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G1315-90006 - Agilent Technologies

1. 3 While releasing the holder with a screwdriver carefully Note remove the holmium oxide filter using a pair of tweezers covered with lint free cloth or tape Do not scratch the holmium oxide filter The holmium oxide filter can be cleaned with alcohol da Oiu and a lint free cloth FOUOOSESOOOOUOoOOO0d 1200 Series DAD and MWD User Manual 129 8 Maintenance 4 While releasing the holder with a screw driver carefully 5 Replace the flow cell cover and fix the six screws insert the holmium oxide filter a eel ILI er Hagai FIT Hf J H Ceea n ee SSI ma n H a Next steps 6 Perform a Wavelength Verification and Recalibration on page 138 to check the proper
2. i lE 1200 Series DAD and MWD User Manual 101 8 Maintenance 3 When replacing the vis lamp assure that the vis lamp is inserted as shown flat edge towards the deuterium lamp opposite as in the figure OOOOOO OOOO OES0oOR a oe fu ar00 i _ OC00TtE000TH 4 Insert the lamp Fix the screws and reconnect the lamp to connector F DOLI C DULI a Doomoo auoogu00 H OgpooggaqgoagaH I uh Next steps 7 Reset the lamp counter as described in the user interface documentation 8 Turn the lamp ON and give the lamp 10 minutes to warm up 9 Performa Wavelength Verification and Recalibration on page 138 ora Holmium Oxide Test on page 139 to check the correct positioning of the UV lamp 10 Perform an intensity test see Intensity Test on page 136 102 1200 Series DAD and MWD User Manual Exchanging a Flow Cell When required repair Tools required Parts required Maintenance 8 If an application needs a different type of flow cell or the flow cell needs Two 1 4 inch wrenches for capillary connections Standard flow cell 10 mm 13 pl 120 bar G1315 60022 Semi micro flow cell 6 mm 5 pl 120 bar G1315 60025 Micro flow cell 3 mm 2 pl 120 bar G1315
3. Use narrow slit e g 1 nm if your analytes have narrow absorbance linearity bands and for high concentrations Use a wide slit e g 16 nm to detect very low concentrations Optimizing spectral acquisition DAD only Select spectra acquisition mode according to your needs Set the spectral wavelength range for colorless samples 190 400 nm is sufficient Set step to 4 nm for normal use set small step and slit width if high resolution of spectra with fine structure is wanted Typical column Typical peak Recommended flow cell length width T lt 5 cm 0 025 min Micro or Semi nano 10 cm 0 05 min Semi micro High pressure flow cell flow cell for pressures 20 cm 0 1 min Standard above 100 bar flow cell gt 40 cm 0 2 min Typical flow 0 01 0 2 0 2 0 4 0 4 0 4 1 2ml min 0 01 5 rate ml min ml min ml min ml min Internal column diameter 0 5 1 mm 2 1 mm 3 0 mm 4 6 mm 70 Figure 25 Choosing a Flow Cell in HPLC 1200 Series DAD and MWD User Manual How to optimize the Detector 5 Optimizing for Sensitivity Selectivity Linearity and Dispersion Flow Cell Path Length Lambert Beer s law shows a linear relationship between the flow cell path length and absorbance I Absorbance logT log C d where T is the transmission defined as the quotient of the intensity of the transmitted light I divided by the intensity of the incident light Io is the ext
4. DADI A Sigmi 4 Reeet0 100 ONR O NON D mau 4 Figure 7 Typical Chromatogram with UV detector Optimization of the System The settings used for this analysis are specific for this purpose For other applications the system can be optimized in various ways Please refer to the section Optimizing the Detector on page 66 or Optimizing in the your module s Reference Manual 1200 Series DAD and MWD User Manual Using the Detector 4 Preparing the HPLC System 1 Turn on the Agilent ChemStation PC and the monitor 2 Turn on the 1200 series HPLC modules 3 Start the Agilent ChemStation software B 02 01 If the pump autosampler thermostatted column compartment and detector are found the ChemStation screen should look like shown in Figure 8 The System status is red Not Ready System status E meat eee La T eu Ahe Urea DEFAULT ro Dotai X eseesessees g On line plot window details window Figure 8 Initial ChemStation screen Method and Run Control 1200 Series DAD and MWD User Manual 49 4 Using the Detector 4 Turn on the detector lamp pump and autosampler by clicking the System On button or the buttons below the module icons on the graphical user interface GUI After some time the pump thermostatted column compartment and detector module will turn to green 2 P Tumen the uv lamp of the DAD Module 7 Ce E Vasa
5. 9 Parts and Materials for Maintenance Overview of Maintenance Parts 144 Standard Flow Cell 146 Semi Micro Flow Cell 148 Micro Flow Cell 150 High Pressure Flow Cell 152 Prep Flow Cell SST 154 Prep Flow Cell Quartz 156 Nano Flow Cells 158 Accessory Kit 162 This chapter provides information on parts for maintenance Agg Agilent Technologies 143 9 Parts and Materials for Maintenance Overview of Maintenance Parts 00O O 0000 144 1200 Series DAD and MWD User Manual Parts and Materials for Maintenance 9 Table 16 Maintenance Parts Item Description Part Number 1 Front cover 5065 9982 2 Control Module G1323B or G1323 67001 Instant Pilot G4208A G4208 67001 3 Flow Cells with ID tag For details see Standard Flow Cell on page 146 Semi Micro Flow Cell on page 148 Micro Flow Cell on page 150 High Pressure Flow Cell on page 152 or Nano Flow Cells on page 158 4 Capillary column detector 380 mm lg 0 17 i d for details see Accessory Kit on page 162 G1315 87311 5 Zero dead volume fitting 5022 6515 6 Flow cell door G1315 68707 Screw special for flow cell door 5022 2112 7 Holmium oxide filter 79880 22711 8 Longlife Deuterium lamp 2140 0813 9 Tungsten lamp G1103 60001 10 Leak funnel 5041 8388 11 Leak funnel holder 5041 8389 12 Clip 5041 8387 13 Corrugated tubing 120 mm lg re order 5 m 5062 2463
6. CAUTION A CAUTION notice denotes a haz ard It calls attention to an operat ing procedure practice or the like that if not correctly performed or adhered to could result in damage to the product or loss of important data Do not proceed beyond a CAUTION notice until the indicated conditions are fully understood and met A WARNING notice denotes a hazard It calls attention to an operating procedure practice or the like that if not correctly per formed or adhered to could result in personal injury or death Do not proceed beyond a WARNING notice until the indicated condi tions are fully understood and met 1200 Series DAD and MWD User Manual In This Manual 1200 Series DAD and MWD User Manual This manual contains information for using your Diode Array and Multiple Wavelength Detectors Introduction to the Detector This chapter gives an introduction to the detector instrument overview and internal connectors Site Requirements and Specifications This chapter gives information on environmental requirements physical and performance specifications Installing the Detector This chapter describes the installation of the detector Using the Detector This chapter provides information on how to set up the detector for an analysis and explains the basic settings How to optimize the Detector This chapter provides information on how to optimize the detector Troubleshooting and Diagnostics Th
7. Exposure day Effective Irradiance 8 hours 0 1 uW cm 10 minutes 5 0 uW cm Typically the radiation values are much smaller than these limits Table 30 UV Radiation Typical Values Position Effective Irradiance Lamp installed 50 cm distance Average 0 016 W cm Lamp installed 50 cm distance Maximum 0 14 W cm 1200 Series DAD and MWD User Manual 173 A Appendix Solvent Information 174 Observe the following recommendations on the use of solvents Flow Cell Avoid the use of alkaline solutions pH gt 9 5 which can attack quartz and thus impair the optical properties of the flow cell Prevent any crystallization of buffer solutions This will lead into a blockage damage of the flow cell If the flow cell is transported while temperatures are below 5 degree C it must be assured that the cell is filled with alcohol Aqueous solvents in the flow cell can built up algae Therefore do not leave aqueous solvents sitting in the flow cell Add small of organic solvents e g Acetonitrile or Methanol 5 Solvents Brown glass ware can avoid growth of algae Always filter solvents small particles can permanently block the capillaries Avoid the use of the following steel corrosive solvents e Solutions of alkali halides and their respective acids for example lithium iodide potassium chloride and so on e High concentrations of inorganic acids like nitric acid sulfuric acid especially at higher temperatu
8. 1 1 GB hee coStBH 888388 E J Figure 9 Turning on the HPLC Module 50 1200 Series DAD and MWD User Manual Using the Detector 4 5 Purge the pump For more information see Priming and Purging the System on page 44 6 Allow the detector to warm up of at least 60 minutes to provide a stable baseline see example in Figure 10 and Table 8 For reproducible chromatography the detector and lamp should be on for at least one hour Otherwise the detector baseline may still drift depending on the environment DAD A Sataa Romano 1 TES Rann SS a Figure 10 Stabilization of Baseline both lamps turned on at the same time Table 8 Baseline drift after lamp turn on example from Figure 10 Time minutes Drift mAU hr 17 20 2 6 27 30 0 8 37 40 0 4 47 50 0 2 57 60 lt 0 2 1200 Series DAD and MWD User Manual 51 4 Using the Detector 7 For the isocratic pump fill the solvent bottle with the mixture of HPLC grade bi distilled water 80 and acetonitrile 70 For binary and quaternary pumps you can use separate bottles 8 Click on the Load Method button and select DEF_LC M and press OK Alternative double click on the method in the method window The default LC method parameters are transferred into the 1200 modules Load Method System 2 x hog None Fer Cx Carel foEF_LC M co chem32 2 nethods OFMOCAL M OGALSTST M OGCALAS M DGCALOO1 M OGCALO
9. 3 Attach the copper foil to a convenient and exposed electrical ground Figure 38 Using the ESD Strap 1200 Series DAD and MWD User Manual 97 7 Maintenance and Repair 98 1200 Series DAD and MWD User Manual Agilent 1200 Series Diode Array and Multiple Wavelength Detectors User Manual 8 Maintenance Overview of Maintenance 100 Exchanging a Lamp 101 Exchanging a Flow Cell 103 Maintenance of Standard Semi Micro or Micro Flow Cell 106 Maintenance of High Pressure Flow Cell 111 Replacing Capillaries on a Standard Flow Cell 114 Replacing Capillaries on a Semi Micro Micro and High Pressure Flow Cell 119 Nano Flow Cell Replacing or Cleaning 123 Cleaning or Exchanging the Holmium Oxide Filter 128 Correcting Leaks 131 Replacing Leak Handling System Parts 132 Replacing the Interface Board 133 Replacing the Detector s Firmware 134 Tests amp Calibrations 135 Intensity Test 136 Wavelength Verification and Recalibration 138 Holmium Oxide Test 139 Cell Test ChemStation only 141 This chapter describes the maintenance of the detector and the required tests ot Agilent Technologies 99 8 Maintenance Overview of Maintenance Table 15 Simple Repairs The following pages describe maintenance simple repairs of the detector that can be carried out without opening the main cover Procedure Typical Frequency Notes Deuterium lamp or tungsten lamp exchange Flow cell exchange Flow cell parts
10. 56 1200 Series DAD and MWD User Manual Using the Detector 4 14 Select the menu item RunControl gt Sample Info and enter information about this application Figure 16 on page 57 Press OK to leave this screen KLI M Sample Info x Offline Data Analysis Operator Name Wolfgang Run Sequence peste Pause Sequence Path JE CHEMSTATION 2 DATAS x Subdirectory JISOTEST Resume Sequence Resume Injection Manual Filename Stop RunfInject Sequence Fs 1s0 01D i Prefix Counter 7 p QI DGCALAS M m Sample Parameters Location vial 1 blank run if no entry Sample Name fisocratic test sample Sample Amount 0 Multiplier fi ISTD Amount fo Dilution fi Comment Isocratic test sample 1 ul 30 70 H2O Acetonitrile 1 5 ml min l Run Method OK Cancel Help Figure 16 Sample Information 15 Fill the content of an isocratic standard sample ampoule into a vial and seal the vial with a cap and place the vial into autosampler tray position 1 1200 Series DAD and MWD User Manual 57 4 58 Using the Detector Running the Sample and Verifying the Results 1 To start a run select the menu item RunControl gt Run Method 2 This will start the 1200 modules and the online plot on the Agilent ChemStation will show the resulting chromatogram DADI A Sipa RetG00 100 1101 4 10 400003 D ral 4 Figure 17 Chromatogram with lsocratic
11. Based on these data the built in processors calculate absorbance data with linear intervals 1 0 2 0 between data points This results in an excellent wavelength accuracy and instrument to instrument reproducibility The micro slit system makes use of the mechanical properties of silicon combined with the precise structuring capabilities of bulk micro machining It combines the required optical functions slit and shutter in a simple and compact component The slit width is directly controlled by the micro processor of the instrument and can be set as method parameter The combination of dispersion and spectral imaging is accomplished by using a concave holographic grating The grating separates the light beam into all its component wavelengths and reflects the light onto the photodiode array 1200 Series DAD and MWD User Manual Introduction to the Detector 1 Diode Array The diode array is a series of 1024 individual photodiodes and control circuits located on a ceramic carrier With a wavelength range from 190 950 nm the sampling interval is lt 1 nm 1200 Series DAD and MWD User Manual 15 1 Introduction to the Detector Early Maintenance Feedback EMF 16 Maintenance requires the exchange of components which are subject to wear or stress Ideally the frequency at which components are exchanged should be based on the intensity of usage of the detector and the analytical conditions and not on a predefined time
12. 120000 Accumulated UV lamp Lowest intensity in Lowest intensity in Lowest intensity n Lowest intensity in 3158 03010353 lfgang 11 2005 36 20 on time range 190nm range 221nm range 351nm range 50lnm 220nm 350nm 500nm 50nm 600 800 Wavelength nm Specification vvv gt 2000 5000 2000 4000 Highest intensity in range 190nm 350nm lt 450000 Highest intensity in range 700nm 950 lt 3 00000 Highest intensity for the D2 alpha line lt 1200000 Figure 42 Intensity Test Results report 1200 Series DAD and MWD User Manual cts cts cts ets cts cts Measured 8 66 h 19677 20612 17300 27853 96368 83955 175830 angqaqaqgqaoaa Arnarr vec aeeunuuvae Result Passed Passed Passed Passed Passed Passed Passed 137 8 138 Wavelength Verification and Recalibration The detector uses the alpha 656 1 nm and beta 486 nm emission lines of the deuterium lamp for wavelength calibration The sharp emission lines enable more accurate calibration than is possible with holmium oxide When verificaation is started the 1 nm slit is moved into the light path automatically and the gain is set to zero To eliminate effects due to absorbing solvents the test should be done with bubble free degassed HPLC water in the flow cell If a deviation is found and displayed it can be recalibrated by pressing Adjust The deviations are tracked in the Calibration
13. 14 Teflon Tubing flexible i d 0 8 mm flow cell to waste see Accessory Kit on page 162 5062 2462 Cable CAN to Agilent 1200 Series modules 0 5 m 5181 1516 Cable CAN to Agilent 1200 Series modules 1 m 5181 1519 LAN Communication Interface Board G1369A Cross over network cable shielded 3 m long for point to point connection Twisted pair network cable shielded 7 m long for hub connections Analog cable BNC to general purpose spade lugs Interface board BCD BCD external contacts G1369 60001 5023 0203 5023 0202 01046 60105 G1351 68701 1200 Series DAD and MWD User Manual 145 9 Parts and Materials for Maintenance Standard Flow Cell 146 Table 17 Standard Flow Cell Parts Item Description Part Number A wo N Standard flow cell assembly 10 mm 13 yl maximum pressure 120 bar 12 MPa with I D tag Window screw Spring washers pack of 10 Compression washer Window holder Quartz window Gasket BACK Teflon 2 3 mm hole outlet side Gasket FRONT Teflon 1 3 mm hole inlet side Window assembly comprises item 1 2 3 4 5 Capillary IN 0 17 mm 590 mm lg including heat exchanger Capillary OUT 0 17 mm 200 mm lg Clamp unit Screw M 2 5 4 mm lg for cell body clamp Zero dead volume fitting Cell repair kit STD includes window screw kit 4mm kexagonal wrench and seal kit Seal kit BACK for STD flow cell qty 12 of item 6 Seal kit FRONT for STD flow cell qty 12 of it
14. 20 and the pressure offset to 50 The X axis range should be 15 minutes Press OK to exit this screen Ln nnn SCC ee G 2 Pl ie mg Ma LTD O Edit Signal Plot DEFAULT O Available Signals Selected Signals uaternary Pump Flow DAD Signal 254 4 Reference 360 aternary Pump A 4d Quaternary Pump Pressure Add gt 2 uaternary Pump B aes uaternary Pump oC D uaternary Pump D E atah lad olumn Thermostat Temperature le E116Bhee 5 L olumn Thermostat Temperature ri AD B Signal 254 16 Reference lt r Window DAD A Signal 254 4 Reference 360 100 x axis 15 J min Type acquired y axis range 1 mau J draw zero line J auto y adjust Offset 20 4 Fraction Colector __ p Method Settings J Show Fraction collection ticks J Use method settings Apply to Method OK Cancel Help Figure 14 Edit Signal Plot Window 1200 Series DAD and MWD User Manual 55 4 Using the Detector The Online Plot Figure 15 on page 56 shows both the pump pressure and the detector absorbance signals Pressing Adjust the signals can be reset to the offset value and Balance would do a balance on the detector Figure 15 Online Plot Window 13 If both baselines are stable set the Y range for the detector signal to 100 mAU If you start with a new UV lamp for the first time the lamp may show initial drift for some NOTE time burn in effect
15. 60024 High pressure flow cell 6 mm 1 7 pl 400 bar G1315 60015 Nano flow cell refer to Nano Flow Cell Replacing or Cleaning on page 123 Preparations Turn the lamp s off Remove the front cover 1 Press the release buttons and remove the front cover to have access to the flow cell area 2 Press the release button and open the flow cell door QO0ooo0000C0o00 7 ar _OOUOUU0S0000000 OOOO EORR0H00eES ogg oggggroooiddy LEEI Sex Sail N 5 oon OOOO OCC OOOROEO000 oooga _ DOOROeO0000e 1200 Series DAD and MWD User Manual 103 8 Maintenance Note Depending on the system setup the inlet capillary might be routed directly from the module above or below to the cell and not to the capillary holder 3 Disconnect the cell inlet and the cell outlet capillary and the waste capillary from the capillary holder a booeone J 4 Loosen the thumb screw and remove the flow cell outlet capillary bottom with the union 104 1200 Series DAD and MWD User Manual Maintenance 8 Note The label attached to the flow cell provides information on part number path length volume and maximum pressure If you want to replace flow cell parts see Maintenance of Standard Semi Micro or Micro Flow Cell on page 106 Maintenance of
16. Array and Multiple Wavelength Detectors User Manual 6 Troubleshooting and Diagnostics Overview of the Detector s Indicators and Test Functions 88 Status Indicators 89 User Interfaces 91 Agilent LC Diagnostic Software 92 This chapter gives an overview about the troubleshooting and diagnostic features and the different user interfaces ee Agilent Technologies 87 6 Troubleshooting and Diagnostics Overview of the Detector s Indicators and Test Functions 88 Status Indicators The detector is provided with two status indicators which indicate the operational state prerun run and error states of the detector The status indicators provide a quick visual check of the operation of the detector see page 51 Error Messages In the event of an electronic mechanical or hydraulic failure the detector generates an error message in the user interface For each message a short description of the failure a list of probable causes of the problem and a list of suggested actions to fix the problem are provided see Error Information in the Service Manual Test Functions A series of test functions are available for troubleshooting and operational verification after exchanging internal components see Test Functions in the Service Manual Wavelength Verification Recalibration Wavelength recalibration is recommended after exchange of lamps and flow cells maintenance of flow cells repair of internal
17. D J20 fie faso F100 nm f Meautedbames e feso fie feso fioo 34 nm Mouv T Vis stop and post time can be set if required depending on the application the lamps aa Peakwidth Responseti poe pace espores can be selected one or both Store None 7 p gt 0 1 min 2s x i A peak width depends on the peaks in the Rangs 90 to 4007 mm E Sit chromatogram see page 64 Ste B 0 nm EER I Prerun anm z autobalance to zero absorbance on the Threshold 1 007 msu J Postrun analog output plus offset at begin and or end of run m Margin for negative Absorbance Timetable Total Lines 0 CEP T mechanical slit width can be changed Cancel Hep for further optimization see page 65 margin for negative absorbance see page 66 DAD Timetable System 2 x Line Time Sig Sam Bw Ref Bw Store Spectra Thresho e time table for programmable actions during the run imen Appena tut co _Baste Table C Graphic Carcel Heo Figure 13 Detector parameter default 54 1200 Series DAD and MWD User Manual Using the Detector 4 11 Pump the water acetonitrile 30 70 mobile phase through the column for 10 minutes for equilibration 12 Click the button and select Change to open the Signal Plot information Select the Pump Pressure and the DAD A Signal 254 4 as signals Change the Y range for the DAD to 1 mAU and the offset to
18. High Pressure Flow Cell on page 111 or Nano Flow Cell Replacing or Cleaning on page 123 6 Insert the flow cell while pressing the flow cell holder 7 Insert the flow cell capillaries into the union holder top is inlet bottom is outlet Tighten the thumb screw and Reconnect the waste tubing bottom to the union Note To check for leaks establish a flow and observe the flow cell outside of the cell compartment and all capillary connections l DZ inlet y it outlet 6 waste thumbscrew Next steps correct positioning of the flow cell 9 Replace the front cover 8 Perform a Wavelength Verification and Recalibration on page 138 or a Holmium Oxide Test on page 139 to check the 1200 Series DAD and MWD User Manual 105 8 Maintenance Maintenance of Standard Semi Micro or Micro Flow Cell When required If the flow cell needs repair due to leaks or contaminations reduced light throughput Tools required Two 1 4 inch wrenches for capillary connections hexagonal key 4 mm Tooth picks Parts required For parts see Standard Flow Cell on page 146 Semi Micro Flow Cell on page 148 or Micro Flow Cell on page 150 Preparations Turn the flow off Remove the front cover Remove the flow cell see Exchanging a Flow Cell on page 103 The gaskets used in the standard and semi micro micro flow cell are different 106
19. Part Number Prep Flow Cell Quartz 0 3 mm 20 bar 2 MPa G1315 60017 completely assembled includes all items below Prep Flow Cell Quartz 0 06 mm 20 bar 2 MPa G1315 60018 completely assembled includes all items below 1 PTFE tubing 2 m length 0 8 mm i d 0 D 1 6 mm G1315 67301 with fittings for flow cell assembled see note below 1 PTFE tubing 80 cm length 0 5 mm i d o D 1 6 mm G1315 67302 with fittings for flow cell assembled see note below 2 Fitting fingertight 2 pk 0100 1516 3 Cell housing G1315 27705 4 Handle for clamp unit G1315 84901 5 Clamp unit G1315 84902 6 Screw M 2 5 4 mm lg for cell body clamp 0515 1056 7 Quartz body Prep Cell 0 3 mm G1315 80004 7 Quartz body Prep Cell 0 06 mm G1315 80003 The flow cell comes with two tubings 0 8 mm i d and one 0 5 mm i d so that the combination at the flow cell could be either 0 8 0 8 or 0 5 0 8 inlet outet Standard is 0 8 0 8 Depending on the system pressure lt 30 ml min or bandbroadening the inlet tubing might be changed to 0 5 mm 1200 Series DAD and MWD User Manual Parts and Materials for Maintenance 9 Figure 55 Prep Flow Cell Quartz Parts 1200 Series DAD and MWD User Manual 157 9 Parts and Materials for Maintenance Nano Flow Cells The following kits are available Table 23 Nano flow cell kits Description Part number 500 nl Flow Cell Kit G1315 68724 includes Flow cell assembly 10 mm 500 nl 5 MPa completely as
20. What you will need 1200 system Column Standard Pump plus degassing Autosampler Detector standard flow cell installed Agilent ChemStation B 02 01 and above or Instant Pilot G4208 A 01 01 and above optional for basic operation or Control Module G1323B B 04 02 and above optional for basic operation System should be correctly set up for LAN communication with the Agilent ChemStation Zorbax Eclipse XDB C18 4 6 x 150 mm 5 um Part No 993967 902 or Part No 5063 6600 Part No 01080 68704 0 15 wt dimethylphthalate 0 15 wt diethylphthalate 0 01 wt biphenyl 0 03 wt o terphenyl in methanol 1200 Series DAD and MWD User Manual Conditions Using the Detector 4 A single injection of the isocratic test standard is made under the conditions given in Table 7 Table 7 Conditions Flow Stoptime Solvent Temperature Wavelength Injection Volume 1 5 ml minute 8 minutes 100 30 water 70 Acetonitrile Ambient sample 254 nm 4 nm bandwidth reference 360 nm 100 nm bandwidth Tul 1200 Series DAD and MWD User Manual 47 4 48 Using the Detector Typical Chromatogram A typical chromatogram for this analysis is shown in Figure 7 The exact profile of the chromatogram will depend on the chromatographic conditions Variations in solvent quality column packing standard concentration and column temperature will all have a potential effect on peak retention and response
21. allow optimum integration and display of your peaks and to minimize disk space required to store chromatograms and spectra DAD1 A Sig 254 4 Ref off NOISE INM1NM D mAU F Unfiltered iu A EN AAE T T T T T T T T 05 4 15 2 25 3 36 mir DAD1 A Sig 254 4 Ref off NOISE INMOOS D mAU F o4 Response ime 0 05 min on OT OLN ATA a aati 044 T TT T T T T T T 0 05 4 1 6 2 2 5 3 35 mi DAD1 A Sig 254 4 Ref off NOISE INMO1 D mAU J o4 Response time 0 1 min P a 0 44 aaa aaa aaa 0 5 4 15 2 2 5 3 35 mit lot Figure 27 Influence of Response Time on Signal and Noise Figure 13 lists the filter choices of the detector To get optimum results set peak width as close as possible to a narrow peak of interest in your chromatogram Response time will the be approximately 1 3 of the peak width resulting in less than 5 peak height reduction and less than 5 additional peak dispersion Decreasing the peak width setting in the detector will result in less than 5 gain in peak height but baseline noise will increase by a factor of 1 4 for a factor of 2 response time reduction Increasing peak width response time by factor of two from the recommended setting over filtering will reduce peak height by about 20 and reduce baseline noise by a factor of 1 4 This gives you the best possible signal to noise ratio but may affect peak resolution 1200 Series DAD and MWD User Manual 73 5 74 How
22. and 500 nl Preparations Turn the lamp s off Remove the front cover Remove the flow cell see Exchanging a Flow Cell on page 103 For details refer to the technical that comes with the nano flow cell kit The quartz block can be cleaned with alcohol DO NOT touch the inlet and outlet windows at the quartz block 1200 Series DAD and MWD User Manual 123 8 Maintenance 1 Disconnect the capillaries from the capillary holder and 2 Unscrew the cell body from the holder remove the flow cell 3 Unscrew the capillaries from the flow cell DO NOT use the 4 Using for example a toothpick press on the plastic part adapter at this time and slide the quartz body out of the cell housing 124 1200 Series DAD and MWD User Manual Maintenance 8 5 The quartz body and the cell seal assembly can be separated for cleaning purpose 6 This figure shows the correct holding of the quartz body and the cell seal assembly 7 Replace the cell seal assembly onto the quartz body Always use a new seal assembly to exclude damage during disassembling 8 Slide the quartz body completely into the cell body to the front stop use for example a toothpick 1200 Series DAD and MWD User Manual 125 8 Maintenance 9 Insert the flow cell capillaries and tighten them fingertight Use the wrench and torque adapter as described on page 127 and tighten the fittings a
23. detector LAN to LC ChemStation location depends on detector Figure 4 Recommended Stack Configuration Rear View 34 1200 Series DAD and MWD User Manual Installing the Detector 3 Installing the Detector Preparations Locate bench space Provide power connections Unpack the detector Parts required Detector Power cord for other cables see below ChemStation and or Control Module G1323B or Instant Pilot G4208A 1 Install the LAN interface board in the detector if required see Replacing the Interface Board on page 133 Place the detector in the stack or on the bench in a horizontal position Ensure the line power switch at the front of the detector is OFF Status indicator poa 7 green yellow red ai 7 wll Line power switch with green light Figure 5 Front View of Detector 1200 Series DAD and MWD User Manual 35 3 Installing the Detector 4 Connect the power cable to the power connector at the rear of the detector 5 Connect the CAN cable to other Agilent 1200 Series modules 6 Ifa Agilent ChemStation is the controller connect the LAN connection to the LAN interface board in the detector If a Agilent 1200 DAD MWD EFLD is in the system the LAN should be connected to the DAD MWD FLD due to higher data load 7 Connect the analog cable s optional 8 Connect the APG remote cable optional for non Agilent 1200 Series instruments 9 Turn ON power by pushing the button at
24. function of the holmium oxide filter 7 Insert the flow cell see Exchanging a Flow Cell on page 103 8 Replace the front cover 9 Turn ON the flow 130 1200 Series DAD and MWD User Manual Correcting Leaks When required Tools required Parts required Preparations Maintenance If a leakage has occurred in the flow cell area or at the heat exchanger or at the capillary connections Tissue Two 1 4 inch wrenches for capillary connections None Remove the front cover 1 Use tissue to dry the leak sensor area and the leak pan 2 Observe the capillary connections and the flow cell area for leaks and correct if required Leak sensor assembly Figure 40 Observing for Leaks 3 Replace the front cover 1200 Series DAD and MWD User Manual 131 8 Maintenance Replacing Leak Handling System Parts When required If the parts are corroded or broken Tools required None Parts required Leak funnel 5061 3388 Leak funnel holder 5041 8389 Leak tubing 120 mm 0890 1711 Preparations Remove the front cover 1 Pull the leak funnel out of the leak funnel holder 2 Pull out the leak funnel with the tubing 3 Insert the leak funnel with the tubing in its position 4 Insert the leak funnel into the leak funnel holder Leak funnel holder Leak funnel Leak tubing Figure 41 Replacing Leak Handling System Parts 5 Replace the front c
25. handling and safety data sheet by the solvent vendor especially when toxic or hazardous solvents are used 1200 Series DAD and MWD User Manual 167 A Appendix Safety Symbols Table 28 shows safety symbols used on the instrument and in the manuals Table 28 Safety Symbols Symbol Description The apparatus is marked with this symbol when the user should refer to the instruction manual in order to protect the apparatus against damage g Indicates dangerous voltages Indicates a protected ground terminal a Indicates eye damage may result from directly viewing the light produced by the deuterium lamp used in I this product WARNING A warning alerts you to situations that could cause physical injury or damage to the equipment Do not proceed beyond a warning until you have fully understood and met the indicated conditions CAUTION A caution alerts you to situations that could cause a possible loss of data Do not proceed beyond a caution until you have fully understood and met the indicated conditions 168 1200 Series DAD and MWD User Manual Appendix A Lithium Batteries Information Danger of explosion if battery is incorrectly replaced Replace only with the same or equivalent type recommended by the equipment manufacturer Lithium batteries may not be disposed off into the domestic waste Transportation of discharged Lithium batteries through carriers regulated by IATA ICAO ADR RID IMDG is not allow
26. interval The early maintenance feedback EMF feature monitors the usage of specific components in the instrument and provides feedback when the user selectable limits have been exceeded The visual feedback in the user interface provides an indication that maintenance procedures should be scheduled EMF Counters The detector provides two EMF counters for the lamps The counters increment increments with lamp use and can be assigned a maximum limit which provides visual feedback in the user interface when the limit is exceeded The counters can be reset to zero after the lamp is exchanged The detector provides the following EMF counters e Deuterium Lamp On Time e Tungsten Lamp On Time 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 lamp burn time is dependent on the requirements for the analysis high or low sensitivity analysis wavelength etc therefore the definition of the maximum limits need to be determined based on the specific operating conditions of the instrument 1200 Series DAD and MWD User Manual Introduction to the Detector 1 Setting the EMF Limits The setting of the EMF limits must be optimized over one or two maintenance cycles Initially no EMF limit should be set When instrument performance indicates maintenance is necessary take note of the values displayed by lamp counters Enter th
27. l a Figure 19 Online Spectra Window 2 Change the absorbance and wavelength range according your needs 60 1200 Series DAD and MWD User Manual P ECs CHEM3 BATCH QI DeaLs QI DECAL QI DGCAL QI pecat QI DGCAL QI DGCAL QI DECAL Pl neces 1200 Seri Using the Detector Analog Output Settings To change the Output Range of the analog outputs see Control Settings on page 59 1 To change the offset and the attenuation select Analog Outputs 2 Change the ranges for absorbance and wavelength according your needs DAD Analog Outputs System 2 xj 4 Set up Pump Set up Injector Set up Column Thermostat Set up DAD Signals More Pump More Injector More Column Thermostat More DAD Analog Outp Control Configuration Run Recovery Output 1 Output 2 Zero Offset E 5 Attenuation 1000 m uU 1000 m u N System On Zero Offset Limits 1 to 99 in steps of 1 System OFF Attenuation Limits 0 98 to 2000 mAU at discrete values for either 100 mV or 1 V full scale Revisions amp Serial s Columns Configure 1100 1200 Access Figure 20 Analog Output Settings 3 Change the values if required es DAD and MWD User Manual 61 4 Using the Detector Spectrum Settings To change the Spectra settings open 1 To change the Spectra settings select Setup Detector Signals 2 In the secti
28. the amplitude of the temperature changes to below 1 C hour 1 8 F hour Turbulences around one minute or less can be ignored Do not store ship or use your detector under conditions where temperature fluctuations could cause condensation within the detector Condensation will damage the system electronics If your detector was shipped in cold weather leave it in its box and allow it to warm up slowly to room temperature to avoid condensation 1200 Series DAD and MWD User Manual 23 2 Site Requirements and Specifications NOTE The G1315B DAD and G1365B MWD are designed to operate in a typical electromagnetic environment EN61326 1 where RF transmitters such as mobile phones should not be used in close proximity 24 1200 Series DAD and MWD User Manual Site Requirements and Specifications 2 Physical Specifications Table 1 Physical Specifications Type Weight Dimensions width x depth x height Specification 11 5 kg 26 lbs 345 x 435 x 140 mm 13 5 x 17 x 5 5 inches Comments Line voltage Line frequency Power consumption G1315B 65B 100 240 VAC 10 50 or 60 Hz 5 300 VA 125 W 427 BTU Wide ranging capability Maximum Ambient operating temperature Ambient non operating temperature Humidity Operating altitude Non operating altitude Safety standards IEC CSA UL EN 0 55 C 32 131 F 40 70 C 4 158 F lt 95 at 25 40 C 77 104 F Up
29. the lower left hand side of the detector The status LED should be green Interface board gt Analog signal 3c APG remote RS 232C Figure 6 Rear View of Detector 36 D gt 2 2 pi JC T 2 2 Security lever gt umn gt Sz M CAN Configuration switch 1200 Series DAD and MWD User Manual Installing the Detector 3 The detector is turned on when the line power switch is pressed and the green indicator lamp is illuminated The detector is turned off when the line power switch is protruding and the green light is OFF WARNING To disconnect the detector from line unplug the power cord The power supply still uses some power even if the power switch at the front panel is turned OFF The detector was shipped with default configuration settings 1200 Series DAD and MWD User Manual 37 3 Installing the Detector Flow Connections to the Detector Preparations Detector is installed in the LC system Parts required Other modules Parts from accessory kit see Detector Accessory Kit Contents on page 31 Two wrenches 1 4 5 16 inch for capillary connections When working with solvents please observe appropriate safety procedures for example goggles safety gloves and protective clothing as described in the material handling and safety data sheet supplied by the solvent vendor especially when toxic or hazardous solvents are used 38 The flow cell is ship
30. to 2000 m 6500 ft Up to 4600 m 14950 ft Installation category Il pollution degree 2 For indoor use only Non condensing For storing the detector 1200 Series DAD and MWD User Manual 25 2 Site Requirements and Specifications Performance Specifications Table2 Performance Specifications Agilent 1200 Series DAD and MWD Type Specification Comments Detection type 1024 element photodiode array Light source Wavelength range Short term noise ASTM Single and Multi Wavelengt Drift Linear absorbance range Wavelength accuracy Wavelength bunching Slit width Diode width Flow cells Control and data evaluation Analog outputs Deuterium and tungsten lamps 190 950 nm 1 x 10 AU at 254 and 750 nm See on page 27 2 x 10 AU hr at 254 nm See on page 27 gt 2 AU upper limit See on page 27 1nm Self calibration with deuterium lines verification with holmium oxide filter 1 400 nm Programmable in steps of 1 nm 1 2 4 8 16 nm Programmable slit lt 1inm Standard 13 ul volume 10 mm cell path length and 120 bar 1760 psi pressure maximum Semi Micro 5 pl volume 6 mm cell path length and 120 bar 1760 psi pressure maximum Micro 2 ul volume 3 mm cell path length and 120 bar 1760 psi pressure maximum High pressure 1 7 ul volume 6 mm cell path length and 400 bar 5880 psi pressure maximum 80 nano 0 08 pl volume 10 mm cell path length and 50 bar 725 psi pressure maximum 500 nan
31. wavelength caffeine shows exactly the same absorbance as at 222 nm When the absorbance values are subtracted from each another any indication of the presence of caffeine is eliminated In the same way hydrochlorothiazide can be suppressed if caffeine is to be quantified In this case the wavelength is set to 204 nm and the reference wavelength to 260 nm Figure 34 shows the chromatographic results of the peak suppression technique The trade off for this procedure is a loss in sensitivity The sample signal decreases by the absorbance at the reference wavelength relative to the signal wavelength Sensitivity may be decreased by as much as 10 30 1200 Series DAD and MWD User Manual 83 5 How to optimize the Detector Hydrochlorothiazide Hydrochlorothiazide and caffeine and caffeine Wavelength 204 nm Wavelength 222 nm No reference No reference Hydrochlorothiazide Caffeine suppressed suppressed Wavelength 204 nm Wavelength 222 nm Reference 260 nm Reference 282 nm Time min Time min Figure 34 Peak Suppression Using Reference Wavelength Ratio Qualifiers for Selective Detection of Compound Classes Ratio qualifiers can be used where in a complex sample only one particular class needs to be analyzed a parent drug and its metabolites in a biological sample for example Another example is the selective analysis of derivatives after pre or postcolumn derivatization Specifying a signal ratio that is typical for the sample clas
32. 0 nm 1nm 453 7 nm 452 7 454 7 nm 1nm 536 7 nm 535 7 537 7 nm 1nm The test is evaluated by the instrument and the measured maxima are displayed automatically The test fails if one or more of the maxima lies outside of the limits see Figure 44 on page 140 The evaluation report is only on the Agilent ChemStation available Test Failed Probable Causes e Absorbing solvent or air bubble in flow cell Incorrect calibration e Dirty or contaminated flow cell e Dirty or contaminated optical components achromat windows e Old or non Agilent lamp 1200 Series DAD and MWD User Manual 139 140 Maintenance Suggested Actions Ensure the flow cell is filled with water Y Recalibrate see Wavelength Verification and Recalibration on page 138 and repeat the test V Run the cell test see Cell Test ChemStation only on page 141 If the test fails exchange the flow cell windows V Clean optical components with alcohol and lint free cloth V Exchange the UV lamp l Number Wavelength nm Holmium Filter Test Results pecifica a Measured Res 361 nm z 6 nm 453 nm 452 453 1 nm 536 nm 535 7 536 4 nm Figure 44 Holmium Oxide Test Results report 1200 Series DAD and MWD User Manual Maintenance 8 Cell Test ChemStation only The cell test measures the intensity of the deuterium and tungsten lamps over the full wavelength range 190 950 nm once with the flow cell
33. 1 Teflon Tubing flexible i d 0 8 mm flow cell to waste 5062 2462 2m re order 5m Corrugated tubing to waste re order 5 m 5062 2463 1 2m Fitting male PEEK 0100 1516 2 Capillary column detector G1315 87311 1 380 mm lg 0 17 mm i d includes Ferrule front SST 0100 0043 2 Ferrule back SST 0100 0044 2 Fitting SST 79814 22406 2 Hex key set 1 5 mm Wrench open end 1 4 5 16 inch Wrench open end 4 mm 8710 0641 8710 0510 8710 1534 1200 Series DAD and MWD User Manual 31 3 Installing the Detector Optimizing the Stack Configuration If your detector is part of a complete Agilent 1200 Series system you can ensure optimum performance by installing the following configuration This configuration optimizes the system flow path ensuring minimum delay volume 32 1200 Series DAD and MWD User Manual Vacuum degasser Pump 0000 oo000 o000 0 Autosampler Column compartment Detector Figure 3 Recommended Stack Configuration Front View 1200 Series DAD and MWD User Manual Installing the Detector Local User Interface 3 33 3 installing the Detector Remote cable CAN Bus cable to local user interface Q o o o CAN Bus cable AC power Analog detector signal 1 or 2 outputs per
34. 1200 Series DAD and MWD User Manual Maintenance 8 1 Use a4 mm hex key to unscrew the window assembly and remove the gasket from the cell body Do not mix the gasket 6 and 7 different hole diameter Note If you want to replace the gasket only continue with step 7 Do not mix the gasket 6 and 7 They are different for standard and semi mirco flow cell 2 Use a tooth pick to remove the quartz window from the window assembly Note If the washers fall out of the window assembly they must be inserted in the correct order with the Teflon ring to prevent any leaks from the flow cell window 1200 Series DAD and MWD User Manual 107 8 Maintenance Noor wohr Orientation of Flow Cell Parts NOTE Gaskets 6 and 7 have different hole diameters window screw spring washers compression washer window holder quartz window gasket light in gasket light out 8 items 2 3 4 and 5 9 10 inlet capillary 11 outlet capillary 12 holder window screw contains flow cell body 3 Assemble the washers and the window assembly in correct order Correct orientation of spring washers 2 is required 108 1200 Series DAD and MWD User Manual Maintenance 5 Press the Teflon ring into the window assembly 6 Press the window assembly onto the new or cleaned quartz window 7 Insert a new gasket
35. 1200 Series DAD and MWD User Manual www agilent com In This Book This manual contains technical reference information about the Agilent 1200 Series diode array detectors The manual describes the following e introcduction and specifications e installation e using and optimizing e troubleshooting e maintenance and repair e parts identification e safety and related information Agilent Technologies 2006 Printed in Germany Edition 02 2006 G1315 90006 Agilent Technologies
36. 3 4 5 and 6 2 Seal ring 79883 27101 3 Quartz window 1000 0953 4 Compression washer 79883 28802 5 Spring washers pack of 10 5062 8553 6 Window screw 79883 22404 7 Capillary IN 0 12 mm 290 mm Ig including heat exchanger G1315 87325 8 Capillary OUT 0 12 mm 200 mm lg G1315 87306 9 Clamp unit G1315 84901 Screw M 2 5 4 mm lg for cell body clamp unit 0515 1056 Capillary 0 12 mm 150 mm lg G1315 87312 Capillary 0 17 mm 380 mm lg G1315 87311 High Pressure Cell Repair Kit includes 1 quartz window 79883 68700 1 compression washer 5 spring washers 2 seal rings 152 1200 Series DAD and MWD User Manual Parts and Materials for Maintenance 9 Figure 52 High Pressure Flow Cell Parts 1200 Series DAD and MWD User Manual 153 9 Parts and Materials for Maintenance Prep Flow Cell SST 154 For more details on the Preparative Flow Cells refer to the technical note that comes with the flow cells Table 21 Prep Flow Cell SST Parts Item Description Part Number AeA wo N 9a Prep Flow Cell SST 3 mm 4 pl 120 bar 12 MPa with I D Tag completely assembled includes all items below Window screw Spring washers pack of 10 Compression washer Window holder Quartz window Gasket BACK FEP Teflon 2 3 mm hole outlet side Seal kit BACK for STD flow cell qty 12 Gasket FRONT FEP Teflon 1 3 mm hole inlet side Seal kit FRONT for STD flow cell qty 12 Window assembly compr
37. 3 exchanging alamp 101 exchanging holmium oxide filter 128 introduction 94 of the detector 93 99 replacing firmware 134 replacing leak handling system 132 replacing semi micro flow cell capillaries 119 replacing STD flow cell capillaries 114 the high pressure flow cell 111 the standard semi micro flow cell 106 using the ESD strap 97 warnings and cautions 94 response time peak width 73 S safety information on lithium batteries 169 standards 25 sample and reference wavelength 74 selectivity optimization 82 semi micro flow cell dimensions 72 site requirements 22 slit width 77 specifications analog outputs 26 communications 27 diode width 26 flow cell 26 GLP features 27 linear range 26 noise and drift ASTM 26 programmable slit width 26 safety and maintenance 27 wavelength accuracy bunching 26 wavelength range 26 spectra acquisition 80 settings 63 spectrograph 13 14 diodes per nm 14 stack configuration 32 34 front view 32 rear view 34 standard flow cell dimensions 72 T test functions 88 tests intensity 136 time table 54 troubleshooting error messages 88 status indicators 88 89 U unpacking 30 using EMF 20 uv radiation 173 V variable entrance slit 14 verification and recalibration of wavelength 138 WwW wavelength accuracy and bunching 26 range 26 recalibration 88 verification and recalibration 138 weight and dimensions 25
38. 337 Waldbrenn 10 129729 9 v Voltage range Made in Germany mava sne Hower consumption frequency Figure 2 Rear View of Detector Electrical Connections and Label 1200 Series DAD and MWD User Manual 19 1 Introduction to the Detector Instrument Layout 20 The industrial design of the detector incorporates several innovative features It uses Agilent s E PAC concept for the packaging of electronics and mechanical assemblies This concept is based upon the use of expanded polypropylene EPP layers of foam plastic spacers in which the mechanical and electronic boards components of the detector are placed 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 e virtual elimination of fixing screws bolts or ties reducing the number of components and increasing the speed of assembly disassembly e the plastic layers have air channels molded into them so that cooling air can be guided exactly to the required locations e the plastic layers help cushion the electronic and mechanical parts from physical shock and e the metal inner cabinet shields the internal electronics from electromagnetic interference and also helps to reduce or eliminate radio frequency emissions from the instrument itself 1200 Series DAD and MWD User Manual Agilent 1200 Series Diode Array and Multiple Wavelength Detectors User Manual 2 Site Requirements and Speci
39. 6 7 and the window assembly 1 into the cell body Assure that the gasket is on the bottom Do not mix the gasket 6 and 7 different hole diameter 8 Using a 4 mm hex key tighten the window screw hand tight plus a quarter turn 1200 Series DAD and MWD User Manual 8 Maintenance Next steps 10 Reconnect the capillaries see Exchanging a Flow Cell on page 103 11 Perform a leak test 12 Insert the flow cell 13 Replace the front cover 14 Perform a Wavelength Verification and Recalibration on page 138 or a Holmium Oxide Test on page 139 to check the correct positioning of the flow cell 110 1200 Series DAD and MWD User Manual Maintenance 8 Maintenance of High Pressure Flow Cell When required If the flow cell needs repair due to leaks or contaminations reduced light throughput Tools required Two 1 4 inch wrenches for capillary connections hexagonal key 4 mm Tooth picks Parts required For parts see High Pressure Flow Cell on page 152 Preparations Turn the flow off Remove the front cover Remove the flow cell see Exchanging a Flow Cell on page 103 All descriptions in this procedure are based on the default orientation of the cell as it is manufactured The heat exchanger capillary and the cell body can be fixed mirror symmetrically to have both capillaries routed to the bottom or to the top depending on the routing of the capillaries to the colum
40. AD and MWD User Manual Site Requirements and Specifications 2 Never operate your instrumentation from a power outlet that has no ground connection Never use a power cord other than the Agilent Technologies power cord designed for your region Never use cables other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations CAUTION Bench Space The detector dimensions and weight see Table 1 allows you to place the detector on almost any desk or laboratory bench It needs an additional 2 5 cm 1 0 inches of space on either side and approximately 8 cm 3 1 inches in the rear for air circulation and electric connections If the bench should carry a Agilent 1200 Series system make sure that the bench is designed to bear the weight of all modules The detector should be operated in a horizontal position Environment Your detector will work within the specifications at ambient temperatures and relative humidity described in Table 1 ASTM drift tests require a temperature change below 2 C hour 3 6 F hour over one hour period Our published drift specification refer also to Performance Specifications on page 26 is based on these conditions Larger ambient temperature changes will result in larger drift Better drift performance depends on better control of the temperature fluctuations To realize the highest performance minimize the frequency and
41. Agilent 1200 Series Diode Array and Multiple Wavelength Detectors G1315B G1365B User Manual tii Agilent Technologies Notices Agilent Technologies Inc 2006 No part of this manual may be reproduced in any form or by any means including elec tronic storage and retrieval or translation into a foreign language without prior agree ment and written consent from Agilent Technologies Inc as governed by United States and international copyright laws Manual Part Number 61315 90006 Edition Edition 02 2006 Printed in Germany Agilent Technologies Hewlett Packard Strasse 8 76337 Waldbronn Manual Structure The User Manual G1315 90006 English and its localized versions contain a subset of the Service Manual and is shipped with the detector in printed matter Latest versions of the manuals can be obtained from the Agilent web The Service Manual G1315 90106 English contains the complete information about the Agilent 1200 Series Diode Array and Multiple Wavelength Detectors It is available as Adobe Reader file PDF only Warranty The material contained in this docu ment is provided as is and is sub ject to being changed without notice in future editions Further to the max imum extent permitted by applicable law Agilent disclaims all warranties either express or implied with regard to this manual and any information contained herein including but not limited to the imp
42. Cleaning or exchange Holmium oxide filter Cleaning or exchange Leak sensor drying Leak handling System replacement If noise and or drift exceeds your application limits or lamp does not ignite If application requires a different flow cell type If leaking or if intensity drops due to contaminated flow cell windows If contaminated If leak has occurred If broken or corroded An intensity test should be performed after replacement A holmium or wavelength calibration test should be performed after replacement A pressure tightness test should be done after repair A holmium or wavelength calibration test should be performed after replacement Check for leaks Check for leaks 100 1200 Series DAD and MWD User Manual Maintenance 8 Exchanging a Lamp When required If noise or drift exceeds application limits or lamp does not ignite Tools required Screwdriver POZI 1 PT3 Parts required Longlife Deuterium lamp 2140 0813 without black cover Longlife Deuterium lamp 5181 1530 with black cover Tungsten lamp G1103 60001 Preparations Turn the lamp s off CAUTION If the detector has been in use the lamp may be hot If so wait five minutes until lamp cools down 1 Press the release buttons and remove the front cover to 2 Disconnect lamp from the connector and unscrew the have access to the lamp area lamp and remove the lamp Do not touch the glass bulb with your fingers
43. History diagnosis buffer in the detector DAD Wavelength Calibration G1315B x Calibration history Deviation D2 Alpha D2 Beta Time Date 0 01nm O 04nm 13 19 31 16 03 2005 D2 Alpha EEEE PELE EE 1 2mm 656 1nm 1 2nm D2 Beta PUUerpi 1 2nm 486 0nm 1 2nm 3 EANN Reset history D2 Alpha line deviation is 0 00nm D2 Beta line deviation is 0 00nm OK Cancel Help Calibration settings equal to measured ones No calibration necessary Figure 43 Wavelength Verification and Recalibration Wavelength calibration should be done e after maintenance or change of the flow cell e lamp exchange or e after a major repair like processor board or optical unit exchange After calibration the holmium oxide test see Holmium Oxide Test on page 139 provides verification of wavelength accuracy at three additional wavelengths 1200 Series DAD and MWD User Manual Maintenance 8 Holmium Oxide Test The holmium oxide test uses three characteristic absorbance maxima of the built in holmium oxide filter to verify wavelength accuracy see also Wavelength Verification and Recalibration on page 138 When the test is started the 1 nm slit is moved into the light path automatically To eliminate effects due to absorbing solvents the test should be done with water in the flow cell See also Declaration of Conformity for HOX2 Filter on page 176 Holmium Oxide Test Evaluation Limits 361 0 nm 360 0 362
44. Q2M zj Oriver mesere 5 E Eke EG mate Obs cer sc z ime Ub oo lt 2 dja aa Bsgu Socoowmemos LA eam MN Oe ea a eras l Sm SIO i i Q vemoca2 m W Ocustst W ooceaasm W OaM oxcacgem U OAIM Uoan W aM Q ocan DEFAULT O Q ostmrs n Q oacerst Q vocrtest m W ow orst m Q DGEAKT M W ogo Q corres W behets owocem Edea Q iS0CRAM Q sonoru EIE Figure 11 Loading Default LC Method eo8b Be 88 eS8 52 1200 Series DAD and MWD User Manual Using the Detector 4 9 Click on the module icons Figure 12 and open the Setup of these modules Figure 13 on page 54 shows the detector settings do not change the detector parameters at this time i AE Set up DAD Signals j of Control E Set up Pump amay Data Curves Fu Online Spectra a Control Not Ready Information B Not Ready Information Help Help B Si 1 2 min Figure 12 Open the module menu 10 Enter the pump parameters mentioned under Conditions on page 47 1200 Series DAD and MWD User Manual 53 4 Using the Detector DAD Signals System 2 d x r Signals m Time Store Sample Bw Reference Bw Stoptime 2 Pump 4 aa up to 5 signals A to E with individual Am E je 380 foo S om x nomi wavelength settings can be selected B I fest fie fso 100 nm Postime Of f min spectrum settings see page 62 c fao fs feo foo Sf nm
45. SST qty 2 6 Fitting SST qty 2 Hex key set 1 5 mm Wrench open end 1 4 5 16 inch Wrench open end 4 mm G1315 68705 5062 2463 5062 2462 0100 1516 G1315 87311 0100 0043 0100 0044 79814 22406 8710 0641 8710 0510 8710 1534 For item number refer to Figure 57 and Figure 58 162 1200 Series DAD and MWD User Manual Parts and Materials for Maintenance 9 Figure 57 Waste Tubing Parts 9 0 This end is 3 pre installed aces Figure 58 Inlet Capillary Column Detector Parts 1200 Series DAD and MWD User Manual 163 9 Parts and Materials for Maintenance 164 1200 Series DAD and MWD User Manual Agilent 1200 Series Diode Array and Multiple Wavelength Detectors User Manual A Appendix General Safety Information 166 Lithium Batteries Information 169 The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC 170 Radio Interference 171 Sound Emission 172 UV Radiation 173 Solvent Information 174 Declaration of Conformity for HOX2 Filter 176 Agilent Technologies on Internet 177 This chapter provides safetey and other general information ee Agilent Technologies 165 A Appendix General Safety Information 166 The following general safety precautions must be observed during all phases of operation service and repair of this instrument Failure to comply with these precautions or with specific warnings elsewhere in this manual violates safety standards of design manufa
46. SWNIR wavelength range The image of the filament of the tungsten lamp is focused on the discharge aperture of the deuterium lamp by means of a special rear access lamp design which allows both light sources to be optically combined and share a common axis to the source lens The achromat source lens forms a single focused beam of light through the flow cell Each cell room and lamp are separated by a quartz window which can be cleaned or replaced In the spectrograph light is being dispersed onto the diode array by a holographic grating This allows simultaneous access to all wavelength information Cell suppor window Tungsten lamp Flow cell Coupling lens Spectro len Deuterium lamp Slit Achromat source lens Diode array Holmium oxide filter Grating Figure 1 Optical System of the Detector Lamps The light source for the UV wavelength range is a deuterium lamp with a shine through aperture As a result of plasma discharge in low pressure deuterium gas the lamp emits light over the 190 nm to approximately 800 nm 1200 Series DAD and MWD User Manual 13 1 Introduction to the Detector Achromat Source Lens Holmium Oxide Filter Cell Support Window Flow Cell Compartment Spectrograph Variable Entrance 14 Slit System Grating wavelength range The light source for the visible and SWNIR wavelength range is a low noise tungsten lamp This lamp emits light over the wavelength range 470 950
47. Test Sample Information about using the Data Analysis functions can be obtained from the Using your ChemStation manual supplied with your system 1200 Series DAD and MWD User Manual Using the Detector 4 Special Settings of the Detector In this chapter special settings of the G1315B DAD and G1365B MWD are described based on the Agilent ChemStation B 02 01 Control Settings setup a DAD Controlo X lt Lamps turn on and off of UV of Control p Lamps gt Error Method and Vis lamp fy Online Spectra Uv Vis J Take current method At Power On automatic lamp on Not Ready Information Help icc a ten at power on off off Analog Output Range Error Method take error method or current method in case of an Seo On Output 1 Output 2 error EEEN D IY Analog Output Range can be 1 Giy set to either 100 mV or 1 V full J Tum Vis lamp on scale see also Analog Output Settings on page 61 penin Automatic Turn On lamps can Tum required lames on ak be programmed detector must be on for this Date ddimm ppyy 28 12 2005 Help online help Time 2 47 42 hhimm ss gt p P OK Cancel Help Figure 18 Detector control settings 1200 Series DAD and MWD User Manual 59 4 Using the Detector Online Spectra 1 To view the online spectra during the run select Online Spectra a l J J
48. When the test is started the 1 nm slit is moved into the light path automatically and the gain is set to zero To eliminate effects due to absorbing solvents the test should be done with water in the flow cell The shape of the intensity spectrum is primarily dependent on the lamp grating and diode array characteristics Therefore intensity spectra will differ slightly between instruments Figure 42 on page 137 shows a typical intensity test spectrum Intensity Test Evaluation The Agilent ChemStation evaluates four spectral ranges automatically and displays the limits for each range the measured intensity counts and passed or failed for each spectral range see Figure 42 on page 137 The evaluation report is only on the Agilent ChemStation available Test Failed Probable Causes e Absorbing solvent or air bubble in flow cell e Dirty or contaminated flow cell e Dirty or contaminated optical components achromat windows e Old or non Agilent lamp 1200 Series DAD and MWD User Manual Suggested Actions V Ensure the flow cell is filled with water and free from air bubbles Maintenance 8 Y Run the cell test see Cell Test ChemStation only on page 141 If the test fails exchange the flow cell windows V Clean optical components with alcohol and lint free cloth V Exchange the lamp Instrument 61 Serial Number DE Operator Wo Date 24 Time 14 Intensity Plot Intensity counts 160000 140000
49. ady state when it is waiting for a specific condition to be reached or completed for example immediately after changing a set point or while a self test procedure is running An error condition is indicated when the status indicator is red An error condition indicates the detector has detected an internal problem which affects correct operation of the detector Usually an error condition requires attention e g leak defective internal components An error condition always interrupts the analysis A red blinking indicator indicates that the module is in resident mode e g during update of main firmware 1200 Series DAD and MWD User Manual Troubleshooting and Diagnostics 6 User Interfaces Depending on the user interface the available tests vary All test descriptions are based on the Agilent ChemStation as user interface Some descriptions are only available in the Service Manual Table 14 Test Functions avaible vs User Interface Test ChemStation Instant Pilot Control Module G4208A G1323B Selftest Yes No No Filter Yes No No Slit Yes No Yes D A Converter Yes No No Test Chromatogram Yes C No Yes Wavelength Calibration Yes Yes M Yes Lamp Intensity Yes Yes D Yes Holmium Yes Yes D Yes Cell Yes Yes D No Dark Current Yes Yes D No C via command M section Maintenance D section Diagnose The Agilent Control Module G1323B does not do any calculations So there will be no reports generated wit
50. aring measurements with internal or external standards To check photometric accuracy of the Agilent 1200 DAD MWD it is necessary to have more precise information on path lengths of the flow cells The correct response is expected response correction factor Please find below the details of the Agilent 1200 DAD MWD flow cells Table 12 Correction factors for Agilent 1200 DAD MWD flow cells Flow cell type Cell volume Part number Path length Path length Correction nominal actual factor Standard flow cell 13 pl G1315 60022 10mm 9 80 0 07 mm 10 9 8 Semi micro flow cell 5 ul G1315 60025 6mm 5 80 0 07 mm 6 5 8 Micro flow cell 2 pl G1315 60024 3mm 3 00 0 05 mm 3 3 0 07 mm 500 Nanoliter flow cell kit 0 5 pl G1315 68724 10 mm 10 00 0 02 mm 10 10 80 Nanoliter flow cell kit 0 08 pl G1315 68716 6mm 6 00 0 02 mm 6 6 High Pressure flow cell 1 7 pl G1315 60015 6mm 5 75 0 07 mm 6 5 75 72 1200 Series DAD and MWD User Manual How to optimize the Detector 5 Peak width response time Response time describes how fast the detector signal follows a sudden change of absorbance in the flow cell The detector uses digital filters to adapt response time to the width of the peaks in your chromatogram These filters do not affect peak area nor peak symmetry When set correctly such filters reduce baseline noise significantly see Figure 27 but reduce peak height only slightly In addition these filters reduce the data rate to
51. bert law 71 bench space 23 beta and alpha line 138 bunching of wavelength 26 C cable connecting APG remote 34 connecting CAN 34 connecting GPIB 34 connecting the ChemStation 34 connecting the power 34 cell support windows 13 choosing a flow cell 70 correction factors for flow cells 72 D delivery checklist 30 dimensions and weight 25 diode array 14 15 diode width 26 disposal of mercury 171 E early maintenance feedback EMF 20 electrical connections description of 18 location of connectors 19 EMF early maintenance feedback 20 entrance slit 14 environment 23 ESD electrostatic discharge strap 97 exchanging see repairs F features safety and maintenance 27 firmware updates 134 flow cell 13 choosing one 70 correction factors 72 path length 72 front view of module 35 G GLP features 27 grating 14 high pressure flow cell dimensions 72 holmium oxide declaration of conformity 176 filter 13 humidity 25 information on uv radiation 173 1200 Series DAD and MWD User Manual installation accessory kit 31 bench space 23 delivery checklist 30 environment 23 flow connections 38 of flow cell and capillaries 38 of the detector 35 physical specifications 25 power considerations 22 power cords 22 site requirements 22 unpacking 30 instrument layout 20 intensity test 136 internet 177 introduction optical system diagram 13 optical sy
52. by 90 as shown below from its end 116 1200 Series DAD and MWD User Manual Maintenance 8 9 Insert the capillary into the hole between fixing screw and the inlet fitting 10 The capillary lays in the grove and should be tied around the body in the grove 5 times 11 Insert the fixing screw so that the capillary cannot leave the grove Screw Inlet capillary with heat 12 Carefully insert the I D tag into the new heat exchanger Shown is the default orientation See Note on page 114 1200 Series DAD and MWD User Manual 117 8 Maintenance 13 Fix the heat exchanger to the clamp unit and the flow cell J14 Fix the inlet capillary to the flow cell body handtight first body to the heat exchanger Then do a 1 4 turn with a 4 mm wrench 15 Check for a centered holder vs hole If required adjust with Next steps the holder screws 16 Reconnect the capillaries see Exchanging a Flow Cell on page 103 17 Perform a leak test 18 Insert the flow cell 19 Replace the front cover 20 Perform a Wavelength Verification and Recalibration on page 138 or a Holmium Oxide Test on page 139 to check the correct positioning of the flow cell 118 1200 Series DAD and MWD User Manual Maintenance 8 Replacing Capillaries on a Semi Micro Micro and High Pressur
53. components and ona regular basis to ensure correct operation of the detector The detector uses the deuterium alpha and beta emission lines for wavelength calibration see Wavelength Verification and Recalibration on page 138 Diagnostic Signals The detector has several signals internal temperatures that can be used for diagnosing baseline problems see Diagnostic Signals in the Service Manual 1200 Series DAD and MWD User Manual Troubleshooting and Diagnostics 6 Status Indicators Two status indicators are located on the front of the detector The lower left indicates the power supply status the upper right indicates the detector status Status indicator green yellow red KYA CN Line power switch with green light Figure 37 Location of Status Indicators Power Supply Indicator The power supply indicator is integrated into the main power switch When the indicator is illuminated green the power is ON 1200 Series DAD and MWD User Manual 89 6 90 Troubleshooting and Diagnostics Detector Status Indicator The detector status indicator indicates one of four possible detector conditions When the status indicator is OFF and power switch light is on the detector is in a prerun condition and is ready to begin an analysis A green status indicator indicates the detector is performing an analysis run mode A yellow indicator indicates a not ready condition The detector is in a not re
54. cture and intended use of the instrument Aligent Technologies assumes no liability for the customer s failure to comply with these requirements General This is a Safety Class I instrument provided with terminal for protective earthing and has been manufactured and tested according to international safety standards This instrument is designed and certified as a general purpose laboratory instrument for research and routine application only It is not certified for in vitro or medical applications Operation Before applying power comply with the installation section Additionally the following must be observed Do not remove instrument covers when operating Before the instrument is switched on all protective earth terminals extension cords auto transformers and devices connected to it must be connected to a protective earth via a ground socket Any interruption of the protective earth grounding will cause a potential shock hazard that could result in serious personal injury Whenever it is likely that the protection has been impaired the instrument must be made inoperative and be secured against any intended operation Make sure that only fuses with the required rated current and of the specified type normal blow time delay and so on are used for replacement The use of repaired fuses and the short circuiting of fuseholders must be avoided 1200 Series DAD and MWD User Manual CAUTION Appendix A The operator of
55. e Flow Cell When required If the capillary is blocked Tools required Two 1 4 inch wrenches for capillary connections Wrench 4 mm for capillary connections Screwdriver Pozi 1 PT3 Parts required For parts see Semi Micro Flow Cell on page 148 Micro Flow Cell on page 150 or High Pressure Flow Cell on page 152 Preparations Turn the lamp s off Remove the front cover Remove the flow cell see Exchanging a Flow Cell on page 103 All descriptions in this procedure are based on the default orientation of the cell as it is manufactured The heat exchanger capillary and the cell body can be fixed mirror symmetrically to have both capillaries routed to the bottom or to the top depending on the routing of the capillaries to the column The fittings at the flow cell body are special types for low dead volumes and not compatible with other fittings When retightening the fittings make sure that they are carefully tightened handtight plus 1 4 turn with a wrench Otherwise damage of the flow cell body or blockage may result 1200 Series DAD and MWD User Manual 119 8 Maintenance Identify the inlet and outlet capillaries Inlet capillary Outlet capillary 2 After replacing the outlet capillary fix it handtight first Then do a 1 4 turn with a 4 mm wrench 3 To replace the inlet capillary use a 4 mm wrench for the fitting Unscrew the cell body from the heat exchanger and t
56. e absorbance in the sample wavelength band minus the average absorbance of the reference wavelength band Signal A in the detector default method is set to sample 250 100 reference 360 100 that is the average absorbance from 200 300 nm minus the average absorbance from 300 400 nm As all analytes show higher absorbance at 200 300 nm than at 300 400 nm this signal will show you virtually every compound which can be detected by UV absorbance Many compounds show absorbance bands in the spectrum Figure 28 shows the spectrum of anisic acid as an example To optimize for lowest possible detectable concentrations of anisic acid set the sample wavelength to the peak of the absorbance band that is 252 nm and the sample bandwidth to the width of the absorbance band that is 30 nm A reference of 360 100 is adequate Anisic acid does not absorb in this range If you work with high concentrations you may get better linearity above 1 5 AU by setting the sample wavelength to a valley in the spectrum like 225 nm for anisic acid Anisic acid sample wavelength 252 nm Reference bandwidth 100 nm 30 nm bandwidth Absorbance mAU Reference wavelength 360 Wavelength nm Figure 28 Optimization of Wavelength Setting 1200 Series DAD and MWD User Manual 75 5 76 How to optimize the Detector A wide bandwidth has the advantage of reducing noise by averaging over a wavelength range compared to a 4 nm bandwidt
57. e relevant spectral information Range Only the wavelength range where the compounds in your sample absorb contains information that is useful for purity checks and library searches Reducing the spectrum storage range saves disk space Step Most substances have broad absorbance bands Display of spectra peak purity and library search works best if a spectrum contains 5 to 10 data points per width of the absorbance bands For anisic acid the example used before a step of 4 nm would be sufficient However a step of 2 nm gives a more optimal display of the spectrum Threshold Sets the peak detector Only spectra from peaks higher than threshold will be stored when a peak controlled storage mode is selected 1200 Series DAD and MWD User Manual How to optimize the Detector 5 Margin for Negative Absorbance The detector adjusts its gain during balance such that the baseline may drift slightly negative about 100 mAU In some special case for example when gradient with absorbing solvents are used the baseline may drift to more negative values Only for such cases increase the margin for negative absorbance to avoid overflow of the analog to digital converter 1200 Series DAD and MWD User Manual 81 5 How to optimize the Detector Optimizing Selectivity 82 Quantifying Coeluting Peaks by Peak Suppression In chromatography two compounds may often elute together A conventional dual signal detector can only detec
58. eaning 123 Cleaning or Exchanging the Holmium Oxide Filter 128 1200 Series DAD and MWD User Manual Correcting Leaks 131 Replacing Leak Handling System Parts 132 Replacing the Interface Board 133 Replacing the Detector s Firmware 134 Tests amp Calibrations 135 Intensity Test 136 Wavelength Verification and Recalibration 138 Holmium Oxide Test 139 Cell Test ChemStation only 141 9 Parts and Materials for Maintenance A Appendix Overview of Maintenance Parts 144 Standard Flow Cell 146 Semi Micro Flow Cell 148 Micro Flow Cell 150 High Pressure Flow Cell 152 Prep Flow Cell SST 154 Prep Flow Cell Quartz 156 Nano Flow Cells 158 Accessory Kit 162 General Safety Information 166 Lithium Batteries Information 169 The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC 170 Radio Interference 171 1200 Series DAD and MWD User Manual Sound Emission 172 UV Radiation 173 Solvent Information 174 Declaration of Conformity for HOX2 Filter 176 Agilent Technologies on Internet 177 Index 1200 Series DAD and MWD User Manual 10 1200 Series DAD and MWD User Manual Agilent 1200 Series Diode Array and Multiple Wavelength Detectors User Manual 1 Introduction to the Detector Introduction to the Detector 12 Optical System Overview 13 Early Maintenance Feedback EMF 16 Electrical Connections 18 Instrument Layout 20 This chapter gives an introduction to the detector instrument ove
59. ect column using the zero dead volume fittings of the detector For small column i d e g 1 mm the inlet capillary of the micro flow cell can be connected directly to the column Setting the peak width response time Use peak width according Figure 25 as starting point Set the peak width close to the width of a narrow peak of interest in your chromatogram Setting wavelength and bandwidth Sample wavelength Never miss a peak by the use of a browser wavelength like 250 nm with 100 nm bandwidth Select specific wavelength with reduced bandwidth if you need selectivity e g 250 10 nm and 360 100 nm as reference wavelength Set the sample wavelength to a peak or valley in the spectrum to get best linearity for high concentrations Reference wavelength Select the reference wavelength with broad bandwidth 30 100 nm wavelength range where your analytes have little or no absorbance e g sample at 254 nm reference at 320 nm peak resolution versus sensitivity chromatographic resolution peak resolution versus sensitivity versus disk space sensitivity versus selectivity sensitivity versus linearity baseline drift due to RI effects 1200 Series DAD and MWD User Manual 69 5 How to optimize the Detector Table 11 Parameter Optimization Overview continued Impact 5 Setting the slit width Use 4 nm slit for normal applications spectral resolution sensitivity and
60. ed Discharged Lithium batteries shall be disposed off locally according to national waste disposal regulations for batteries Lithiumbatteri Eksplosionsfare ved fejlagtig handtering Udskiftning ma kun ske med batteri af samme fabrikat og type Lever det brugte batteri tilbage til leverand ren Lithiumbatteri Eksplosionsfare Ved udskiftning benyttes kun batteri som anbefalt av apparatfabrikanten Brukt batteri returneres appararleverandoren Bij dit apparaat zijn batterijen geleverd Wanneer deze leeg zijn moet u ze niet weggooien maar inleveren als KCA 1200 Series DAD and MWD User Manual 169 A Appendix The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC Abstract The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC adopted by EU Commission on 13 February 2003 is introducing producer responsibility on all Electric and Electronic appliances from 13 August 2005 This product complies with the WEEE Directive 2002 96 EC marking requirements The affixed label indicates that you must not discard this electrical electronic product in domestic household waste Product Category With reference to the equipment types in the WEEE Directive Annex this product is classed as a Monitoring and Control instrumentation product Do not dispose off in domestic household waste To return unwanted products contact your local Agilent office or see www agilent com fo
61. ed on the default orientation of the cell as it is manufactured The heat exchanger capillary and the cell body can be fixed mirror symmetrically to have both capillaries routed to the bottom or to the top depending on the routing of the capillaries to the column The fittings at the flow cell body are special types for low dead volumes and not compatible with other fittings When retightening the fittings make sure that they are carefully tightened handtight plus 1 4 turn with a wrench Otherwise damage of the flow cell body or blockage may result 114 1200 Series DAD and MWD User Manual Maintenance 8 1 Identify the inlet and outlet capillaries To replace the inlet 2 After replacing the outlet capillary fix it handtight first capillary continue with step 3 Then do a 1 4 turn with a 4 mm wrench Outlet 3 To replace the inlet capillary use a 4 mm wrench for the 4 Unscrew the cell body from the heat exchanger and the fitting heat exchanger from the clamp unit 1200 Series DAD and MWD User Manual 115 8 Maintenance 5 Use a small flat screw driver to carefully lift off the I D tag 6 Unscrew the fixing screw and unwrap the inlet capillary Shown is the default orientation See Note on page 114 from the grove in the flow cell body Screw Inlet capillary with heat exchanger 7 Take the new inlet capillary and bend it 90 about 35 mm 8 Bend the capillary again
62. em 7 Window screw kit includes 2 quartz windows 2 compression washers 2 window holders 2 window screws and 10 spring washers G1315 60022 79883 22402 5062 8553 79883 28801 79883 22301 1000 0488 See kits below See kits below No part number G1315 87321 G1315 87302 G1315 84910 0515 1056 5022 2184 G1315 68712 G1315 68711 G1315 68710 79883 68703 1200 Series DAD and MWD User Manual NOTE Gaskets 6 and 7 have different hole diameters Parts and Materials for Maintenance 9 Figure 46 Standard Flow Cell Parts 1 window screw 2 spring washers 3 compression washer 4 window holder 5 quartz window 6 Gasket Figure 47 Orientation of Spring Washers 1200 Series DAD and MWD User Manual 147 9 Parts and Materials for Maintenance Semi Micro Flow Cell 148 Table 18 Semi Micro Flow Cell Parts Item Description Part Number A wo N Semi micro flow cell assembly 6 mm 5 ul maximum pressure 120 bar 12 MPa with I D tag Window screw Spring washers pack of 10 Compression washer Window holder Quartz window Gasket BACK Teflon 1 8 mm hole outlet side Gasket FRONT Teflon 1 3 mm hole inlet side Window assembly comprises item 1 2 3 4 5 Capillary IN 0 17 mm 310 mm lg including heat exchanger Capillary OUT 0 12 mm 200 mm lg Capillary OUT 0 17 mm 200 mm lg Clamp unit Screw M 2 5 4 mm lg for cell body clamp Zero dead volume f
63. en Spectra are taken at the apex and baselines of the peak Spectra are taken at the apex baselines upslope and downslope of the peak All spectra within the peak are taken The three spectra acquisition types mentioned above are also referred to as peak controlled spectra acquisition The peak detection is done by the detector firmware based on the threshold and peakwidth parameters you set for the DAD If you want to use peak controlled spectra storage make sure that you set these parameters to recognize all the peaks of interest The integration algorithm also includes peak detection based on the threshold and peakwidth parameters set in the integration events Spectra are taken continuously as for All but only every second spectrum is stored other spectra are discarded This reduces the amount of data storage necessary Spectra are taken continuously depending on the setting of the Peakwidth Eight spectra are acquired per Peakwidth The acquisition time for one spectrum is slightly less than the Peakwidth divided by 8 that is greater than or equal to 0 01s and less than or equal to 2 55s If there are no peaks in Signal A there are no spectra You cannot process spectra present in other signals Range defines the wavelength range for spectral storage Limits 190 to 950 nm in steps of 1 nm for both low and high values The high value must be greater than the low value by at least 2 nm Step defines the wavelength resoluti
64. er Manual How to optimize the Detector 5 Furthermore the absorbance is no longer strictly linear with concentration for wavelengths at a steep slope of a compound s spectrum Substances with fine structures and steep slopes like benzene are very rare In most cases the width of absorbance bands in the spectrum is more like 30 nm as with anisic acid see Figure 28 In most situations a slit width of 4 nm will give the best results Use a narrow slit 1 or 2 nm if you want to identify compounds with fine spectral structures or if you need to quantify at high concentrations gt 1000 mAU with a wavelength at the slope of the spectrum Signals with a wide bandwidth can be used to reduce baseline noise Because digital bandwidth is computed as average of absorbance there is no impact on linearity Use a wide 8 or 16 nm slit when your sample contains very small concentrations Always use signals with bandwidth at least as wide as the slit width 1200 Series DAD and MWD User Manual 79 5 80 How to optimize the Detector Optimizing Spectral Acquisition DAD only Storage of all spectra consumes a lot of disk space It is very useful to have all spectra available during optimization of a method or when analyzing unique samples However when running many samples of the same type the large size of data files with all spectra may become a burden The detector provides functions to reduce the amount of data yet retaining th
65. ese values or values slightly less than the displayed values as EMF limits and then reset the EMF counters to zero The next time the EMF counters exceed the new EMF limits the EMF flag will be displayed providing a reminder that maintenance needs to be scheduled 1200 Series DAD and MWD User Manual 17 1 Introduction to the Detector Electrical Connections e The GPIB connector is used to connect the detector with a computer The address and control switch module next to the GPIB connector determines the GPIB address of your detector The switches are preset to a default address which is recognized once after power is switched on e The CAN bus is a serial bus with high speed data transfer The two connectors for the CAN bus are used for internal Agilent 1200 Series module data transfer and synchronization e Two independent analog outputs provide signals for integrators or data handling systems e The interface board slot is used for external contacts and BCD bottle number output or LAN connections e The REMOTE connector may be used in combination with other analytical instruments from Aligent Technologies if you want to use features such as start stop common shut down prepare and so on e With the appropriate software the RS 232C connector may be used to control the detector from a computer through a RS 232C connection This connector is activated and can be configured with the configuration switch next to the GPIB connecto
66. fications Site Requirements 22 Physical Specifications 25 Performance Specifications 26 This chapter gives information on environmental requirements physical and performance specifications ot Agilent Technologies 21 2 Site Requirements and Specifications Site Requirements A suitable environment is important to ensure optimal performance of the detector Power Consideration The detector power supply has wide ranging capabilities and accepts any line voltage in the range mentioned in Table 1 Consequently there is no voltage selector in the rear of the detector There are also no externally accessible fuses because automatic electronic fuses are implemented in the power supply To disconnect the detector from line unplug the power cord The power supply still uses some power even if the power switch on the front panel is turned off Shock hazard or damage of your instrumentation can result if the devices are connected to a line voltage higher than specified CAUTION Make sure to have easy access to the power cable of the instrument in order to disconnect the instrument from line 22 Power Cords Different power cords are offered as options with the detector The female end of all power cords is identical It plugs into the power input socket at the rear of the detector The male end of each power cord is different and designed to match the wall socket of a particular country or region 1200 Series D
67. h the baseline noise is reduced by a factor of approximately 2 5 whereas the signal is about 75 of a 4 nm wide band The signal to noise ratio for a 30 nm bandwidth is twice that for a 4 nm bandwidth in our example Bandwidth 30 nm 12 nm 4nm Figure 29 Influence of Bandwidth on Signal and Noise Because the detector averages absorbance values that are calculated for each wavelength using a wide bandwidth does not negatively impact linearity The use of a reference wavelength is highly recommended to further reduce baseline drift and wander induced by room temperature fluctuations or refractive index changes during a gradient An example of the reduction of baseline drifts is shown in Figure 30 for PTH amino acids Without a reference wavelength the chromatogram drifts downwards due to refractive index changes induced by the gradient This is almost completely eliminated by using a reference wavelength With this technique PTH amino acids can be quantified in the low picomole range even in a gradient analysis 1200 Series DAD and MWD User Manual How to optimize the Detector 5 Lud a og e 1 pmol each I cc J q T Gn a a EE a Wavelength 267 nm Reference 380 nm Wavelength 267 nm No reference Time min Grad 0 02 m KH2P04 ACN from 12 ACN to 45 ACN in 12 min Figure 30 Gradient Analysis of PTH Amino Acids 1 pmol each with and without Refer ence Slit Width The detector ha
68. h passed failed information 1200 Series DAD and MWD User Manual 91 6 Troubleshooting and Diagnostics Agilent LC Diagnostic Software 92 The Agilent LC diagnostic software is an application independent tool that provides troubleshooting capabilities for the Agilent 1200 Series modules It provides for all 1200 Series LC the possibility of a first guided diagnostic for typical HPLC symptoms and a status report stored as Adobe Acrobat pdf or as a printable file to assist users evaluating the instrument state At the introduction following modules will be fully supported by the software including module tests and calibrations as well as injector steps and maintenance positions e Agilent 1200 Series binary pump SL G1312B e Agilent 1200 Series high performance autosampler SL G1367B e Agilent 1200 Series thermostatted column compartment SL G1316B e Agilent 1200 Series diode array detector SL G1315C With further releases of the diagnostic software all Agilent 1200 Series HPLC modules will be fully supported This diagnostic software provides tests and diagnostic features that may differ from the descriptions in this manual For details refer to the help files provided with the diagnostic software 1200 Series DAD and MWD User Manual Agilent 1200 Series Diode Array and Multiple Wavelength Detectors o User Manual ee Maintenance and Repair e e e i Introduction into Repairing the Detector 94 Wa
69. he heat exchanger from the clamp unit 120 1200 Series DAD and MWD User Manual Maintenance 8 5 Use a small flat screw driver to carefully lift off the I D tag Carefully insert the I D tag into the new heat exchanger Shown is the default orientation See Note on page 114 Shown is the default orientation See Note on page 114 7 Fix the new heat exchanger to the clamp unit and the heat B Fix the inlet capillary to the flow cell body handtight first exchanger to the cell body Then do a 1 4 turn with a 4 mm wrench 1200 Series DAD and MWD User Manual 121 8 Maintenance 9 Check for a centered holder vs hole If required adjust with the holder screws Next steps 10 Reconnect the capillaries see Exchanging a Flow Cell on page 103 11 Perform a leak test 12 Insert the flow cell 13 Replace the front cover 14 Perform a Wavelength Verification and Recalibration on page 138 or a Holmium Oxide Test on page 139 to check the correct positioning of the flow cell 122 1200 Series DAD and MWD User Manual Maintenance 8 Nano Flow Cell Replacing or Cleaning When required If parts are contaminated or leaky Tools required Screwdriver POZI 1 PT3 Two 1 4 inch wrenches for capillary connections Parts required For parts identification refer to Nano Flow Cells on page 158 80 nl
70. inction coefficient which is a characteristic of a given substance under a precisely defined set of conditions of wavelength solvent temperature and other parameters C is the concentration of the absorbing species usually in g l or mg l and d is the path length of the cell used for the measurement Therefore flow cells with longer path lengths yield higher signals Although noise usually increases little with increasing path length there is a gain in signal to noise ratio For example in Figure 26 the noise increased by less than 10 but a 70 increase in signal intensity was achieved by increasing the path length from 6 mm to 10 mm When increasing the path length the cell volume usually increases in our example from 5 13 ul Typically this causes more peak dispersion As Figure 26 demonstrates this did not affect the resolution in the gradient separation in our example As a rule of thumb the flow cell volume should be about 1 3 of the peak volume at half height To determine the volume of your peaks take the peak width as reported in the integration results multiply it by the flow rate and divide it by 3 1200 Series DAD and MWD User Manual 71 5 How to optimize the Detector Analysis of pesticide standard g 6 mm optical path length E 10 mm optical path lengt 3 lt Time min Figure 26 Influence of Cell Path Length on Signal Height Traditionally LC analysis with UV detectors is based on comp
71. installed and once with the flow cell removed The resulting intensity ratio is a measure of the amount of light absorbed by the flow cell The test can be used to check for dirty or contaminated flow cell windows When the test is started the 1 nm slit is moved into the light path automatically and the gain is set to zero To eliminate effects due to absorbing solvents the test should be done with water in the flow cell This test should be performed inititially with a new detector flow cell The values should be kept for later reference comparison Cell Test Evaluation The Agilent ChemStation calculates the intensity ratio automatically The intensity ratio typically between 0 5 and 0 7 for new standard flow cells is dependent on the degree of contamination of the flow cell windows and on the type of flow cell used Figure 45 Cell Test Results report with no flow cell inserted This test can be used for the standard flow cells only The nano flow cells will give very low values due to their design 1200 Series DAD and MWD User Manual 141 8 Maintenance Test Failed low ratio value Probable Causes e Absorbing solvent or air bubble in flow cell e Dirty or contaminated flow cell Suggested Actions Y Ensure the flow cell is filled with water and free from air bubbles Exchange the flow cell windows 142 1200 Series DAD and MWD User Manual Agilent 1200 Series Diode Array and Multiple Wavelength Detectors User Manual
72. is chapter gives an overview about the troubleshooting and diagnostic features and the different user interfaces Maintenance and Repair This chapter provides general information on maintenance and repair of the detector Maintenance This chapter describes the maintenance of the detector and the required tests Parts and Materials for Maintenance This chapter provides information on parts for maintenance Appendix This chapter provides safetey and other general information 1200 Series DAD and MWD User Manual Contents 1 introduction to the Detector Introduction to the Detector 12 Optical System Overview 13 Early Maintenance Feedback EMF 16 EMF Counters 16 Using the EMF Counters 16 Electrical Connections 18 Instrument Layout 20 2 Site Requirements and Specifications Site Requirements 22 Physical Specifications 25 Performance Specifications 26 3 Installing the Detector Unpacking the Detector 30 Damaged Packaging 30 Delivery Checklist 30 Optimizing the Stack Configuration 32 Installing the Detector 35 Flow Connections to the Detector 38 4 Using the Detector Setting up an Analysis 44 1200 Series DAD and MWD User Manual Before Using the System 44 Requirements and Conditions 46 Optimization ofthe System 48 Preparing the HPLC System 49 Running the Sample and Verifying the Results 58 Special Settings of the Detector 59 Control Settings 59 Online Spectra 60 Analog Output Settings 61 Spec
73. ises item 1 2 3 4 5 Window screw kit includes 2 quartz windows 2 compression washers 2 window holders 2 window screws and 10 spring washers Cell repair kit semi micro includes window screw kit 4 mm kexagonal wrench and seal kits Capillary SST 250 mm length 0 5 mm i d 0 D 0 9 mm with fittings for flow cell assembled Fitting FRONT Fitting BACK Fitting 1 16 re order 10 pk Cell body G1315 60016 79883 22402 5062 8553 79883 28801 79883 22301 1000 0488 G1315 68711 G1315 68710 No part number 79883 68703 G1315 68713 G1315 87305 5062 2418 G1315 27706 1200 Series DAD and MWD User Manual Parts and Materials for Maintenance 9 Table 21 Prep Flow Cell SST Parts continued Item Description Part Number 11 Handle for clamp unit G1315 84901 12 Clamp unit G1315 84902 13 Screw M 2 5 4 mm lg for cell body clamp 0515 1056 5 3 2 __ 0 0 05 A 1 6 NOTE Gaskets 6 and 7 have different hole diameters Figure 53 Prep Flow Cell SST Parts 1 window screw 2 spring washers i 3 compression washer 4 window holder 5 quartz window 6 Gasket Figure 54 Orientation of Spring Washers 1200 Series DAD and MWD User Manual 155 9 Parts and Materials for Maintenance Prep Flow Cell Quartz 156 For more details on the Preparative Flow Cells refer to the technical note that comes with the flow cells Table 22 Prep Flow Cell Quartz Parts Item Description
74. itting Cell repair kit semi micro includes window screw kit 4 mm kexagonal wrench and seal kits Seal kit BACK for semi micro flow cell qty 12 of item 6 Seal kit FRONT for semi micro flow cell qty 12 of item 7 Window screw kit includes 2 quartz windows 2 compression washers 2 window holders 2 window screws and 10 spring washers G1315 60025 79883 22402 5062 8553 79883 28801 79883 22301 1000 0488 See kits below See kits below No part number G1315 87319 G1315 87306 G1315 87302 G1315 84910 0515 1056 5022 2184 G1315 68713 79883 68702 G1315 68710 79883 68703 1200 Series DAD and MWD User Manual Parts and Materials for Maintenance 9 NOTE Gaskets 6 and 7 have different hole diameters Figure 48 Semi Micro Flow Cell Parts 1 window screw 2 spring washers CINTA 3 compression washer gt Ny 4 window holder 5 quartz window 6 Gasket Figure 49 Orientation of Spring Washers 1200 Series DAD and MWD User Manual 149 9 Micro Flow Cell 150 Parts and Materials for Maintenance Table 19 Micro Flow Cell Parts Item Description Part Number A wo N Micro flow cell assembly 3 mm 2 ul maximum pressure 120 bar 12 MPa Window screw Spring washers pack of 10 Compression washer Window holder Quartz window Gasket BACK Teflon 1 8 mm hole outlet side Gasket FRONT Teflon 1 3 mm hole inlet side Window assembly comprises item 1 2 3 4 5 Ca
75. kit One side is already factory assembled Pre assembled 6 Connect the newly assembled fitting of the capillary to the top fitting holder and the other end to the column 7 Assemble the waste tubing from the accessory kit 8 Connect the waste tubing to the bottom fitting holder and a waste tubing to the leak outlet y oO Sach E a 40 1200 Series DAD and MWD User Manual Installing the Detector 3 9 Remove the flow cell and establish a flow and observe for leaks 10 Insert the flow cell close the cover and replace the front cover The installation of the detector is now complete The detector should be operated with the front cover in place to protect the flow cell area against strong drafts from the ouside and to cover the deuterium lamp Some types of the Agilent deuterium lamps show a light ring during operation This is not harmful refer to UV Radiation on page 173 1200 Series DAD and MWD User Manual 41 3 Installing the Detector 42 1200 Series DAD and MWD User Manual Agilent 1200 Series Diode Array and Multiple Wavelength Detectors User Manual 4 Using the Detector Setting up an Analysis 44 Before Using the System 44 Requirements and Conditions 46 Optimization of the System 48 Preparing the HPLC System 49 Running the Sample and Verifying the Results 58 Special Setti
76. lary Inlet 100 pm pre mounted to cell includes Inlet capillary 300 mm long 100 um i d with pre fixed ferrules 4 and fittings 3 plus one PEEK Fitting FT 5 PEEK coated fused silica capillary Outlet 100 um pre mounted to cell includes Outlet capillary 120 mm long 100 pm i d with pre fixed ferrules 4 and fitting 3 plus one PEEK Fitting FT 5 PEEK coated fused silica capillary Inlet 50 pm alternative includes Inlet capillary 400 mm long 50 um i d with pre fixed ferrules 4 and fittings 3 plus one PEEK Fitting FT 5 PEEK coated fused silica capillary Outlet 50 um alternative includes Outlet capillary 120 mm long 50 um i d with pre fixed ferrules 4 and fitting 3 plus one PEEK Fitting FT 5 Cell Housing 500 nl Cell Seal Assembly 500 nl Quartz Body 500 nl Sealing Kit includes items 10 12 QTY 2 and 7 QTY 5 1200 Series DAD and MWD User Manual G1315 68724 G1315 87333 61315 87338 61315 87323 G1315 87328 G1315 27703 G1315 87101 G1315 80001 G1315 68715 Parts and Materials for Maintenance 9 Table 26 lists the specific parts for the 80 nl flow cell Table 26 Specific 80 nl Flow Cell Parts Item Description Part Number 80 nl Flow Cell Kit G1315 68716 1 PEEK coated fused silica capillary Inlet 50 pm G1315 87323 pre mounted to cell includes Inlet capillary 400 mm long 50 um i d with pre fixed ferrules 4 and fittings 3
77. lied warranties of merchantability and fitness for a par ticular purpose Agilent shall not be liable for errors or for incidental or consequential damages in connec tion with the furnishing use or per formance of this document or of any information contained herein Should Agilent and the user have a separate written agreement with warranty terms covering the material in this document that conflict with these terms the warranty terms in the sep arate agreement shall control Technology Licenses The hardware and or software described in this document are furnished under a license and may be used or copied only in accor dance with the terms of such license Restricted Rights Legend Software and technical data rights granted to federal government customers include only those rights customarily provided to end user Customers of Software Agilent provides this customary commercial license in Software and technical data pursuant to FAR 12 211 Technical Data and FAR 12 212 Computer Software and for Department of Defense purchases DFARS 252 227 7015 Technical Data Commercial Items and DFARS 227 7202 3 Rights in Commercial Computer Software or Computer Software Documentation If a federal government or other public sector Customer has a need for rights not conveyed under these terms it must negotiate with Agilent to establish acceptable terms in a written agreement executed by all relevant parties Safety Notices
78. lternately 10 Reassemble the flow cell body to the holder 11 Re install the flow cell and connect the capillaries to the union holder Next Steps 12 Perform a leak test with the flow cell outside of the detector 13 If no leak is observed install the flow cell and you are ready to work 14 Make sure that the flow cell assembly is inserted correctly and fits perfectly in the optical unit especially when PEEK capillaries are used The cell body can be fitted in two positions to allow the capillaries routed upwards or downwards depending on where the column is located Route the capillaries directly column inlet and waste assembly outlet 126 1200 Series DAD and MWD User Manual Wrench Maintenance 8 With the instrument accessory kit comes a 4 mm wrench and with the Sealing Kit a special adapter Both together work as a torque wrench with pre defined torque maximum allowed torque for the cell fittings is 0 7 Nm It can be used to tight the capillary fittings at the flow cell body The wrench has to be plugged into the adapter as shown in Figure 39 Adapter DO NOT press down more than shown here Figure 39 Wrench plus Torque Adapter 1200 Series DAD and MWD User Manual 127 8 Maintenance Cleaning or Exchanging the Holmium Oxide Filter When required If holmium oxide filter is contaminated Tools
79. n The variation in Measured Wavelength depends on the different Optical Bandwidth Agilent Technologies guarantees the trace ability of the specified absorbance maxima to a National Instinate of Standards amp Technology NIST Holnimm Oxide Sobution Qandard with a lot to lot tolerance of 0 3 rm The wavelength calibration filter built into the Agilent Technologies UV VIS detectors is made of this material and meets these specifications E is therefore suitable for wavelength calibration of these detectors vrithin the specified wavelength accuracy of the respective dete ctor over its vravelength rimge Juary 13 2006 Date P N 89550 90501 _ Revision E y Agilent Techaologies WMATA ILA OA A A DININ uit 176 1200 Series DAD and MWD User Manual Appendix A Agilent Technologies on Internet For the latest information on products and services visit our worldwide web site on the Internet at http www agilent com Select Products Chemical Analysis It will provide also the latest firmware of the Agilent 1200 Series modules for download 1200 Series DAD and MWD User Manual 177 A Appendix 178 1200 Series DAD and MWD User Manual Index A accessory kit 31 162 accuracy of wavelength 26 achromat source lens 13 Agilent on internet 177 algea information 174 alpha and beta line 138 array 14 ASTM environmental conditions 23 ASTM reference 27 battery safety information 169 Beer Lam
80. n 1200 Series DAD and MWD User Manual 111 8 Maintenance 1 Use a4 mm hex key to unscrew the window assembly 1 If you want to replace the gasket only continue with and remove the gasket 2 from the cell body step 7 on page 109 2 Use a tooth pick to remove the quartz window from the Note window assemsly If the washers fall out of the window assembly they must be inserted in the correct order with the Teflon ring to prevent any leaks from the flow cell window 112 1200 Series DAD and MWD User Manual Maintenance 8 Orientation of Flow Cell Parts 1 window assembly contains 6 items 2 3 4 5 and 6 2 seal ring D quartz window compression washer spring washers window holder window screw inlet capillary outlet capillary OI PAO Bw 3 Follow the procedure Maintenance of Standard Semi Micro or Micro Flow Cell on page 106 for reassembling 1200 Series DAD and MWD User Manual 113 8 Maintenance Replacing Capillaries on a Standard Flow Cell When required If the capillary is blocked Tools required Two 1 4 inch wrenches for capillary connections Wrench 4 mm for capillary connections Screwdriver Pozi 1 PT3 Parts required For parts see Standard Flow Cell on page 146 Preparations Turn the lamp s off Remove the front cover Remove the flow cell see Exchanging a Flow Cell on page 103 All descriptions in this procedure are bas
81. n Overview 69 Optimizing for Sensitivity Selectivity Linearity and Dispersion 71 Optimizing Selectivity 82 This chapter provides information on how to optimize the detector ee Agilent Technologies 67 5 How to optimize the Detector Optimizing the Detector Performance 68 The detector has a variety of parameters that can be used to optimize performance Depending on whether signal or spectral data need to be optimized different settings are recommended The following sections describe optimization for e signal sensitivity selectivity and linearity e spectral sensitivity and resolution DAD only and e disk space required for storing data The information in this chapter should be seen as a basic introduction to diode array detector technics Some of these technics may not be available in the instrument firmware or the user interfaces Control Module and or Agilent Chemstation How to Get the Best Detector Performance The information below will guide you on how to get the best detector performance Follow these rules as a start for new applications It gives rules of thumb for optimizing detector parameters 1200 Series DAD and MWD User Manual Optimization Overview Table 11 Optimization Overview How to optimize the Detector 5 Parameter Impact 1 4 Selection of flow cell Choose flow cell according to used column see Figure 25 Connection of flow cell For flow rates from 0 5 ml min conn
82. ngs of the Detector 59 Control Settings 59 Online Spectra 60 Analog Output Settings 61 Spectrum Settings 62 Peakwidth Settings 64 Slit Settings 65 Margin for Negative Absorbance Settings 66 Optimizing the Detector 66 This chapter provides information on how to set up the detector for an analysis and explains the basic settings ee Agilent Technologies 43 4 Using the Detector Setting up an Analysis This chapter can be used for e preparing the system e to learn the set up of an HPLC analysis and e to use it as an instrument check to demonstrate that all modules of the system are correctly installed and connected It is not a test of the instrument performance e to learn about special settings Before Using the System Solvent Information Observe recommendations on the use of solvents in chapter Solvents in the pump s reference manual Priming and Purging the System When the solvents have been exchanged or the pumping system has been turned off for a certain time for example overnight oxygen will re diffuse into the solvent channel between the solvent reservoir vacuum degasser when available in the system and the pump Solvents containing volatile ingredients will slightly lose these Therefore priming of the pumping system is required before starting an application 44 1200 Series DAD and MWD User Manual Using the Detector 4 Table 5 Choice of Priming Solvents for Different Purposes Acti
83. nm The achromat receives the light from both lamps and focuses it so that the beam passes through the flow cell The holmium oxide filter is electromechanically actuated During the holmium filter test it moves into the light path The cell support window assembly separates the holmium filter area from the flow cell area The optical unit has a flow cell compartment for easy access to flow cells A variety of optional flow cells can be inserted using the same quick simple mounting system The flow cell can be removed to check the optical and electronic performance of the detector without having influences from the flow cell The spectrograph material is ceramic to reduce thermal effects to a minimum The spectrograph consists of the spectrograph lens the variable entrance slit the grating and the photodiode array with front end electronics The spectrograph lens refocuses the light beam after it has passed through the flow cell The sampling interval of the diode array is lt 1 nm over the wavelength range 190 950 nm Depending on the wavelength this varies from 1 0 to 1 25 diodes per nanometer for example a diode every 0 8 to 1 nm For a small wavelength range the small non linearity could be neglected With the wavelength range from 190 950 nm a new approach is required to achieve wavelength accuracy over the full range Each spectograph is calibrated individually The calibration data is stored in the spectrograph on an EEPROM
84. o 0 5 pl volume 10 mm cell path length and 50 bar 725 psi pressure maximum See Optimization Overview on page 69 Agilent ChemStation for LC Recorder integrator 100 mV or 1 V output range 0 001 2 AU two outputs 26 1200 Series DAD and MWD User Manual Site Requirements and Specifications 2 Table2 Performance Specifications Agilent 1200 Series DAD and MWD continued Type Specification Comments Communications Controller area network CAN GPIB RS 232C APG Remote ready start stop and shut down signals LAN optional Safety and maintenance Extensive diagnostics error detection and display through control module and ChemStation leak detection safe leak handling leak output signal for shutdown of pumping system Low voltages in major maintenance areas GLP features Early maintenance feedback EMF for continuous tracking of instrument usage in terms of lamp burn time with user settable limits and feedback messages Electronic records of maintenance and errors Verification of wavelength accuracy with built in holmium oxide filter Housing All materials recyclable ASTM Standard Practice for Variable Wavelength Photometric Detectors Used in Liquid Chromatography Reference conditions cell path length 10 mm response time 2 s flow 1 ml min LC grade Methanol slit width 4 nm Linearity measured with caffeine at 265 nm For environmental conditions refer to Environment
85. on Spectrum click on the drop down list and chose a parameter Table 9 on page 63 shows the possible parameters 3 Change the Range Step width and Threshold according to your needs QE Set up DAD Signals DAD Signals System 2 DAD Signals System 2 of Control r Signals r Signals fu Online Spectra Store Sample Bw Reference Bw Store Sample Bw Reference Bw Not Ready Information Aam fa54 j4 feo foo 4 nm AW 254 J4 faeo fioo 4 nm aie e f fs feo fio nm e fie f0 fioo 2 nm c Jao fe feo fico Sf nm c Jao Je feso fico Sf nm D J20 fie Jaso froo Sf nm D 230 fie 360 100 2 nm E feo fie Jaso fioo 3 nm e fas fie faeo foo Sf rm CSpectrum gt Spectrum Store None Store Every 2nd spectrum None Range Apex Baselines Range 130 to 400 nm Apex Slopes B aselines Step Allin peak Step 2 0 nm Every 2nd spectrum Threshold Timetable Total Lines 0 Cancel Help Figure 21 Spectra Settings Threshold J100 m u 62 1200 Series DAD and MWD User Manual Table9 Spectrum Settings Using the Detector 4 Store None Apex Baselines Apex Slopes Baselines All in Peak Every 2nd spectrum All Range Step Threshold Defines at which points on signal A spectra will be taken and saved Signal A is used to control the peak controlled spectra acquisition the other signals have no influence on spectra acquisition No spectra are tak
86. on for spectral storage Limits 0 10 to 100 00 nm in steps of 0 1 nm The threshold is the height in mAU of the smallest expected peak The peak detector ignores any peaks which are lower than the threshold value and does not save spectra Limits 0 001 to 1000 00 mAU in steps of 0 001 mAU Usable for modes Apex Baselines Apex Slopes Baselines and All in Peak 1200 Series DAD and MWD User Manual 63 4 Using the Detector Peakwidth Settings ons EE Set up DAD Signals of Control fy Online Spectra Not Ready Information Help Peakwidth Responsetime gt 0 1 min 2s 7 lt 0 01 min 0 1 s gt 0 01 min 0 2 s gt 0 03 min 0 5 s gt 0 05 min 1s gt 01 mn f2 sl gt 0 2 min 4s gt 0 4 min 8s gt 0 85 min 16s r Margin for negative Absorbance Figure 22 64 Do not use peak width shorter than necessary To change the Peakwidth settings select Setup Detector Signals In the section Peakwidth Responsetime click on the drop down list Change the Peakwidth according to your needs Peakwidth enables you to select the peak width response time for your analysis The peak width is defined as the width of a peak in minutes at half the peak height Set the peak width to the narrowest expected 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 o
87. on page 23 1200 Series DAD and MWD User Manual 27 2 Site Requirements and Specifications 28 1200 Series DAD and MWD User Manual Agilent 1200 Series Diode Array and Multiple Wavelength Detectors User Manual 3 Installing the Detector Unpacking the Detector 30 Optimizing the Stack Configuration 32 Installing the Detector 35 Flow Connections to the Detector 38 This chapter describes the installation of the detector ee Agilent Technologies 29 3 installing the Detector Unpacking the Detector Damaged Packaging If the delivery packaging shows signs of external damage please call your Agilent Technologies sales and service office immediately Inform your service representative that the detector may have been damaged during shipment CAUTION If there are signs of damage please do not attempt to install the detector Delivery Checklist Ensure all parts and materials have been delivered with the detector The delivery checklist is shown below Please report missing or damaged parts to your local Agilent Technologies sales and service office Table3 Detector Checklist Description Quantity Detector 1 Power cable 1 Flow cell As ordered User Manual 1 Accessory kit see Table 4 1 30 1200 Series DAD and MWD User Manual Detector Accessory Kit Contents Table4 Accessory Kit Contents Installing the Detector 3 Description Part Number Quantity Accessory kit G1315 68705 CAN cable 0 5 m 5181 1516
88. over 132 1200 Series DAD and MWD User Manual Maintenance 8 Replacing the Interface Board When required For all repairs inside the detector or for installation of the board Part required Interface board BCD G1351 68701 with external contacts and BCD outputs LAN Communication Interface board G1369A or G1369 60001 Tools required None Preparations Turn off the lamp Switch off the detector and disconnect the power cable Assure you have a access to the rear of the module 1 Install the ESD strap Move the power lock across the 2 If required unscrew and remove the interface board Place power inlet the board on the ESD kit ESD strap EDUBUD 7 power lock 3 If required insert the interface board and fix the screws Next steps Remove the ESD strap Reinstall the module into the stack 1200 Series DAD and MWD User Manual 133 8 Maintenance Replacing the Detector s Firmware When required If new version solves problems of currently installed version or after exchange of the detector main board VWM the version on board is older than previous installed one Tools required LAN RS 232 Firmware Update Tool or Instant Pilot G4208A or Control Module G1323B Parts required Firmware tools and documentation from Agilent web site Preparations Read update documentation provided with the Firmware Update Tool The installation of older firmware might be necessary e to keep all sy
89. ped with a filling of isopropanol also recommended when the instrument and or flow cell is shipped to another location This is to avoid breakage due to subambient conditions The heat exchanger capillary and the cell body can be fixed mirror symmetrically to have both capillaries routed to the bottom or to the top depending on the routing of the capillaries to the column For details see Replacing Capillaries on a Standard Flow Cell on page 114 1200 Series DAD and MWD User Manual Installing the Detector 3 1 Press the release buttons and remove the front cover to gain access to the flow cell area 2 Press the release button and open the flow cell door OOO 3 Insert the flow cell and install the capillaries to the capillary holder top is inlet bottom is outlet sR SEER 4 If another Agilent 1200 Series module is positioned on top of the detector route the tubing assembly waste from the accessory kit behind the capillary holder and connect the top end to the other module s waste outlet To other module To waste 1200 Series DAD and MWD User Manual 39 3 Installing the Detector 5 Assemble the column detector capillary from the accessory
90. pillary IN 0 12 mm 310 mm lg including heat exchanger Capillary OUT 0 12 mm 200 mm lg Capillary OUT 0 17 mm 200 mm lg Clamp unit Screw M 2 5 4 mm lg for cell body clamp Zero dead volume fitting Cell repair kit semi micro includes window screw kit 4 mm kexagonal wrench and seal kits Seal kit BACK for semi micro flow cell qty 12 of item 6 Seal kit FRONT for semi micro flow cell qty 12 of item 7 Window screw kit includes 2 quartz windows 2 compression washers 2 window holders 2 window screws and 10 spring washers G1315 60024 79883 22402 5062 8553 79883 28801 79883 22301 1000 0488 See kits below See kits below No part number G1315 87339 G1315 87306 G1315 87302 G1315 84910 0515 1056 5022 2184 G1315 68713 79883 68702 G1315 68710 79883 68703 1200 Series DAD and MWD User Manual NOTE Parts and Materials for Maintenance Gaskets 6 and 7 have different hole diameters Micro Flow Cell Parts Figure 50 1 window screw 2 spring washers 3 compression washer 4 window holder 5 quartz window 6 Gasket Figure 51 Orientation of Spring Washers 1200 Series DAD and MWD User Manual 9 151 9 Parts and Materials for Maintenance High Pressure Flow Cell Table 20 High Pressure Flow Cell Parts Item Description Part Number High pressure flow cell assembly 6 mm 1 7 ul G1315 60015 maximum pressure 400 bar 40 MPa Window assembly comprises items 2
91. plus one PEEK Fitting FT 5 2 PEEK coated fused silica capillary Outlet 50 um G1315 87328 pre mounted to cell includes Outlet capillary 120 mm long 50 um i d with pre fixed ferrules 4 and fitting 3 plus one PEEK Fitting FT 5 1 PEEK coated fused silica capillary Inlet 25 pm G1315 87313 alternative includes Inlet capillary 200 mm long 25 pm i d with pre fixed ferrules 4 and fittings 3 plus one PEEK Fitting FT 5 2 PEEK coated fused silica capillary Outlet 25 um G1315 87318 alternative includes Outlet capillary 600 mm long 25 um i d with pre fixed ferrules 4 and fitting 3 plus one PEEK Fitting FT 5 11 Cell Housing 80 nl G1315 27704 12 Cell Seal Assembly 80 nl G1315 42301 13 Quartz Body 80 nl G1315 80002 Sealing Kit 80 nl cell includes items 10 12 QTY 2 7 G1315 68725 QTY 5 and Lite Touch Sleeves QTY 5 1200 Series DAD and MWD User Manual 161 9 Parts and Materials for Maintenance Accessory Kit This kit contains some accessories and tools needed for the installation and repair of the detector Table 27 Accessory Kit Parts Item Description Part Number Accessory kit Corrugated tubing to waste re order 5 m 1 Teflon Tubing flexible i d 0 8 mm flow cell to waste re order 5m 2 Fitting male PEEK Oty 1 3 Capillary column detector 380 mm lg 0 17 i d includes items 4 5 and 6 not assembled 4 Ferrule front SST qty 2 5 Ferrule back
92. r See your software documentation for further information e The power input socket accepts a line voltage of 100 240 V AC 10 with a line frequency of 50 or 60 Hz Maximum power consumption is 300 VA 125 W There is no voltage selector on your detector because the power supply has a wide ranging capability There are no externally accessible fuses because automatic electronic fuses are implemented in the power supply The security lever at the power input socket prevents that the detector cover is taken off when line power is still connected WARNING Never use cables other than the ones supplied by Aligent Technologies to ensure proper functionality and compliance with safety or EMC regulations 18 1200 Series DAD and MWD User Manual Interface board Introduction to the Detector 1 Security lever Analog signals APG remote RS 232C la A f S CAN GPIB serial number DE manufactured in Germany Configuration switch 6 2006 01 week of last major change 00130 number of unit product number G13158 DAD G13158 DAD serial number Serial No Serial No Me CEs bzg safety standards EE Agilent Technologies y 1R6I703 00725 ICES NMB OM deter tf z afa s e f configuration switch oma fofo wamm settings Apiiers Technologies 76
93. r 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 When the All spectrum storage option is selected then spectra are acquired continuously depending on the setting of the peak width The time specified by the peak width is used as a factor in the acquisition of spectra The acquisition time for one spectrum is slightly less than the peak width divided by 8 that is the acquisition time is between 0 1 seconds 20 Hz and 3 2 seconds 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 in Table 10 on page 65 Peakwidth Setting 1200 Series DAD and MWD User Manual Using the Detector 4 Table 10 Peak Width Response Time Data Rate Peak Width min Response Time sec Data Rate Hz lt 0 01 0 1 20 gt 0 01 0 2 20 gt 0 03 0 5 10 gt 0 05 1 0 5 gt 0 10 2 0 2 5 gt 0 20 40 1 25 gt 0 40 8 0 0 62 gt 0 85 16 0 0 31 Slit Settings 1 To change the Slit settings select Setup Detector Signals 2 In the section Slit click on the drop down list 3 Change the Slit width according to your needs The Slit group allows you to select the optical bandwidth of the J Set up DAD Signals En detector the narrower the slit the smaller the optical bandwidth fy Online Spectra of the instrumen
94. r more information 170 1200 Series DAD and MWD User Manual Appendix A Radio Interference Never use cables other than the ones supplied by Aligent Technologies to ensure proper functionality and compliance with safety or EMC regulations Test and Measurement If test and measurement equipment is operated with equipment unscreened cables and 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 1200 Series DAD and MWD User Manual 171 A Appendix Sound Emission Manufacturer 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 lt 70 dB e Sound Pressure Lp lt 70 dB A e At Operator Position e Normal Operation e According to ISO 7779 1988 EN 27779 1991 Type Test 172 1200 Series DAD and MWD User Manual Appendix A UV Radiation This information is only valid for UV lamps without cover e g 2140 0590 and 2140 0813 Emissions of ultraviolet radiation 200 315 nm from this product is limited such that radiant exposure incident upon the unprotected skin or eye of operator or service personnel is limited to the following TLVs Threshold Limit Values according to the American Conference of Governmental Industrial Hygienists Table 29 UV Radiation Limits
95. required Screwdriver POZI 1 PT3 Screwdriver flat blade Two 1 4 inch wrenches for capillary connections A pair of tweezers Parts required Holmium oxide filter 79880 22711 Preparations Turn the lamp s off Remove the front cover Remove the flow cell see Exchanging a Flow Cell on page 103 See also Declaration of Conformity for HOX2 Filter on page 176 The glass tends to build a film on its surface even under normal environmental conditions This is a phenomenon which can be found also on the surface of several other glasses and has something to do with the composition of the glass There is no indication that the film has an influence on the measurement Even in the case of a thick film which scatters the light remarkably no shift of the peak positions is to be expected A slight change in the absorbance might be possible Other components within the light path lenses windows are also changing their behavior over the time 128 1200 Series DAD and MWD User Manual Maintenance 8 1 Unscrew the six screws and remove the flow cell cover 2 If not already in this position move the filter up
96. res replace if your chromatography method allows by phosphoric acid or phosphate buffer which are less corrosive against stainless steel e Halogenated solvents or mixtures which form radicals and or acids for example 2CHC13 Og 2COClg 2ZHCI1 This reaction in which stainless steel probably acts as a catalyst occurs quickly with dried chloroform if the drying process removes the stabilizing alcohol 1200 Series DAD and MWD User Manual Appendix A e Chromatographic grade ethers which can contain peroxides for example THF dioxane di isopropylether such ethers should be filtered through dry aluminium oxide which adsorbs the peroxides e Solutions of organic acids acetic acid formic acid and so on in organic solvents For example a 1 solution of acetic acid in methanol will attack steel e Solutions containing strong complexing agents for example EDTA ethylene diamine tetra acetic acid e Mixtures of carbon tetrachloride with 2 propanol or THF 1200 Series DAD and MWD User Manual 175 A Appendix Declaration of Conformity for HOX2 Filter Declaration of Conformity We herevrith infomm you that the Holmium Oxide Glass Filter Type Hoya HY 1 Part No 79380 22711 meets the folling specification of absorbance maxima positions mema waga ccc pasa Accura aan La lym i C 79853 1050 gam Srm ee ren 418 Sr gam dron G13144 B C 1100 1200 360 8ren aa lrm 418 5 536 4ro
97. rnings and Cautions 95 Cleaning the Detector 96 Using the ESD Strap 97 This chapter provides general information on maintenance and repair of the detector ee Agilent Technologies 93 7 Maintenance and Repair Introduction into Repairing the Detector Simple Repairs The detector is designed for easy repair The most frequent repairs such as lamp change and flow cell change can be done from the front of the detector with the detector in place in the system stack These repairs are described in Maintenance on page 99 Exchanging Internal Parts Some repairs may require exchange of defective internal parts Exchange of these parts requires removing the detector from the stack removing the covers and disassembling the detector The security lever at the power input socket prevents that the detector cover is taken off when line power is still connected These repairs are described in the Service Manual 94 1200 Series DAD and MWD User Manual Maintenance and Repair 7 Warnings and Cautions WARNING To prevent personal injury the power cable must be removed from the instrument before opening the detector cover Do not connect the power cable to the detector while the covers are removed WARNING To prevent personal injury be careful when getting in contact with sharp metal areas WARNING When working with solvents please observe appropriate safety procedures for example goggles safety gloves and protec
98. rview and internal connectors ee Agilent Technologies 1 Introduction to the Detector Introduction to the Detector 12 The detector is designed for highest optical performance GLP compliance and easy maintenance It includes the following features long life deuterium and tungsten lamps for highest intensity and lowest detection limit over a wavelength range of 190 950 nm no loss in sensitivity for up to five wavelengths simultaneous programmable slit from 1 16 nm for complete optimization of sensitivity linearity and spectral resolution optional flow cell cartridges with I D tag standard 10 mm 13 ul semi micro 6 mm 5 ul micro 3 mm 2 ul 80 nl 500 nl 10 mm high pressure 10 mm 1 7 ul and prep cells are available and can be used depending on the application needs easy front access to lamps and flow cell for fast replacement and built in holmium oxide filter for fast wavelength accuracy verification built in temperature control for improved baseline stability G1315B DAD and G1365B MWD For specifications see Performance Specifications on page 26 1200 Series DAD and MWD User Manual Introduction to the Detector 1 Optical System Overview Optical System The optical system of the detector is shown in Figure 1 Its illumination source is a combination of a deuterium arc discharge lamp for the ultraviolet UV wavelength range and a tungsten lamp for the visible VIS and short wave near infrared
99. s a variable slit at the entrance of the spectrograph This is an effective tool to adapt the detector to changing demand of different analytical problems A narrow slit provides spectral resolution for analytes with very fine structures in the absorbance spectrum An example of such a spectrum is benzene The five main absorbance bands fingers are only 2 5 nm wide and just 6 nm apart from each other 1200 Series DAD and MWD User Manual 77 5 How to optimize the Detector Figure 31 Benzene at 1 4 and 16 nm slit width A wide slit uses more of the light shining through the flow cell This gives lower baseline noise as shown in Figure 32 DAD1 A Sig 254 4 Ref off NOISEVINMANM D maAU A o Slit width 1 nm 0 2 0 4 T T T T T T T 41 15 2 25 3 35 min DAD1 A Sig 254 4 Ref off NOISEVINM4NM D mAU Slit width 4 nm 02 0 4 T T T T T T T 0 5 4 15 2 25 3 35 mit DAD1 A Sig 254 4 Ref off NOISE INMIGNM D mAU o Slit width 16 nm z 0 4 eee SS 05 4 15 Zz 25 3 35 mir Figure 32 Influence of the Slit Width on Baseline Noise However with a wider slit the spectrograph s optical resolution its ability to distinguish between different wavelengths diminishes Any photodiode receives light within a range of wavelength determined by the slit width This explains why the fine spectral structure of benzene disappears when using a 16 nm wide slit 78 1200 Series DAD and MWD Us
100. s is one way of selectively plotting only those peaks that are of interest The signal output remains at zero so long as the ratio is out of the user specified ratio range When the ratio falls within the range the signal output corresponds to the normal absorbance giving single clear peaks on a flat baseline An example is shown in Figure 35 and Figure 36 84 1200 Series DAD and MWD User Manual How to optimize the Detector 5 Select 2 characteristic wavelengths Signal WL1 WL2 20 o Terphenyl Biphenyl WL2 WL1 Scaled Wavelength nm Figure 35 Wavelength Selection for Ratio Qualifiers Signals at 250 nm Biphenyl o Terphenyl No selectivity With ratio qualifier 250 222 nm 3 5 20 Time min Figure 36 Selectivity by Ratio Qualifiers In a four component mixture only biphenyl was recorded The other three peaks were suppressed because they did not meet the ratio qualifier criterion and therefore the output was set to zero The characteristic wavelengths 249 nm A and 224 nm Ag were found from the spectra shown in Figure 35 1200 Series DAD and MWD User Manual 85 5 How to optimize the Detector The ratio range was set at 2 2 4 2 2 10 Only when the ratio between 249 and 224 nm was within this range is the signal plotted Of all four peaks only the third fulfilled the criterion Figure 36 The others were not plotted 86 1200 Series DAD and MWD User Manual Agilent 1200 Series Diode
101. sembled includes items 1 2 3 4 10 11 12 13 14 15 and 16 80 nl Flow Cell Kit G1315 68716 includes Flow cell assembly 10 mm 500 nl 5 MPa completely assembled includes items 1 2 3 4 10 11 12 13 14 15 and 16 Figure 56 shows all parts delivered with the nano flow cell kits Figure 56 Content of kits 158 1200 Series DAD and MWD User Manual Parts and Materials for Maintenance 9 Table 24 lists the generic parts for both nano flow cells Table 24 Generic Parts Item Description Part Number 3 Fitting Screw for 4 mm wrench QTY 2 reorder 10 pk 5063 6593 4 Cell ferrules are factory installed 5 PEEK fitting 1 32 not attached to capillaries reorder 10 pk 5065 4422 7 Litetouch ferrules LT 100 1 32 Ferrule and SS lock ring QTY 2 reorder 10 pk 5063 6592 8 Union Adjustment Tool used for item 7 5022 2146 9 ZDV SS Union no fitting OTY 2 5022 2184 10 Torque Adapter G1315 45003 14 Handle for clamp unit G1315 84902 15 Clamp unit G1315 84910 16 Screw M 2 5 4 mm long for cell body clamp 0515 1056 17 Wrench open end 4 mm supplied with standard accessory kit 8710 1534 G1315 68705 of your detector 1200 Series DAD and MWD User Manual 159 9 Parts and Materials for Maintenance Table 25 lists the specific parts for the 500 nl flow cell Table 25 Specific 500 nl Flow Cell Parts Item Description Part Number 160 500 nl Flow Cell Kit PEEK coated fused silica capil
102. stem overview 13 optical unit parts 13 L lamps 13 leaks correcting 131 line voltage and frequency 25 location of connectors 19 M Maintenance 93 99 Mercury disposal 171 Nano Flow Cells 158 nano flow cells dimensions 72 nano flow cells 158 negative absorbance 81 179 Index 0 operation temperature 25 optical system diagram 13 overview 13 optimization choosing a flow cell 70 detector performance 68 for sensitivity selectivity linearity dispersion 71 how to get the best performance 68 margins for negative absorbance 81 of selectivity 82 of the system 48 overview 69 peak width 73 sample and reference wavelength 74 slit width 77 spectra acquisition 80 P Parts 143 parts identification 143 accessory kit 162 high pressure micro flow cell 152 main assemblies 145 nl flow cells 158 overview 144 semi micro flow cell 148 standard flow cell 146 peak width response time 73 performance optimization 68 photometric accuracy 72 physical specifications 25 humidity 25 line voltage and frequency 25 operation temperature 25 power consumption 25 safety standards 25 weight and dimensions 25 power considerations 22 power consumption 25 power cords 22 programmable slit width 26 180 rear view of module 36 recalibration of wavelength 88 repairs cleaning the instrument 96 correction leaks 131 definition of 94 exchanging a flow cell 10
103. stems on the same validated revision or e if third part control software requires a special version To upgrade downgrade the detector s firmware the following steps have to be performed 1 Download the module s firmware the LAN RS 232 FW Update Tool Version 2 10 or above and the documentation from the Agilent web http www chem agilent com scripts cag_firmware asp 2 Load the firmware into the detector as described in the documentation The G1315B DAD and the G1365B MWD require firmware revision A 04 x or above main and resident 134 1200 Series DAD and MWD User Manual Maintenance Tests amp Calibrations The following tests are required after maintenance of lamps and flow cells e Intensity Test on page 136 e Wavelength Verification and Recalibration on page 138 e Holmium Oxide Test on page 139 e Cell Test ChemStation only on page 141 1200 Series DAD and MWD User Manual 135 8 136 Intensity Test The test is for the standard flow cells 10 mm and 6 mm pathlength only The nano flow cells 80 nl and 500 nl cannot be run with this test due to its low volume The intensity test measures the intensity of the deuterium and tungsten lamps over the full wavelength range 190 950 nm Four spectral ranges are used to evaluate the intensity spectrum The test is used to determine the performance of the lamps and optics see also Cell Test ChemStation only on page 141
104. t but the lower its sensitivity The smaller the Not Ready Information optical bandwidth the higher the spectral resolution Help To set the slit width display the drop down list and select an appropriate slit width from the list Slit 4nm 4 nm E Figure 23 Slit Settings 1200 Series DAD and MWD User Manual 65 4 Using the Detector Margin for Negative Absorbance Settings 1 To change the settings select Setup Detector Signals 2 In the section Margin for Negative Absorbance change the value according to your needs CETSet up DAD Signals Use this field to modify the detector s signal handling to increase Peep ae the margin for negative absorbance Use this option if for example your solvent gradient produces a decreasing baseline fy Online Spectra absorbance and for GPC analyses Not Ready Information i Limits 100 to 4000 mAU Margin for negative Absorbance fi o0 mAU Figure 24 Margin for Negative Absorbance The higher the value the greater the baseline noise Set this value only if you expect negative absorbance greater than 100 mAU Optimizing the Detector Additional theoretical information can be found in chapter How to optimize the Detector on page 67 66 1200 Series DAD and MWD User Manual Agilent 1200 Series Diode Array and Multiple Wavelength Detectors User Manual 5 How to optimize the Detector Optimizing the Detector Performance 68 Optimizatio
105. t and quantify both compounds independently from each other if their spectra do not overlap However in most cases this is highly unlikely With a dual channel detector based on diode array technology quantifying two compounds is possible even when both compounds absorb over the whole wavelength range The procedure is called peak suppression or signal subtraction As an example the analysis of hydrochlorothiazide in the presence of caffeine is described If hydrochlorothiazide is analyzed in biological samples there is always a risk that caffeine is present which might interfere chromatographically with hydrochlorothiazide As the spectra in Figure 33 show hydrochlorothiazide is best detected at 222 nm where caffeine also shows significant absorbance It would therefore be impossible with a conventional variable wavelength detector to detect hydrochlorothiazide quantitatively when caffeine is present 1200 Series DAD and MWD User Manual How to optimize the Detector 5 WL1 204 nm caffeine WL2 222 nm hydrochlorothiazide WL3 260 nm reference to suppress hydrochlorothiazide WL4 282 nm reference to suppress caffeine WL3 WL4 WLI w2 Wavelength nm Figure 33 Wavelength Selection for Peak Suppression With a UV visible detector based on a diode array and the correct choice of a reference wavelength setting quantitative detection is possible To suppress caffeine the reference wavelength must be set to 282 nm At this
106. this instrument is advised that if the equipment is used in a manner not specified in this manual the protection provided by the equipment may be impaired Some adjustments described in the manual are made with power supplied to the instrument and protective covers removed Energy available at many points may if contacted result in personal injury Any adjustment maintenance and repair of the opened instrument under voltage should be avoided as much as possible When inevitable this should be carried out by a skilled person who is aware of the hazard involved Do not attempt internal service or adjustment unless another person capable of rendering first aid and resuscitation is present Do not replace components with power cable connected Do not operate the instrument in the presence of flammable gases or fumes Operation of any electrical instrument in such an environment constitutes a definite safety hazard Do not install substitute parts or make any unauthorized modification to the instrument Capacitors inside the instrument may still be charged even though the instrument has been disconnected from its source of supply Dangerous voltages capable of causing serious personal injury are present in this instrument Use extreme caution when handling testing and adjusting When working with solvents please observe appropriate safety procedures e g goggles safety gloves and protective clothing as described in the material
107. tive clothing as described in the material handling and safety data sheet supplied by the solvent vendor especially when toxic or hazardous solvents are used CAUTION Electronic boards and components are sensitive to electronic discharge ESD In order to prevent damage always use an ESD protection when handling electronic boards and components see Using the ESD Strap on page 97 Eye damage may result from directly viewing the light produced by the deuterium lamp used in this product Always turn the deuterium lamp off before removing it 1200 Series DAD and MWD User Manual 95 7 Maintenance and Repair Cleaning the Detector The detector case should be kept clean Cleaning should be done with a soft cloth slightly dampened with water or a solution of water and mild detergent Do not use an excessively damp cloth allowing liquid to drip into the detector WARNING Do not let liquid drip into the detector It could cause shock hazard and it could damage the detector 96 1200 Series DAD and MWD User Manual Maintenance and Repair 7 Using the ESD Strap Electronic boards are sensitive to electronic discharge ESD In order to prevent damage always use an ESD strap when handling electronic boards and components 1 Unwrap the first two folds of the band and wrap the exposed adhesive side firmly around your wrist 2 Unroll the rest of the band and peel the liner from the copper foil at the opposite end
108. to optimize the Detector Table 13 Peak Width Response Time Data Rate Peak Width minutes Response Time seconds Data Rate Hz lt 0 01 0 1 20 gt 0 01 0 2 20 gt 0 03 0 5 10 gt 0 05 1 0 5 gt 0 10 2 0 2 5 gt 0 20 4 0 1 25 gt 0 40 8 0 0 62 gt 0 85 16 0 0 31 Sample and Reference Wavelength and Bandwidth The detector measures absorbance simultaneously at wavelengths from 190 to 950 nm Two lamps provide good sensitivity over the whole wavelength range The deuterium discharge lamp provides the energy for the UV range 190 to 400 nm and the tungsten lamp emits light from 400 to 950 nm for the visible and short wave near infrared If you know little about the analytes in your sample use both lamps and store all spectra over the full wavelength range This provides full information but fills up your disk space rather quickly Spectra can be used to check a peak s purity and identity Spectral information is also useful to optimize wavelength settings for your chromatographic signal The detector can compute and store at run time up to 8 signals with these properties e sample wavelength the center of a wavelength band with the width of sample bandwidth BW and optionally e reference wavelength the center of a wavelength band with the width of reference bandwidth 1200 Series DAD and MWD User Manual How to optimize the Detector 5 The signals comprises a series of data points over time with the averag
109. trum Settings 62 Peakwidth Settings 64 Slit Settings 65 Margin for Negative Absorbance Settings 66 Optimizing the Detector 66 5 How to optimize the Detector Optimizing the Detector Performance 68 Optimization Overview 69 Optimizing for Sensitivity Selectivity Linearity and Dispersion 71 Flow Cell Path Length 71 Peak width response time 73 Sample and Reference Wavelength and Bandwidth 74 Slit Width 77 Optimizing Spectral Acquisition DAD only 80 Margin for Negative Absorbance 81 Optimizing Selectivity 82 Quantifying Coeluting Peaks by Peak Suppression 82 Ratio Qualifiers for Selective Detection of Compound Classes 84 1200 Series DAD and MWD User Manual 6 Troubleshooting and Diagnostics Overview of the Detector s Indicators and Test Functions 88 Status Indicators 89 Power Supply Indicator 89 Detector Status Indicator 90 User Interfaces 91 Agilent LC Diagnostic Software 92 7 Maintenance and Repair Introduction into Repairing the Detector 94 Simple Repairs 94 Exchanging Internal Parts 94 Warnings and Cautions 95 Cleaning the Detector 96 Using the ESD Strap 97 8 Maintenance Overview of Maintenance 100 ExchangingaLamp 101 Exchanging a Flow Cell 103 Maintenance of Standard Semi Micro or Micro Flow Cell 106 Maintenance of High Pressure Flow Cell 111 Replacing Capillaries on a Standard Flow Cell 114 Replacing Capillaries on a Semi Micro Micro and High Pressure Flow Cell 119 Nano Flow Cell Replacing or Cl
110. vity Solvent Comments After an installation lsopropanol Best solvent to flush air out of the When switching between reverse lsopropanol phase and normal phase both times After an installation Ethanol or Methanol To clean the system when using buffers Bidistilled water After a solvent change Bidistilled water After the installation of normal phase Hexane 5 lsopropanol seals P N 0905 1420 system Best solvent to flush air out of the system Alternative to lsopropanol second choice if no lsopropanol is available Best solvent to re dissolve buffer crystals Best solvent to re dissolve buffer crystals Good wetting properties The pump should never be used for priming empty tubings never let the pump run dry Use a syringe to draw enough solvent for completely filling the tubings to the pump inlet before continuing to prime with the pump 1 Open the purge valve of your pump by turning it counterclockwise and set flow rate to 3 5 ml min 2 Flush all tubes with at least 30 ml of solvent 3 Set flow to required value of your application and close the purge valve Pump for approximately 10 minutes before starting your application 1200 Series DAD and MWD User Manual 45 4 46 Using the Detector Requirements and Conditions What You Will Need Table 6 lists the items you need to have for the set up of the analysis Some of these are optional not required for the basic system Table 6

G1315-90006 - Agilent Technologies

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