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Agilent Technologies G7121A Скачать руководство пользователя страница 94

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Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual

5

Optimizing the Detector

Method Development

Step 1: Check the LC System for Impurities

A critical issue in trace level fluorescence detection is to have an LC system free
of fluorescent contamination. Most contaminants derive from impure solvents.
Taking a fluorescence scan is a convenient way to check the quality of the solvent
in a few minutes. This can be done, for example, by filling the FLD cuvette
directly with the solvent for an offline measurement even before the start of a
chromatographic run. The result can be displayed as an isofluorescence plot or a
three-dimensional plot. Different colors reflect different intensities.

on page 94 shows a sample of slightly impure water which was

planned for use as mobile phase. The area where fluorescence of the
contaminated water sample can be seen is between the stray light areas: the first-
and second-order Raleigh stray light and Raman stray light.

Since "excitation" and "emission" wavelength are the same for Raleigh stray light,
the area of first-order Raleigh stray light is visible in the left upper area of the
diagram. The Raman bands of water are seen below the first-order Raleigh stray
light. Since the cut-off filter cuts off light below 280 nm, the second-order Raleigh
stray light starts above 560 nm.

Stray light acts in the same way as impurities in that it simulates background
noise. In both cases, a higher noise level and therefore a higher limit of detection
are obtained. This indicates that high sensitivity measurements should be done
away from wavelength settings that have a high stray light background.

A pure water sample
was put into the flow
cell. Spectra were
recorded at 5 nm
step sizes.

Figure 29

Isofluorescence plot of a mobile phase

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Содержание G7121A

Страница 1: ...Agilent Technologies Agilent InfinityLab LC Series 1260 Infinity II Fluorescence Detectors User Manual ...

Страница 2: ...e described in this document are furnished under a license and may be used or copied only in accordance 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 sub contract Software is delivered and licensed as Commercial computer software as defined in DFAR 252 227 7014 June 1995 or as a commercial item as defined in ...

Страница 3: ...formation on environmental requirements physical and performance specifications 3 Using the Fluorescence Detector This chapter explains the essential operational parameters of the module 4 Preparing the Module This chapter provides information on how to set up the module for an analysis and explains the basic settings 5 Optimizing the Detector This chapter provides information on how to optimize t...

Страница 4: ...ntenance of the detector 10 Parts for Maintenance and Repair This chapter provides information on parts for maintenance and repair 11 Identifying Cables This chapter provides information on cables used with the Agilent InfinityLab LC Series modules 12 Hardware Information This chapter describes the detector in more detail on hardware and electronics 13 Appendix This chapter provides safetey and ot...

Страница 5: ...te Requirements and Specifications 41 Site Requirements 42 Physical Specifications 45 Performance Specifications 46 3 Using the Fluorescence Detector 53 Magnets 54 Turn on off 55 Status Indicators 57 Instrument Configuration 58 Set up the Detector with Agilent Open Lab ChemStation 60 The Detector User Interface 61 Detector Control Settings 63 Method Parameter Settings 64 Advanced Settings 65 Acqui...

Страница 6: ...ation 121 Finding the Best Wavelengths 122 Finding the Best Signal Amplification 124 Changing the Xenon Flash Lamp Frequency 130 Selecting the Best Response Time 132 Reducing Stray Light 135 6 Troubleshooting and Diagnostics 137 Available Tests vs User Interfaces 138 Agilent Lab Advisor Software 140 Diagnostic Signals 141 Monitoring of Additional Signals 144 7 Error Information 147 What Are Error ...

Страница 7: ...Maintenance 205 Introduction to Maintenance 206 Warnings and Cautions 207 Overview of Maintenance 209 Cleaning the Module 210 Remove and Install Doors 211 Exchanging a Flow Cell 213 How to use the Cuvette 217 Flow Cell Flushing 218 Correcting Leaks 219 Replace Leak Handling System Parts 221 Replacing Module Firmware 223 Tests and Calibrations 224 10 Parts for Maintenance and Repair 225 Overview of...

Страница 8: ... Connections 247 Interfaces 250 Setting the 6 bit Configuration Switch 258 Instrument Layout 262 Early Maintenance Feedback 263 13 Appendix 265 General Safety Information 266 Waste Electrical and Electronic Equipment WEEE Directive 272 Radio Interference 273 Sound Emission 274 Solvent Information 275 Agilent Technologies on Internet 276 ...

Страница 9: ...ence Detector FLD Spectra 13 How the Detector Operates 15 Raman Effect 18 Optical Unit 19 Reference System 26 Analytical Information From Primary Data 27 Fluorescence Detection 27 Phosphorescence Detection 28 Processing of Raw Data 29 System Overview 32 Operating Principle 32 Leak and Waste Handling 33 Bio inert Materials 38 This chapter gives an introduction to the detector and instrument overvie...

Страница 10: ...n to the Detector Table 1 Detector versions Version Description G7121A Introduced as 1260 Infinity II FLD without spectra and multi signal capabilities Maximum data rate is 74 Hz G7121B SPECTRA Introduced as 1260 Infinity II FLD with spectra and multi signal capabilities Maximum data rate is 148 Hz The G7121B can be converted to G7121A emulation mode ...

Страница 11: ...Infinity II Fluorescence Detector provides highest sensitivity for the analysis of trace level components Time programmable excitation and emission wavelength switching allows you to optimize the detection sensitivity and selectivity for your specific applications High speed detection with up to 74 Hz data rates keeping you pace with the analysis speed of fast LC Figure 1 Overview of the detector ...

Страница 12: ...4 Hz data acquisition rate Long life xenon lamp for highest sensitivity The long life 4000 hours flash lamp lamp reference system and efficient light collection ensure constant lamp energy for maximum excitation of fluorophores Easy front access enables fast inspection or exchange of the flow cell Automatic recognition of all flow cell cartridges provides documentation of instrument parameters and...

Страница 13: ...n to your laboratory This easy to use detector provides quantitative data and fluorescence spectra from a single run Simultaneous multi wavelength detection improves sensitivity and selectivity Use the online spectral information for rapid method optimization and verification of separation quality High speed fluorescence detection with up to 148 Hz data rates keeping pace with the analysis speed o...

Страница 14: ... Simplified optical design for optimized baseline stability Up to 100 resolution gain in fast LC using a 148 Hz data acquisition rate Long life xenon lamp for highest sensitivity The long life 4000 hours flash lamp lamp reference system and efficient light collection ensure constant lamp energy for maximum excitation of fluorophores Easy front access enables fast inspection or exchange of the flow...

Страница 15: ...h with its own excitation process For example when the excitation energy is light the process is called photoluminescence In basic cases the emission of light is the reverse of absorption see Figure 3 on page 15 With sodium vapor for example the absorption and emission spectra are a single line at the same wavelength The absorption and emission spectra of organic molecules in solution produce band...

Страница 16: ...e of the vibrational and rotational sub levels of its ground state emitting light see Figure 4 on page 16 The characteristic maxima of absorption for a substance is its λEX and for emission its λEM Figure 4 Relationship of Excitation and Emission Wavelengths Photoluminescence is the collective name for two phenomena fluorescence and phosphorescence which differ from each other in one characteristi...

Страница 17: ...sphorescence Energy Transitions The molecule must change its spin back again before it can return to its ground state Since the chance of colliding with another molecule with the necessary spin for change is slight the molecule remains in its triplet state for some time During the second spin change the molecule loses more energy by relaxing without radiation The light which is emitted during phos...

Страница 18: ...6 Raman The energy difference between the incident light Ei and the Raman scattered light Es is equal to the energy involved in changing the molecule s vibrational state i e getting the molecule to vibrate Ev This energy difference is called the Raman shift Ev Ei Es Several different Raman shifted signals will often be observed each being associated with different vibrational or rotational motions...

Страница 19: ... excitation condenser lens excitation slit mirror excitation grating flow cell emission condenser lens cut off filter emission slit emission grating and photo multiplier tube are housed in the metal casting inside the detector compartment The fluorescence detector has grating grating optics enabling the selection of both excitation and emission wavelengths The flow cell can be accessed from the fr...

Страница 20: ...al 1 Introduction to the Fluorescence Detector Optical Unit Figure 7 Optical Unit 0 UDWLQJ DVVHPEO 3KRWR 0XOWLSOLHU 7XEH 6OLW 0 UDWLQJ DVVHPEO ORZ HOO 0LUURU 6OLW XWRII ILOWHU 5 LRGH LIIXVHU ODVK ODPS ERDUG HQRQ ODVK ODPS 7ULJJHU 3DFN RQGHQVHU OHQV 0 RQGHQVHU OHQV ...

Страница 21: ...amp can be used until it no longer ignites but the noise level may increase with usage UV degradation especially below 250 nm is significantly higher compared to Visible wavelength range Generally the LAMP ON during run setting or using economy mode will increase lamp life by a magnitude Figure 8 Lamp Energy Distribution vendor data The radiation emitted by the lamp is dispersed and reflected by t...

Страница 22: ...he geometry of the grooves is optimized to reflect almost all of the incident light in the 1st order and disperse it with about 70 efficiency in the ultra violet range Most of the remaining 30 of the light is reflected at zero order with no dispersion Figure 10 on page 23 illustrates the light path at the surface of the grating UDWLQJ LQVLGH 0LUURU ...

Страница 23: ...nge its position and therefore the wavelength during a run For spectra acquisition and multi wavelength detection the grating rotates at 4000 rpm The excitation and emission gratings are similar in design but have different blaze wavelengths The excitation grating reflects most 1st order light in the ultra violet range around 250 nm whereas the emission grating reflects better in the visible range...

Страница 24: ... body Figure 11 Cross Section of Flow Cell The luminescence from the sample in the flow cell is collected at right angles to the incident light by a second lens and passes through a second slit Before the luminescence reaches the emission monochromator a cut off filter removes light below a certain wavelength to reduce noise from 1st order scatter and 2nd order stray light see Figure 10 on page 23...

Страница 25: ...finally so many electrons are generated that a current can be measured The amplification is a function of the voltage at the dynodes and is microprocessor controlled You can set the amplification using the PMTGAIN function Figure 12 Photo multiplier Tube This type of so called side on photo multiplier is compact ensuring fast response conserving the advantages of the short optical path shown in Fi...

Страница 26: ... the excitation EX light transmitted by the flow cell and corrects flash lamp fluctuations and long term intensity drift Because of a non linear output of the diode depending on the EX wavelength the measured data are normalized A diffuser is located in front of the reference diode see Figure 7 on page 20 This diffuser is made of quartz reduces light and allows integral measurement of the light ...

Страница 27: ...stry of your application the luminescence measured by the fluorescence detector will have different characteristics You must decide using your knowledge of the sample what mode of detection you will use Fluorescence Detection When the lamp flashes the fluorescing compounds in your sample will luminesce almost simultaneously see Figure 13 on page 27 The luminescence is short lived therefore the flu...

Страница 28: ...r Analytical Information From Primary Data Phosphorescence Detection An appropriate parameter set will be specified as soon as you chose the phosphorescence detection mode special setpoints under FLD parameter settings Figure 14 Measurement of Phosphorescence QWHQVLW 0HDVXUHPHQW 7LPH VHF ODVK 3KRVSKRUHVFHQFH ...

Страница 29: ...uorescence and Phosphorescence You can improve the signal to noise characteristics by disabling the economy mode The data resolution is 20 bit at a response time of 4 s default which is equivalent to a time constant of 1 8 s and appropriate for standard chromatographical conditions Weak signals may cause errors in quantification because of insufficient resolution Check your proposed PMTGAIN If it ...

Страница 30: ...verted and multiplied into an electronic signal the signal at present analog is tracked and held beyond the photo multiplier After being held the signal is converted by an A to D converter to give one raw data point digital Eleven of these data points are bunched together as the first step of data processing Bunching improves your signal to noise ratio The bunched data shown as larger black dots i...

Страница 31: ... Manual 31 Introduction to the Fluorescence Detector 1 Analytical Information From Primary Data Figure 17 RESPONSETIME Signal to Noise Ratio SRLQWV 5 63216 7 0 JLYHV SRLQWV SHU ER FDU ILOWHU GDWD SRLQWV R FDU LOWHUHG LJK 6 1 UDWR XQFKHG GDWD 6PDOO 6 1 UDWLR ...

Страница 32: ...32 Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 1 Introduction to the Fluorescence Detector System Overview System Overview Operating Principle Figure 18 Optical path of the FLD ...

Страница 33: ...he Internet All leak plane outlets are situated in a consistent position so that all Infinity and Infinity II modules can be stacked on top of each other Waste tubes are guided through a channel on the right hand side of the instrument keeping the front access clear from tubes The leak plane provides leak management by catching all internal liquid leaks guiding them to the leak sensor for leak det...

Страница 34: ...34 Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 1 Introduction to the Fluorescence Detector System Overview Figure 19 Infinity II Leak Waste Concept Flex Bench installation ...

Страница 35: ...Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 35 Introduction to the Fluorescence Detector 1 System Overview Figure 20 Infinity II Single Stack Leak Waste Concept bench installation ...

Страница 36: ...ncept bench installation The waste tube connected to the leak pan outlet on each of the bottom instruments guides the solvent to a suitable waste container Leak Sensor CAUTION Solvent incompatibility The solvent DMF dimethyl formamide leads to corrosion of the leak sensor The material of the leak sensor PVDF polyvinylidene fluoride is incompatible with DMF Do not use DMF ...

Страница 37: ...he Fluorescence Detector 1 System Overview Waste Concept 1 Agilent recommends using the 6 L waste can with 1 Stay Safe cap GL45 with 4 ports 5043 1221 for optimal and safe waste disposal If you decide to use your own waste solution make sure that the tubes don t immerse in the liquid ...

Страница 38: ... which are widely accepted by life science scientists as they are known for optimum inertness to biological samples and ensure best compatibility with common samples and solvents over a wide pH range Explicitly the complete flow path is free of stainless steel and free of other alloys containing metals such as iron nickel cobalt chromium molybdenum or copper which can interfere with biological sam...

Страница 39: ...D G7165A PEEK ceramic sapphire PTFE Bio inert flow cell 8 µL 20 bar pH 1 12 includes Capillary Kit Flow Cells BIO p n G5615 68755 G5615 60005 for Agilent 1260 Infinity II FLD G7121A B PEEK fused silica PTFE Bio inert Heat Exchangers Valves and Capillaries Quick Connect Heat Exchanger Bio inert G7116 60041 for Agilent 1260 Infinity II Multicolumn Thermostat G7116A PEEK steel cladded Bio inert Valve...

Страница 40: ...40 Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 1 Introduction to the Fluorescence Detector Bio inert Materials ...

Страница 41: ...chnologies 2 Site Requirements and Specifications Site Requirements 42 Physical Specifications 45 Performance Specifications 46 Specifications 46 Specification Conditions 51 This chapter provides information on environmental requirements physical and performance specifications ...

Страница 42: ...sult if the devices are connected to a line voltage higher than specified Connect your instrument to the specified line voltage only WARNING Electrical shock hazard The module is partially energized when switched off as long as the power cord is plugged in The cover protects users from personal injuries for example electrical shock Do not open the cover Do not operate the instrument and disconnect...

Страница 43: ...nded purposes can lead to personal injury or damage of electronic equipment Never use a power cord other than the one that Agilent shipped with this instrument Never use the power cords that Agilent Technologies supplies with this instrument for any other equipment Never use cables other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC...

Страница 44: ...ed to bear the weight of all modules The module should be operated in a horizontal position Condensation NOTE Agilent recommends that you install the HPLC instrument in the InfinityLab Flex Bench rack This option helps to save bench space as all modules can be placed into one single stack It also allows to easily relocate the instrument to another Lab CAUTION Condensation within the module Condens...

Страница 45: ...Line voltage 100 240 V 10 Wide ranging capability Line frequency 50 or 60 Hz 5 Power consumption 70 VA 60 W Ambient operating temperature 4 40 C 39 104 F Ambient non operating temperature 40 70 C 40 158 F Humidity 95 r h at 40 C 104 F Non condensing Operating altitude Up to 3000 m 9842 ft Non operating altitude Up to 4600 m 15092 ft For storing the module Safety standards IEC EN CSA UL Installatio...

Страница 46: ... capabilities and spectral data analysis Single wavelength operation RAMAN H2O 500 noise reference measured at signal Ex 350 nm Em 397 nm dark value 450 nm standard flow cell RAMAN H2O 3000 noise reference measured at dark value Ex 350 nm Em 397 nm dark value 450 nm standard flow cell Dual wavelength operation RAMAN H2O 300 Ex 350 nm Em 397 nm standard flow cell RAMAN H2O 300 Ex 350 nm Em 450 nm s...

Страница 47: ...eters Spectrum acquisition Excitation or Emission spectra Scan speed 28 ms per datapoint e g 0 6 s spectrum 200 400 nm 10 nm step Step size 1 20 nm Spectra storage All Wavelength characteristic Repeatability 0 2 nm Accuracy 3 nm setting Flow cells Standard 8 µL volume and 20 bar 2 MPa pressure maximum fused silica block Optional Bio inert 8 µL volume and 20 bar 2 MPa pressure maximum pH 1 12 Micro...

Страница 48: ...ions Controller area network CAN USB ERI ready start stop and shut down signals Safety features and maintenance Leak detection safe leak handling leak output signal for shutdown of the pumping system No hazardous voltages in major maintenance areas Extensive diagnostics error detection and display with Agilent Lab Advisor software GLP features Early maintenance feedback EMF for continuous tracking...

Страница 49: ...alue 450 nm standard flow cell RAMAN H2O 3000 noise reference measured at dark value Ex 350 nm Em 397 nm dark value 450 nm standard flow cell Light source Xenon Flash Lamp normal mode 20 W economy mode 5 W lifetime 4000 h Pulse frequency 296 Hz for single signal mode 74 Hz for economy mode Maximum data rate 74 Hz Excitation monochromator Range settable 200 nm 1200 nm and zero order Bandwidth 20 nm...

Страница 50: ...ted software versions refer to the compatibility matrix of your version of the LC and CE Drivers Communications Controller area network CAN USB ERI ready start stop and shut down signals Safety features and maintenance Leak detection safe leak handling leak output signal for shutdown of the pumping system No hazardous voltages in major maintenance areas Extensive diagnostics error detection and di...

Страница 51: ...nfinity II FLD User Manual 51 Site Requirements and Specifications 2 Performance Specifications Specification Conditions Standard flow cell Standard Photomultiplier Using Agilent Lab Advisor see Raman ASTM Signal to Noise Test on page 173 ...

Страница 52: ...52 Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 2 Site Requirements and Specifications Performance Specifications ...

Страница 53: ...nt Configuration 58 Set up the Detector with Agilent Open Lab ChemStation 60 The Detector User Interface 61 Detector Control Settings 63 Method Parameter Settings 64 Advanced Settings 65 Acquire Spectra G7121B Only 68 Special Settings 69 Time Table 72 Instrument Curves 73 Agilent Local Control Modules 74 This chapter explains the essential operational parameters of the module ...

Страница 54: ...54 Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 3 Using the Fluorescence Detector Magnets Magnets 1 This stack exemplarily shows the magnets positions in the modules ...

Страница 55: ...t InfinityLab LC Series 1260 Infinity II FLD User Manual 55 Using the Fluorescence Detector 3 Turn on off Turn on off 1 2 Power switch On 3 Turn instrument On Off with the control software 4 Power switch Off ...

Страница 56: ...56 Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 3 Using the Fluorescence Detector Turn on off 5 ...

Страница 57: ...ditions Status indicators 1 Idle 2 Run mode 3 Not ready Waiting for a specific pre run condition to be reached or completed 4 Error mode interrupts the analysis and requires attention for example a leak or defective internal components 5 Resident mode blinking for example during update of main firmware 6 Bootloader mode fast blinking Try to re boot the module or try a cold start Then try a firmwar...

Страница 58: ...tion 1 Set the switches of the Configuration switch at the rear of the module a All switches DOWN module uses the default IP address 192 168 254 11 b Switch 4 UP and others DOWN module uses DHCP c Switch 5 UP and others DOWN modules uses STORED address 2 Enter the setup information MAC IP address and or Instrument Name a Agilent OpenLab ChemStation Configure Instrument ...

Страница 59: ...scence Detector 3 Instrument Configuration b Lab Advisor Instrument Overview Add Instrument Instrument Configuration 1 Enter the setup information MAC IP address and or Instrument Name a Agilent OpenLab ChemStation Configure Instrument b Lab Advisor Instrument Overview Add Instrument ...

Страница 60: ...wn with the Agilent OpenLab ChemStation C 01 07 and Driver A 02 14 Figure 22 ChemStation Method and Run Control just detector is shown After successful load of the OpenLab ChemStation you should see the module as an active item in the graphical user interface GUI NOTE This section describes the detector settings only For information on the Agilent OpenLab ChemStation or other 1200 Infinity modules...

Страница 61: ... will change 1 Lamp turn on and off of UV lamp 2 EMF status 3 Detector status 4 Lamp status on off EMF Status shows Run Ready Error state and Not Ready text or Error text Offline gray Ok No Maintenance required green EMF warning Maintenance might be required yellow EMF warning Maintenance required red Important The EMF settings can be accessed via Agilent Lab Advisor The limit s can be changed Bas...

Страница 62: ... Idle green Offline dark gray Standby light gray A right click into the Active Area will open a menu to Show the Control Interface special module settings Show the Method interface similar as via menu Instrument Setup Instrument Method Set Error Method Identify Module Status LED will blink Perform a Balance Switch the lamp on off same as click on button Make Device Ready Turn device off standby NO...

Страница 63: ...ol Settings The figure shows the default settings Lamps can be turned ON OFF Response Multiply Factor 1 Analog Output Range can be set to either 100 mV or 1 Vfull scale for additional settings see Analog Output under Method Parameter Settings on page 64 At Power On automatic lamp on at power on Automatic Turn On automatic detector power on ...

Страница 64: ...the Fluorescence Detector Method Parameter Settings Method Parameter Settings These settings are available via Menu Instrument Set up Instrument Method or via right click into the module s active area does not show the Instrument Curves tab Figure 23 Method parameter settings ...

Страница 65: ...ia Mulitiple Wavelength mode G721B ONLY Zero Order Ex The full spectrum of light from the Xenon lamp illuminates the flow cell Each compound can absorb its characteristic wavelength of light and then emit maximum fluorescence An increased stray light level is inherent in this setting and this will decrease sensitivity signal to noise Zero Order Em Zero order sets the monochromator so that all ligh...

Страница 66: ...90 of the output signal in response to an input step function Limits When you set the peak width in minutes the corresponding response time is set automatically and the appropriate data rate for signal and spectra acquisition is selected as shown Stoptime Posttime The Stoptime is the time where either the complete system stops As Pump Injector or the module if different from system stop time The d...

Страница 67: ...ntrol Settings Zero Offset 1 99 in steps of 1 5 equal to 50 mV5 Attenuation 0 195 1600 LU at discrete values for either 100 mV100 or 1 V full scale Multiple Wavelengths G7121B ONLY You can choose to use three additional wavelengths either excitation or emission wavelengths in your analysis Choose Off to switch off multiple wavelengths or Multi Ex or Multi Em to set up multiple wavelengths See also...

Страница 68: ...n time rate The acquisition time in ms for a single spectrum under the specified conditions is calculated and displayed The acquisition rate depends on the number of additional signals specified the scan range of spectral acquisition the step size wavelength resolution Fit Spectral Range Select Fit Spectral Range to adjust the spectral range so that there is always a difference of at least 25 nm b...

Страница 69: ...ch on the Phosphorescence detection mode the two parameters Delay and Gate are activated The luminescence that you measure with the fluorescence detector has different characteristics depending on the chemistry of your application The characteristic of your sample determines whether you need to use fluorescence detection mode or phosphorescence detection mode Delay Sets a waiting period during whi...

Страница 70: ... of data points per spectrum The time per scan in seconds is calculated automatically and displayed The Lamp section allows you to specify how the lamp is used in an analysis Select Only On During Run to specify that the lamp is switched on only during the run time and is switched off when the run finishes Select Economy Mode to run the lamp in a low frequency low current mode This increases the l...

Страница 71: ...length or PMT gain Append Resets the baseline to the previous baseline position Free Makes no adjustment to the baseline position you may see a shift in baseline position Zero Resets the baseline to the baseline position at the start of the run Note that this may not be 0 LU Luminescence Units depending on the analysis conditions Signal Polarity Select the Signal Polarity you expect from your data...

Страница 72: ...ou may set up time events to change functions with their parameters over the run time Add lines as required Time Limits 0 00 99999 00 min in steps of 0 01 min Via the buttons in the bottom area time table lines can be added removed cut copied pasted or completely cleared Based on the chosen function a certain parameter can be selected ...

Страница 73: ...l that can be used for diagnosing problems These can be baseline problems deriving from lamp wander drift problems due to temperature changes This signals can be used in addition to the normal baseline signal to determine whether correlation to temperature of the lamp These signals are available via the Agilent ChemStation Online Plot Data Signal and or Agilent Lab Advisor Software ...

Страница 74: ...nion gives you maximum flexibility and ease of use to control and monitor your LC system modules Features Complete local control and monitoring of Agilent Infinity II Prime LC Modules Excellent usability and ease of use through a user interface specifically tailored for mobile devices simple intuitive touch enabled and visual controllable High flexibility through a modern Bring your own device app...

Страница 75: ...inity system or a single module from a single point However not for Agilent 1220 Compact LC Mixed system configurations supported e g 1200 Series 1200 Series SL and 1100 Series Excellent readability and usability by large colored display with background light high resolution and contrast Convenient ergonomic operation either handheld or at the stack with newly developed secure attachment Handheld ...

Страница 76: ...76 Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 3 Using the Fluorescence Detector Agilent Local Control Modules ...

Страница 77: ...D User Manual Agilent Technologies 4 Preparing the Module Leak and Waste Handling 78 Waste Concept 79 Before You Start 80 Solvent Information 81 This chapter provides information on how to set up the module for an analysis and explains the basic settings ...

Страница 78: ...perature in the sample compartment at least 25 K below the boiling point of the solvent used Do not operate the instrument in an explosive atmosphere Do not use solvents of ignition Class IIC according IEC 60079 20 1 for example carbon disulfide Reduce the volume of substances to the minimum required for the analysis Never exceed the maximum permissible volume of solvents 8 L in the solvent cabine...

Страница 79: ...the Module 4 Leak and Waste Handling Waste Concept 1 Agilent recommends using the 6 L waste can with 1 Stay Safe cap GL45 with 4 ports 5043 1221 for optimal and safe waste disposal If you decide to use your own waste solution make sure that the tubes don t immerse in the liquid ...

Страница 80: ...esults most of the time But experience shows that baseline noise can be higher lower signal to noise ratio when impurities are in the solvents Flush your solvent delivery system for at least 15 minutes before checking sensitivity If your pump has multiple channels you should also flush the channels not in use For optimal results refer to Optimizing the Detector on page 87 ...

Страница 81: ...r typical HPLC conditions For any special condition please consult the material information section or contact Agilent Disclaimer Subsequent data was collected from external resources and is meant as a reference Agilent cannot guarantee the correctness and completeness of such information Data is based on compatibility libraries which are not specific for estimating the long term life time under s...

Страница 82: ...t uses stainless steel cladded PEEK capillaries in bio inert systems The use of stainless steel cladded PEEK capillaries keeps the flow path free of steel and ensures pressure stability to at least 600 bar If in doubt consult the available literature about the chemical compatibility of PEEK Polyimide Agilent uses semi crystalline polyimide for rotor seals in valves and needle seats in autosamplers...

Страница 83: ...li halides their respective acids for example lithium iodide potassium chloride and so on and aqueous solutions of halogens High concentrations of inorganic acids like nitric acid sulfuric acid and organic solvents especially at higher temperatures replace if your chromatography method allows by phosphoric acid or phosphate buffer which are less corrosive against stainless steel Halogenated solven...

Страница 84: ...ion in anhydrous methanol which can be avoided by adding a small amount of water about 3 Slight corrosion is possible with ammonia 10 Diamond Like Carbon DLC Diamond Like Carbon is inert to almost all common acids bases and solvents There are no documented incompatibilities for HPLC applications Fused silica and Quartz SiO2 Fused silica is used in Max Light Cartridges Quartz is used for classical ...

Страница 85: ...DD copolymer tubings which are used in all Agilent degassers except G1322A are not compatible with fluorinated solvents like Freon Fluorinert or Vertrel They have limited life time in the presence of Hexafluoroisopropanol HFIP To ensure the longest possible life with HFIP it is best to dedicate a particular chamber to this solvent not to switch solvents and not to let dry out the chamber For optim...

Страница 86: ...86 Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 4 Preparing the Module Solvent Information ...

Страница 87: ...ization for Multiple Compounds 108 Setting the Chromatographic Conditions 108 Evaluating The System Background 114 How to collect spectra with modes SPECTRA ALL IN PEAK and APEX SPECTRA ONLY 116 Optimization Overview 119 Design Features Help Optimization 121 Check Performance Before You Start 121 Finding the Best Wavelengths 122 A Real Example 123 Finding the Best Signal Amplification 124 FLD Scal...

Страница 88: ... which takes about 5 minutes 3 You are now ready to change the settings of your detector NOTE Some features for example spectrum acquisition multi wavelength detection described in this chapter are not available on the G1321C FLD When If you want to checkout the detector Parts required p n Description 1 5063 6528 Start up Kit includes 1 LC Column and parts listed below 1 01080 68704 Agilent isocra...

Страница 89: ... Table 6 Chromatographic Conditions Mobile phases A water 35 B Acetonitrile 65 Column OSD Hypersil column 125 mm x 4 mm i d with 5 µm particles Sample Isocratic standard sample 1 10 diluted in methanol Flow rate 1 5 ml min Compressibility A water 46 Compressibility B Acetonitrile 115 Stroke A and B auto Stop time 4 min Injection volume 5 µl Oven temperature 1200 30 C FLD Excitations Emission Wavel...

Страница 90: ... 5 Optimizing the Detector Getting Started and Checkout 2 Set the FLD setpoints according to Figure 26 on page 90 In this example additional excitation wavelenghts B C D are used This will increase the scan time and may lower the performance Figure 26 FLD Parameters ...

Страница 91: ...er Manual 91 Optimizing the Detector 5 Getting Started and Checkout 3 Start the run The resulting chromatograms are shown below Figure 27 Biphenyl Peak With Different Excitation Wavelengths The excitation maxima are around 250 nm QP QP QP QP LSKHQ O SHDN ...

Страница 92: ...r Manual 5 Optimizing the Detector Getting Started and Checkout Observe the Maxima via the Isoabsorbance Plot 1 Load the data file λEX 246 nm λEM 317 nm and open the isoabsorbance plot 2 The maximum λEX will be found around 250 nm Figure 28 Isoabsorbance Plot ...

Страница 93: ... how to benefit from the operation modes during these steps Table 7 Steps for thorough method development Step 1 Check system Step 2 Optimize limits of detection and selectivity Step 3 Set up routine methods Fluorescence scan Find impurities for example in solvents and reagents Determine simultaneously the excitation and emission spectra of a pure compound Signal mode Perform wavelength switching ...

Страница 94: ...The area where fluorescence of the contaminated water sample can be seen is between the stray light areas the first and second order Raleigh stray light and Raman stray light Since excitation and emission wavelength are the same for Raleigh stray light the area of first order Raleigh stray light is visible in the left upper area of the diagram The Raman bands of water are seen below the first orde...

Страница 95: ... that is similar to the fluorescence excitation spectrum see Figure 30 on page 95 and to record the emission spectrum Then with an optimum emission wavelength determined the excitation spectrum is acquired These tasks have to be repeated for each compound using either a fluorescence spectrophotometer or stop flow conditions in LC Usually each compound requires a separate run As a result a set of e...

Страница 96: ...e differences expected in optical design between an LC detector and a dedicated fluorescence spectrophotometer or even between detectors These differences can lead to variations for the optimum excitation and emission wavelengths The Agilent 1260 Infinity Fluorescence Detector offers a fluorescence scan that delivers all spectral information previously obtained with a standard fluorescence spectro...

Страница 97: ...nfinityLab LC Series 1260 Infinity II FLD User Manual 97 Optimizing the Detector 5 Method Development Figure 31 Characterization of a pure compound from a fluorescence scan QP QP QP VWUD OLJKW RUGHU D LV P D LV ...

Страница 98: ...velength Most fluorophores show strong absorption at these wavelengths and the quantum yield is high Excitation is sufficient for collecting emission spectra Table on page 99 contains all emission spectra obtained in a single run from a mix of 15 PNAs This set of spectra is used to set up a timetable for optimum emission wavelengths for all compounds The individual compound spectra in the isofluor...

Страница 99: ... 4 ml min Column temperature 18 C Injection volume 5 µl FLD settings PMT 12 response time 4 s step size 5 nm This shows the isofluorescence plot of emission spectra for 15 PNAs 5 µg ml with a fixed excitation wavelength 260 nm Figure 32 Optimization of the time program for the emission wavelength P VSHFWUD IL HG 8 7LPH PLQ 1DSKWKDOHQH FHQDSKWKHQH OXRUHQH 3KHQDQWKUHQH QWKUDFHQH OXRUDQWKHQH 3 UHQH K...

Страница 100: ...are summarized in Table 10 on page 100 This timetable gives the conditions for optimum detection based on the results of two chromatographic runs Table 10 Timetable for the analysis of 15 polynuclear aromatic hydrocarbons Time min Exitation Wavelength nm Emission Wavelength nm 0 260 350 8 2 260 420 19 0 260 500 1DSKWKDOHQH FHQDSKWKHQH OXRUHQH 3KHQDQWKUHQH QWKUDFHQH OXRUDQWKHQH 3 UHQH HQ D DQWKUDFH...

Страница 101: ...tra available from the diode array detector the fluorescence detector is set to acquire emission spectra with a fixed excitation wavelength in the low UV range The example is taken from the quality control of carbamates Samples are analyzed for the impurities 2 3 diaminophenazine DAP and 2 amino 3 hydroxyphenazine AHP Reference samples of DAP and AHP were analyzed with diode array and fluorescence...

Страница 102: ...m in a second run shows the equivalence of UV spectra and fluorescence excitation spectra An excitation wavelength at 265 nm was used for taking the emission spectrum and an emission wavelength at 540 nm was used for taking the excitation spectrum Figure 34 UV spectrum and fluorescence spectra for 2 3 diaminophenazine DAP 89 VSHFWUD DYHOHQJWK QP 1RUP PLVVLRQ FLWDWLRQ ...

Страница 103: ...e analysis of MBC 2 benzimidazole carbamic acid methylester and impurities DPLQR 2 SKHQD LQH GLDPLQRSKHQD LQH 6WDQGDUG 7LPH PLQ 8QNQRZQ Table 11 Conditions for analysis of DAP and MBC according to figures above Column Zorbax SB 2 x 50 mm PNA 5 µm Mobile phase A water B acetonitrile Gradient 0 minutes 5 10 minutes 15 Flow rate 0 4 ml min Column temperature 35 C Injection volume 5 µl FLD settings PM...

Страница 104: ...alysis Such switching is difficult if compounds elute closely and require a change in excitation or emission wavelength Peaks can be distorted and quantitation made impossible if wavelength switching occurs during the elution of a compound Very often this happens with complex matrices influencing the retention of compounds In spectral mode the FLD can acquire up to four different signals simultane...

Страница 105: ...rate 0 4 mL min Column temperature 22 C Injection volume 2 µL FLD settings PMT 12 response time 4 s The upper trace was received with traditional wavelength switching Figure 36 Simultaneous multi wavelength detection for PNA analysis HQ D DQWKUDFHQH KU VHQH HQ R E IOXRUDQWKHQH HQ R N IOXRUDQWKHQH HQ D S UHQH LEHQ R D K DQWKUDFHQH HQ R J K L SHU OHQH QGHQR FG S UHQH 1DSKWKDOHQH FHQDSKWKHQH OXRUHQH ...

Страница 106: ...s this principle using a PNA analysis The match factor given in the report for each peak indicates the degree of similarity between the reference spectrum and the spectra from a peak A match factor of 1 000 means identical spectra In addition the purity of a peak can be investigated by comparing spectra obtained within a single peak When a peak is calculated to be within the user defined purity li...

Страница 107: ...anthrene em 8 841 8 800 9 328 1 1 76064e 1 1 993 Anthracene em 9 838 10 000 10 353 1 2 15360e 1 1 997 Fluoranthene em 10 439 10 400 10 988 1 8 00754e 2 1 1000 Pyrene em 12 826 12 800 13 469 1 1 40764e 1 1 998 Benz a anthracene em 13 340 13 300 14 022 1 1 14082e 1 1 999 Chrysene em 15 274 15 200 16 052 1 6 90434e 1 1 999 Benzo b fluoranthene em 16 187 16 200 17 052 1 5 61791e 1 1 998 Benzo k fluora...

Страница 108: ...ographic conditions the detector settings are shown in Figure 37 on page 109 Table 14 Chromatographic Conditions Mobile phases A water 50 B Acetonitrile 50 Column Vydac C18 PNA 250 mm x 2 1 mm i d with 5 µm particles Sample PAH 0 5 ng Flow rate 0 4 ml min Compressibility A water 46 Compressibility B Acetonitrile 115 Stroke A and B auto Time Table at 0 min B 50 at 3 min B 60 at 14 5 min B 90 at 22 ...

Страница 109: ...ization for Multiple Compounds Select a Excitation wavelength in the low UV 230 260 nm This will cover nearly all fluorescence in your sample DO NOT select additional emission wavelengths B C D Doing so will increase the scan time and will lower the performance Figure 37 Detector Settings for Emission Scan ...

Страница 110: ...ompounds 1 Wait until the baseline stabilizes Complete the run 2 Load the signal In this example just the time range of 13 min is displayed Figure 38 Chromatogram from Emissions Scan 3 Use the isoabsorbance plot and evaluate the optimal emission wavelengths shown in the table below Figure 39 Isoabsorbance Plot from Emission Scan ...

Страница 111: ...cond run for the evaluation of the optimal excitation wavelength See Figure 40 on page 111 Table 15 Peak Time Emission Wavelength 1 5 3 min 330 nm 2 7 2 min 330 nm 3 7 6 min 310 nm 4 8 6 min 360 nm 5 10 6 min 445 nm 6 11 23 min 385 nm DO NOT select additional excitation wavelengths B C D Doing so will increase the scan time and will lower the performance Figure 40 Detector Settings for Excitation ...

Страница 112: ... the run 6 Load the signal Figure 41 Chromatogram Excitation Scan at Reference Wavelength 260 330 nm 7 Use the isoabsorbance plot and evaluate the optimal excitation wavelengths in this example just in the time range of 13 minutes Figure 42 Isoabsorbance Plot Excitation The table below shows the complete information about emission from Figure 39 on page 110 and excitation maxima ...

Страница 113: ... the Detector 5 Example Optimization for Multiple Compounds Table 16 Peak Time Emission Wavelength Excitation Wavelength 1 5 3 min 330 nm 220 280 nm 2 7 3 min 330 nm 225 285 nm 3 7 7 min 310 nm 265 nm 4 8 5 min 360 nm 245 nm 5 10 7 min 445 nm 280 nm 6 11 3 min 385 nm 270 330 nm ...

Страница 114: ...solvent through your system 2 Set the fluorescence scan range under FLD special setpoints according to your needs 3 Set PMT gain to 16 NOTE The scan time will increase when the range is enlarged With the default values the scan takes about 2 minutes The wavelength range and step number defines the duration Using the maximum range the scan would take approximately 10 minutes Figure 43 FLD Special S...

Страница 115: ...e and take a fluorescence scan After the scan is completed the isoabsorbance scan results appear see Figure 44 on page 115 Figure 44 Fluorescence Scan of Water NOTE A low background will improve the signal to noise see also Reducing Stray Light on page 135 1RWH 7KLV ZKLWH DUHD LV QRUPDOO GDUN EOXH 5DOHLJK VFDWWHULQJ 5DPDQ VFDWWHULQJ RI ZDWHU QG RUGHU RI FXWRII ...

Страница 116: ... CDS with the Fluorescence Detector G7121B In these modes spectra intermittently are not collected into the data file The peak triggered spectra acquisition in the FLD is controlled by two parameters THRS Threshold and PDPW PeakDetector PeakWidth In addition the parameter PKWD Detector PeakWidth only influences the filtering of the chromatogram 1 Set the parameters THRS PDPW and PKWD accordingly t...

Страница 117: ...een there are 2 fields to enter the PKWD Peakwidth Responsetime and the THRS Threshold visible when Multi EX or Multi EM is selected Defaults are PKWD 6 0 2 min THRS 5 000 LU The selected values are fixed during the run THRS and PDPW influence the peak triggered spectra acquisition You can change THRS in the FLD s setup screen PDPW can only be changed with the Peakwidth field in the Timetable ...

Страница 118: ...meter The peak detector works online on the current chromatogram This means that begin apex end of a peak is recognized with delay Additionally the points of spectra are sequentially acquired This means that the acquisition of wide range spectra lasts much longer than the acquisition of a short range spectrum When you have a fast chromatography it is nearly impossible to collect a clean APEX spect...

Страница 119: ...ponse time For most applications a setting of 4 seconds is adequate see Selecting the Best Response Time on page 132 Only for high speed analyses short columns at high flow rates a lower setting is recommended Bear in mind that even if the response time is too high fast peaks will appear a little smaller and broader but retention time and peak areas are still correct and reproducible NOTE Some fea...

Страница 120: ...e spectrum on the baseline is very noisy no light 5 Switching lamp ON only for analysis Unless maximum sensitivity is needed the lamp lifetime can significantly be increased by switching it on just for analysis In contrast to other LC detectors the fluorescence detector equilibrates within seconds after the lamp is switched ON 6 Do not overpressurize the detector flow cell Be aware to not exceed a...

Страница 121: ...performing according to the specifications published by Agilent Technologies Your normal LC grade solvents may give good results most of the time but our experience shows that baseline noise can be higher with LC grade solvents than with fluorescence grade solvents Flush your solvent delivery system for at least 15 minutes before checking sensitivity If your pump has multiple channels you should a...

Страница 122: ...instrument characteristics for example the lamp type and the gratings As most organic molecules absorb best in the ultra violet range the module was designed to give an optimum signal to noise ratio in the 210 nm to 360 nm range of the spectrum To achieve greatest sensitivity the absorbance wavelength of your sample molecule should match the wavelength range for your instrument In other words an e...

Страница 123: ... 240 nm Figure 45 Scan Orthophthalaldehyde Derivative of Alanine When you are looking for the wavelength by scanning scan over the whole range As this example shows a maximum may be found in a completely different wavelength range NOTE When comparing fluorescence excitation spectra directly with DAD spectra or literature based absorbance spectra you should consider large differences in the used op...

Страница 124: ...ratic sample which was diluted 1000 times With increasing PMTGAIN there was an improvement in signal to noise up to 10 Above 10 the noise increased proportionately to the signal with no improvement in signal to noise Figure 46 Finding Best PMTGAIN for Biphenyl The reason for this is the fact that quantification of baselines especially at low background levels is not sufficient for statistically wo...

Страница 125: ...ods This is not a problem in general but could be inconvenient when operating more than one G7121 FLD in the lab Both scaling issues can be resolved Refer to Optimize the PMT Gain Level on page 125 Optimize the PMT Gain Level Start the PMT Gain Test with your operating conditions used method parameter EX EM wavelength solvent flow rate The resulting PMT value will give you the best signal to noise...

Страница 126: ...Gain Level Now check with your highest concentration amount that your highest peak does not clip or overflow If this check is ok you finished the PMT Gain Level optimization Continue with Set your Luminescence Units in LU If the check shows that the highest concentration doesn t fit to the selected range e g by clipping you may decrease the sensitivity of your FLD by gradually decreasing the PMT L...

Страница 127: ...strument to your favorable LU level We recommend not exceeding around 100 LU The parameter of choice is called Scale factor and is applicable by the local controller the Instant Pilot B 02 07 or later In case older revisions are used the Scale factor can be entered using the command line of Agilent ChemStation PRINT SENDMODULE LFLD DMUL x xx Instant Pilot Service Mode FLD then type DMUL x xx and p...

Страница 128: ...aphic peak Raw ADC counts The measured light intensity is limited by the max range of the ADC converter A filter smoothes the peak making it not clearly visible that the max intensity is reached Also peak area and peak height are distorted which leads to poor linearity performance Note that max LU is not a fix number but depends on the intensity of the reference channel New implementation with fir...

Страница 129: ...verflow is visible as a real flat peak in the chromatogram showing the user that the setting of the detector parameter PMT gain or the concentration of the solution is set to high NOTE The transfer of methods 1 1 from one FLD to another may result into the above ADC overflow problem For details see FLD Scaling Range and Operating Conditions on page 125 ...

Страница 130: ...e lamp flash frequency can be changed into the following modes Best sensitivity can be expected with no economy see Figure 48 on page 130 Figure 48 Xenon Flash Lamp Frequency Table 17 Flash Lamp Modes Positioning 296 Hz Standard 560 V 63 mJ 18 8 W 74 Hz Economy 560 V 63 mJ 4 7 W Rotation Multi Ex Em 74 Hz Standard 950 V 180 mJ 13 3 W 74 Hz Economy 560 V 63 mJ 4 7 W 6WDQGDUG FRQRP ...

Страница 131: ...1 Optimizing the Detector 5 Changing the Xenon Flash Lamp Frequency Lamp Life Savings There are three ways to save lamp life switch to lamp on during run without loss of sensitivity switch to economy mode with a certain loss of sensitivity a combination of the above ...

Страница 132: ...ease your signal to noise ratio For example see Figure 49 on page 132 Figure 49 Finding Best Response Time LC fluorescence detectors typically work with response times of 2 or 4 s The default of the module is 4 seconds It is important to know that comparing sensitivity requires using the same response time A response time of 4 s default is equivalent to a time constant of 1 8 s and appropriate for...

Страница 133: ...pected peak in your chromatogram The peak width sets the optimum response time for your detector The peak detector ignores any peaks that are considerably narrower or wider than the peak width setting The response time is the time between 10 and 90 of the output signal in response to an input step function Limits When you set the peak width in minutes the corresponding response time is set automat...

Страница 134: ...nse Time Table 18 Peakwidth Setting Peak Width Data Rate At half height min Response sec Hz ms 0 0016 0 016 148 15 6 9 G7121B 0 003 0 03 74 07 13 5 G7121A B 0 003 0 06 37 04 27 0 0 005 0 12 37 04 27 0 0 01 0 25 37 04 27 0 0 025 0 5 18 52 54 0 0 05 1 0 9 26 108 0 0 1 2 0 4 63 216 0 0 2 4 0 2 31 432 0 0 4 8 0 1 16 864 0 ...

Страница 135: ...the 2nd order light is the limiting factor To explain the effect of such higher order light assume the detector is on but no sample is eluting through the flow cell The lamp sends 1 million photons into the flow cell at for example 280 nm Scattering on the surface of the flow cell and scattering from the molecules of solvent allow 0 1 of this light to leave the cell through the window at right ang...

Страница 136: ...ityLab LC Series 1260 Infinity II FLD User Manual 5 Optimizing the Detector Reducing Stray Light Figure 51 Reducing Stray Light 8 OXRUHVFHQFH DYHOHQJWK QP LWDWLRQ QP 6WUD OLJKW 6HFRQG RUGHU OLJKW QP QR ILOWHU ILOWHU QP ...

Страница 137: ...erfaces 138 Agilent Lab Advisor Software 140 Diagnostic Signals 141 Overview of Diagnostic Signals 141 Baseline Problems Deriving from Xenon Flash Lamp Trigger Pack Assembly 142 Wander Drift Problems Due to Temperature Changes 143 Monitoring of Additional Signals 144 Agilent Lab Advisor 144 Overview about the troubleshooting and diagnostic features ...

Страница 138: ...ots used within these procedures are based on the Agilent Lab Advisor software NOTE Depending on the used interface the available tests and the screens reports may vary Preferred tool should be the Agilent Lab Advisor see Agilent Lab Advisor Software on page 140 Agilent Lab Advisor B 02 08 or later is required The Instant Pilot G4208A supports the G7121A B with B 02 19 or later ...

Страница 139: ...Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 139 Troubleshooting and Diagnostics 6 Available Tests vs User Interfaces Figure 52 The Lab Advisor shows the available tests ...

Страница 140: ... every Agilent 1200 Infinity Series and Agilent InfinityLab LC Series instrument The Lab Advisor Advanced features can be unlocked by purchasing a license key and include real time monitoring of instrument actuals all various instrument signals and state machines In addition all diagnostic test results calibration results and acquired signal data can be uploaded to a shared network folder The Revi...

Страница 141: ...eral signals internal temperatures signal reference signal that can be used to diagnose problems These can be baseline problems deriving from the Xenon flash lamp trigger pack assembly wander drift problems due to temperature changes These signals can be used in addition to the normal baseline signal to determine if there is any correlation to the temperature or voltage current of the lamp ...

Страница 142: ...Problem Verification Perform the following steps to diagnose whether the lamp is the cause of the baseline instability 1 Ensure that the detector has been properly warmed up see Overview of Diagnostic Signals on page 141 2 Flush the flow cell with water and start runs without sample injections blank runs 3 Use a restriction capillary to remove any influence of the column 4 If you see instabilities...

Страница 143: ...ion are unstable laboratory air conditioning systems Other causes include direct sunshine or drafts from open doors and windows These temperature changes may cause baseline wander which can make reproducible integration of trace level peaks difficult or impossible Problem Verification Use the diagnostic signals to find a correlation to ambient changes Normal signal LU Reference signal LU Board Tem...

Страница 144: ... diagnosing problems These can be baseline problems deriving from lamp wander drift problems due to temperature changes For intermittent baseline problems the board temperature should be monitored to see the impact of it to the signal Agilent Lab Advisor Open Instrument Control Select Main signal and board temperature Scale the signals NOTE Measurements should be taken over 60 120 min minimum to i...

Страница 145: ...nual 145 Troubleshooting and Diagnostics 6 Monitoring of Additional Signals Figure 53 Signal plus board temperature Agilent Lab Advisor The board temperature short long term is also stored in the module s memory and can be retrieved via the Module Info Signals ...

Страница 146: ...Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 6 Troubleshooting and Diagnostics Monitoring of Additional Signals Figure 54 Internal board temperature Agilent Lab Advisor Module Info Signals ...

Страница 147: ...n Failed 154 Open Cover 154 Cover Violation 155 ERI Messages 155 Detector Error Messages 156 Lamp Cover Open 156 FLF Board not found 156 ADC Not Calibrated 157 A D Overflow 157 Flash Lamp Current Overflow 158 No light at reference diode despite lamp is on 158 Flash Trigger Lost 159 Wavelength Calibration Failed 159 Wavelength Calibration Lost 160 Flow Cell Removed 160 Motor Errors 161 This chapter...

Страница 148: ...get a notification all LEDs get red and the run will be stopped Depending on the module type this stop is implemented differently For example for a pump the flow will be stopped for safety reasons For a detector the lamp will stay on in order to avoid equilibration time Depending on the error type the next run can only be started if the error has been resolved for example liquid from a leak has be...

Страница 149: ...s exceeded Probable cause Suggested actions 1 The analysis was completed successfully and the timeout function switched off the module as requested Check the logbook for the occurrence and source of a not ready condition Restart the analysis where required 2 A not ready condition was present during a sequence or multiple injection run for a period longer than the timeout threshold Check the logboo...

Страница 150: ... module with a CAN connection to the system Fix the leak in the external instrument before restarting the module 2 Leak detected in an external instrument with a remote connection to the system Fix the leak in the external instrument before restarting the module 3 Shut down in an external instrument with a remote connection to the system Check external instruments for a shut down condition 4 The d...

Страница 151: ... board mounted temperature compensation sensor are used by the leak algorithm to determine whether a leak is present When a leak occurs the leak sensor is cooled by the solvent This changes the resistance of the leak sensor which is sensed by the leak sensor circuit on the main board Probable cause Suggested actions 1 CAN cable disconnected Ensure all the CAN cables are connected correctly Ensure ...

Страница 152: ...thin defined limits If the current increases above the upper limit the error message is generated Probable cause Suggested actions 1 Leak sensor not connected to the Power Switch board Please contact your Agilent service representative 2 Defective leak sensor Please contact your Agilent service representative 3 Leak sensor incorrectly routed being pinched by a metal component Please contact your A...

Страница 153: ...he power switch board in the module has failed open circuit The resistance across the temperature compensation sensor NTC on the power switch board is dependent on ambient temperature The change in resistance is used by the leak circuit to compensate for ambient temperature changes If the resistance across the sensor falls below the lower limit the error message is generated Probable cause Suggest...

Страница 154: ... e g the lamp in the detector are turned off to assure that the module does not overheat inside Open Cover Error ID 0205 The top foam has been removed Probable cause Suggested actions 1 Fan cable disconnected Please contact your Agilent service representative 2 Defective fan Please contact your Agilent service representative 3 Defective main board Please contact your Agilent service representative...

Страница 155: ... with the foam removed the lamps are switched off and the error message is generated ERI Messages Error ID 11120 5 V 11121 25 V The ERI Enhanced Remote Interface provides two error events related to over current situations on the 5 V and 24 V lines Probable cause Suggested actions 1 The top foam was removed during operation Please contact your Agilent service representative 2 Foam not activating t...

Страница 156: ...0 The FLF board could not be found by the main board FLM This message comes together with some other message generated on the FLF board e g Leak Probable cause Suggested actions 1 Lamp cover removed Please contact your Agilent service representative Probable cause Suggested actions 1 FLF board not connected to the FLM board Please contact your Agilent service representative 2 Defective FLF board P...

Страница 157: ...ge comes up during a run it includes the time of occurrence and when it disappears 1200 FLD 1 A D overflow RT is 0 32 min 16 33 24 02 11 99 1200 FLD 1 A D overflow finished RT is 0 67 min 16 33 46 02 11 99 If this condition is present prior to a run the not ready will prevent the system to start the run sequence With firmware revision A 06 11 and above the A D overflow leads into a flat peak in th...

Страница 158: ...gram the user interface shows the module still in Ready Perform a Lamp Intensity Test see Lamp Intensity Test on page 171 first If flat use below steps Revision D Front End Board FLF The flashing of the xenon flash lamp is monitored constantly If the Lamp has not flashed for more than 100 times in series an error is generated and the lamp is turned OFF Probable cause Suggested actions 1 Short circ...

Страница 159: ...nd the instrument stays in a Not Ready condition Probable cause Suggested actions 1 Firmware problem Reboot the detector power cycle 2 Multi Mode Off Please contact your Agilent service representative 3 Defective encoder Please contact your Agilent service representative Probable cause Suggested actions 1 Flash lamp not ignited or position not correct Please contact your Agilent service representa...

Страница 160: ...stays in a Not Ready condition Flow Cell Removed Error ID 6616 6702 6760 The detector has an automatic cell recognition system When the flow cell is removed the lamp is turned off and a NOT READY condition exists If the flow cell is removed during an analysis a SHUT DOWN is generated Probable cause Suggested actions 1 Reset of monochromator settings after exchange Perform a wavelength calibration ...

Страница 161: ...citation or the emissionside If an error occurs do a lamp ignition This will clear the errorand a re initialization of the motors is performed Probable cause Suggested actions 1 Friction too high Exchange the monochromator assembly 2 Defective monochromator assembly Exchange the monochromator assembly Probable cause Suggested actions 1 Encoder cables mixed on FLM board Check encoder connections to...

Страница 162: ...n and the emission grating drives The number of revolutions after switching off the drives is a measure of friction The history may show an increasing friction of the drive s over a length of time Probable cause Suggested actions 1 Encoder defective Replace monochromator assembly 2 Encoder electric defective Replace FLM board 3 Monochromator defective or missing Check for connection to FLF Ex and ...

Страница 163: ...vement of the monochromator The position is lost and the lamp will turn off Motor Speed Too Low Error ID EX 6715 EM 6716 For proper operation the monochromator gratings must run at a certain constant revolution Probable cause Suggested actions 1 Defective monochromator assembly Reset monochromator settings Replace monochromator assembly Probable cause Suggested actions 1 Short mechanical shock re ...

Страница 164: ...Encoder Index Wrong Error ID EX 6719 EM 6720 The actual encoder pattern is checked against a known pattern Probable cause Suggested actions 1 Defective monochromator assembly Re ignite the lamp Replace monochromator assembly Probable cause Suggested actions 1 Encoder was replaced and has a different pattern or no reset of pattern was made Reset pattern via user interface and recalibrate 2 Encoder ...

Страница 165: ...Interpretation of the Results 181 Wavelength Verification and Calibration 182 Wavelength Calibration Process 184 Wavelength Calibration Procedure 187 Excitation and Emission Grating Resistance History 191 D A Converter DAC Test 192 Test Evaluation 193 Dark Current Test 194 Test Failed 195 Using the Built in Test Chromatogram 196 Procedure Using the Agilent Lab Advisor 196 Other Lab Advisor Functio...

Страница 166: ...or shows the available test under Service Diagnostics Table 19 Interfaces and available test functions Interface Comment Available Function Agilent Lab Advisor For functions refer to Function Overview Lab Advisor G7121A Table 20 on page 167 Available functions depend on Product Level Basic Advanced FSE Agilent ChemStation No tests available Adding of temperature lamp signals to chromatographic sig...

Страница 167: ...est Basic Advanced Wavelength Accuracy Test Basic Advanced Calibrations Wavelength Calibration Basic Advanced Tools Diagnostic Buffers Basic Advanced Module Info Basic Advanced Test Chromatogram Basic Advanced Spectral Scan G7121B only Basic Advanced Controls Advanced Method Parameters Analog Output 1 Offset Full Scale Advanced Configuration Remote Pulse Duration s Basic Advanced Analog Output 1 R...

Страница 168: ...tion Identify Module Basic Advanced Special Commands Detector Reset Basic Advanced Clear Error Basic Advanced Statemachines UV Lamp Advanced Signals Signal A LU G7121A allows just Signal A Advanced Signal B LU Advanced Signal C LU Advanced Signal D LU Advanced Photomultiplier raw counts Advanced Reference diode raw counts Advanced Ambient Temperature C Advanced EMF Counters Flash Lamp Life Time Ba...

Страница 169: ...Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 169 Test Functions and Calibration 8 Introduction Figure 55 The Lab Advisor shows the available tests ...

Страница 170: ...finityLab LC Series 1260 Infinity II FLD User Manual 8 Test Functions and Calibration Diagram of Light Path Diagram of Light Path The light path is shown in Figure 56 on page 170 Figure 56 Schematic of the light path ...

Страница 171: ...indow There is no further evaluation of the test Results of this test are stored as lamp history date code intensity Figure 57 Lamp Intensity Test Agilent Lab Advisor UV degradation especially below 250 nm is significantly higher compared to visible wavelength range Generally the LAMP ON during run setting or using economy mode will increase lamp life by a magnitude NOTE The profile can vary from ...

Страница 172: ...f the lamp intensity test if the last one is older than one week are stored as lamp history date code intensity of four different wavelengths 250 nm 350 nm 450 and 600 nm in a buffer The data plot can be retrieved via the diagnostics and provides intensity data over a length of time Figure 58 Lamp Intensity History Agilent Lab Advisor under Module Info ...

Страница 173: ...low cell G1321 60005 G5615 60005 flow of 0 25 mL min of water Table 21 Raman ASTM Signal to Noise Specification Instrument SNR Specification Raman Dark SNR Specification Dual WL G7121A 500 3000 G7121B 500 3000 300 NOTE The Dark and Dual WL values are just additional specifications Only the Raman value is used for the standard instrument checkout NOTE The specification single wavelength at signal c...

Страница 174: ...th at background 3000 according to settings in Table 24 on page 174 Specification dual wavelength 300 according to settings in Table 25 on page 174 Table 23 Settings for Single Wavelength Specifications at signal Time EX EM PMT Baseline 0 350 397 12 Free 20 30 350 450 12 Free Table 24 Settings for Single Wavelength Specifications at background Time EX EM PMT Baseline 0 350 450 14 Free 20 30 350 39...

Страница 175: ...re 60 on page 175 for details Figure 60 Raman ASTM signal noise calculation 397 em 350 ex n noise_rama 450 em 350 ex round mean_backg 397 em 350 ex mean_raman SNR_Raman 450 em 350 ex ground noise_back 450 em 350 ex round mean_backg 397 em 350 ex mean_raman SNR_Dark 0HDQ 5DPDQ XUUHQW 0HDQ DUN XUUHQW DYHUDJH EHWZHHQ DQG PLQXWHV 6701RLVH WLPH WR PLQXWHV DYHUDJH EHWZHHQ DQG PLQXWHV ...

Страница 176: ...oise Test Using the Agilent Lab Advisor 1 Set up the HPLC system and the Lab Advisor 2 Flush the flow cell with clean bi distilled water 3 Start the test in the Lab Advisor Figure 61 Raman ASTM Signal to Noise Test Agilent Lab Advisor In case of failing this test as shown above see Interpretation of the Results on page 177 ...

Страница 177: ... Signal to Noise Test Interpretation of the Results If the test shows low Raman values check for correctly positioned flow cell clean flow cell flush with clean bi distilled water no air bubble s check via fluorescence scan or visual check of cell cuvette solvent inlet filter may create air bubbles in flow cell ...

Страница 178: ... The FLD will change into the multi excitation mode with emission wavelength at 397 nm and scan in the range of the expected maximum of 350 nm 20 nm As result the maxima should be found at 350 nm 3 nm see Figure 62 on page 178 The FLD will change into the multi emission mode with excitation wavelength at 350 nm and scan in the range of the expected maximum of 397 nm 20 nm As result the maxima shou...

Страница 179: ...8 Wavelength Accuracy Test Figure 63 Wavelength Accuracy Test with Lab Advisor If the test fails observe the maxima of the EX or EM side under the Signals tab NOTE If the plots do not have a maximum around EM 397 nm and EX 350 nm 3 nm the test fails Refer to Interpretation of the Results on page 181 ...

Страница 180: ...r Manual 8 Test Functions and Calibration Wavelength Accuracy Test Figure 64 Example of good EX EM maxima If the plots do not have a maximum around EX 397 nm and EX 350 nm 3 nm the test fails see figure below Refer to Interpretation of the Results on page 181 ...

Страница 181: ...e Results If the test fails check for correctly positioned flow cell clean flow cell flush with isopropanol and clean bi distilled water no air bubble s check via fluorescence scan or visual check of cell cuvette solvent inlet filter may create air bubbles in flow cell check optical path for contamination service check alignment of lamp trigger pack assembly service perform a Wavelength Calibratio...

Страница 182: ...ation functionality is used The algorithm is based on evaluating different grating orders and calculating the wavelength scales of both excitation and emission monochromator by applying the fundamental grating equation NOTE New instruments are shipped with a flow cell installed and have a factory wavelength calibration If a flow cell is used other then the installed shipped flow cell verify that t...

Страница 183: ...Functions and Calibration 8 Wavelength Verification and Calibration Table 26 Reasons for doing a Verification or Calibration Verification WL calibration interest X GLP compliance X cell change X lamp change X monochromator change X main board change X optical unit change X ...

Страница 184: ...d with the photomultiplier tube NOTE Prior to a wavelength calibration a wavelength accuracy verification should be performed see Wavelength Accuracy Test on page 178 If the deviation is more than 3 nm the wavelength calibration should be done as described in Wavelength Calibration Procedure on page 187 NOTE The duration of the wavelength calibration is about 20 minutes plus setup time for the cal...

Страница 185: ...Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 185 Test Functions and Calibration 8 Wavelength Calibration Process Figure 66 Wavelength Calibration Agilent Lab Advisor ...

Страница 186: ...on rotation scan full circle of scans depends on the required PMT gain 1 minute per scan 61 s variable 6 n Em rotation scan full circle instrument profile 9 s 6 n Em rotation scan full circle instrument profile 9 s 6 n Em rotation scan full circle instrument profile 9 s 6 n Em rotation scan full circle instrument profile 9 s 7 Emission rotation scan high resolution part I 44 s 8 Emission rotation ...

Страница 187: ...n After 10 minutes the solution is ready When If application requires or see Table 27 on page 186 Tools required Description Laboratory balance Parts required p n Description G7121 68001 FLD Calibration Kit contains parts below 5063 6597 Calibration Sample Glycogen 9301 1446 Syringe 9301 0407 Needle 5190 5111 Syringe filter 0 45 µm 100 pk 0100 1516 Fitting male PEEK 2 pk NOTE New instruments are s...

Страница 188: ...ringe with Sample Filter h Lift the needle tip and carefully eject approximately 0 5 ml to remove air out of the syringe and to flush the needle i Add the PEEK fitting to the needle tip and fix both at the flow cell inlet j Slowly inject about 0 2 ml and wait for about 10 seconds to inject another 0 1 ml This will assure that the cell is filled properly 3 Wavelength Calibration a From the user int...

Страница 189: ...ctions and Calibration 8 Wavelength Calibration Procedure b If a deviation is displayed press Yes to adjust to new values The history table will be updated Figure 68 Wavelength Calibration Agilent Lab Advisor Figure 69 Calibration History Agilent Lab Advisor under Module Info ...

Страница 190: ... using the Wavelength Accuracy Test a Refit the capillary to the flow cell b Follow the procedure Wavelength Accuracy Test NOTE Rinse the flow cell with pure water at a minimum of 1 5 mL min to get rid of the Glycogen from the cell and the capillaries When organic solvent is sequentially applied without rinsing a blockage of capillaries may occur ...

Страница 191: ...l test It provides the resistance history of the excitation and the emission grating drives The number of revolutions after switching off the drives is a measure of friction The history may show an increasing friction of the drive s over a length of time The history data contains the data time information and the number of turns The data plot can be retrieved via the diagnostics and provides turn ...

Страница 192: ...nal of approximately 50 mV if the zero offset of the analog output is set to the default value of 5 which can be plotted on an integrator A continuous square wave with an amplitude of 10 μV and a frequency of approximately 1 cycle 24 seconds is applied to the signal The amplitude of the square wave and the peak to peak noise are used to evaluate the DAC test Running the test with Agilent Lab Advis...

Страница 193: ...st can be started via the command line 1 To start the test TEST DAC 1 Reply RA 00000 TEST DAC 1 2 To stop the test TEST DAC 0 Reply RA 00000 TEST DAC 0 Test Evaluation The noise on the step should be less than 3 μV Probable cause Suggested actions 1 Bad cable or grounding problem between detector and external device Check or replace the cable 2 Defective detector main board Please contact your Agi...

Страница 194: ...st measures the PMT signal with maximum and minimum gain while the lamp is OFF It also reads the signal of the reference diode The resulting values two via reference diode and two from PMT are shown in a table and checked against reasonable limits see below Figure 73 Dark Current Test Agilent Lab Advisor Figure 74 Dark Current History Agilent Lab Advisor ...

Страница 195: ...260 Infinity II FLD User Manual 195 Test Functions and Calibration 8 Dark Current Test Test Failed Probable cause Suggested actions 1 Defective PMT Exchange the PMT 2 Defective reference diode or A D converter Exchange the FLF board ...

Страница 196: ... stop is executed either by means of a stop time or manually Procedure Using the Agilent Lab Advisor This procedure works for all Agilent 1200 Infinity detectors DAD MWD VWD FLD and RID The example figure is from the RID detector 1 Assure that the default LC method is loaded via the control software 2 Start the Agilent Lab Advisor software B 01 03 SP4 or later and open the detector s Tools selecti...

Страница 197: ...on 8 Using the Built in Test Chromatogram 6 To start a test chromatogram enter in the command line STRT Figure 75 Test Chromatogram with Agilent Lab Advisor 7 To stop the test chromatogram enter in the command line STOP NOTE The test chromatogram is switched off automatically at the end of a run ...

Страница 198: ...eral tab shows information about the module s firmware and options in a two column table Signals The Signals tab shows the plots of the available diagnostic signals from the module The signal plots that are available are module dependent where available both short term and long term plots are displayed for a signal Tables The Tables tab allows you to display the contents of all available diagnosti...

Страница 199: ...ual 199 Test Functions and Calibration 8 Other Lab Advisor Functions Figure 77 Module Info Signals Includes signal for Board Temperature long short term Number of Revolutions of EX EM Gratings Dark current of Reference Diode and PMT Figure 78 Module Info Tables ...

Страница 200: ...Change The Board Check and Change tool allows you to set the module type and serial number after a main board has been changed After boot of the FLD normally the Module Type is automatically entered based on installed optical unit In this case just the module Serial Number has to be entered same on the module label Figure 79 Module Info Tables ...

Страница 201: ...libration 8 Other Lab Advisor Functions Instrument Control The Instrument Control screen provides controls for each of the modules in the selected system Module information and status are displayed click to display the module s controls or to hide them Figure 80 Instrument Control Overview ...

Страница 202: ...C Series 1260 Infinity II FLD User Manual 8 Test Functions and Calibration Other Lab Advisor Functions EMF Early Maintenance Feature The EMFs screen allows you to view and manage the EMF counters for all modules in all systems ...

Страница 203: ...nctions Spectral Scan G7121B only The Spectral Scan tool for the fluorescence detector FLD allows you to scan a spectrum over a specified wavelength range in a specified mode and export the data to a csv comma separated values file that can be used in other applications for example Microsoft Excel Figure 81 Spectral Scan ...

Страница 204: ...m and the excitation scan range Emission Scan You specify an excitation wavelength Ex and the emission scan range Scan Parameters Flash Lamp On Switches on the FLD flash lamp Sample Scan Scans the sample spectrum over the specified wavelength range at the specified resolution and in the selected mode You specify the wavelength range in the from and to fields and the resolution in the step field Yo...

Страница 205: ...ns 207 Overview of Maintenance 209 Cleaning the Module 210 Remove and Install Doors 211 Exchanging a Flow Cell 213 How to use the Cuvette 217 Flow Cell Flushing 218 Correcting Leaks 219 Replace Leak Handling System Parts 221 Replacing Module Firmware 223 Tests and Calibrations 224 This chapter provides general information on maintenance of the detector ...

Страница 206: ... Manual 9 Maintenance Introduction to Maintenance Introduction to Maintenance The module is designed for easy maintenance Maintenance can be done from the front with module in place in the system NOTE There are no serviceable parts inside Do not open the module ...

Страница 207: ... sheet supplied by the vendor and follow good laboratory practice The volume of substances should be reduced to the minimum required for the analysis Do not operate the instrument in an explosive atmosphere WARNING Eye damage by detector light Eye damage may result from directly viewing the UV light produced by the lamp of the optical system used in this product Always turn the lamp of the optical...

Страница 208: ...ications to the products failure to comply with procedures in Agilent product user guides or use of the products in violation of applicable laws rules or regulations Use your Agilent products only in the manner described in the Agilent product user guides CAUTION Safety standards for external equipment If you connect external equipment to the instrument make sure that you only use accessory units ...

Страница 209: ...hange If application requires a different flow cell type or if defective Complete Assembly A wavelength calibration check should be performed after replacement If the flow cell is removed and inserted then a quick calibration check is performed If this fails you must do a wavelength recalibration see Wavelength Verification and Calibration on page 182 Flow cell flushing If flow cell is contaminate...

Страница 210: ... case clean use a soft cloth slightly dampened with water or a solution of water and mild detergent WARNING Liquid dripping into the electronic compartment of your module can cause shock hazard and damage the module Do not use an excessively damp cloth during cleaning Drain all solvent lines before opening any connections in the flow path ...

Страница 211: ...l Doors Remove and Install Doors Parts required p n Description 5067 5737 Door left 5067 5736 Door right NOTE The figures shown in this procedure exemplarily show the Infinity II Multisampler module The principle of how to remove and or install doors works in the same way for all Infinity II modules ...

Страница 212: ...r Manual 9 Maintenance Remove and Install Doors 1 Press the release buttons and pull the front door out 2 For the Installation of the front door Insert the hinges into their guides and move the door in until the release buttons click into their final position ...

Страница 213: ...H 1 9 5 G1321 60015 Flow cell 4 µL 20 bar pH 1 9 5 G5615 60005 Bio inert flow cell 8 µL 20 bar pH 1 12 includes Capillary Kit Flow Cells BIO p n G5615 68755 G1321 60007 FLD Cuvette Kit 8 µL 20 bar Preparations Turn off the flow CAUTION Sample degradation and contamination of the instrument Metal parts in the flow path can interact with the bio molecules in the sample leading to sample degradation ...

Страница 214: ...ration If a flow cell is used other then the installed shipped flow cell verify that the installed flow cell is fixed very tight perform a wavelength calibration using Glycogen part of FLD Calibration Kit G7121 68001 NOTE Before storing the flow cell flush it with iso propanol and close it with Plastic fittings 0100 1259 1 Open the doors of the module 2 Disconnect the capillaries from the flow cel...

Страница 215: ...w cell If defective leaky the flow cell has to be replaced completely 5 Insert the flow cell and tighten the thumb screws strongly NOTE If the cell is not completely in its end position and tightened strongly this could result into a wavelength shift NOTE The label attached to the flow cell provides information on part number cell volume and maximum pressure The cell type will be automatically det...

Страница 216: ...tem the fluorescence detector should be the last detector in the flow path except for evaporative detectors like LC MSD Otherwise the back pressure generated by the other detector may overload the flow cell and will lead to a defective cell maximum pressure is 20 bar 2 MPa Always use the outlet capillary set supplied with the accessory kit 8 Check for leaks establish a flow and observe the flow ce...

Страница 217: ... bore capillary connections for easier injections with a syringe identification lever for cell auto recognition system 1 Install the cuvette instead of the standard flow cell 2 Connect the waste tubing to the outlet of the cuvette 3 Use the syringe see Cuvette Kit on page 229 to inject the compound 4 Setup the parameters for the Fluorescence Scan under Special Setpoints 5 Select Take Fluorescence ...

Страница 218: ... nitric acid 65 tubings to waste WARNING Dangerous concentration of nitric acid The nitric acid flushing procedure is not an infallible remedy for a dirty cell It is to be used as a last attempt to salvage the cell before cell replacement Note that the cell is a consumable item Give proper attention to safety NOTE Aqueous solvents in the flow cell can built up algae Algae do fluoresce Therefore do...

Страница 219: ...19 Maintenance 9 Correcting Leaks Correcting Leaks Figure 82 Correcting leaks ORZ HOO UHD HDN 6HQVRU UHD When If a leakage has occurred in the flow cell area or at the capillary connections Tools required Description Tissue Wrench 1 4 inch for capillary connections ...

Страница 220: ...1260 Infinity II FLD User Manual 9 Maintenance Correcting Leaks 1 Open the doors 2 Use tissue to dry the leak sensor area 1 3 Observe the capillary connections and the flow cell area 2 for leaks and correct if required 4 Close the doors ...

Страница 221: ...bly i d 6 mm o d 9 mm 1 2 m to waste Preparations Turn off the module Disconnect the power cable Disconnect the hydraulic connection from the flow Remove the module from the stack and place it on the working bench 1 Remove the doors see Remove and Install Doors on page 211 2 Lift the cover levers at the rear of the module until you can lift the cover 3 Remove the cover and place it on the bench 4 ...

Страница 222: ... it together with the tubing Next Steps 6 Install the Leak Adapter by pressing it into the Main Cover 7 Slide the Main Cover into its front position 8 Press the main cover down at the rear until the levers click 9 Insert the Tubing 2 ca 85 mm required for replacement between Leak Adapter outlet and Leak Pan 10 Install the Doors see Remove and Install Doors on page 211 ...

Страница 223: ...c Information There is no specific information for this module When The installation of newer firmware might be necessary if a newer version solves problems of older versions or to keep all systems on the same validated revision The installation of older firmware might be necessary to keep all systems on the same validated revision or if a new module with newer firmware is added to a system or if ...

Страница 224: ...inity II FLD User Manual 9 Maintenance Tests and Calibrations Tests and Calibrations The following tests are required after maintenance of lamps and flow cells Lamp Intensity Test on page 171 Wavelength Verification and Calibration on page 182 ...

Страница 225: ...ityLab LC Series 1260 Infinity II FLD User Manual Agilent Technologies 10 Parts for Maintenance and Repair Overview of Maintenance Parts 226 Kits 228 This chapter provides information on parts for maintenance and repair ...

Страница 226: ...226 Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 10 Parts for Maintenance and Repair Overview of Maintenance Parts Overview of Maintenance Parts Figure 83 Overview of Maintenance Parts ...

Страница 227: ...18 300 mm long part of Capillary kit for 0 12 mm id p n G1316 68716 OR G5615 60005 Bio inert flow cell 8 µL 20 bar pH 1 12 includes Capillary Kit Flow Cells BIO p n G5615 68755 OR G5615 68755 Capillary Kit Flow Cells BIO includes Capillary PK 0 18 mm x 1 5 m and PEEK Fittings 10 PK p n 5063 6591 2 5067 5737 Door left 3 5067 5736 Door right 4 5043 1013 Tubing Clip 5 5043 0856 Leak Adapter 6 5063 65...

Страница 228: ...87311 Capillary ST 0 17 mm x 380 mm S S Column to detector includes ST ferrule front ST ferrule back and ST fitting 5063 6527 Tubing assembly i d 6 mm o d 9 mm 1 2 m to waste see item 6 in Figure 83 on page 226 5181 1516 CAN cable Agilent module to module 0 5 m 5043 1013 Tubing Clip see item 4 in Figure 83 on page 226 5500 1155 Tube Connector 90 degree ID 6 4 p n Description 5063 6597 Calibration ...

Страница 229: ...ittings 10 PK p n 5063 6591 G5615 68755 includes Cuvette Kit p n Description 0890 1763 Capillary PK 0 18 mm x 1 5 m 5063 6591 PEEK Fittings 10 PK p n Description G1321 60007 FLD Cuvette Kit includes Flow cell 8 µL 20 bar pH 1 9 5 5062 2462 Tube PTFE 0 7 mm x 5 m 1 6 mm od 79814 22406 ST Fitting 0100 0043 ST front ferrule 0100 0044 ST back ferrule 0100 1516 Fitting male PEEK 2 pk 9301 0407 Needle 9...

Страница 230: ...230 Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 10 Parts for Maintenance and Repair Kits ...

Страница 231: ... Manual Agilent Technologies 11 Identifying Cables Cable Overview 232 Analog Cables 234 Remote Cables 236 CAN LAN Cables 240 Agilent Module to PC 241 USB Cables 242 This chapter provides information on cables used with the Agilent InfinityLab LC Series modules ...

Страница 232: ...Description 35900 60750 Agilent 35900A A D converter 01046 60105 Analog cable BNC to general purpose spade lugs p n Description 5188 8029 ERI to general purpose 5188 8044 Remote Cable ERI ERI 5188 8045 Remote Cable APG ERI 5188 8059 ERI Extension Cable 1 2 m 5061 3378 Remote Cable to 35900 A D converter 01046 60201 Agilent module to general purpose 5188 8057 Fraction Collection ERI remote Y cable ...

Страница 233: ...le shielded 7 m for point to point connection p n Description RS232 61601 RS 232 cable 2 5 m Instrument to PC 9 to 9 pin female This cable has special pin out and is not compatible with connecting printers and plotters It s also called Null Modem Cable with full handshaking where the wiring is made between pins 1 1 2 3 3 2 4 6 5 5 6 4 7 8 8 7 9 9 5181 1561 RS 232 cable 8 m p n Description 5188 805...

Страница 234: ...log Cables One end of these cables provides a BNC connector to be connected to Agilent modules The other end depends on the instrument to which connection is being made Agilent Module to 35900 A D converters p n 35900 60750 35900 Pin Agilent module Signal Name 1 Not connected 2 Shield Analog 3 Center Analog ...

Страница 235: ... Cables 11 Analog Cables Agilent Module to BNC Connector Agilent Module to General Purpose p n 8120 1840 Pin BNC Pin Agilent module Signal Name Shield Shield Analog Center Center Analog p n 01046 60105 Pin Pin Agilent module Signal Name 1 Not connected 2 Black Analog 3 Red Analog ...

Страница 236: ...tension Cable 1 2 m D Sub15 pin male female p n 5188 8029 pin Color code Enhanced Remote Classic Remote Active TTL 1 white IO1 START REQUEST Low 2 brown IO2 STOP Low 3 green IO3 READY High 4 yellow IO4 POWER ON High 5 grey IO5 NOT USED 6 pink IO6 SHUT DOWN Low 7 blue IO7 START Low 8 red IO8 PREPARE Low 9 black 1wire DATA 10 violet DGND 11 grey pink 5V ERI out 12 red blue PGND 13 white green PGND 1...

Страница 237: ...s 5188 8045 ERI to APG Connector D_Subminiature 15 pin ERI Connector D_Subminiature 9 pin APG p n 5188 8045 Pin ERI Signal Pin APG Active TTL 10 GND 1 1 Start Request 9 Low 2 Stop 8 Low 3 Ready 7 High 5 Power on 6 High 4 Future 5 6 Shut Down 4 Low 7 Start 3 Low 8 Prepare 2 Low Ground Cable Shielding NC ...

Страница 238: ... provides a Agilent Technologies APG Analytical Products Group remote connector to be connected to Agilent modules The other end depends on the instrument to be connected to Table 29 5188 8057 ERI to APG and RJ45 p n 5188 8057 Pin ERI Signal Pin APG Active TTL Pin RJ45 10 GND 1 5 1 Start Request 9 High 2 Stop 8 High 3 Ready 7 High 4 Fraction Trigger 5 High 4 5 Power on 6 High 6 Shut Down 4 High 7 ...

Страница 239: ...d 2 Brown 2 Brown Prepare run Low 3 Gray 3 Gray Start Low 4 Blue 4 Blue Shut down Low 5 Pink 5 Pink Not connected 6 Yellow 6 Yellow Power on High 7 Red 7 Red Ready High 8 Green 8 Green Stop Low 9 Black 9 Black Start request Low p n 01046 60201 Wire Color Pin Agilent module Signal Name Active TTL White 1 Digital ground Brown 2 Prepare run Low Gray 3 Start Low Blue 4 Shut down Low Pink 5 Not connect...

Страница 240: ...to be connected to Agilent modules CAN or LAN connectors CAN Cables LAN Cables p n Description 5181 1516 CAN cable Agilent module to module 0 5 m 5181 1519 CAN cable Agilent module to module 1 m p n Description 5023 0203 Cross over network cable shielded 3 m for point to point connection 5023 0202 Twisted pair network cable shielded 7 m for point to point connection ...

Страница 241: ...C p n Description RS232 61601 RS 232 cable 2 5 m Instrument to PC 9 to 9 pin female This cable has special pin out and is not compatible with connecting printers and plotters It s also called Null Modem Cable with full handshaking where the wiring is made between pins 1 1 2 3 3 2 4 6 5 5 6 4 7 8 8 7 9 9 5181 1561 RS 232 cable 8 m ...

Страница 242: ...LD User Manual 11 Identifying Cables USB Cables USB Cables To connect a USB Flash Drive use a USB OTG cable with Mini B plug and A socket p n Description 5188 8050 USB A M USB Mini B 3 m PC Module 5188 8049 USB A F USB Mini B M OTG Module to Flash Drive ...

Страница 243: ...ar view of the module 248 Information on Instrument Serial Number 249 Interfaces 250 Overview Interfaces 252 ERI Enhanced Remote Interface 255 USB Universal Serial Bus 257 Setting the 6 bit Configuration Switch 258 Special Settings 260 Instrument Layout 262 Early Maintenance Feedback 263 This chapter describes the detector in more detail on hardware and electronics ...

Страница 244: ... properties are the complete communication capabilities CAN LAN USB and RS 232 memory management ability to update the firmware of the main system Main System Its properties are the complete communication capabilities CAN LAN USB and RS 232 memory management ability to update the firmware of the resident system In addition the main system comprises the instrument functions that are divided into co...

Страница 245: ...uct number for example 1315B for the G1315B DAD R the firmware revision for example A for G1315B or B for the G1315C DAD VVV is the revision number for example 650 is revision 6 50 XXX is the build number of the firmware For instructions on firmware updates refer to section Replacing Firmware in chapter Maintenance or use the documentation provided with the Firmware Update Tools Figure 84 Firmware...

Страница 246: ...their initial firmware revision For example a G1315C DAD SL cannot be downgraded below firmware revision B 01 02 or to a A xx xx Some modules can be re branded e g G1314C to G1314B to allow operation in specific control software environments In this case the feature set of the target type is used and the feature set of the original one is lost After re branding e g from G1314B to G1314C the origin...

Страница 247: ... LAN connector may be used to control the module from a computer through a LAN connection This connector is activated and can be configured with the configuration switch With the appropriate software the USB connector may be used to control the module from a computer through a USB connection The power input socket accepts a line voltage of 100 240 VAC 10 with a line frequency of 50 or 60 Hz Maximu...

Страница 248: ... II FLD User Manual 12 Hardware Information Electrical Connections Rear view of the module Figure 85 Rear view of detector example shows a G7114A B VWD electrical connections and label 1 RQILJXUDWLRQ VZLWFK QDORJ RXWSXW 5 1 3RZHU VRFNHW 86 0LQL 3RUW ...

Страница 249: ...al number information on the instrument labels provide the following information CCYWWSSSSS Format CC country of manufacturing DE Germany JP Japan CN China YWW year and week of last major manufacturing change e g 820 could be week 20 of 1998 or 2008 SSSSS real serial number CCXZZ00000 Format CC Country of manufacturing DE Germany JP Japan CN China X Alphabetic character A Z used by manufacturing Z...

Страница 250: ...rd RS 232 Analog APG A ERI E Special Pumps G7104A C 2 No Yes Yes 1 A G7110B 2 Yes Yes No No E G7111A B G5654A 2 Yes Yes No No E G7112B 2 Yes Yes No No E G7120A 2 No Yes Yes 1 A G7161A B 2 Yes Yes No No E Samplers G7129A B C 2 Yes Yes No No E G7167B C G5667A 2 Yes Yes No No E G7157A 2 Yes Yes No No E Detectors G7114A B 2 Yes Yes No 1 E G7115A 2 Yes Yes No 1 E G7117A B C 2 Yes Yes No 1 E G7121A B 2 ...

Страница 251: ...No No No Requires a host module with on board LAN with minimum FW B 06 40 or C 06 40 or with additional G1369C LAN Card G1364E F G5664B 2 Yes Yes No No E THERMOSTAT for G1330B Others G7116A B 2 No No No No No Requires a HOST module via CAN G7122A No No No Yes No A G7170B 2 No No No No No Requires a host module with on board LAN with minimum FW B 06 40 or C 06 40 or with additional G1369C LAN Card ...

Страница 252: ...her on board LAN nor an interface slot for a LAN card e g G1170A Valve Drive or G4227A Flexible Cube These are hosted modules and require a Host module with firmware B 06 40 or later or with additional G1369C LAN Card USB The USB interface replaces the RS 232 Serial interface in new FUSION generation modules For details on USB refer to USB Universal Serial Bus on page 257 Analog Signal Output The ...

Страница 253: ...OWN the system s critical parts in case any module detects a serious problem To detect whether all participating modules are switched on or properly powered one line is defined to summarize the POWER ON state of all connected modules Control of analysis is maintained by signal readiness READY for next analysis followed by START of run and optional STOP of run triggered on the respective lines In a...

Страница 254: ... injection Receiver is any module performing run time controlled activities 3 READY H System is ready for next analysis Receiver is any sequence controller 4 POWER ON H All modules connected to system are switched on Receiver is any module relying on operation of others 5 Not used 6 SHUT DOWN L System has serious problem for example leak stops pump Receiver is any module capable to reduce safety r...

Страница 255: ...e electronics use ERI This interface is already used in the Agilent Universal Interface Box 2 UIB2 ERI Description The ERI interface contains eight individual programmable input output pins In addition it provides 24 V power and 5 V power and a serial data line to detect and recognize further add ons that could be connected to this interface This way the interface can support various additional de...

Страница 256: ...rolled 1 Wire Data Future Use This serial line can be used to read out an EPROM or write into an EPROM of a connected ERI device The firmware can detect the connected type of device automatically and update information in the device if required Pin Enhanced Remote 1 IO 1 START REQUEST 2 IO 2 STOP 3 IO 3 READY 4 IO 4 POWER ON 5 IO 5 NOT USED 6 IO 6 SHUT DOWN 7 IO 7 START 8 IO 8 PREPARE 9 1 wire DAT...

Страница 257: ... Use Available by firmware command defined turn on off For devices that need higher power Class 0 0 5 A maximum 12 W Class 1 1 0 A maximum 24 W Class 2 2 0 A maximum 48 W Class depends on hosting module s internal power overhead If a connected device requires more power the firmware detects this overcurrent detection and provides the information to the user interface Fuse used for safety protectio...

Страница 258: ...ectronics Switch settings provide configuration parameters for LAN and instrument specific initialization procedures All modules with FUSION electronics Default is ALL switches DOWN best settings Default IP address for LAN 192 168 254 11 For specific LAN modes switches 4 5 must be set as required For boot resident cold start modes switches 1 2 or 6 must be UP Figure 87 Location of Configuration sw...

Страница 259: ...T COM settings are tak en from non volatile memory 0 n a 2 2 not assigned Always keep these switches on position 0 off n a LAN Init Mode n a Use Default IP Address3 3 Default IP Address is 192 168 254 11 0 0 0 0 0 Use Stored IP Address 0 0 0 1 0 Use DHCP to request IP Address4 4 Host Name will be the MAC address 0 0 1 0 0 Test 1 System n a n a n a ColdStart Boot Main System Keep Data 0 0 0 0 0 Boo...

Страница 260: ...cold start can be used to bring the module into a defined mode with default parameter settings Boot Main System Revert to Default Data The instrument will boot to main mode and changes to the module s default parameter May be also required to load resident firmware into the module Boot Resident System Revert to Default Data The instrument will boot to resident mode and changes to the module s defa...

Страница 261: ...guration Switch Table 33 Boot Resident Forced Coldstart SW1 SW2 SW3 SW4 SW5 SW6 Init Mode 1 0 0 0 0 0 Boot Main System Keep Data 1 1 0 0 0 0 Boot Resident System Keep Data 1 0 0 0 0 1 Boot Main System Revert to Default Data 1 1 0 0 0 1 Boot Resident System Revert to Default Data Note The setting 0 down is essential ...

Страница 262: ...This pack is then housed in a metal inner cabinet which is enclosed by a plastic external cabinet The advantages of this packaging technology are virtual elimination of fixing screws bolts or ties reducing the number of components and increasing the speed of assembly disassembly the plastic layers have air channels molded into them so that cooling air can be guided exactly to the required location...

Страница 263: ... zero after the required maintenance procedure Using the EMF Counters The user settable EMF limits for the EMF Counters enable the early maintenance feedback to be adapted to specific user requirements The useful maintenance cycle is dependent on the requirements for use Therefore the definition of the maximum limits need to be determined based on the specific operating conditions of the instrumen...

Страница 264: ...264 Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 12 Hardware Information Early Maintenance Feedback ...

Страница 265: ... Ground the Instrument 267 Do Not Operate in an Explosive Atmosphere 268 Do Not Remove the Instrument Cover 268 Do Not Modify the Instrument 268 In Case of Damage 268 Solvents 269 Safety Symbols 270 Waste Electrical and Electronic Equipment WEEE Directive 272 Radio Interference 273 Sound Emission 274 Solvent Information 275 Agilent Technologies on Internet 276 This chapter provides safetey and oth...

Страница 266: ...es assumes no liability for the customer s failure to comply with these requirements Safety Standards This is a Safety Class I instrument provided with terminal for protective earthing and has been manufactured and tested according to international safety standards General Do not use this product in any manner not specified by the manufacturer The protective features of this product may be impaire...

Страница 267: ...d compliance with safety or EMC regulations Make all connections to the unit before applying power NOTE Note the instrument s external markings described under Safety Symbols on page 270 WARNING Missing electrical ground Electrical shock If your product is provided with a grounding type power plug the instrument chassis and cover must be connected to an electrical ground to minimize shock hazard T...

Страница 268: ...RNING Presence of flammable gases or fumes Explosion hazard Do not operate the instrument in the presence of flammable gases or fumes WARNING Instrument covers removed Electrical shock Do Not Remove the Instrument Cover Only Agilent authorized personnel are allowed to remove instrument covers Always disconnect the power cables and any external circuits before removing the instrument cover WARNING ...

Страница 269: ...below the boiling point of the solvent used Do not operate the instrument in an explosive atmosphere Do not use solvents of ignition Class IIC according IEC 60079 20 1 for example carbon disulfide Reduce the volume of substances to the minimum required for the analysis Never exceed the maximum permissible volume of solvents 8 L in the solvent cabinet Do not use bottles that exceed the maximum perm...

Страница 270: ...ains fluorinated greenhouse gas refrigerant according to the Kyoto protocol For specifications of refrigerant charge capacity carbon dioxide equivalent CDE and global warming potential GWP see instrument label Flammable Material For Sample Thermostat which uses flammable refrigerant consult Agilent Information Center User Manual before attempting to install or service this equipment All safety pre...

Страница 271: ...ould damage TVs and laptops computer hard drives credit and ATM cards data storage media mechanical watches hearing aids and speakers Keep magnets at least 25 mm away from devices and objects that could be damaged by strong magnetic fields Indicates a pinching or crushing hazard Indicates a piercing or cutting hazard Table 34 Symbols WARNING A WARNING alerts you to situations that could cause phys...

Страница 272: ...ment WEEE Directive This product complies with the European WEEE Directive marking requirements The affixed label indicates that you must not discard this electrical electronic product in domestic household waste NOTE Do not dispose of in domestic household waste To return unwanted products contact your local Agilent office or see http www agilent com for more information ...

Страница 273: ...d to provide optimized protection against radio interference All cables are in compliance with safety or EMC regulations Test and Measurement If test and measurement equipment is operated with unscreened cables or used for measurements on open set ups the user has to assure that under operating conditions the radio interference limits are still met within the premises ...

Страница 274: ...r s Declaration This statement is provided to comply with the requirements of the German Sound Emission Directive of 18 January 1991 This product has a sound pressure emission at the operator position 70 dB Sound Pressure Lp 70 dB A At Operator Position Normal Operation According to ISO 7779 1988 EN 27779 1991 Type Test ...

Страница 275: ... and nitric acid especially at higher temperatures if your chromatography method allows replace by phosphoric acid or phosphate buffer which are less corrosive against stainless steel halogenated solvents or mixtures which form radicals and or acids for example 2CHCl3 O2 2COCl2 2HCl This reaction in which stainless steel probably acts as a catalyst occurs quickly with dried chloroform if the dryin...

Страница 276: ...s 1260 Infinity II FLD User Manual 13 Appendix Agilent Technologies on Internet 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 ...

Страница 277: ... E electrical connections descriptions of 247 electronic waste 272 EMF early maintenance feedback 263 emission condenser lens 19 emission grating 19 emission monochromator 23 emission slit 19 error messages A D Overflow 157 ADC Not Calibrated 157 compensation sensor open 153 compensation sensor short 153 fan failed 154 Flash Lamp Current Overflow 158 Flash Trigger Lost 159 FLF Board not found 156 ...

Страница 278: ...firmware 223 materials bio inert 38 message A D Overflow 157 ADC Not Calibrated 157 encoder index not found 162 Flash Lamp Current Overflow 158 Flash Trigger Lost 159 FLF Board not found 156 Flow Cell Removed 160 ignition without cover 154 154 Lamp Cover Open 156 Motor Encoder Index Wrong 164 motor friction too high 162 motor messages 161 motor or encoder not found 161 Motor Position Lost 163 moto...

Страница 279: ...on 266 standards 45 symbols 270 selecting peakwidth 132 response time 132 serial number information 249 249 set up detector 60 settings peakwidth 133 response time 133 shutdown 150 site requirements 41 power cords 43 solvents 275 sound emission 274 special interfaces 254 special settings boot main 260 boot resident 260 specification physical 45 specifications analog outputs 50 47 communications 50...

Страница 280: ...al Index W warnings and cautions 207 waste electrical and electronic equipment 272 wavelength calibration procedure 184 187 wavelength calibration 184 wavelength shift of spectra 120 wavelength recalibration 170 WEEE directive 272 weight 45 X xenon flash lamp 19 21 ...

Страница 281: ...Agilent InfinityLab LC Series 1260 Infinity II FLD User Manual 281 Index ...

Страница 282: ...This manual contains technical reference information about the Agilent 1260 Infinity II Fluorescence Detector G7121A and the Agilent 1260 Infinity II Fluorescence Detector Spectra G7121B introduction and specifications using and optimizing troubleshooting and diagnose maintenance parts identification hardware information safety and related information ...

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