SofTA 2300 ELSD. Installation and Operation Guide

Transcription

1 Installation and Operation Guide SofTA 2300 ELSD Copyright 2016 Teledyne Technologies Incorporated. All rights reserved. April 2016

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3 Installation and Operation Guide Safety Warnings SofTA 2300 ELSD Safety SofTA 2300 ELSD Installation and Operation Guide Safety Warnings Safety Before installing, operating, or maintaining this equipment, it is imperative that all hazards and preventive measures are fully understood. While specific hazards may vary according to location and application, take heed in the following general warnings: WARNING LASER SAFETY: This ELSD is a Class I laser product with a Class IIIa (3R) embedded laser. The embedded laser beam is less than 5 mw, 650 nm, and is collimated. There are no user adjustments for power, alignment, or collimation. Under use conditions, the laser is never visible, and presents no special safety considerations. As required by regulation, the back panel has the following laser related labels, in addition to a serial number label. Internal components may have the following label: Never remove a component with this label. Doing so could expose a user to a potentially hazardous laser beam. Always refer service to qualified personnel. WARNING Do not use gases that support combustion with combustible solvents. Always use inert gases.to avoid inhaling harmful organic solvents and other materials, the detector exhaust should not be allowed to enter the laboratory atmosphere. DANGER To avoid burns, turn off the power at least 30 minutes before performing maintenance procedures. 3

4 Installation and Operation Guide Safety Warnings WARNING Liquids associated with this instrument may be classified as carcinogenic, biohazard, flammable, or radioactive. Should these liquids be used, it is highly recommended that this application be accomplished in an isolated environment designed for these types of materials in accordance with federal, state, and local regulatory laws, and in compliance with your company s chemical/hygiene plan in the event of a spill. WARNING Avoid hazardous practices! If you use this instrument in any way not specified in this manual, the protection provided by the instrument may be impaired. WARNING If you are using flammable solvents or chemicals with this system, vapor concentration levels may exceed the maximum exposure levels as recommended by OSHA Guide To reduce those levels to a safe exposure, Teledyne Isco recommends that you place the system in a laboratory hood designed for the purpose of ventilation. This hood should be constructed and operated in accordance with federal state and local regulations. In the event of a solvent or chemical spill, your organization should have a plan to deal with these mishaps. In all cases, use good laboratory practices and standard safety procedures. Hazard Severity Levels This manual applies Hazard Severity Levels to the safety alerts. These three levels are described in the sample alerts below. CAUTION Cautions identify a potential hazard, which if not avoided, may result in minor or moderate injury. This category can also warn you of unsafe practices, or conditions that may cause property damage. WARNING Warnings identify a potentially hazardous condition, which if not avoided, could result in death or serious injury. 4

5 Installation and Operation Guide Safety Warnings DANGER DANGER limited to the most extreme situations to identify an imminent hazard, which if not avoided, will result in death or serious injury. Hazard Symbols The equipment and this manual use symbols used to warn of hazards. The symbols are explained in the table below. Hazard Symbols Warnings and Cautions The exclamation point within the triangle is a warning sign alerting you of important instructions in the instrument s technical reference manual. The lightning flash and arrowhead within the triangle is a warning sign alerting you of dangerous voltage inside the product. Symboles de sécurité Ce symbole signale l existence d instructions importantes relatives au produit dans ce manuel. Ce symbole signale la présence d un danger d électocution. Warnungen und Vorsichtshinweise Das Ausrufezeichen in Dreieck ist ein Warnzeichen, das Sie darauf aufmerksam macht, daß wichtige Anleitungen zu diesem Handbuch gehören. Der gepfeilte Blitz im Dreieck ist ein Warnzeichen, das Sei vor gefährlichen Spannungen im Inneren des Produkts warnt. Advertencias y Precauciones Esta señal le advierte sobre la importancia de las instrucciones del manual que acompañan a este producto. Esta señal alerta sobre la presencia de alto voltaje en el interior del producto. 5

6 Installation and Operation Guide Safety Warnings For Additional Information Technical assistance for the 2300 ELSD can be obtained from: Teledyne Isco 4700 Superior St. Lincoln NE Phone: (800) or (402) Fax: (402)

7 Table of Contents Section 1 Introduction 1.1 Introduction ELSD Principle of Operation Operation Section 2 Site Requirements 2.1 Site Requirements Power Connection Gas Considerations Exhaust Venting Drain Requirements Location of the Detector Environment Section 3 Installing the Detector 3.1 Installing the Detector What You Will Need Unpacking Connections Installation Qualification Checklists Overview Installation Qualification (IQ) for SofTA ELSDs Operational Qualification (OQ) for SofTA ELSDs Performance Qualification (PQ) for SofTA ELSDs Overall Detector Performance for SofTA ELSDs Section 4 Using the Detector 4.1 Using the Detector Turning On and Off the Detector Touch Screen Menu Structure Start Up Procedure Shut Down Procedure Section 5 Optimizing Performance 5.1 Optimizing Performance Choosing Operating Conditions Mobile Phase Considerations QC Test Conditions Tuning Examples

8 Table of Contents Section 6 Troubleshooting 6.1 Troubleshooting Chromatography Problems Troubleshooting Baseline Noise Appendix A Specifications A.1 Specifications A-1 8

9 Section 1 Introduction 1.1 Introduction ELSD Principle of Operation ELSDs are near universal detectors primarily used in High Performance Liquid Chromatography (HPLC), though they have been used successfully in other types of chromatography as well. Their principal requirement is that the analyte be less volatile than the mobile phase. An ELSD cannot detect highly volatile analytes. However, most analytes of interest are less volatile than the eluting solvents. Evaporative light scattering detectors are complementing ultraviolet (UV) detection because they can detect most analytes, even those that do not absorb UV radiation, are stable during gradient elutions, and respond to the relative mass of the analyte (an important feature that is useful when detecting unknown materials). The ELSD is superior to the refractive index detector (RID) as it can be use with gradient chromatography, it is not susceptible to ambient temperature changes, and it does not produce negative peaks which can be difficult to quantify. The ELSD does not respond to the mobile phase disruption seen as solvent front peaks in the void volume with UV and RI detectors so early eluting analytes can be easily quantified. Mass spectrometry (MS) detection is also a nearly universal detector, but its high cost and complexity have kept it from being widely used. In fact, the operation requirements of MS closely match that of the ELSD. This allows the less expensive and less complicated ELSD to be used as a method development detector for methods to be used on the MS systems. 1-1

10 Section 1 Introduction Figure 1-1 The Model 2300 detector The Model 2300 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 USB port. The output can be sent to a recorder or data station Operation The SofTA ELSD employs a unique method of detection. The process involves the nebulization of the column eluent and transforms it into an aerosol cloud. As this cloud travels through a heated zone within the instrument, the more volatile mobile phase evaporates leaving a smaller cloud of analyte particles. These particles pass through a beam of light and scatter some of the light which is converted into an electronic signal. Nebulization Nebulization transforms the liquid phase leaving the column into an aerosol cloud of fine droplets. The size and uniformity of the droplets are extremely important in achieving sensitivity and reproducibility. The SofTA ELSD uses a special concentric flow nebulizer and a constant flow of an inert gas to ensure a narrow droplet size distribution. Our nebulizer is constructed entirely from PTFE, which accumulates fewer deposits than either glass or stainless steel. To handle flow rates and mobile phases common in HPLC, all ELSDs need a way to divert part of the aerosol cloud to waste. Softa uses a patent pending Thermo-Split technology. Our Thermo-Split chamber combines a gentle bend with temperature controlled walls. When the aerosol exiting the nebulizer encounters a cool environment, it partially condenses into larger particles whose momentum carries them into the wall and down the drain. 1-2

11 Section 1 Introduction Figure 1-2 Thermo-Split with cooling The temperature of the spray chamber may be elevated. As the aerosol traverses the chamber, it partially evaporates and shifts the particle size distribution low enough for essentially all the particles to negotiate the bend. These operating conditions may be useful for special applications. Under these conditions, a majority of the aerosol particles pass through the chamber and are carried into the evaporative zone. Figure 1-3 Thermo-Split with heating 1-3

12 Section 1 Introduction Evaporation Detection After passing through the nebulization chamber, the aerosol cloud is propelled through the heated evaporation tube assisted by the carrier gas. In the evaporation tube, the solvent is volatilized to produce particles or droplets of pure analyte. The temperature of the drift tube is set at the temperature required to evaporate the solvent. The temperature is kept as low as possible to avoid particle shape distortion, evaporation of the analyte, or when working with thermally-sensitive compounds. The particles emerging from the evaporation tube enter the optical cell where the sample particles pass through the light emitted by a low-power laser. The particles scatter the light which is detected by a silicon photodiode located at a 90 angle from the laser. A light trap is located opposite the laser to collect the light not scattered by particles. The quantity of light detected is proportional to the solute concentration and solute particle size distribution. The photodiode produces a signal which is sent to the outputs for collection. Figure 1-4 Detection process 1-4

13 Section 2 Site Requirements 2.1 Site Requirements A suitable environment is important to ensure optimum performance of the Model 2300 ELSD Power Connection The ELSD operates at either 120 V or 240 V, 50/60 Hz. Confirm that the SofTA ELSD is configured for the correct voltage before plugging it into line voltage. WARNING Incorrect line voltage at the instrument could result in a shock hazard or damage to your instrumentation. Do not connect devices to a line voltage higher than specified Gas Considerations A supply of clean, filtered, oil free inert clean gas (typically Nitrogen) regulated to 65 ± 5 psi is required to operate the detector. The gas supply needs to be free of particles as particles will create background noise in the chromatograms. Pure gas is not required as the gas is only used as a carrier. Argon, Helium, or Nitrogen are acceptable. Do not use gases that support combustion Exhaust Venting The ELSD should be located close to a fume hood or other ventilation device. The exhaust from the detector must be directed into a fume hood or exhaust vent. The potentially hazardous exhaust of evaporated solvent and sample must not be allowed to enter the laboratory atmosphere. Alternatively, connect tubing and direct exhaust to a cooled collection vessel for later disposal. CAUTION Exhausts could cause personal injury or laboratory pollution. Make sure the instrument is installed and the exhaust is vented according to local regulatory authorities for health and safety requirements. Any appropriate accessory like solvent filters should be disposed according to local environmental requirements Drain Requirements The instrument s internal P trap must be full during operation. Restricting the drain port with liquid is very important to ensure detection sensitivity. Direct the outlet of the tube to a collection vessel located so that the solvent can flow freely from the trap into a collection vessel. Make sure that the liquid container is appropriate for the solvents used. 2-1

14 Section 2 Site Requirements Location of the Detector The detector should be placed in an area that is free from drafts or significant temperature changes. Access to the power supply used to disconnect the device 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. The detector should be spaced so that there is about 3 inches (~8 cm) of space on the right side to allow good flow from the fan Environment This instrument has been designed for following conditions: Use inside buildings in clean and dry laboratories. Ambient temperature from 15 to 29 F. Maximum humidity of 90% non-condensing. Maximum variations for main power voltage: 10% from nominal voltage. 2-2

15 Section 3 Installing the Detector 3.1 Installing the Detector What You Will Need In addition to the ELSD and its accompanying accessories, you will need: Gas Supply: Clean, dry, inert gas regulated to 65 ± 5 psi is needed for nebulization. Either Argon or Nitrogen is acceptable. Do not use gases that support combustion. Exhaust Device: The carrier gas containing volatilized mobile phase and sample components will exit the ELSD. You should provide a means of removing this from the laboratory. The ELSD should be located close to a fume hood or other ventilation device. Tubing, fittings, and tools to connect your HPLC system to the ELSD Unpacking Carefully unpack the accessory items included with the ELSD shipment and confirm that all the items are present. SofTA ELSDs come in a high-quality shipping container engineered to avoid damage in transit. Save the shipping container and packaging for future shipments. CAUTION If there are signs of damage, please do not attempt to install the ELSD. Inspection by authorized and qualified personnel is required to determine if the instrument is damaged. Notify Teledyne Isco or your local sales and service office about any damages. 3-1

16 Section 3 Installing the Detector Connections Power Connection Exhaust The ELSD operates at either 120 V or 240 V, 50/60 Hz. Confirm that the SofTA ELSD is configured for the correct voltage before plugging it into line voltage. Plug the modular power cord provided into the power input module on the back of the detector. The ELSD exhaust is a ½ O.D. aluminum tube. Use ½ I.D. tubing to connect to a fume hood during operation. In high-humidity environments, or when higher flow rates of mobile phase are being used, it is possible to condense liquid in the exhaust line. Install the vent tube so that it cannot become blocked or bent. Avoid the use of long tubes especially if installed with an upward slope. Any condensation in the tube could flow back into the detector. Avoid loops or bends in the tubing which can trap condensation. Alternatively, connect tubing and direct exhaust to the optional exhaust trap kit (part no ) or a cooled collection vessel for later disposal. CAUTION Blocked exhaust tubing, either from bends or condensation blockage, will result in poor and/or inconsistent results. WARNING The exhaust coming from the detector contains evaporated solvent and sample. Many HPLC solvents like acetonitrile or methanol are toxic and flammable. Verify that no tube damage or inappropriate installation could allow a gas leak in the laboratory. 3-2

17 Section 3 Installing the Detector Communication I/O Connections An output jack can be found on the back of the ELSD for connection to a chart recorder, computing integrator, or computerized data system. The maximum signal output is 1 V. Figure 3-1 I/O Connection port USB Pin 1 and Pin 2: A contact closure output. Pins are open when error conditions exist. Pin 3 and Pin 4: A contact closure input to turn on/off gas remotely. Gas control toggles with electrical continuity between these two pins. Maintain connectivity for a minimum of 500 ms. A subsequent toggle command within 2 seconds may be ignored. Pin 5 and Pin 6: A TTL output for instrument status. A logical high indicates the detector is in standby. Pin 7 and Pin 8: A contact closure input to reset baseline remotely. Maintain connectivity for a minimum of 500 ms. A subsequent toggle command within 2 seconds may be ignored. Figure 3-2 USB connection port The Model 2300 should be connected to a Windows computer for access of its full list of functions. Connect the USB port of a Model 2300 to a powered-up computer before powering up the unit. This will prevent the computer mistaking the 2300 as a serial device. 3-3

18 Section 3 Installing the Detector Gas Connections Fluid Connections If the 2300 has to be powered up before or at the same time with its control computer, the automatic serial device searching function of the computer should be disabled. Consult your IT department for how to disable this function on your computer. Connect a supply of clean, dry, inert gas regulated to 60 to 75 psi to the GAS INLET port on the back of the unit. The internal gas regulator maintains the gas pressure at the factory set value displayed on the front panel. Figure 3-3 Fluid connection ports Connect the outlet from your column to the 1 / 16 Valco bulkhead LIQUID INLET on the side of the unit (Figure 3-3). Use Teflon or PEEK tubing of sufficient length, 1 / 16 O.D., and an I.D. in the range of.005 to.010. The instrument is supplied with a nut and ferrule for the Valco bulkhead. The liquid inlet located on the left side of the detector by default but can be ordered configured for the right or reconfigured by an authorized and qualified personnel. CAUTION The length and volume between the column outlet and the detector inlet should be kept as short as possible to avoid unnecessary band broadening. A larger I.D. will contribute to band broadening, but has the advantage of plugging less frequently. In the event of a split flow configuration (common with a mass spec), the ELSD is quite versatile. However, please keep flow to the ELSD in the.5 ml/min. to 1 ml/min. range if possible. Flow must be measured by removing the nebulizer and allowing it to spray into a graduated collection vessel for an appropriate time. Also, please use the smallest internal bore T when configuring the external plumbing. Internal bore T s are now available with.006 construction. 3-4

19 Section 3 Installing the Detector WARNING The handling of solvents and reagents can hold health risks. Nebulization Chamber Drain Liquid leaks could cause personal injury, laboratory contamination, or negatively affect the detector performance. Ensure that all liquid connections are tight and not leaking. Solvents may flow from fitting when released. Please observe appropriate safety procedures as described in the material handling and safety data sheet supplied by the solvent vendor especially when toxic or hazardous solvents are used. The instrument s internal P trap must be full during operation. Restricting the drain port with liquid is very important to ensure detection sensitivity. Pump mobile phase at 1 ml/min without gas flow for 10 minutes or until you see liquid coming out of the drain. Alternatively, you may introduce 10 ml of mobile phase into the drain using a squirt bottle or syringe. Place the ¼ stainless steel drain adapter with flexible tubing provided in the accessory kit on the drain port located on the side of the instrument. The drain port is located on the left side of the detector by default but can be ordered configured for the right or reconfigured by an authorized and qualified personnel. Direct the outlet of the tube to a collection vessel located so that the solvent can flow freely from the trap into a collection vessel. Make sure that the liquid container is appropriate for the solvents used. Watch the liquid level in the vessel during operation and empty when full. If the solvent that you are using is not compatible with elastometric tubing, replace it with PTFE tubing or any material that is compatible with the solvent. 3-5

20 Section 3 Installing the Detector 3.2 Installation Qualification Checklists Overview This qualification procedure will document that the SofTA ELSD is operating as designed. This documentation is important when working in a regulated environment to demonstrate to various auditors that the detector is working properly. This process is called qualification or validation. Qualification can be divided into the following steps: Installation Qualification (IQ) documents that the correct detector was received and installed properly. Operational Qualification (OQ) tests that the detector meets specifications in the user environment. OQ generally requires specialized test detectors that have been calibrated against NIST standards. Performance Qualification (PQ) tests that the system performs the selected application correctly. The original test report is shipped with each detector to certify that the detector passed the final test. 3-6

21 Section 3 Installing the Detector Installation Qualification (IQ) for SofTA ELSDs Model Number: Detector Serial Number: Location of the Detector: Step Description Yes No 1 Is the detector the same as specified on the purchase order? 2 Has the detector been inspected for damages and found unharmed? 3 Has the required documentation identified by model number and date been supplied? 4 Have details of all services and utilities required to operate the detector been provided? 5 Have methods and instructions for user maintenance been provided along with a contact point for service and spare parts? 6 Is the selected environment suitable for the detector (i.e. is adequate room provided for installation, operation, and servicing)? Have appropriate solvent services and utilities (electricity, Nitrogen gas, ventilation, solvent waste recovery, etc.) been provided? 7 Has health, safety, and environmental information relating to the operation of this detector been provided? Installer Initials Operator Initials The undersigned individual, certified by the manufacturer, completed the manufacturer s procedure for the proper Installation Qualification of this detector. Name: Title/Affiliation: Signature: Date: 3-7

22 Section 3 Installing the Detector Operational Qualification (OQ) for SofTA ELSDs Set-up Model Number: Detector Serial Number: Location of the Detector: Turn on the detector. Confirm the following settings are accurate: Full Scale Gain Modify if necessary. Set the drift tube temperature to 65 C and the spray chamber temperature to 20 C. Wait until the temperature stabilizes (approx. 30 minutes). 1 V Normal Calibration 100% Drift tube SET TEMPERATURE Drift tube DISPLAYED TEMPERATURE The display should read 65 C ± 1 C. PASS FAIL Spray chamber SET TEMPERATURE Spray chamber DISPLAYED TEMPERATURE The display should read 20 C ± 1 C. PASS FAIL Gas Sensor Apply air or Nitrogen pressure monitored by a regulator and checked with a pressure gauge. Set the pressure to 65 psi. Record the gas pressure value displayed on the ELSD. ELSD Value The display should read 50 ± 10 psi. PASS FAIL Baseline Noise Connect a digital voltmeter calibrated with an NIST-traceable standard to the analog output of the ELSD. The meter must also be able to record a minimum and maximum voltage with a response time of at least 10 ms (Fluke 45 or equivalent). Fill the ELSD drain tube with HPLC Grade Water. Occasionally, liquid will be aspirated from the inlet line or drain which will result in a spike in the signal. The test should be restarted if this occurs. Set the detector filter to RC 5. Reset the baseline. Monitor the output signal for 2 minutes. Record the lowest voltage and the highest voltage. Calculate the difference. 3-8

23 Section 3 Installing the Detector Maximum Value Minimum Value Difference mv mv mv The difference should be less than 2.5 mv. PASS FAIL Baseline Drift Set the detector filter to RC 5. Autozero the detector to change the signal to about 20 mv, and monitor the signal for 30 minutes. Initial Signal Value Final Signal Value Difference mv mv mv The baseline drift should be less than 2.5 mv. PASS FAIL The undersigned individual, certified by the manufacturer, completed the manufacturer s procedure for the proper Operational Qualification of this detector. Name: Title/Affiliation: Signature: Date: 3-9

24 Section 3 Installing the Detector Performance Qualification (PQ) for SofTA ELSDs Model Number: Detector Serial Number: Location of the Detector: Note Before this procedure is performed, it is necessary to complete the Installation Qualification and the Operational Qualification. Sensitivity Set the HPLC System and ELSD conditions to: 1 ml/min of 50/50 Water/Methanol. Add a backpressure regulator or a length of small ID tubing between the pump outlet and the injector inlet. Ideally, this will maintain 1000 psi pressure on the pump check valves enough to minimize baseline pulsation on the ELSD. Full Scale Gain The injection standard (Sodium Benzoate) should be delivered in the smallest practical injection volume. This volume will be system dependent with some autosamplers more reproducible than others. (1 or 2 µl injection volume is recommended.) Alternatively, a 1 or 2 µl manual injector can be used. Enter the conditions into the detector. Start the HPLC pump and the gas flow and wait for temperatures to reach the set points. When the temperatures reach equilibrium, reset the baseline to about 20 mv. Monitor the ELSD signal for stability. The signal should be stable within.25 mv. Connect a digital voltmeter calibrated with an NIST traceable standard to the analog output of the ELSD. (Alternatively, a calibrated Chromatography Data System can be used.) When the signal is stable, inject the 100 ng sample three times. Record the maximum signal for each peak. 1 V Normal Calibration 100% Spray chamber 10 C Drift tube 65 C Filter 0 Injection Standard 100 ng Sodium Benzoate Peak 1 Height Peak 2 Height Peak 3 Height Average Peak Height mv mv mv mv 3-10

25 Section 3 Installing the Detector The Average Peak Height should be at least 1.5 mv. PASS FAIL Reproducibility Load the method labeled Low Flow Organic (55C SC/ 70C DT/ FLT5/ low gain). Set the HPLC system to deliver 0.5 ml/min of Methanol, and inject 1000 ng of Sodium Benzoate from the smallest injection volume the system Autosampler will reliably deliver. (1 or 2 µl injections are recommended.) Alternatively, a 1 or 2 µl manual injector can be used. Peak Area 1 Peak Height 1 Peak Area 2 Peak Height 2 Peak Area 3 Peak Height 3 Peak Area 4 Peak Height 4 Peak Area 5 Peak Height 5 Peak Area 6 Peak Height 6 Peak Area% RSD Peak Height % RSD The reproducibility of six injections should be no greater than 2.0% RSD (area) or 2.0% RSD (height). PASS FAIL The undersigned individual, certified by the manufacturer, completed the manufacturer s procedure for the proper Performance Qualification of this detector. Name: Title/Affiliation: Signature: Date: 3-11

26 Section 3 Installing the Detector Overall Detector Performance for SofTA ELSDs After the Installation Qualification, Operational Qualification, and Performance Qualification procedures have been completed, the Overall Detector Performance document should be completed to verify the completion of all tests. Model Number: Detector Serial Number: Location of the Detector: Procedure Date Pass Fail Installation Qualification (IQ) Date Operational Qualification (OQ) Date Performance Qualification (PQ) Date The undersigned individual, certified by the manufacturer, completed the certification for this detector. Name: Title/Affiliation: Signature: Date: Customer Name/LC Operator: Signature: Date: 3-12

27 Section 4 Using the Detector 4.1 Using the Detector Turning On and Off the Detector Figure 4-1 The 2300 ELSD The power button is touch-sensitive. After plugging in a live power cord, the power button will be red. The detector is in power-up standby. Briefly touching the power button will turn the detector on. 4-1

28 Section 4 Using the Detector Button Color Illumination Status Solid Red Detector has power, in power-up standby Flashing Blue to White Not ready Solid White Power up complete, Ready for operation Flashing Red Fatal Error 1. Turn unit off immediately 2. Check for abnormal smell 3. Wait 30 seconds 4. Power up unit again 5. If still flashing red, turn off and contact qualified service technician To power off the unit, touch the power button for 5 to 10 seconds and the unit will be powered off. Wait 10 seconds or longer before powering the unit up again Touch Screen The Model 2300 ELSD is equipped with a hidden touch-sensitive area in front of the vacuum fluorescent display. Figure 4-2 Touch sensitive options Touching [Home] button always brings the display back to the HOME screens. 4-2

29 Section 4 Using the Detector Menu Structure Generally both [<=] and [Next] buttons function as the [Next] button to bring up the next screen, and both [=>] and [Enter] buttons function as the [Enter] button to accept the confirmation and take the actions displayed on the screen. Precise touching of a button is not critical. On some screens, touching [<=] or [=>] button will open more screens. Some screens, for example the method screens, use arrow symbols to prompt you to touch [<=] or [=>] button to bring up more screens. But due to the limitation of the screen size, some other screens do not show arrow symbols to prompt getting more screens. These screens do not display [Next] or [Enter] action prompts either. In general, though the hidden buttons are not always labeled by legends on the display due to the size limitation of the display panel, they are present and functional. Users can touch the areas where the buttons normally are in order to access the actions they want to take. Figure 4-3 Menu structure flowchart Initial Power Up Screens After powering up, the 2300 will automatically enter the operation standby mode. Follow the prompt on the screen to exit the standby mode. 4-3

30 Section 4 Using the Detector SofTA ELS Detector Model 2300 SofTA ELS Detector Rev E.1.1 Standby [Touch to exit] Home Screen After exiting the standby mode, the display enters the HOME screen and operation run mode. In run mode, the detector will heat up or cool down its spray chamber and drift tube to their set temperatures. It will also open the gas valve and power the laser. The top line of the HOME screen will always display actual output voltage in mv ##.###mv ##C/##C/FLT#/XG The bottom line cycles to provide additional information. ##.###mv ##C/##C/FLT#/XG Set point values of spray chamber, drift tube, filter type, and weight and gain. ##.###mv ##.#C/##.#C/#.#psi Actual values of spray chamber, drift tube, and regulated gas pressure. The actual temperatures can be differentiated from the set points by the display of the additional decimal point. 4-4

31 Section 4 Using the Detector ##.###mv Touch for more Touch for more... is a reminder that additional menu screens are available. ##.###mv Standby Timeout:#### Standby Timeout Menu Screens Services Standby Timeout can be used to close the gas valve when the detector is not detecting peaks to save gas. The timeout countdown timer counts at 1 second interval. The countdown counts will be replaced by ---- to indicate the gas valve has been shut off before the timer counts down to zero. If the gas valve is opened again through front panel operations, the timer is reset and starts countdown again. The feature can be disabled in the service menu described below. At the HOME screens, touching the display will bring up menu screens. The detector will decide which menu screen to bring up first depending on which HOME screen has been touched. But, you can always browse through all of the available menu screens. The display will automatically return to home screens after a few minutes if the display is not touched. The menu screens available include: Service Return Home Reset Baseline Select a Method Enter Standby Enter Services? [Next] [Yes] The services available include: Gas Control Laser Control Operational Readings Operational Parameters Set Timeout Functions Self-diagnostics 4-5

32 Section 4 Using the Detector Gas Control Start Gas Control? [Next] [Yes] Turn off gas? [Esc] [Yes] Turn on gas? [Esc] [Yes] Laser Control Start Laser Control? [Next] [Yes] Turn on Laser? [Esc] [Yes] Turn off Laser? [Esc] [Yes] Operational Readings Show OP Readings? [Next] [Yes] SC/DT/Gas ##C/##C/#.#psi OC/ET/Laser ##C/##C/LaserOn[Off] 4-6

33 Section 4 Using the Detector Baseline Offset #.###mv +5 V -5 V #.#V -#.#V [Continue] [home] Operational Parameters Show OP Parameters? [Next] [Yes] SC/DT/Filter/Gain ##C/##C/XXT#/XG [OC/ET/Cal/LaserPOT ##C/##C/###%/### Set Timeout Functions The Model 2300 has implemented a group of timeout functions. At timeout, it will automatically shut off the gas valve to preserve the gas tank from running out or set the detector into standby which will shut off everything. The timeout timer will be reset by the detector having detected peaks or by user operations on the touch sensitive display. CAUTION The timeout functions may not perform as described. If the function fails, the gas will flow continuously until the source is depleted. Use of this feature is at the user s discretion. Set Timout Func? [Next] [Yes] 4-7

34 Section 4 Using the Detector Disable Timout Func [Next] [Yes] GasCtrlTimout:10 min [Next] [Yes] GasCtrlTimout:30 min [Next] [Yes] GasCtrlTimout:1 hr [Next] [Yes] GasCtrlTimout:2 hr [Next] [Yes] Self-diagnostics Start Self-diagnosis [Next] [Yes] I am not ready [Home] [More] XXX/XXX/XXX/XXX not ready Followed by more details and suggestions for correction. 4-8

35 Section 4 Using the Detector I am ready [Home] [More] flat slope cls pk tr ## # # This screen is used for diagnostics during service. Go Home? [Next] [Yes] Return to Home Return to Home? [Next] [Yes] Reset Baseline Reset Baseline? [Next] [Yes] Select a Method The Model 2300 can store 10 methods. Four are set at factory. Six can be set through computer control software. Select a Method [Next] [Yes] <= UNIVERAL => 10C/65C/FLT2/HG <= HI FLO AQUEOUS => 20C/85C/FLT2/HG 4-9

36 Section 4 Using the Detector <= LOW FLO ORGANIC => 55C/70C/FLT5/LG <= VOLATILES => 10C/45C/BFT1/HG <= USR Method # => ##C/##C/XXT#/XG Enter Standby Enter Standby? [Next] [Yes] Filters The Model 2300 has implemented two sets of filters, the RC filters and baseline filters. RC filters work well in most applications. Baseline filters reduce the filtering strength when the detector has detected a peak to output (such as a peak unaffected by filtering which is higher and narrower for fast chromatography). Baseline filters are not recommended for detecting very wide and slow-changing peaks Start Up Procedure If this is the first time you have operated the detector, it is very important that you reproduce the QC tests found in Installation Qualification Checklists (3.2) before beginning your analysis. 1. Make all connections (gas, liquid, power, communications, exhaust, and drain) as described in Connections on page Turn on the power to the ELSD. Allow the system to run through the start-up sequence, and then touch the screen to exit standby and enter the HOME screen. 3. Select the desired method. If developing a new method, UNIVERAL (10 C/65C/FLT2/HG). 4. Allow the drift tube and spray chamber temperatures to reach thermal equilibrium as indicated by the blinking of the corresponding status lights. 5. While waiting for the temperatures to reach the setpoints, fill the drain. Pump mobile phase at 1 ml/min without gas flow for 10 minutes or until you see liquid coming out of the drain. Alternatively, you may introduce 10 ml of mobile phase into the drain using a squirt bottle or syringe. Stop the pump. 4-10

37 Section 4 Using the Detector 6. When temperatures reach setpoints, turn on the regulated gas flow, and gradually increase the gas pressure to 65 ± 5 psi. If you use an on-off valve to turn on the gas, make sure the upstream gas pressure is below 70 psi before you turn on the valve. Gas pressure higher than 70 psi may permanently damage the detector. Re-start the pump. 7. A * may be blinking at the upper right corner indicating the detector is not ready yet. Briefly touching * will bring up screens to explain why the detector is not ready for detection. 8. When the detector is ready, begin the data collection system and monitor the baseline. 9. Autozero the detector to rest the baseline to approx. 20 mv. Repeated autozeros may be necessary until the baseline stabilizes. 10. When the baseline is stable (variation is less than 1.0 mv) and the * has disappeared, the detector is ready for use. 11. Inject your standard or sample and begin analysis Shut Down Procedure 1. Stop the flow of mobile phase to the system. 2. Allow gas to flow through the detector for at least 5-minutes to remove the remaining vapor. 3. If there are no leaks between your gas source and the ELSD, you do not have to turn the gas off at the source. 4. If the detector is going to be used again the same day or the next morning, turn off the laser from the menu. If the detector will not to be used for more than 24 hours, place the detector in standby by selecting that function from the menu screens or turn the power off. 4-11

38 Section 4 Using the Detector 4-12

39 Section 5 Optimizing Performance 5.1 Optimizing Performance Choosing Operating Conditions Thermo-Split Spray Chamber Temperature The drift tube temperature and the Thermo-Split spray chamber temperature are selected to provide the maximum detector response with minimum baseline noise. The temperatures are selected based on the solvent volatility and mobile phase flowrate. Some experimentation will be required to optimize the ELSD. There are two issues to consider when optimizing operating temperatures: 1. A high drift tube temperature will more effectively evaporate the mobile phase. 2. A lower drift tube temperature will minimize the decomposition of thermally labile compounds and the evaporation of semi-volatile compounds. When setting the ELSD temperatures for a new method, select UNIVERAL (10C/65C/FLT2/HG). These temperatures should then be adjusted for the best signal-to-noise ratio during method optimization. For the best performance, a mobile phase that is highly organic and volatile requires an ambient or elevated spray chamber temperature and moderately high drift tube temperature. When highly aqueous or high boiling point organic mobile phases with the detector, the best performance will be at sub-ambient spray chamber temperatures and moderate drift tube temperatures. The Thermo-Split spray chamber can operate from 10 C to 60 C. The spray chamber temperature controls the vapor phase split ratio. For an easily evaporated mobile phase, the split ratio can be set low. To achieve this, the Thermo-Split chamber is heated. As the aerosol traverses the chamber, it partially evaporates and shifts the particle size distribution low enough for essentially all the particles to negotiate the bend. When highly organic mobile phases are used, the Thermo-Split chamber is used at ambient or elevated temperatures. Under these conditions a majority of the aerosol particles pass through the chamber and are carried into the evaporative zone. For difficult to evaporate mobile phases, or high flow rates, the split ratio needs be high so the Thermo-Split chamber is cooled. When the aerosol exiting the nebulizer encounters a cooled environment, it partially condenses into larger particles whose 5-1

40 Section 5 Optimizing Performance Drift Tube Temperature Mobile Phase Flowrate Gradient Separations Suggested Operating Temperatures momentum carries them into the wall and down the drain. By making the walls suitably cold, 80% of an aqueous stream can be diverted away from the evaporative zone. The drift tube temperature can be set from ambient to 120 C. The drift tube temperature is set at a temperature high enough to evaporate the mobile phase and not vaporize the analyte. A higher drift tube temperature may give result in a quieter baseline but smaller peak. The drift tube temperature should always be higher than the spray chamber temperature but only as high as needed to achieve a quiet baseline. A drift tube temperature setting that is too high could vaporize the analyte and cause a loss of sensitivity. If the analyte is thermally labile, use a lower temperature to improve sensitivity. However, there will be a point where the temperature is not high enough to evaporate the mobile phase and the increase in noise will negate the increase in signal. Optimize for the best signal-to-noise ratio. Mobile phase flowrate will also affect the optimum temperature set point. The higher the flowrate of an aqueous mobile phase, the lower the spray chamber temperature will need to be. High flowrates of volatile mobile phases may require a higher drift tube temperature. The ELSD may perform best at sub-ambient spray chamber temperatures if the flowrate is extremely high even for volatile mobile phases. For gradient separations, select the system temperatures required for the least volatile segment of the gradient program. 1.0 ml/min Drift Tube Temperature, C 90/10 Water/Methanol /10 Methanol/Water /50 Methanol/Water Acetonitrile Methanol Water Spray Chamber Temperature, C Mobile Phase Considerations Selecting a solvent High purity mobile phase solvents with low boiling points are recommended for use with the ELSD. Solvents should be spectral or HPLC grade. Dirty or contaminated solvents will cause baseline noise and drift, blocked fluid paths, and a build up in the detector. All solvents used should have less than 1 ppm of residue after evaporation and be filtered to less than 0.45 µm. Solvents can be evaluated by pumping them directly into the detector and comparing the noise to other known solvents. We have found that not all HPLC grade solvents are acceptable for 5-2

41 Section 5 Optimizing Performance Mobile Phase Flowrate and Composition Buffer Compatibility Column Pre-Treatment use with an ELSD. Preservatives commonly used in Tetrahydrofuran (THF) will increase the noise level. If unstabilized, THF is used. Ensure that it is fresh as THF can contain peroxides that can increase noise and are potentially explosive if taken to dryness. The recommended flowrate for the ELSD is 0.25 ml/min to 3 ml/min. The mobile phase flowrate will affect baseline noise. In general, more baseline noise will be generated by the higher flowrate of a mobile phase. The ELSD will operate with common HPLC solvents that are volatile enough to form a vapor under the operating conditions. This includes common HPLC solvents such as water, methanol, acetonitrile, acetone, isopropyl alcohol, and THF. Normal phase solvents such as dichloromethane and hexane may also be used. Note that solvents with higher boiling points will generally result in more baseline noise. These should be used in limited percentages or at a lower flowrates. The ELSD is not compatible with mobile phase modifiers that are not volatile such as salts. Some modifiers are volatile and can be used. These include, but are not limited to acetic acid, trifluoroacetic aid (TFA), formic acid, triethylamine, and ammonia. The concentration of buffer in the mobile phase should be as low as possible. Chromatographic columns may introduce particles into the mobile phase which may lead to increased noise and blocked fluid paths. It is recommended that the chromatographic column be flushed with at least 10 column volumes before it is connected to the ELSD QC Test Conditions Please refer to the QC report in Installation Qualification Checklists (3.2) for the exact conditions used to test your ELSD. The general conditions are: 1.0 ml/min of 50/50 Water/Methanol Spray chamber 10 C Drift tube 65 C Filter 0 Injected Standard: 1000 ng Sodium Benzoate in water Tuning Examples The SofTA ELSD performs best when the conditions are optimized to provide the best signal-to-noise ratio for the separation. In many applications, the detection limit requirements are high enough that the universal operating conditions, UNIVERAL (10C/65C/FLT2/HG), provides acceptable results. There are applications where optimization of the conditions will improve the results considerable. 1. The mobile phase may be difficult to evaporate, high aqueous or volatile buffer content or high flow rates. 2. The required detection limit is low, 100 ng or lower. 3. The analyte is semi-volatile. 5-3

42 Section 5 Optimizing Performance ELSD Conditions to Optimize Filter Type and Weight Drift Tube Temperature Optimization Please note that the filter should not be used to compensate for poor evaporation of the mobile phase. The spray chamber and drift tube temperatures should be optimized for the best signal-to-noise before a filter is applied. In these examples, raising the drift tube temperature would eliminate all of the noise. The drift tube temperature and the Thermo-Split spray chamber temperature are selected to provide the maximum detector response with minimum baseline noise. The temperatures are selected based on the solvent volatility and mobile phase flow rate. Some experimentation will be required to optimize the ELSD. When setting the ELSD temperatures for a new method, select 15 C for spray chamber temperature and 65 C for drift tube temperature. These temperatures should then be adjusted for the best signal-to-noise ratio during method optimization. For the best performance, a mobile phase that is highly organic and volatile requires an ambient or elevated spray chamber temperature and moderately high drift tube temperature. With highly aqueous or high boiling point organic mobile phases, the best performance will be at sub-ambient spray chamber temperatures and moderate drift tube temperatures. There are two filter types to choose from: RC (FLT) and Baseline (BFT.) Filter weight is the level of baseline noise filtration. OFF indicates no filtration. 10 is maximum filtration. In most cases, select FLT for baseline filtering. The FLT setting applies an RC filter to the entire signal. For high-speed chromatography, less than a 5 second peak width, select BFT 1 or turn the weight to 0. If the peak widths are 5 to 30 seconds, use either the FLT or BFT filter with a weight of 1 to 10. For peak widths greater than 30 seconds, select the FLT setting with a weight of 1 to 10. In this example (Figure 5-1), a difficult to evaporate mobile phase was selected, 1 ml/min water. The spray chamber was set at 15 C and the drift tube was varied. At time 0, the drift tube was 65 C. The drift tube cooled to 65 C by time 5 and returned to 60 after 6 minutes. 5-4

43 Section 5 Optimizing Performance Figure 5-1 This example illustrated how a 5 C temperature change dramatically increases or decreases the baseline noise Spray Chamber Temperature Optimization In this example (Figure 5-2), an easy to evaporate mobile phase was selected, 0.5 ml/min Methanol. The drift tube temperature was held constant at 40 C. The spray chamber temperature was increased from 10 C to 60 C. You can see the effects of reducing the spray chamber temperature in the image below. Figure 5-2 Effects of reducing Spray chamber temperature 5-5

44 Section 5 Optimizing Performance Semi-Volatile Analyte Detection ELSDs have inherent limitations when looking for volatile and semi-volatile analytes. An ELSD functions by evaporating the mobile phase and more volatile analytes tend to evaporate simultaneously with the mobile phase. While the use of an ELSD might not be ideal for these analytes, there are some techniques that an analytical chemist can use to get the best results possible. Generally, the strategy for semi-volatiles is to run the instrument as cold as possible. Recall from previous discussion that the downside of a cold drift tube would be increased noise from poorly evaporated mobile phase. However, signal-to-noise ratio (S/N) is really what an analytical chemist is optimizing. Even though baseline noise might increase with a cold instrument, the signal for the analyte often increases to a greater extent. A drift tube that is even a few degrees colder can sometimes produce significantly better results. The following example (Figure 5-3) shows what happens to 1000 ng Methyl Paraben sensitivity when the drift tube is raised from 55 C to 65 C. Even though baseline noise is somewhat higher at 55 C, S/N is much improved. Figure 5-3 Effect of raising drift tube temperature on Methyl Paraben sensitivity The Model 2300 ELSD also has a very unique way of analyzing for semi-volatile compounds. The spray chamber and drift tube of the Model 2300 have been specifically designed to take full advantage of the unique thermal transfer properties of Helium. Replacing the Nitrogen gas source with Helium can dramatically improve ELSD response to troublesome compounds. In many instances, sensitivity for a semi-volatile analyte can approach that of a non-volatile one. The next example (Figure 5-4) shows sensitivity of a moderate amount of Methyl Paraben (1000 ng) vs. 5-6

45 Section 5 Optimizing Performance Sodium Benzoate, a non-volatile. Nitrogen is used as the nebulizer gas. The mobile phase is 50/50 Methanol/Water at 1 ml/min. Instrument conditions have been optimized at 10/55/FLT1 Figure 5-4 Sensitivity of Methyl Paraben vs. Sodium Benzoate with Nitrogen gas Next (Figure 5-5), the nebulizer gas is changed from Nitrogen to Helium, and the instrument conditions are changed to 10/32/FLT1. Again, 1000 ng injections of Sodium Benzoate are compared to 1000 ng injections of Methyl Paraben. Figure 5-5 Sensitivity of Methyl Paraben vs. Sodium Benzoate with Helium gas 5-7

46 Section 5 Optimizing Performance Filter Type Of course, Helium is more expensive than Nitrogen, and so Helium would not be the best choice for routine analyses. But for those occasions when an ELSD simply must be used for semi-volatile compounds, the Model 2300 in conjunction with Helium can prove very capable. In this example (Figure 5-6) both filter types, Baseline and RC, are compared to no filtering. The mobile phase was 1 ml/min water. A length of wide ID tubing was inserted between the injection valve and the detector to simulate broad peaks. The spray chamber and drift tube temperatures were selected to provide significant noise for the example, SC 30, DT 56. Figure 5-6 Baseline and RC filtering vs. no filtering Filter Weight The baseline filter weight 5 slightly improves the baseline noise translated to the peaks. The RC filter 5 significantly improves the baseline and peak noise. In these examples, the effect of the filter weight for both types is shown. Again, the spray chamber and drift tube temperatures, SC 30, DT 53, were deliberately selected to provide significant noise for the example. The mobile phase was 1 ml/min water. The analyte was 100 ng NaBenzoate. This example (Figure 5-7) shows the Baseline filter at filter 0 and

47 Section 5 Optimizing Performance Figure 5-7 Baseline filter 0 vs 5 The baseline is considerably improved with filter 5. The next example (Figure 5-8) shows the effect of filter weight on the RC or FLT Filter type. The conditions are the same, but the concentration is 1000 ng instead of 100 ng. Figure 5-8 Effect of filter weight on the RC or FLT Filter type This example shows that, while a larger filter weight with RC filter type decreases the noise, it also broadens and decreases the peak heights. 5-9

48 Section 5 Optimizing Performance 5-10

49 Section 6 Troubleshooting 6.1 Troubleshooting Keep in mind that the source of the apparent detector problem may lay within the chromatography or other instruments within the system as well as the detector. Most detector problems are easy to correct. If you are unable to correct a problem, please contact your local Teledyne Isco representative or contact Teledyne Isco directly. To expedite your request for service, please have the following available when you call or Detector serial number Problem symptom(s) Spray chamber and drift tube temperature Mobile phase and flowrate Filter setting System operating pressure System configuration (other equipment in the system) Error Message/Symptom Gas Control Error Gas has been turned off for nebulizer protection Main board stops responding SC temperature is not ready DT temperature is not ready Possible Causes Gas may have been shut off by remote control or auto-protection circuit Gas may have been shut off by auto-protection circuit Corrective Action Check I/O for contact closure Check source pressure Reduce pressure to less than 80 psi Gas may have been shut off Check source pressure by auto-protection circuit Reduce pressure to less than 70 psi Electrical error Turn the detector off, wait 30 sec, then back on Contact Teledyne Isco or your local dealer Spray chamber temperature setting has changed and detector is adapting to new setting. Temperature reading is negative Temperature is not changing Temperature reading is not normal, for example 140 C Drift tube temperature setting has changed and detector is adapting to new setting Wait for thermal equilibrium Contact Teledyne Isco or your local dealer Contact Teledyne Isco or your local dealer Contact Teledyne Isco or your local dealer Wait for thermal equilibrium 6-1

50 Section 6 Troubleshooting OC temperature is not ready ET temperature is not ready Signal level is very low. Please reset the baseline Signal level does not change Baseline level is too high Temperature reading is negative your local dealer Contact Teledyne Isco or Temperature is not changing Contact Teledyne Isco or your local dealer Temperature reading is not Contact Teledyne Isco or normal, for example 140 C your local dealer Drift tube temperature setting has changed and detec- Wait for thermal equilibrium tor is adapting to new setting Temperature reading is negative your local dealer Contact Teledyne Isco or Temperature is not changing Contact Teledyne Isco or your local dealer Temperature reading is not Contact Teledyne Isco or normal, for example 140 C your local dealer Drift tube temperature setting has changed and detec- Wait for thermal equilibrium tor is adapting to new setting Temperature reading is negative your local dealer Contact Teledyne Isco or Temperature is not changing Contact Teledyne Isco or your local dealer Temperature reading is not Contact Teledyne Isco or normal, for example 140 C your local dealer Detector signal has changed Autozero the detector Baseline offset is too high Laser is not operational Mobile phase is not being evaporated. Check detector settings and mobile phase composition Check laser number. If very low, replace laser Check for correct flow and leaks No liquid or gas entering the flow path Detector signal has changed Autozero the detector Detector optics are saturated Stop Mobile phase flow, allow the system to dry with gas flow only. Elevating the drift tube temperature may assist the drying process Gas pressure is too high Gas source is providing too much pressure Reduce supply gas to less than 70 psi Gas pressure is too low Gas source is not providing enough much pressure Increase supply gas to more than 60 psi Gas Pressure is not stable Gas supply is variable at the Check gas supply source Gas pressure has just been changed Wait for pressure to equilibrate Gas has been turned off Gas solenoid is closed Open solenoid via front panel or I/O connection Laser is turned off Laser turned off on front panel or I/O Detector inoperative, power button is not lit Fuse blown Check that the front panel is operational, if not replace No power at the outlet Bad internal power supply of circuit board the fuse Check the outlet by connecting another electrical unit known to be in working order and see if it operates Contact Teledyne Isco or your local dealer 6-2

51 Section 6 Troubleshooting Chromatography Problems Front panel fails to illuminate Fuse blown Contact Teledyne Isco or your local dealer Analog output incorrect Baseline too high or too low Autozero the detector Mobile phase is not being evaporated Decrease the spray chamber temperature and/or increase the drift tube temperature Optical are wet Stop mobile phase flow, increase drift tube temperature, wait until signal decreases Fumes detected Error Message/Symptom Baseline drift Exhaust is not connected or restricted Drain tube is not properly connected Possible Causes Mobile phase mixing inconsistent Ensure the exhaust hose from the detector runs downward, towards the floor and is unobstructed. Properly attach and route the drain tube Corrective Action Correct mobile phase composition or use premixed mobile phase Wait for drift tube temperature to reach equilibrium Drift tube temperature not stable Baseline noise (repetitive) Air in mobile phase or pump Degass mobile phase. Purge pump to remove air Pump pulsations Incorporate a pulse dampener into system. Baseline noise (irregular) Improper volatilization of mobile phase Inconsistent or low gas flow rate Contaminated mobile phase Column leaking packing material or shedding bonded phase Air trapped in system Leaks Correct pump check valves or pump seals problems. Increase backpressure on pump if very low Correct spray chamber and drift tube temperature If correct decrease spray chamber by 5 C and increase drift tube by 5 C Check gas flow setting and gas source Use only high purity HPLC solvents and low concentrations of volatile mobile phase additives Remove column from system If noise disappears, replace column, change column type, and flush thoroughly before connecting to the detector Degas mobile phase and purge air from system Check system for leaks Check pump for leaks or unusual noise 6-3

52 Section 6 Troubleshooting Spiking No gas flow No mobile phase flow No peaks detected Improper volatilization of mobile phase Contaminated mobile phase Gas flow set too low Detector dirty Gas source not flowing or too low I/O connection closing solenoid Blocked nebulizer Gas filter capacity exceeded Error condition triggered I/O to shut off pump Flow interrupted or obstructed Leak Pump not programmed correctly Blocked nebulizer Sample is volatile at detector conditions Sample being retained on column Sample concentration too low Baseline too high Filter value set too high Correct spray chamber and drift tube temperature If correct decrease spray chamber by 5 C and increase drift tube by 5 C Use only high purity HPLC solvents and low concentrations of volatile mobile phase additives Increase gas flow at the source Call Teledyne Isco or your local dealer Check source Remove contact closure to I/O connector Call Teledyne Isco or your local dealer Call Teledyne Isco or your local dealer Clear error Remove contact closure to I/O connector Check mobile phase levels in reservoirs Check flow throughout system Make sure mobile phase inlet filter is clean Check system for loose fittings Check pump for leaks or unusual noise Check pump settings Call Teledyne Isco or your local dealer Check to see if temperatures are set correctly for the mobile phase Modify method to use a more volatile mobile phase. Remove column Connect injector directly to detector and inject into mobile phase Peak should be seen immediately Inject a higher concentration Autozero detector Select a lower filter value 6-4

53 Section 6 Troubleshooting Change in peak height or loss of sensitivity Flow interrupted or obstructed Nebulizer blocked Leak Detector temperature set points have changed Calibration setting has changed Filter value has changed Injected sample has changed Detector output or system input settings have changed Leak Check mobile phase levels in reservoirs Check flow throughout system Make sure mobile phase inlet filter is clean Call Teledyne Isco or your local dealer Check system for loose fittings Check pump for leaks or unusual noise Confirm that detector parameters are correct Confirm that Calibration setting is correct Confirm the filter value is correct Confirm dilution and injection volume are the same Evaluate sample degradation Confirm the output value has not changed Check system for loose fittings especially between column and detector Check pump for leaks or unusual noise Nebulizer not operating properly Call Teledyne Isco or your local dealer Broad peaks Leak Check system for loose fittings especially between column and detector Check pump for leaks or Tubing between column and detector too long or too large an ID unusual noise Use a short piece of to 0.01 ID tubing Straight baseline, no peaks No pump flow Check pump for proper operation No gas flow Check gas source Check to see if detector is in stand by Detector is in stand by Exit stand by mode Detector not zeroed Autozero detector Improper connection between detector and data collection system Check data cable connection No sample injected Leak Check injection and sample vial Check system for loose fittings especially between column and detector Flat topped peaks Baseline is not zeroed Autozero the detector Sample concentration Select low in gain setting, exceeds detector output reduce the CAL setting 6-5

54 Section 6 Troubleshooting Troubleshooting Baseline Noise There are two readings that need to be observed and recorded at each step to determine if the noise is improving. Signal: The top line of the HOME screen will always display actual output voltage in mv. Record the fluctuations as the difference between the largest and smallest number over a 3 minute period. This is the noise level. ##.###mv ##C/##C/FLT#/XG Offset: This number found on the Baseline Offset screen in the operational parameters menu indicates the voltage offset by the autozero. Reset the baseline after each change to rest this number. Baseline Offset #.###mv Troubleshooting Steps First, does the detector indicate it is READY? If there is a * at the upper right corner, touch the ESC key for information regarding the condition of the detector and recommendations to correct errors. Correct the error. See if that solves the baseline noise problem. 1. Turn on the gas and mobile phase. Allow the detector to stabilize. Reset the baseline. Record the Noise level and Offset. 2. Check that there is free flow of vapor out the exhaust tube. There should not be condensed liquid between the detector and the exit of the tube directing the vapor to your exhaust system. 3. Check that there is liquid in the detector "P" trap. Liquid should be coming out of the drain port, never vapor. Fill the trap though the drain if required. 4. Turn the mobile phase off, but leave the gas on and allow the detector to stabilize. Reset the baseline. Record the Noise level and Offset. If the numbers are similar to that in Step 1, continue with the next step. If these numbers are significantly smaller than the numbers from Step 1, the noise is being generated when the mobile phase is not being fully evaporated. See troubleshooting noise generated with mobile phase flowing below for additional steps. 5. Turn off the gas, and allow the detector to stabilize. Reset the baseline. Record the Noise level and Offset. If the numbers are similar, continue to the next step. If these numbers are significantly smaller than the numbers from Step 1, the gas supply may contain moisture or particulates or the gas 6-6

55 Section 6 Troubleshooting Troubleshooting noise generated with mobile phase flowing filter may have reached capacity. Replace the gas supply and gas filter. Deposits inside the detector due to years of use without service or improper use can float inside the detector and be an additional source of noise. If this is the case, contact your local representative for service. The noise can be the result of many sources. Follow the directions below to systematically locate the source of the noise: Column Effects HPLC columns can cause noise for various reasons. 1. Replace the HPLC column with enough tubing to create the required pump backpressure. 2. Allow the detector to stabilize and reset the baseline. 3. Record the Noise level and Offset. Do the numbers decrease significantly from Step 1 above? If so, replace the HPLC column. Detector temperatures not set to evaporate mobile phase 1. Confirm that the detector temperatures are set as recommended. 2. If the noise level and baseline number are still large, try increasing the DT temperature by 5 degree increments until the noise improves. 3. If your analyte is semi-volatile, you may need to reduce the SC temperature instead of raising the DT temperature to eliminate noise to avoid volatilizing the analyte. Mobile phase contains residue upon evaporation 1. Stop the flow of Solvent B. If you pre-mix the mobile phase, replace the pre-mix with bottles of the individual solvents. 2. Allow the detector to stabilize and reset the baseline. 3. Record the Noise level and Offset. 4. Stop the flow of Solvent A and start Solvent B. 5. Allow the detector to stabilize and reset the baseline. 6. Record the Noise level and Offset. Does one of the solvents have larger numbers than the other? If so, that solvent may have contaminates that create residue upon evaporation. Replace the solvent with a higher quality. Mobile Phase additives If the mobile phase contains non-solvent components, and they are equally present in both Solvent A and Solvent B, the test above may not be conclusive. 1. Remove the additives from the solvents. 2. Allow the detector to stabilize and reset the baseline. 3. Record the Noise level and Offset. 6-7

56 Section 6 Troubleshooting Do the numbers decrease significantly? If so, replace the additive with one of a higher quality. Consider LCMS grade additives. ELSD contaminated by deposits of analyte or mobile phase contamination. The ELSD should be serviced once a year by a SofTA qualified technician. During this service, O-ring integrity, drain line condition, and many other parameters relating to safe operation are checked. If the detector has not been serviced regularly, buildup of analyte, mobile phase contamination, or additives can build up in the detector and create significant noise as the mobile phase and vapor dissolve them. Call your local representative for service. 6-8

57 Appendix A Specifications A.1 Specifications Dimensions Weight Display User Interface 7.5 W x 15 D x 12 H 21 lbs 2 line x 20 character per line VFD Touch-sensitive invisible keys Evaporative Zone Temperature Ambient to 120 C Thermo-Split TM spray chamber Temperature Liquid Flow Rate Gas Requirements Gas Consumption Gain Settings 10 C to 60 C 0.2 ml/min to 5 ml/min 65 psi Nitrogen or other inert gas ~3 SLPM Normal, low Operating Conditions Intended for indoor use only, 15 C to 29 C and <90% R.H. non condensing Electrical Requirements Wetted Materials Light Source Detector Output Signal Interface Nominal 120 VAC, 50/60 Hz or nominal 240 VAC, 50/60 Hz; 600 watts Stainless steel, glass (lenses only), anodized aluminum, PTFE 650 nm laser diode, <5 mw Hermetically sealed photo-diode/operational amplifier 0-1 VDC USB, contact closure, software driver (DataApex Clarity) A-1

58 Appendix A Specifications A-2

59 Index A anchored frame instructions, 2-4 assistance, technical, 1-6 B body example, 1-1 instructions, 2-2 bulleted example, 1-1 instructions, 2-2 bulleted2 example, 1-1 instructions, 2-2 C callouts instructions, 2-5 caution example, 1-4 instructions, 2-3 D danger example, 1-4 instructions, 2-3 F figure heading example, 1-5 instructions, 2-5 footnote example, 1-3 H heading 1 example, 1-1 instructions, 2-1 heading 2 example, 1-1 instructions, 2-2 heading runin instructions, 2-2 help, contact information, 1-6 I images, in sidehead area, 2-4 indented instructions, 2-3 indented2 instructions, 2-3 M margin anchor, 2-4 marginalia example, 1-3 instructions, 2-2 N note example, 1-2 instructions, 2-3 numbered example, 1-2 instructions, 2-3 numbered1 example, 1-2 instructions, 2-3 S section example, 1-1 instructions, 2-1 sidehead images, 2-4 small cells example, 1-5 instructions, 2-5 substep example, 1-2 instructions, 2-3 T table example, 1-4 instructions, 2-6 table footnote example, 1-5 technical assistance, 1-6 Title example, 1-1 title instructions, 2-1 Index-1

60 Index W warning example, 1-1 instructions, 2-3 Index-2

61 Compliance Statements CE Declaration of Conformity

62

63 Warranty Component Name LCD Display (none on RF4X) Circuit boards Wiring Internal Cables Line Cord Stepper Motor Deuterium lamp Valve Body Name and amount of Hazardous Substances or Elements in the product (Pb) (Hg) Hazardous Substances or Elements (Cd) (Cr(VI)) (PBB) (PBDE) O O O O O O X O O O O O O O O O X O O O O O X O O O O O X O X O O O X O O O X O O O O O O O X O Name and amount of Hazardous Substances or Elements in the product O: ST/ O: Represent the concentration of the hazardous substance in this component s any homogeneous pieces is lower than the ST/ standard limitation. X ST/ ( X ) X: Represent the concentration of the hazardous substance in this component s at least one homogeneous piece is higher than the ST/ standard limitation. (Manufacturer may give technical reasons to the X marks) The Environmentally Friendly Use Period (EFUP) was determined through experience. ( ) A B The date of Manufacture is in code within the serial number. The first three numbers are the year of manufacture (207 is year 2007) followed by a letter for the month. "A" is January, "B" is February and so on. CombiFlash Hazmat Table Rev. B

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65 Teledyne Isco One Year Limited Factory Service Warranty* This warranty exclusively covers Teledyne Isco instruments, providing a one-year limited warranty covering parts and labor. Any instrument that fails during the warranty period due to faulty parts or workmanship will be repaired at the factory at no charge to the customer. Teledyne Isco s exclusive liability is limited to repair or replacement of defective instruments. Teledyne Isco is not liable for consequential damages. Teledyne Isco will pay surface transportation charges both ways within the 48 contiguous United States if the instrument proves to be defective within 30 days of shipment. Throughout the remainder of the warranty period, the customer will pay to return the instrument to Teledyne Isco, and Teledyne Isco will pay surface transportation to return the repaired instrument to the customer. Teledyne Isco will not pay air freight or customer s packing and crating charges. This warranty does not cover loss, damage, or defects resulting from transportation between the customer s facility and the repair facility. The warranty for any instrument is the one in effect on date of shipment. The warranty period begins on the shipping date, unless Teledyne Isco agrees in writing to a different date. Excluded from this warranty are normal wear; expendable items such as charts, ribbon, lamps, tubing, and glassware; fittings and wetted parts of valves; and damage due to corrosion, misuse, accident, or lack of proper maintenance. This warranty does not cover products not sold under the Teledyne Isco trademark or for which any other warranty is specifically stated. No item may be returned for warranty service without a return authorization number issued by Teledyne Isco. This warranty is expressly in lieu of all other warranties and obligations and Teledyne Isco specifically disclaims any warranty of merchantability or fitness for a particular purpose. The warrantor is Teledyne Isco, 4700 Superior, Lincoln, NE 68504, U.S.A. * This warranty applies to the USA and countries where Teledyne Isco does not have an authorized dealer. Customers in countries outside the USA, where Teledyne Isco has an authorized dealer, should contact their Teledyne Isco dealer for warranty service. Before returning any instrument for repair, please call, fax, or the Teledyne Isco Service Department for instructions. Many problems can often be diagnosed and corrected over the phone, or by , without returning the instrument to the factory. Instruments needing factory repair should be packed carefully, and shipped to the attention of the service department. Small, non-fragile items can be sent by insured parcel post. PLEASE BE SURE TO ENCLOSE A NOTE EXPLAINING THE PROBLEM. Shipping Address: Teledyne Isco - Attention Repair Service 4700 Superior Street Lincoln, NE USA Mailing Address: Teledyne Isco PO Box Lincoln, NE USA Phone: Repair service: (800) (lab instruments) (866) (samplers & flow meters) Sales & General Information: (800) (USA & Canada) Fax: (402) IscoService@teledyne.com February 28, 2012 P/N Rev G