 4 Cleaning the Detector After each session and before ending HPLC use the whole HPLC system including the ELSD should be cleaned in order to ensure good performance For the ELSD preventive maintenance consists in cleaning the detector before shutting down after last analyses Let mobile phase or solvent flow to flush particles which could remain Eventually increase temperature in order to dissolve possible deposit 1 2 3 Stop the mobile phase flow but let gas flow to
26. also be in a different module e g volumes injected by the autosampler might be too low or dead volumes in capillary connections cause peak broadening If the stray light test gives signals which are too low V Use the instrument keyboard for checking if the life time of the light source has been exceeded If needed please contact your Agilent service representative for a replacement V Set the stray light to the original value See The Light source Normalization Screen on page 102 A Noisy Signal is Observed High noise is often due to a lack of maintenance The detector should be cleaned on a routine basis If the signal is noisy apply noise tests in order to ensure that the high noise is not due to the mobile phase or the column Then 1 Doa general inspection as described in General Inspection on page 82 2 Clean the nebulizer as described in Cleaning the Nebulizer on page 86 3 Exchange the nebulizer Agilent 1260 Infinity ELSD User Manual Agilent 1260 Infinity Evaporative Light Scattering Detector User Manual e00 ee Ogee Maintenance e i e j General Inspection 82 Cleaning and Decontamination 83 Adjusting Stray Light Settings 89 This chapter describes e The maintenance procedures that should be performed by the operator on a routine basis see General Inspection on page 82 e Cleaning and decontamination procedures that should be performed to maintain instrumen
27. ant temperature changes Do not place it near air conditioning vents windows ovens etc 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 The detector should be placed close to the outlet of the column to minimize extra column band broadening which will reduce the resolution of the chromatogram Environment This instrument has been designed for following conditions e Use inside buildings e Altitude up to 2000 meters e Ambient temperature from 5 C to 40 C e Maximum humidity of 80 for temperatures under 31 C with linear decrease to 50 at 40 C e Maximum variations for main power voltage 10 from nominal voltage e Transitory overvoltage of class II e Pollution degree 2 Agilent 1260 Infinity ELSD User Manual 23 2 Site Requirements and Specifications Specifications 24 Table1 Specifications ELSD Detection Light Source Temperature Range Gas Flow Control Gas Consumption Gas pressure Eluent Flow Rate Instrument Control Operating Parameters Control Signal Drift Signal Noise Signal Output Inputs High Sensitivity Photomultiplier Selected High Efficiency Blue LED Ambient to 100 C Manual and computer controlled nebulization gas flow and auxiliary gas flow Less than 4 l min for the Large Flow Nebulizer less than 3 l min for all other nebulizers 3 5 bar 51 p
28. anual Troubleshooting and Diagnostics 6 Troubleshooting Based on Instrument Signals In all cases of troubleshooting for the detector signal noise tests as described in Perform the Noise Tests on page 69 should be run in order to distinguish detector problems from application or system problems and in order to acquire parameters under defined conditions No Signal is Observed If no signal is observed V Make sure that the light source is on Y Run astray light test and check whether the stray light signal for gain 12 is in the range of 100 mV to 150 mV V Check that the gas valve is opened and the pressure regulator is opened if any A pressure display that is greater than 0 indicates that there is gas flow V If the stray light test passes and the gas flow is on the nebulizer is probably blocked Clean the nebulizer as described in Cleaning the Nebulizer on page 86 Y If cleaning the nebulizer does not solve the problem replace the nebulizer Agilent 1260 Infinity ELSD User Manual 79 6 80 Troubleshooting and Diagnostics Decrease in Sensitivity If noise tests did not show that the problem could be caused by the application or system a decrease in the sensitivity is often caused by the nebulizer main cause or the detector sensing section If noise tests pass but the signal heights areas for known standards are too low the nebulizer might need to be replaced Please note that the root cause might
29. any technical data Safety Notices CAUTION A CAUTION notice denotes a haz ard It calls attention to an operat ing procedure practice or the like that if not correctly performed or adhered to could result in damage to the product or loss of important data Do not proceed beyond a CAUTION notice until the indicated conditions are fully understood and met A WARNING notice denotes a hazard It calls attention to an operating procedure practice or the like that if not correctly per formed or adhered to could result in personal injury or death Do not proceed beyond a WARNING notice until the indicated condi tions are fully understood and met Agilent 1260 Infinity ELSD User Manual In This Guide This manual is designed to describe the installation operation maintenance and basic troubleshooting of the G4218A Agilent Evaporative Light Scattering Detector It includes 1 introduction 2 Site Requirements and Specifications Site requirements and specifications of the Agilent 1260 Infinity Evaporative Light Scattering Detector 3 Installing the Detector Installation of the detector 4 Using the System This chapter 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 down the unit 5 Optimizing Performance This chapter shows how to optimize the detector to achieve bes
30. aring solvents the most critical parameter is the dry residue this parameter should be less than 1 ppm to maximize the sensitivity of the detector The mobile phase should not contain non volatile solvent modifiers or buffers Volatile solvent modifiers e g CF COOH CH3COOH C2H5 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 contains BHT as a Stabilizer The parts of the detector which are in contact with the solvent and sample are made of PTFE stainless steel and glass Make sure that the solvents are compatible to these materials Depending on the mobile phase nature and flow rate the suggested gas pressure 3 5bars 51psi may have to be adjusted in order to optimize the background noise and the signal to noise ratio Sample Pre Treatment If the sample contains insoluble particles it should be filtered through a 0 2 um or 0 45 um filter before injection Agilent 1260 Infinity ELSD User Manual Optimizing Performance 5 Column Treatment The chromatographic column typically contains micro particles which are used to separate the compounds of interest In some cases such particles may be eluted from a column and enter the detector which may increase in the noise The degradation of column packing material depends on a variety of factors including the particle size type of column packing and the nature of the
31. asurement conditions Table 4 Measurement conditions for test run Sample Caffeine standard 250 pg ml in water p n G4218 85000 Solvent Isocratic 80 water 20 Acetonitrile Flow 1 ml minute Injection volume 20 pl Column Eclipse XDB C18 4 6x150 5u Analytical TCC temperature 40 C ELSD temperature 40 C ELSD pressure 3 5 bar 51 psi ELSD gain 7 System Agilent 1260 Infinity Standard LC System with Standard Degasser Binary Pump High Performance Autosampler Thermostatted Column Compartment and Diode Array Detector After turning on the detector allow 15 minutes for equilibration Monitor the temperature using the Data System online signal display or the instrument display As soon as the target temperature is reached and stable start a method run using conditions listed in Table 4 Agilent 1260 Infinity ELSD User Manual Using the System 4 A chromatogram similar to the one in the following figure can be expected Please note that retention time peak area and peak shape can vary and depend on the individual HPLC system configuration mV 100 r 0 1 2 3 4 m Figure 14 Typical chromatogram for caffeine see measurement conditions in text Agilent 1260 Infinity ELSD User Manual 55 4 Using the System Powering Down and Shutting Down the System If desired some or all functions of the system can be powered down at the end of an automated series of analyses These power down features are described in det
32. d easily during the method optimization process If you suspect that the compound of interest is thermally labile a lower temperature could be used to improve the sensitivity by reducing the thermal decomposition 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 35 C and 60 C see Installation Test Procedure on page 54 Table 4 It is clear that the use of a low temperature provides significantly better sensitivity for volatile and thermally sensitive compounds 60 Agilent 1260 Infinity ELSD User Manual Optimizing Performance Selecting the Optimum Temperature A Di E 2s 25 7 7 285 27 ma Figure 15 Chromatogram of Caffeine at Various Temperatures The minimum temperature that can be used depends on the flow rate and the nature of the mobile phase Agilent 1260 Infinity ELSD User Manual 61 5 Optimizing Performance Optimizing the Mobile Phase 62 Particulate matter in the mobile phase will increase the background and the noise The purity of the solvent is a critical issue in the noise In general filtering of the solvent is not recommended as the solvent may extract contaminants from the filter In most cases distilled water and HPLC grade solvents are satisfactory If you are comp
33. determine the problem V It is important to recognize that in almost all cases there are several possible causes for a problem As an example an increase in the noise of the chromatogram could be due to a defective nebulizer the slow elution of very tightly retained material from the column a unsuitable mobile phase non volatile buffers high dry residue amount an increase in pump pulsation the solvent e g improper degassing or high residue after evaporation the pump e g a defective check valve the detector e g an electronic problem the gas supply e g particles in the gas line V Check the nebulizer The flow from the nebulizer should be fine and homogeneous If it is not the nebulizer the needle or the PTFE tube may be obstructed with foreign material It is rather unlikely that two problems occur at the same time The role of the troubleshooting activities is to determine the cause of the problem In this discussion we will assume that the operator has determined that other components of the system are operating properly Agilent 1260 Infinity ELSD User Manual Troubleshooting and Diagnostics 6 WARNING Destruction of the nebulizer by disassembly Disassembling the nebulizer will destroy it gt Do not disassemble the nebulizer The control panel and system electronics do not contain any replaceable components If the suggestions provided in this chapter do not remedy the problem contact your Agilent ser
34. does not return to normal the solvent contains a residue material and is not suitable for use with the detector V If the noise of the system 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 90 C and pump solvent at the rate of 2 ml min using a gas pressure of 3 5 bar 51 psi 70 Agilent 1260 Infinity ELSD User Manual Troubleshooting and Diagnostics 6 Noise Test Procedures This section describes some procedures which can be helpful for troubleshooting and help the user to distinguish detector issues from issues caused by other elements contributing to the measurement result like solvents or the column 4 tests are described which add more of these elements step by step so their contribution can be measured V The electronic noise test measures the signal created by the detector without irradiation and solvent flow V The background noise stray light test measures the signal measured by the detector while the light source is on but no sample or solvent flow through the detector V The solvent noise tests measures the signal created by pure solvent flowing through the detector while no column is installed V The column noise test measures the signal created by pure solvent flowing through the column and the detector Agilent 1260 Infinity ELSD User Manual 71 6 Troubleshooting and Diagnostics Preliminary Activit
35. e normal manner Deliver a mobile phase that will dissolve the contaminant at a flow rate of 1 ml min If you do not know what the contaminant is ethanol or acetone are good choices 3 Access the temperature adjustment mode via the control panel 4 Push and hold the button for 5 s This will automatically set the temperature to 100 C Maintain the flow and temperature for at least 3 hours Clean the exterior of the detector with a rag saturated with isopropanole Removing the Nebulization Chamber and Nebulizer If the nebulizer shall be stored afterwards please flush the nebulizer and shut down the system as described in Powering Down and Shutting Down the System on page 56 1 Turn off the LC pump or any other mobile phase flow 2 Turn off the ELS detector 3 Stop the gas flow using the external pressure regulator Agilent 1260 Infinity ELSD User Manual Maintenance 7 Cleaning and Decontamination Figure 17 Replacing the nebulization chamber For subsequent steps see Figure 17 4 Remove the capillary connection to the mobile phase 5 Remove the connection to pressurized gas by pushing on the white inlet see arrow 6 Remove the drain tube Agilent 1260 Infinity ELSD User Manual 85 7 Maintenance CAUTION 7 Remove the small black plastic nut at the rear and carefully lift out the nebulization chamber 8 Remove the large black plastic nut at the top and remove the nebulizer I
36. e mains line V Check that the mains line voltage corresponds to the detector voltage version V Disconnect the system from the power line and check the mains fuses The mains fuses are in the vicinity of the power line socket as shown in Figure 1 Pry the plastic cover off the fuse holder to access the fuses and inspect them If a fuse s is blown replace it with one of the same rating T3 15 AL 230V part number G4218 68005 B B Gas inlet 1V Output REMOTE MAIN POWER 0 eae O O RS 232 Ext events Ext Autozero Fuses Figure 16 Control Panel 68 Agilent 1260 Infinity ELSD User Manual Troubleshooting and Diagnostics 6 Perform the Noise Tests Repeat the tests described in Installation Test Procedure on page 54 and compare the observed data to the limits defined for these values For example if the Electronic Noise test see Electronic Noise Test on page 73 Background Noise test see Background Noise Stray Light Test on page 74 and Solvent Noise test see Solvent Noise Test on page 75 provide results which are well within limits but the Column Noise test see Column Noise Test on page 77 fails or is significantly different from values obtained for comparable measurement conditions same system setup solvent column it is likely that the problem is due to the column e g highly retained compounds are being eluted Agilent 1260 Infinity ELSD User Manual 69 6 Troubleshooting
37. e siphon and ensure that the end of the tube is not immersed in the collected liquid Make sure that the liquid container is appropriate for the solvents used 3 Ensure that no siphon liquid leak could affect detector performances or create a laboratory pollution If the solvent that you are using is not compatible with Tygon e g THF use instead a piece of PTFE tubing or any material that you know is compatible with your solvent WARNING Toxic and hazardous solvents The handling of solvents and reagents can hold health risks gt When opening capillary or tube fittings solvents may leak out gt Please observe appropriate safety procedures for example goggles safety gloves and protective clothing as described in the material handling and safety data sheet supplied by the solvent vendor especially when toxic or hazardous solvents are used 46 Agilent 1260 Infinity ELSD User Manual Installing the Detector 3 Connecting the Nebulization Gas to the Nebulizer Attach the nebulization gas tube coming out of the front panel to the nebulizer gas inlet fitting located on the nebulizer side as shown in Figure 11 Connecting the Column 1 Attach the nebulization gas tube coming out of the front panel to the nebulizer gas inlet fitting located on the nebulizer side as shown in Figure 11 Powering Up the Unit 1 Place the ON OFF switch to the OFF position and plug the unit into the wall 2 Turn on the unit via the ON
38. ed by linearly interpolating to the concentration of organic phase in the solvent e g a mixture of half and organic solvent 50 50 should provide a signal of approximately less than 105 mV 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 A different supply of solvent should be obtained and employed MS grade solvents or lowest dry residue lt HPLC grade solvent should be used for this test The purity of the solvent is critical for a low background noise 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 1 ppm to maximize the sensitivity of the detector Mass spectroscopy grade solvents have a very low dry residue Agilent 1260 Infinity ELSD User Manual 75 6 76 Troubleshooting and Diagnostics The mobile phase should not contain non volatile solvent modifiers Volatile solvent modifiers e g CFs COOH CHCOOH CoH 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 normally contains BHT as a stabilizer When filtering solvents only use filters recommended for the use with HPLC systes as the solvent may extract contaminants from other filters Stainless s
39. el Use the button followed by the button to turn the LED on and the button followed by the button to turn it off The hours field indicates the number of hours that the LED has been in use The lifetime of the LED is approximately 5000 h When this period has been reached a message will be displayed after powering up the unit that the maximum usage of the lamp has been exceeded To reset the field move the cursor to the Reset Time Elapsed field and validate by pressing The Reset Time Elapsed field should be validated with only when you change the lamp When you press the 7 button the Light source Normalization screen Figure 26 will be presented The Light source Normalization Screen Stray Light percent Value 100 Figure 26 The Light Source Normalization Screen The intensity of the light source and consequently the measured signals will decrease over time After replacing the light source the signal intensity may also change Agilent 1260 Infinity ELSD User Manual The Control Panel 9 In case your application requires to have constant signal intensities this option can be used for adjusting the level of the signal intensity Use either stray light measurements see Background Noise Stray Light Test on page 74 or peak areas measured under defined conditions for doing this adjustment Example Your reference measurement gave a stray light value of 120 mV Your actual stray ligh
40. es required for the instrument service Agilent 1260 Infinity ELSD User Manual Agilent 1260 Infinity Evaporative Light Scattering Detector User Manual 10 Appendix Safety Information 108 Solvent Information 112 The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC 113 Lithium Batteries Information 114 Agilent Technologies on Internet 115 ee Agilent Technologies 107 10 Appendix Safety Information 108 The following general safety precautions must be observed during all phases of operation service and repair of this instrument Failure to comply with these precautions or with specific warnings elsewhere in this manual violates safety standards of design manufacture and intended use of the instrument Agilent Technologies assumes no liability for the customer s failure to comply with these requirements General This is a Safety Class I instrument provided with terminal for protective earthing and has been manufactured and tested according to international safety standards The Agilent 1260 Infinity modules are designed and certified as a general purpose laboratory instrument for research and routine application only It is not certified for in vitro or medical applications Operation Before applying power comply with the installation section Additionally the following must be observed Before the instrument is switched on all protective earth terminals extension cords auto transf
41. f you want to store the nebulizer clean the nebulizer as described in Cleaning the Nebulizer on page 86 and protect all openings using Parafilm or similar polymer foils For re installing the nebulizer or replacing it by a new one or different type reverse the order of previous steps as described in Installing the Nebulizer and Nebulization Chamber Assembly on page 43 In case the black gas tubing has been removed make sure you are using the correct orientation where the white filter is at the lower end Avoid leaks at all connections and check for leakages when you turn on the pump again Damage of flow cells When using the ELSD with another detector using a flow cell which is not compatible with the back pressure generated by the chosen ELSD nebulizer can destroy the flow cell gt See Nebulizers for the G4218A Agilent Evaporative Light Scattering Detector on page 94 for nebulizer back pressures and compare them to the specifications of your flow cell Cleaning the Nebulizer Damage of the nebulizer rare part Improper handling of the nebulizer can damage the part gt Handle the nebulizer with care and do not disassemble the rear part of the nebulizer which is protected by the colored thermal seal 86 To remove the nebulizer from the system 1 Turn off the HPLC system 2 Disconnect the nebulizer liquid inlet from the column 3 Disconnect the gas inlet from the nebulizer Agilent 1260 Infin
42. ffected by the absorbtion characteristics of the solvent thus solvents which absorb UV radiation can be used Aptent chnatging 1208 Sones ELSO Figure 1 The Agilent 1260 Infinity Evaporative Light Scattering Detector 10 Agilent 1260 Infinity ELSD User Manual Introduction 1 The detector is controlled via the keypad and digital display on the front panel Alternatively the system can be controlled by an external computer using the RS 232 port The output can be sent to a recorder or data station The detector includes a nebulizer evaporation tube and detector head The evaporation tube is located in an oven to assist in the evaporation of the solvent Agilent 1260 Infinity ELSD User Manual 11 1 Introduction Principle of Operation 12 There are three steps in the operation of the detector 1 nebulization of the eluent 2 evaporation of the solvent and 3 detection of the compound s of interest Figure 2 NEBULIZATION gt EVAPORATION gt DETECTION Figure 2 Schematic Diagram of an Evaporative Light Scattering Detector Nebulization involves the conversion of the eluent into a fine mist The mist is passed through an evaporator to vaporize solvent In the detector unit the mist is irradiated by a light source and scattered light is measured by a photomultiplier PM The degree of light scattering is related to the concentration of the compound of interest in the sample A cros
43. filter solvents small particles can permanently block the capillaries Avoid the use of the following steel corrosive solvents e Solutions of alkali halides and their respective acids for example lithium iodide potassium chloride and so on e High concentrations of inorganic acids like nitric acid sulfuric acid especially at higher temperatures replace if your chromatography method allows by phosphoric acid or phosphate buffer which are less corrosive against stainless steel e Halogenated solvents or mixtures which form radicals and or acids for example 2CHCl Oo 2COCl 2HCI This reaction in which stainless steel probably acts as a catalyst occurs quickly with dried chloroform if the drying process removes the stabilizing alcohol e Chromatographic grade ethers which can contain peroxides for example THF dioxane di isopropylether such ethers should be filtered through dry aluminium oxide which adsorbs the peroxides e Solutions of organic acids acetic acid formic acid and so on in organic solvents For example a 1 solution of acetic acid in methanol will attack steel e Solutions containing strong complexing agents for example EDTA ethylene diamine tetra acetic acid e Mixtures of carbon tetrachloride with 2 propanol or THF The ELSD waste and exhaust tubes are made of Tygon 2375 PVC which has limited chemical compatibility to ethers like THF halogenated organic solvents ketones and other non
44. ich 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 sure that an adapter is used to connect the third wire to ground Agilent 1260 Infinity ELSD User Manual Installing the Detector 3 Installing the Nebulizer and Nebulization Chamber Assembly Installing the Nebulizer and Nebulization Chamber Assembly Parafilm is used to cover various openings inside the compartment nebulizer and nebulization chamber to prevent dust particles from entering the system during shipment The installed Nebulizer Nebulization chamber assembly is shown in Figure 11 To Column Bulkhead Nebulizer Gas inlet Fitting Nut Nebulization chamber Bracket Nut Siphon Overflow Figure 11 Installing the nebulizer nebulization chamber assembly Agilent 1260 Infinity ELSD User Manual 43 3 Installing the Detector Installing the Nebulizer and Nebulization Chamber Assembly To install the assembly 1 Remove the Parafilm on all detector openings and on the nebulizer cell these coatings are used to prevent dust particles from entering the system during shipment 2 Position the nebulization chamber as shown in Figure 11 and tighten the black nut at the bottom The nebulization chamber is in the correct position if it is in contact with the back wall as shown in Figure 12 Figure 12 Fixing the Tip of Nebulization chamber
45. ies CAUTION Before starting the tests for new instruments or after storage flush detector with water at a flow of 1 ml min for at least 15 minutes Before running any of the following tests make sure that the signal behavior after gain changeuses option free in ChemStation method settings which is used for getting an uncorrected signal level after a gain change which is done in subsequent procedures Also do not set a signal offset after autozero Both settings can cause wrong measurement results The following activities should be performed 1 Power up the instrument 2 Set the gain to 1 and the offset to 0 mV The Signal screen should indicate 000 or a very small signal 3 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 4 Provide gas to the detector and adjust the pressure to 3 5 bar 51 psi If the pressure is less than 3 bar 44 psi an error message will be presented indicating that the detector is not ready Damage of pressure sensor Pressures above 4 5 bar 67 psi can damage the pressure sensor of the detector The gas consumption for the large nebulizer is 4 l min and 3 l min for all other nebulizers The target pressure for most applications is 3 5 bar 51 psi gt Do not switch on the gas flow suddenly as this may cause the exceeding of maximum pressure The use of a separate pres
46. ilent Technologies 95 9 The Control Panel The Control Panel The Control Panel Figure 19 includes a digital display and a number of buttons that are used to enter data Figure 19 The Control Panel 96 Agilent 1260 Infinity ELSD User Manual The Control Panel 9 The Digital Display The digital display presents information about the present status of the detector and is used for controlling its measurement 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 After the system has completed the initialization procedures the Status screen Figure 20 will be presented The signal should be close to zero Signal Temp Press Gain 001 26 C 3 5b 1 Figure 20 The Status Screen The user interface is provided via a series of screens see The User Interface on page 98 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 22 are used to enter control parameters The control panel is disabled while the instrument is controlled externally via an RS232 connection Chemstation etc The keys on the control panel are used to provide the following functions 1 unit If you keep the key pressed the rate of change of the parametter will increase used to i
47. in the day field when the screen is accessed The day can be changed via the or key and the next previous field can be accessed via the key Press to validate any changes When you press the button the Total Lifetime Elapsed screen Figure 30 will be presented The Total Lifetime Elapsed Screen Total Lifetime Elapsed HHHHHHHH hrs Figure 30 The Total Lifetime Elapsed Screen The Total Lifetime Elapsed information screen indicates the usage of the detector and cannot be edited When you press the 7 button the Serial Number screen Figure 31 will be presented The Serial Number Screen Serial Number FRAA700001 Figure 31 The Serial Number Screen The Serial Number screen cannot be edited When you press the button the Firmware screen Figure 32 will be presented Agilent 1260 Infinity ELSD User Manual 105 9 106 The Control Panel The Firmware Screen Firmware Version 1 3 Firmware Date MM YY Figure 32 The Firmware Screen This information screen presents the firmware version and date where MM is the month and YY the year The Firmware screen cannot be edited When you press the V button the Factory Method Code screen Figure 33 will be presented The Factory Method Code Screen Factory Method Code Authorized persons only Figure 33 The Factory Method Code Screen The Factory Method Code screen is used by the service engineer to access procedur
48. ing and fire hazard The exhaust coming from the detector contains evaporated solvent and sample Many HPLC solvents like acetonitrile or methanol are toxic and inflammable gt Verify that no tube damage or inappropriate installation could allow a gas leak in laboratory CAUTION Contamination of detector If gas from the hood enters the detector it is possible that foreign material from the hood could contaminate the detector gt Avoid negative pressure between the detector and the fume hood CAUTION 3 Install the vent tube so that it cannot become blocked or bent Bias of measurement results Turbulences in the glass cell siphon liquid spilled into the heating tube or contamination of the detector gt Make sure the negative pressure of hood or exhaust line is not too strong such that no air is drawn in from the leak drain 4 The vacuum must be moderate to avoid turbulence in the glass cell siphon 5 Using the drain bottle assembly G1946 60111 and included tubing is recommended as a condensation trap and for pressure compensation Avoid long tube installations in upward direction creating condensation dropping back into the detector Avoid loops or bends in the tubing which form condensation traps causing bad measurement results Agilent 1260 Infinity ELSD User Manual 37 3 installing the Detector Electrical Connections All electrical connections are made via the supply panel Figure 8 1 Use the R
49. ing the mobile phase and samples Avoid air oxygen or reactive or inflammable gases in order to avoid the risk of burnings or explosions Do not use solvents which could inflame at temperatures reached by the detector 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 siphon overflow tube must contain liquid at all times When working with solvents please observe appropriate safety procedures e g goggles safety gloves and protective clothing as described in the material handling and safety data sheet by the solvent vendor especially when toxic or hazardous solvents are used The gas pressure should not exceed 4 5 bar 67 psi Make sure that the gas flow is maintained while the mobile phase flows through the system If the gas flow is interrupted for extended periods of time organic solvents could possibly damage the pressure sensor and or the photosensor Agilent 1260 Infinity ELSD User Manual 109 10 Appendix 110 Repair Do not open the cover of the rear part of this instrument Access to and repair of internal parts is restricted to Agilent service and service providers authorized by Agilent and certified for this instrument For internal parts even if the instrument is grounded there is a potential shock hazard that could result in serious personal injury Make sure that only fuses with the required rated current and of the
50. ity ELSD User Manual Maintenance 7 4 Remove the nebulizer from the detector taking care to never pull on the rear connection tubing 5 Remove the nebulizer black nut and seal 6 Remove the gas inlet quick fitting To clean the nebulizer 1 Fill an ultrasonic bath with approximately 2 cm of the appropriate solvent which depends on the nature of the material that is present in the nebulizer In most cases ethanol is a satisfactory solvent 2 Place the nebulizer vertically in the solvent bath The nebulizer outlet should be placed at the bottom Agilent 1260 Infinity ELSD User Manual 87 7 Maintenance Cleaning and Decontamination maximum liquid level 3 Clean the nebulizer for approximately 30 minutes with the solvent and then replace the solvent with water and clean for an additional 30 minutes To fix up the nebulizer 1 Fix the gas inlet fitting on the nebulizer see Installing the Nebulizer and Nebulization Chamber Assembly on page 43 2 Fix the nebulizer on the nebulization chamber 3 Test the nebulizer to ensure that it is working properly If the performance cannot be improved by cleaning the nebulizer a new nebulizer might be required 88 Agilent 1260 Infinity ELSD User Manual Maintenance 7 Adjusting Stray Light Settings You may set the desired stray light using the instrument keyboard Agilent 1260 Infinity ELSD User Manual 89 7 Maintenance Adjusting Stray Light Settings
51. l leakage of hazardous solvents Leakage of hazardous solvents could cause personal injury or laboratory pollution by pressed out liquid gt Make sure all flow connections to and inside the detector are tight gt After having switched on the LC pump for several minutes verify that there are no leaks gt In case of any leak switch off the pump immediately and remove the liquid 52 Y Ensure that the siphon is filled with liquid at all times The overflow from the siphon should be collected in a suitable waste container by using the drain tube V Never exceed a gas pressure of 4 5 bar 67 psi Agilent 1260 Infinity ELSD User Manual Using the System 4 Y Avoid the use of solvent or samples that could corrode the detector The mobile phase is in contact with glass and PTFE tubing and the evaporation tube is constructed from stainless steel See Solvent Information on page 112 Agilent 1260 Infinity ELSD User Manual 53 4 Using the System Installation Test Procedure 54 In order to verify the correct operation of the instrument this test procedure is used During this test a standard sample is run under defined conditions and the result is inspected visually by comparing it to a reference measurement This test is not intended as a replacement for a full IQ or OQ procedure which is available as a service from Agilent and provides quantitative results For running the test please use the following me
52. l the detector Inspection by Agilent is required to evaluate if the instrument is in good condition or damaged gt Notify your Agilent sales and service office about the damage gt An Agilent service representative will inspect the instrument at your site and initiate appropriate actions gt It is recommended that the shipping carton be retained as it can be used if it should become necessary to transport the system 32 Table3 Parts required Quantity Part Number Description 1 See Table 9 Nebulizer 1 G4218 40000 Nebulization chamber glass 1 G4218 90000 Operator s Manual 1 G4218 6800x Accessory Kit consists of 1 Power cable 1 Auto zero cable 1 Signal cable 1 RS 232 cable 1 External event cable 6 mm 0 D gas tubing 2 meters 1 meter sets 1 set of replacement fuses Agilent 1260 Infinity ELSD User Manual Installing the Detector 3 Installing the Detector Gas Supply The unit is connected to the gas supply via the 6 mm plastic tubing supplied using the fitting on the upper left corner of the supply panel on the back of the detector Figure 7 Gas inlet 1V Output REMOTE MAIN POWER of fo Ext events Ext Autozero eye Figure 8 Supply Panel Agilent 1260 Infinity ELSD User Manual 33 3 Installing the Detector Inserting the gas inlet tube Ensure that Parafilm is removed from exhaust tube and drain tubing before installing unit 1 The tubing should be cut
53. ment depending on Filter Filter 0S Filter 1S Filter 2S Signal height 124 mV 122 mV 110 mV Noise ASTM 3 2 mV 1 1 mV 0 7 mV Peak width at 50 height 2 5 seconds 2 5 seconds 2 8 seconds S N 37 110 157 Signal to noise ratio is multiplied by 3 when choosing Filter 1S without any peak broadening effect If Signal to Noise ratio is more important than resolution a Filter 2S or higher can be set to improve sensitivity even more Agilent 1260 Infinity ELSD User Manual s Agilent 1260 Infinity Evaporative Light Scattering Detector o User Manual ee 6 Troubleshooting and Diagnostics e E e j Basics of Troubleshooting 66 Specific Detector Troubleshooting 70 Noise Test Procedures 71 Troubleshooting Based on Diagnostic Tests 78 Troubleshooting Based on Instrument Signals 79 This chapter describes troubleshooting activities which are useful for determining the cause of wrong or unexpected s S Agilent Technologies 65 6 Troubleshooting and Diagnostics Basics of Troubleshooting 66 The Agilent 1260 Infinity Evaporative Light Scattering Detector is designed to be used with a liquid chromatography system For troubleshooting it is important to distinguish between detector issues and issues with other parts of the system which might affect detector output Troubleshooting refers to the task of finding the reason for an abnormal response from the system and the following guidelines should be used to
54. mobile phase e g a high pH may degrade silica based columns After you have installed a new column it is recommended that you pump mobile phase through it for a few minutes before connecting it to the detector This will flush out micro particles that may reside in the column from the manufacturing or shipment process It is suggested that you perform the Column Noise test see Column Noise Test on page 77 to obtain the baseline signal value for the column Agilent 1260 Infinity ELSD User Manual 63 5 Optimizing Performance Optimizing the Filter 64 The Digital Filter see The Noise Filter Pressure Unit Screen on page 101 allows maximizing signal to noise ratio by filtering noise The filter length should be optimized according to peak shape and more specifically to peak width The following table suggests some Filter settings depending on peak width Table 5 Digital Filter suggestion versus Peak Width Peak width at 50 Seconds Suggested filter Seconds lt 1 second 0 second 2 1 4 2 6 4 8 6 gt 10 8 and higher These suggested values can be optimized depending on your specific chromatography by decreasing the filter length if peaks are poorly resolved e g if Rs lt 1 5 or increasing Filter when optimizing Signal to Noise ratio Example Comparison of digital filters using the SOP test injection of 5 ppm glucose at gain 12 Peak width at half height is 2 5S Table 6 Sensitivity improve
55. ncrease the present value of a user settable parameter e g the offset by by 1 unit If you keep the key pressed the rate of change of the parameter will used to decrease the present value of a user settable parameter e g the offset increase at sets the value of the parameter that you have edited OK Agilent 1260 Infinity ELSD User Manual 97 9 98 The Control Panel sets the present intensity for the detector to zero cap is used to power the LED in the detector When the LED is lit the keyboard LED lg immediately above the button will be illuminated changes the active line on the display to the next previous line or the next previous screen D D moves the cursor on the display to the next previous field The User Interface The Status screen Figure 20 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 keystrokes within a few seconds Each parameter change must be validated with or the change will not be applied The Status Screen The Status screen Figure 20 displays the current measurement values of the detector This screen cannot be edited but the desired offset can be set via the Offset screen Figure 21 the temperature and gain can be set via the Temp Gain screen Figure 22 and the pressure units can be selected via the Noise Filter Pressure
56. nstalling the Detector Additional Options Use the RS 232 cable as a standard connection to a control PC with Agilent Data System for control and data evaluation All electrical connections are made via the supply panel Figure 10 Gas inlet 1V Output REMOTE MAIN POWER C o Jo Ext events Ext Autozero os Figure 10 Supply Panel Connecting the Recorder Integrator If a recorder or integrator is employed connect the recorder input to the 1 V output terminal on the rear panel of the detector Figure 10 and to the appropriate socket on the recorder integrator Connecting the External Autozero If the external autozero function shall be used plug the cable that is supplied into the Ext Autozero socket on the detector Figure 10 and to the appropriate socket on the controlling device Use a contact closure or TTL signal for auto zeroing the detector 40 Agilent 1260 Infinity ELSD User Manual Installing the Detector 3 Do not install the autozero cable if not needed An unintentional short circuit of the loose ends will result in a contiuous zero signal If a TTL signal is used please make sure to use the correct polarity identified on the cable Refer to External Auto zeroing of the Detector on page 51 to operate external autozero contact closure Connecting the External Events Cable If the external events functions shall be used plug the cable that is supplied into the appropriate socket on
57. nutes is recommended for achieving good temperature stabilization The stabilization time for detector temperatures close to the ambient temperature is longer than for high temperatures Agilent 1260 Infinity ELSD User Manual 99 9 100 The Control Panel When you press the 7 button the Gain field can be edited The gain range is from 1 12 each increment of one unit increases the gain by a factor of 2 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 is 1 2048 After setting the desired gain press the button for navigating to the Autozero offset screen Figure 23 The Autozero offset Screen Output Signal Value After AZ xxx mV Figure 23 The Autozero offset Screen This screen is used to allow the signal to reach the desired value when performing an autozero by keyboard software control or external contact closure 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 and buttons and validated by the x button After you have set the desired autozero offset press the 7 button for navigating to the Noise Filter Pressure Unit screen Figure 24 Agilent 1260 Infinity ELSD User Manual The Control Panel 9 The Noise Filter Pressure Unit Screen Filter 1S
58. ormers and devices connected to it must be connected to a protective earth via a ground socket Any interruption of the protective earth grounding will cause a potential shock hazard that could result in serious personal injury Whenever it is likely that the protection has been impaired the instrument must be made inoperative and be secured against any intended operation The operator of this instrument is advised that if the equipment is used ina manner not specified in this manual the protection provided by the equipment may be impaired Maintain a well ventilated laboratory If the mobile phase or sample contains volatile substances ensure that the laboratory is ventilated well such that no flammable or nocious vapors can accumulate Agilent 1260 Infinity ELSD User Manual Appendix 10 Do not operate the instrument in the presence of flammable gases or fumes Operation of any electrical instrument in such an environment constitutes a definite safety hazard The exhaust from the detector must be vented into a fume hood exhaust line or similar installation Make sure that the exhaust gas does not escape into the laboratory Take in consideration any solvent filter that could be required by your local environmental laws Potential leakage of hazardous liquids Make sure all flow connections to and inside the detector are tight After switching on the LC pump verify that there are no leaks Use only inert gases nitrogen for nebuliz
59. peration Colored seal Nebulizer To HPLC system Nebulization chamber Inlet for nitrogen To evaporator and detection To waste Siphon Figure 5 The Nebulizer Introduction 1 Principle of Operation Evaporation of the Solvent A heated tube is used to evaporate the solvent The exit of the heated tube leads directly into the detector cell In liquid chromatography water and organic solvents with low boiling points are typically employed e g methanol acetonitrile A typical mobile phase for a reverse phase separation using evaporative light scattering detection might be methanol water 60 40 while a typical mobile phase for normal phase separation might be hexane If acids bases and salts are used to modify mobile phase to provide the desired separation they should 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 C2H5 3N NH4zOAc HCOOH CH3COOH CF3 COOH and HNO3 Detection The carrier gas transports the microparticles from the heating tube into the detection chamber Figure 6 Detection chamber Photomultiplier Light trap Figure 6 The Detection Chamber Agilent 1260 Infinity ELSD User Manual 17 1 18 Introduction The detector chamber contains a light emitting diode LED and a photomultiplier that is positioned at an
60. r 51 psi Set the temperature to 50 C Wait for stable temperature Switch on the light source Set the Gain to 1 Set the offset to 0 mV Wait 15 minutes for stabilization and record the signal level on oO ol FF amp N Increase the gain to 12 and monitor the signal The expected level is typically 100 mV to 150 mV 74 Agilent 1260 Infinity ELSD User Manual Troubleshooting and Diagnostics 6 Solvent Noise Test To determine the solvent noise 1 ao oo A Ww N Make sure the siphon is filled and the bulkhead is blocked with a blank nut or Parafilm to avoid a Venturi effect Switch on the gas flow at 3 5 bar 51 psi Set the temperature to 50 C Wait for stable temperature For now keep off the HPLC pump Switch on the light source Set the gain to 12 and monitor monitor the signal Do not autozero the detector The signal may be negative Bypass the column of your HPLC system 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 1 ml min Monitor the baseline for a few minutes If water is used as the solvent the signal should be 10 mV or less Higher values could be observed if non HPLC grade water is used which may have a higher non volatile residue If an organic solvent is used the signal should be 200 mV or less For mixtures of water and organic solvents the expected signal can be estimat
61. r Down Options Mode Photomultiplier LED Heating Gas flow General Off Off Off Off Standby Off Off On Off Cleaning Off Off On On To select the desired Power Down mode use the or key to access the desired mode and then press to validate the selection 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 the detector can be powered down in two ways External event cable power down contact closure The detector will stay in the power down mode chosen while the contact remains closed It comes back in normal mode when the contact closure is released e Power down screen Press the 7 button to access the power down screen then press again the 7 button to place the cursor on the Power down activate line Validate with to put the detector in power down mode To leave the power down mode release the contact closure if power down has been activated by external event or press any key if power down has been activated from the Power down screen Agilent 1260 Infinity ELSD User Manual The Control Panel 9 When the cursor is on the Power down activate line pressing the 7 button will present the Date Time screen Figure 29 will be presented The Date Time Screen Date 01 05 04 Time 14 33 21 Figure 29 The Date Time Screen The Date format is MM DD YY and the cursor will be
62. r Manual Agilent 1260 Infinity Evaporative Light Scattering Detector User Manual 3 Installing the Detector Lifting and Carrying the Module 28 Unpacking the System 30 Installing the Detector 32 Electrical Connections 38 Additional Options 40 Installing the Nebulizer and Nebulization Chamber Assembly 43 This chapter describes how the laboratory should be prepared to optimize the performance of the Agilent 1260 Infinity 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 4 for start up procedures a Agilent Technologies 27 3 Installing the Detector Lifting and Carrying the Module CAUTION Safe transport Module is heavy gt To ensure safe transport and avoid bodily injury make sure that the module is lifted by two persons Once the module is unpacked ensure that no cables or tubing are connected when you carry the module The module should be lifted by the bottom e g place your hands under the module Two persons are needed to ensure easy transport and avoid bodily injury Figure 7 28 Agilent 1260 Infinity ELSD User Manual Installing the Detector 3 er 4 n had Carrying the Module Apne Technangies 1290 Series B50 Do NOT Lift Here Do NOT Lift Here Carry Here Carry Here Figure 7 Carrying the System Agilent 1260 Infinity ELSD U
63. s lines for sufficient time might take days and consider additional filters Pure gas is not required as the gas is only used as a carrier for the solid sample particles CAUTION Damage of pressure sensor Pressures above 4 5 bar 67 psi can damage the pressure sensor of the detector The gas consumption for the large nebulizer is 4 l min and 3 l min for all other nebulizers The target pressure for most applications is 3 5 bar 51 psi gt Do not switch on the gas flow suddenly as this may cause the exceeding of maximum pressure The use of a separate pressure regulator is highly recommended A filter 0 01 pm and manometer part number G4218 60100 is available as an option Replacement filter cartridges are available as part number G4218 40150 Power Cords Your detector is delivered with a power cord which matches the wall socket of your particular country or region The plug on the power cord which connects to the rear of the instrument is identical for all types of power cord WARNING Electric Shock The absence of ground connection and the use of an unspecified power cord can lead to electric shock or short circuit gt Never operate your instrumentation from a power outlet that has no ground connection gt Never use a power cord other than the power cord designed for your region Agilent 1260 Infinity ELSD User Manual 21 2 Site Requirements and Specifications Use of unsupplied cables The use of cable
64. s sectional view of the system is presented in Figure 3 Agilent 1260 Infinity ELSD User Manual Introduction 1 Principle of Operation Detection Nebulization Figure 3 Schematic display of principle measurement steps in the ELSD of the Detector Agilent 1260 Infinity ELSD User Manual 13 1 14 Introduction Nebulization The eluent from the chromatograph is nebulized by the inlet gas typically nitrogen At the outlet of the nebulizer the aerosol travels through a chamber Large droplets in the aerosol go to a siphon while the fine mist moves to the evaporation tube The overall design of the nebulizer is shown in Figure 1 and the nebulization chamber is shown in Figure 5 Five different nebulizers are available for optimizing the performance of the detector at different HPLC flow rates see Table 9 The user should select the nebulizer to best match the flow rate that will be used with the separation when the detector is ordered the optimal range for each nebulizer is indicated in Table 9 Additional nebulizers are available from Agilent Technologies and can be easily installed see Installing the Nebulizer and Nebulization Chamber Assembly on page 43 Agilent 1260 Infinity ELSD User Manual Introduction Principle of Operation mae Peek inlet Colored Seal Gas inlet Figure 4 Design of the Nebulizer Agilent 1260 Infinity ELSD User Manual 1 15 Introduction Principle of O
65. s which haven t been supplied by Agilent Technologies can lead to damage of the electronic components or personal injury gt Never use cables other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations Exhaust venting and drain requirements CAUTION The exhaust from the detector must 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 potentially hazardous exhaust of evaporated solvent and sample must not be allowed to enter the laboratory atmosphere and any appropriate accessory like solvent filters should be disposed according to local environmental requirements The drain tube must be directed to a suitable solvent The user is responsible for decontamination or recycling of any residue regarding to local environmental requirements Toxic and hazardous vapors Exhausts could cause personal injury or laboratory pollution gt Make sure the instrument is installed and the exhaust is vented according to local regulatory authorities for health and safety requirements 22 Agilent 1260 Infinity ELSD User Manual Site Requirements and Specifications 2 Location of the Detector in the Laboratory All components of the system e g HPLC pumps detector should be located on a robust table The detector should be placed in an area that is free from drafts or signific
66. ser Manual 29 3 Installing the Detector Unpacking the System CAUTION Defective on arrival problems If there are signs of damage please do not attempt to install the module Inspection by Agilent is required to evaluate if the instrument is in good condition or damaged gt Notify your Agilent sales and service office about the damage gt An Agilent service representative will inspect the instrument at your site and initiate appropriate actions gt It is recommended that the shipping carton be retained as it can be used if it should become necessary to transport the system 30 Table 2 Components Shipped with the Agilent 1260 Infinity Evaporative Light Scattering Detector Quantity Part Number Description 1 G4218 20000 Standard Flow Nebulizer 1 G4218 40000 Nebulization chamber glass 1 G4218 90000 Operator s Manual 1 G4218 85000 Caffeine Standard 250 pg ml in water 1 G4218 6800x Accessory Kit consists of 1 Power cable 1 Auto zero cable 1 Signal cable 1 RS 232 cable 1 External event cable 6 mm O D gas tubing 2 meters 1 meter sets 1 set of replacement fuses 1 Drain tube including seal Ferrule and fitting 1 G1946 60111 Drain bottle assembly 1 G1312 67305 Capillary 600 x 0 17 mm Agilent 1260 Infinity ELSD User Manual Installing the Detector 3 Table 2 Components Shipped with the Agilent 1260 Infinity Evaporative Light Scattering Detector Quantity Part Number Description 1 50
67. si typical min 3 bar 44 psi to max 4 5 bar 67 psi Micro Flow Nebulizer 5 pl min to 50 pl min Semi Micro Flow Nebulizer 40 l min to 1 ml min Standard Flow Nebulizer 500 l min to 2 ml min Large Flow Nebulizer 1 5 to 5 ml min Rapid Resolution Nebulizer 0 2 to 1 4 ml min Microprocessor with stand alone manual keypad or Windows based PC control see specifications for your instrument control software Liquid Crystal Digital Panel Less than 1 mV h lt 1mvV 0 1000 mV Analog RS 232 Digital Remote Autozero Contact Closure or TTL Signal Remote Powerdown Mode Contact Closure or TTL Signal Agilent 1260 Infinity ELSD User Manual Site Requirements and Specifications 2 Table1 Specifications ELSD Power Down Mode General Standby Cleaning Zero Control Manual Auto Zero and Remote Auto Zero Interface RS 232 1 0 Serial Output Power 100 V 2 1 A 115 V 60 Hz 1 8 A or 230V 50 Hz 1 7 A Dimensions 250 mm 10 W x 450 mm 18 H x 550mm 22 D Weight 18 5 kg 40 Ib Conditions for drift and noise Eluent Flow 0 ml min Evaporator Temperature 50 C Nebulizer Gas Nitrogen pressure 3 5 bar Gain 12 Noise measured as mean value for 10 segments of 1 minute For each segment the difference of the highest minus the lowest peak is calculated and divided by 2 Agilent 1260 Infinity ELSD User Manual 25 2 Site Requirements and Specifications Specifications 26 Agilent 1260 Infinity ELSD Use
68. splay 97 E electrical connections 38 electronic noise 73 electronic noise test 73 environmental conditions 23 evaporation of the solvent 17 exhaust requirements 22 external auto zero 51 F Factory Method Code 106 Factory Method Code screen 106 filter 64 Firmware 106 G gas considerations 21 supply 33 Gas Valve screen 103 glass tube 45 installation 32 installation test procedure 54 installing the nebulizer 43 integrator 40 L LED screen 102 lifting the system 28 ocation of the detector in the laboratory 23 M maintenance 81 manual auto zeroing 51 mobile phase optimizing 62 nebulization 14 nebulizer cleaning 86 gas 33 installation 43 Noise Filter Pressure Unit 101 Agilent 1260 Infinity ELSD User Manual Index noise tests 69 71 73 74 75 77 0 Offset screen 99 optimizing mobile phase 62 perfomance 59 temperature 60 P power cord 38 requirements 20 up the detector 47 Power Down screen 103 powering down 56 preparing the system for operation 50 Pressure Unit Noise Filter Pressure Unit screen 101 Principle of Operation 12 recorder integrator connecting 40 routine operation 52 RS 232 port 38 S safety information 108 on lithium batteries 114 safety symbols 110 sample pre treatment 62 117 screens Z Date Time 105 External Power Down 103 Factory Method Code 106 Firmware 106 Gas Valve 103 LED 10
69. sure regulator is highly recommended A filter 0 01 pm and manometer part number G4218 60100 is available as an option Replacement filter cartridges are available as part number G4218 40150 72 5 Press the button The signal should read close to zero and remain constant 6 Set the noise filtering to 1S see The Noise Filter Pressure Unit Screen on page 101 Agilent 1260 Infinity ELSD User Manual Troubleshooting and Diagnostics 6 Electronic Noise Test For this test do not turn on the light source Do not turn on the HPLC pump no solvent flow To determine the electronic noise 1 Make sure the siphon is filled and the bulkhead is blocked with a blank nut or Parafilm to avoid a Venturi effect Switch on the gas flow at 3 5 bar 51 psi Set the temperature to 50 C Wait for stable temperature Set the Gain to 12 At gain 12 monitor the signal for a period of 5 min The variation in the signal should be less than 2 mV there may be some spiking of the signal ao FF WwW N 6 Record the level and autozero the detector again Agilent 1260 Infinity ELSD User Manual 73 6 Troubleshooting and Diagnostics Background Noise Stray Light Test For this test the HPLC pump must be off no solvent flow To determine the background noise 1 Make sure the siphon is filled and the bulkhead is blocked with a blank nut or Parafilm to avoid a Venturi effect Switch on the gas flow at 3 5 ba
70. t results 6 Troubleshooting and Diagnostics This chapter 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 7 Maintenance Introduction to maintenance and repair instructions on simple routine repair procedures Agilent 1260 Infinity ELSD User Manual 3 10 Parts for Repair Detailed illustrations and lists for identification of parts and materials for repair The Control Panel This chapter describes the role of the various controls and displays on the detector Appendix This chapter contains safety information Agilent 1260 Infinity ELSD User Manual Contents 1 Introduction 9 The Low Temperature Evaporative Light Scattering Detector 10 Principle of Operation 12 Nebulization 14 Evaporation of the Solvent 17 Detection 17 2 Site Requirements and Specifications 19 Site Requirements 20 Power Considerations 20 Gas Considerations 21 Power Cords 21 Exhaust venting and drain requirements 22 Location of the Detector in the Laboratory Environment 23 Specifications 24 3 Installing the Detector 27 Lifting and Carrying the Module 28 Unpacking the System 30 Installing the Detector 32 Gas Supply 33 Vent the Exhaust Line to a Fume Hood 37 Electrical Connections 38 Additional Options 40 Agilent 1260 Infinit
71. t measurement gives a value of 140 mV To re adjust the Stray Light use the Light Source Normalization screen and enter a value of 85 120 140 100 using the and buttons then validate by pressing This will result in the stray light re adjustment Adjustable range is from 70 to 130 If the calculated percentage is out of this range please contact your Agilent service representative When you press the 7 button the Gas Valve screen Figure 27 will be presented The Gas Valve Screen Gas Valve Open Prog Time 0 mm Off Figure 27 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 or key to select On and press When you press the 7 button the External Shutdown screen Figure 28 will be presented The Power Down Screen The Power Down Mode screen Figure 28 is used to indicate which features should be shut down upon receipt of a power down signal from an external source e g a personal computer or an HPLC pump or from the menu Agilent 1260 Infinity ELSD User Manual 103 9 104 The Control Panel Power down Mode General Activate Figure 28 The Power Down Screen The three options provided for external shutdown are summarized in Table 10 Table 10 Powe
72. t performance see Cleaning and Decontamination on page 83 ee Agilent Technologies 81 7 Maintenance General Inspection 82 The Agilent 1260 Infinity Evaporative Light Scattering Detector is designed for a low level of maintenance activities Maintenance activities are normally the responsibilities of the user If preventive maintenance activities are followed the system provides high sensitivity measurements without intervention of the operator Y Ensure that the detector is installed according to the site requirements see Site Requirements on page 20 in a clean laboratory environment away from sources of heat or ventilation V Make sure that the detector exhaust is vented to a fume hood or lab exhaust line V Check that the detector is kept clean There should be no foreign material on the joints in the nebulizer the glass tube etc V Make sure the siphon of the nebulization chamber is filled with solvent Ensure that the gas flow does no bubble through the siphon and the liquid level is not fluctuating If this should be the case check for a correct installation of the exhaust tube and make sure the vacuum applied there is neither to strong nor to weak V Check the nebulizer The flow from the nebulizer should be fine and homogeneous If it is not the nebulizer the needle or the PTFE tube may be obstructed with foreign material Clean the nebulizer as described in Cleaning the Nebulizer on page 86
73. teel filters should be preferred over glass filters and others HPLC grade solvents or lowest dry residue solvent should be used for this test If changing the solvent source does not solve the problem it may be necessary to decontaminate the system as described in General Inspection on page 82 or clean the nebulizer as described in Cleaning the Detector on page 83 Agilent 1260 Infinity ELSD User Manual Troubleshooting and Diagnostics 6 Column Noise Test It is recommended that a specific column is reserved for this test This column should not be used for routine analyses If a column is used for a number of different separations it is possible that some compounds can be tightly bound to it and slowly eluted over time When tightly bound compounds are slowly eluted from the column excessive noise will be observed To determine the column noise 1 Turn off the pump and connect the column 2 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 applied depends on to the column ID and is indicated in the following table Table 7 Flow rate versus Column diameter indication Column ID mm Flow Rate pl min 46 1000 2 1 208 1 0 47 0 8 30 0 32 48 3 Set the gain to 12 and monitor the baseline for a few minutes A suitable column will provide a baseline
74. that is 20 50 mV above the solvent baseline If the instrument fails the Column Noise test it is most likely the fault of the column rather than the instrument Obtain a new column and repeat the test If the mobile phase contains acidic modifiers e g CF COOH disconnect the detector and wash the HPLC system for 12 h before starting to analyze unknown samples This wash should be performed after the column noise test is completed but does not need not be performed after each analysis Agilent 1260 Infinity ELSD User Manual 77 6 Troubleshooting and Diagnostics Troubleshooting Based on Diagnostic Tests If the actions described in the noise test procedures see Noise Test Procedures on page 71 do not solve the problems of failing tests following actions should be performed by the service Electronic Noise Test In case the electronic noise is higher than expected see A Noisy Signal is Observed on page 80 Stray Light Test In case the intensity of the stray light background noise is out of the expected range see No Signal is Observed on page 79 Failed Run of Standard Sample In case the noise tests have passed but the peak area for a standard is lower than previous reference measurements check the nebulizer see Cleaning the Nebulizer on page 86 make sure you are using a fresh sample and consider running the test using a backpressure loop instead of a column 78 Agilent 1260 Infinity ELSD User M
75. vice representative If the digital display does not illuminate when the system is powered up turn the unit off and inspect the main fuses If necessary replace the fuses with some of the same rating as the original fuse 3 15 AT time delayed part number G4218 68005 for units of all voltages The fuses are located inside the main power module on the rear panel Figure 8 A set of replacement fuses is delivered in starting kit If the fuses are not blown or if the replacement fuses blow up contact your Agilent service representative Initial Troubleshooting Activities V Make sure that the instrument and all components of the system are grounded properly V Ensure that the liquid level in the siphon is appropriate and that there is no liquid accumulating in the nebulization chamber V Check that the gas pressure is sufficient 3 5 bar target 3 bar minimum 4 5 bar maximum and is stable The gas filter should be clean and in place V Ensure that the flow rate of the pump is constant and check that there are no leaks in the chromatographic system Agilent 1260 Infinity ELSD User Manual 67 6 Troubleshooting and Diagnostics If there is no response from the system If there is no response from the system when the unit is powered up e g the fan does not rotate the display does not light up the light source is off etc it is likely that the instrument is not getting power V Check that the unit is plugged into an activ
76. y ELSD User Manual 23 Contents Installing the Nebulizer and Nebulization Chamber Assembly 43 Connecting the Siphon Overflow 46 Connecting the Nebulization Gas to the Nebulizer 47 Connecting the Column 47 Powering Up the Unit 47 Using the System 49 Preparing the System for Operation 50 Auto zeroing the Detector 51 Manual Auto zeroing of the Detector 51 External Auto zeroing of the Detector 51 Routine Operation of the System 52 Installation Test Procedure 54 Powering Down and Shutting Down the System 56 Optimizing Performance 59 Selecting the Optimum Temperature 60 Optimizing the Mobile Phase 62 Sample Pre Treatment 62 Column Treatment 63 Optimizing the Filter 64 Troubleshooting and Diagnostics 65 Basics of Troubleshooting 66 Initial Troubleshooting Activities 67 If there is no response from the system 68 Perform the Noise Tests 69 Specific Detector Troubleshooting 70 Noise Test Procedures 71 Preliminary Activities 72 Agilent 1260 Infinity ELSD User Manual Contents Electronic Noise Test 73 Background Noise Stray Light Test 74 Solvent Noise Test 75 Column Noise Test 77 Troubleshooting Based on Diagnostic Tests 78 Electronic Noise Test 78 Stray Light Test 78 Failed Run of Standard Sample 78 Troubleshooting Based on Instrument Signals 79 No Signalis Observed 79 Decrease in Sensitivity 80 A Noisy Signal is Observed 80 7 Maintenance 81 General Inspection 82 Cleaning and Decontamination 83

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