Waters 2489 Overview And Maintenance Manual Download Page 15

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June 11, 2015, 715004752 Rev. A

Page 15

Theory and Principles of
Operation

This chapter summarizes the 2489 UV/Visible Detector features and describes

the theory and principles of operation.

See also:

Appendix B

for system specifications and

Appendix C

for information

on high-performance liquid chromatography (HPLC) solvent considerations.

1.1 Detector description

The 2489 UV/Vis detector is a two-channel ultraviolet/visible (UV/Vis) detector

designed for high-performance liquid chromatography (HPLC) applications.

Figure 1–1:

2489 UV/Vis detector

The detector can operate as a stand-alone unit (with an integrator) or as an integral

part of a Waters chromatography system.

Summary of Contents for 2489

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Page 3: ...with or arising from its use For the most recent revision of this document consult the Waters Web site waters com Trademarks ACQUITY Empower MassLynx PIC THE SCIENCE OF WHAT S POSSIBLE and Waters are...

Page 4: ...bol is used to find out the nature of the potential hazard and any actions which have to be taken Considerations specific to the 2489 UV Visible detector Power cord replacement hazard Waters contact i...

Page 5: ...ent so that it is difficult to disconnect the power cord Equipment misuse notice If equipment is used in a manner not specified by its manufacturer protections against personal injury inherent in the...

Page 6: ...are the same or very similar vary the location of the QC samples in the trays Ensure that QC sample results fall within an acceptable range and evaluate precision from day to day and run to run Data c...

Page 7: ...dustrial Scientific and Medical ISM instruments requirements Group 1 products apply to intentionally generated and or used conductively coupled radio frequency energy that is necessary for the interna...

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Page 9: ...ymbols v Audience and purpose vi Intended use of the 2489 UV Visible detector vi Calibrating vi Quality control vi EMC considerations vii Canada spectrum management emissions notice vii ISM Classifica...

Page 10: ...or off 36 2 6 6 Connecting the detector to the 2695 separations module 36 2 6 7 Connecting the detector to a chromatography data workstation using an e SAT IN module 38 2 6 8 Connecting the detector t...

Page 11: ...87 3 3 6 Replaying a spectrum 88 3 3 7 Scanning using the cuvette 88 3 3 8 Scanning using a flow cell and a syringe 90 3 3 9 Conserving lamp life 90 3 3 10 Shutting down the detector 92 4 Maintaining...

Page 12: ...s 115 5 2 3 Service diagnostic tests 120 5 3 Troubleshooting 121 5 3 1 Diagnostic tests 121 5 3 2 Power surges 121 5 3 3 Hardware troubleshooting 121 5 3 4 Lamp troubleshooting 123 A Safety Advisories...

Page 13: ...quality 141 C 1 4 Preparation checklist 141 C 1 5 Water 142 C 1 6 Using buffers 142 C 1 7 Tetrahydrofuran 142 C 2 Solvent miscibility 142 C 2 1 How to use miscibility numbers 143 C 3 Buffered solvent...

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Page 15: ...ns and Appendix C for information on high performance liquid chromatography HPLC solvent considerations 1 1 Detector description The 2489 UV Vis detector is a two channel ultraviolet visible UV Vis de...

Page 16: ...lity Method editing and storage Supports basic method programming storage and retrieval from the front panel Full diagnostic capability Supports built in diagnostic tools to optimize functionality and...

Page 17: ...heel Shutter wavelength calibration filter and second order filter Entrance slit Blazed plane holographic diffraction grating Beamsplitter Sample and reference photodiodes Waters TaperSlit Flow Cell i...

Page 18: ...ted just ahead of the flow cell diverts a portion of the light to a reference photodiode 3 When you enter a new wavelength through the detector s front panel or through Empower or MassLynx software th...

Page 19: ...haracteristics The standard analytical inert and LC MS cells have a path length of 10 mm The semi prep and microbore cell path length is 3 mm The autopurification cell path length is 1 0 mm A variable...

Page 20: ...a value The filter constant is determined by the sampling rate If you select Other you can enter a value However the value you enter is rounded up or down to a value based on the sampling rate The fi...

Page 21: ...elength Verification Failure message The detector verifies rather than recalibrates on startup to avoid errors arising from residual materials left in the flow cell or the cuvette when required Requir...

Page 22: ...atioPlot difference plot and MaxPlot functions Note that these modes are for local control of the detector only Please check the Empower and MassLynx online help for additional control information in...

Page 23: ...put channels and corresponds to the AU value where the analog outputs become saturated at full scale values Absorbance units full scale AUFS vary from 0 0001 to 4 000 AUFS Tip Changing the sensitivity...

Page 24: ...essage that any data collected for the wavelength above 370 nm can contain inaccuracies because of possible UV light interference second order effects 1 3 2 1 Chart out selection modes When operating...

Page 25: ...so the results displayed or charted are of the sample only To obtain a spectrum of a sample using the detector run a zero scan first followed by a sample scan Typically you run the zero scan using pur...

Page 26: ...or analyte at two wavelengths The RatioPlot divides absorbances at two selected wavelengths and plots the resulting ratio on a data system over one output channel channel A Use the RatioPlot when dete...

Page 27: ...ces and also how to handle solvents Before installing the detector ensure that the required components are present none of the shipping containers or unpacked items are damaged 2 2 Unpacking and inspe...

Page 28: ...Ensure that the instrument serial number found on the nameplate on the rear panel or inside the front door of the detector corresponds to the number on the instrument integrity certificate 2 3 Select...

Page 29: ...he previously added module in the system stack and slide it backward until its rear alignment pin rests in the rear alignment slot on the previously added module Power Grounded ac 100 to 240 Vac 50 to...

Page 30: ...outlet must be common and connected near the system To connect to the electricity source Recommendation Use a line conditioner or an uninterruptible power supply UPS for optimum long term input voltag...

Page 31: ...tic tests while the lamp LED blinks green The lamp LED shows steady green when the lamp is ignited 2 6 Connecting signal cables See also Waters Ethernet Instrument Getting Started Guide The following...

Page 32: ...ject Start1 TTL contact closure Configurable input to initiate sequencing of time programmed events Defines the start of a run typically an injection and resets and starts the run time clock to 0 00 m...

Page 33: ...nditions Operating mode you select for the detector stand alone or remote control Types of instruments that make up your HPLC system This section describes the input output I O and digital signal conn...

Page 34: ...length range of 190 to 700 nm To connect Ethernet cables from the detector to chromatography data software 1 Connect the single receptacle end of the Ethernet cable to your chromatography data worksta...

Page 35: ...6 2695 separations module connections to the detector for starting a method Note If the injector is an 2695 separations module running in Ethernet mode the inject start cable should not be connected H...

Page 36: ...low Figure 2 7 2695 separations module connections to the detector for turning the lamp on or off 2 6 6 Connecting the detector to the 2695 separations module You can connect the detector to the 2695...

Page 37: ...n the detector at the start of an injection make the connections summarized in the table and illustrated in the figure below Table 2 4 2695 separations module connections to the detector to generate a...

Page 38: ...n the following hardware Ethernet card Ethernet satellite interface e SAT IN module 2 6 7 1 e SAT IN module The Waters e SAT IN module shown in the figure below translates analog signals from devices...

Page 39: ...3 mm 1 8 inch from one end of the e SAT IN 9 pin connector exposing the white and black wires 2 Connect the other end of the cable to the Channel 1 or Channel 2 connectors on the front panel of the e...

Page 40: ...and configuration guide The following table summarizes the detector connections to the e SAT IN module Table 2 6 Detector connections to a e SAT IN module 2489 UV Vis detector I e SAT IN connector Pin...

Page 41: ...detector to the integrator do not exceed the input voltage rating of the integrator The analog output signal is transmitted via the cable provided in the 2489 UV Vis detector startup kit Make the con...

Page 42: ...he detector s two programmable switches SW1 or SW2 and program the timed event threshold or ratio at the front panel You can also connect the fraction collector to trigger a chart mark event input eac...

Page 43: ...bed in Verifying that the detector operates properly on page 67 Install a waste reservoir connected to the drain tube and located next to the rubber foot on the front lower left hand section of the de...

Page 44: ...the inlet tubing to the column outlet and then to the flow cell inlet 2 Connect the outlet tubing to the flow cell outlet and route it to a waste container Notice To avoid damaging the flow cell or t...

Page 45: ...rption of dissolved oxygen for systems using vacuum degassers run the solvent degasser continuously when operating the detector at wavelengths less than 230 nm 2 7 5 Making tubing connections in ACQUI...

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Page 47: ...eps three times and runs a series of startup diagnostic tests If it passes all startup diagnostic tests the Startup Diagnostics screen appears with OK following each test Figure 3 1 Startup Diagnostic...

Page 48: ...stic test failure If one or more of the internal startup diagnostic tests show a failing result the detector beeps and displays an error message For serious errors it displays the word Error in bracke...

Page 49: ...able below You can use the A B key to toggle between absorbance screens for channels A and B 3 2 1 1 Absorbance and message icons The absorbance screens and message screens in the detector program dis...

Page 50: ...lected channel Lamp on Indicates the lamp is turned on Lamp off Indicates the lamp is turned off Shift off Blank Shift off Shift on Indicates the shift is on for one key press Single wavelength Indica...

Page 51: ...and Clear Field Local method number Indicates the detector is not controlled by a data system The icon s cursive m and the current method number or an asterisk indicate current conditions are not stor...

Page 52: ...d item and then press Enter For the number 10 select 0 and then press Enter To go to the end of a choice list select For entries numbered 11 or 12 scroll to the desired item on the choice list and the...

Page 53: ...t the border of the active field is thickened highlighted Using the arrow keys you move the highlighted board to the field you want to activate Up moves up or left Down moves down or right On screens...

Page 54: ...Selects the corresponding number from a choice list 0 9 See descriptions for specific shifted numeric keys 1 See 0 9 above Lamp Displays a lamp use statistics for the currently installed lamp and prov...

Page 55: ...f the message text Enters a decimal point Also positions the cursor at the last entry in a list Some edit fields accept negative number entries Use this function to invert the sign of the number in an...

Page 56: ...nable disable inputs Enable disable external events Tip The parameters you enter into the secondary function fields become part of the current method conditions and are stored when you store the metho...

Page 57: ...the absorbance and the output voltage The output voltage reaches full scale 2 V when the current absorbance attains the AUFS value Caution Changing the sensitivity AUFS setting affects the 2 V output...

Page 58: ...Volts out Larger absorbance A or B 2 V AUFS of selected channel RatioPlot A B Charts the ratio of absorbances at two wavelengths Theoretically the ratio is constant for a pure chromatographic peak an...

Page 59: ...lected channel for scaling Volts out Absorbance difference A B 2 V AUFS of selected channel Filter time constant Adjusts the noise filter time constant to achieve the optimum signal to noise ratio wit...

Page 60: ...ro sets the signal level to zero Selecting to baseline maintains the previous baseline level when the wavelength change is made The default is to zero Table 3 1 Detector functions continued Function D...

Page 61: ...nm 254 nm AUFS 1 Numeric AUFS 0 0001 to 4 0000 2 0000 Analog rate 2 of 4 or 2 of 5 Choice Hz 10 20 40 80 1 or 2 10 1 Filter time constant 2 of 4 or 2 of 5 Numeric Sec Slow Normal Fast Other 0 0125 to...

Page 62: ...set to auto T1 represents the left hand side of the trace or ending time to be displayed The default is 30 T2 represents the right hand side of the trace or starting time The default is 0 The figure b...

Page 63: ...nctions 3 2 5 1 Configuring the detector To change default configurations use the Configuration screens Press the CONFIGURE key Shift DIAG The first of three Configuration screens appears Tip Other fu...

Page 64: ...ou determine the response of the channel using the enable chart mark function explained in the table on page 61 and shown in the figure on page 60 High Create chart mark s when contact closure changes...

Page 65: ...ne pulse period in a rectangular wave or pulse train is as specified in this field range is 0 2 to 60 seconds The figure below shows the difference between a single pulse and a rectangular wave Figure...

Page 66: ...l of the detector before the external system takes control Recommendation To prevent reabsorption of dissolved oxygen run the solvent degasser continuously when operating the detector at wavelengths l...

Page 67: ...inearity cuvette kits and system qualification tool for the detector For spare parts details see the Waters Quality Parts Locator on the Services Support page of the Waters web site Tip Before you pum...

Page 68: ...ow cell is contaminated the lamp needs to be replaced an air bubble is in the flow cell To record the sample and reference beam energies 1 Ensure that the detector is connected to the workstation 2 Fl...

Page 69: ...e calibration procedure and briefly displays a series of initialization messages similar to those you saw at startup see page 47 If calibration is successful the detector beeps three times and display...

Page 70: ...e dual wavelength mode offers the following functions Absorbance A and B MaxPlot RatioPlot A B Difference plot A B To change from single to dual wavelength mode 1 From the absorbance or HOME screen wh...

Page 71: ...ge 57 To obtain a RatioPlot 1 Be sure the detector is operating in dual wavelength mode see the previous discussion 2 From the absorbance screen press Next to reach screen 3 of 5 3 In the Data out fie...

Page 72: ...ditions so that the original recalled method is no longer in effect causing the method number to change to an asterisk The operating parameters at the time of system shutdown are restored however any...

Page 73: ...een 4 Chart mark 10 of full scale Does not apply Does not apply Yes 5 Polarity 1 2 Yes 6 Auto zero Does not apply Does not apply Yes 7 Lamp 1 Off 2 On Off No 8 Switch 1 1 High 2 Low 3 Pulse 4 Rect wav...

Page 74: ...the time field is active 10 Press HOME to return to the absorbance screen and then press Run Stop 11 Press Reset Tips If the detector is under the control of an external device the Inject Start progra...

Page 75: ...e the table above 5 When the To field is active press Enter to display the options in the table shown above or press the number corresponding to the threshold parameter you are programming 6 To set th...

Page 76: ...retrieve and press Enter Result A brief message Retrieving method n appears When the Method choice list reappears the method number icon incorporates the method number you specified see the table on...

Page 77: ...estore the previous conditions 3 Press 4 Store method and enter a storage location number Tip To clear other stored methods repeat this step until you clear all desired methods When you press HOME the...

Page 78: ...nding wavelength Scanning ends at this wavelength Pace Rate of scanning in nanometers min Determines how fast the scan is output and data are acquired The scan data are acquired at the highest possibl...

Page 79: ...he figures below show a 190 nm to 600 nm scan of erbium standard in a cuvette at 200 nm min with tick marks specified every 20 nm and the same scan without tick marks AUFS The sensitivity setting used...

Page 80: ...e zero scan The most recent zero scan performed or retrieved remains current until you perform or retrieve another scan The zero scan should be appropriate for the subsequent sample scans you perform...

Page 81: ...parameters appear on the first New scan screen for both a zero and a sample scan You can return to screen 1 to review the parameters for either scan type by pressing Next from the Run screen screen 3...

Page 82: ...to the figure on page 79 which shows two scans of anthracene overlaid one at 100 nm min and one at 1000 nm min The higher the value you enter in the Pace field the lower the resolution of the scan 6...

Page 83: ...ro scan To run a sample scan 1 Set up the zero or reference scan by following the steps in the zero scan procedure on To run a sample scan on page 83 2 On the first New scan screen press 1 Sample Scan...

Page 84: ...graphically Figure 3 27 Sample erbium scan graphical display Tip To return to the Scan choice list at the end of the scan press SCAN Shift Chart mark 5 Press Next Result Doing so displays as many as...

Page 85: ...r more scaling parameters press Enter to reformat the graphical display 10 Once the scan reappears press Next to display the four highest peaks of the scaled scan Figure 3 30 Four highest peaks from t...

Page 86: ...rk 2 Press 2 Store last scan Tip When you select Store last scan you are storing the zero scan and the sample scan as a pair Sample scan 200 nm to 400 nm 0 001 AU to 0 5 AU anthracene Zoom of sample s...

Page 87: ...ace Displays the pace for the selected spectrum 4 Press Enter to exit the information screen and return to the Scan choice list 3 3 4 Reviewing a stored spectrum Once you store a spectrum you can retr...

Page 88: ...og connector to the chart or A D device in a data collection system Once you retrieve a spectrum for playback the detector displays it graphically and you can adjust the AUFS Sample energy is not char...

Page 89: ...conditions 3 3 7 1 Before you begin Recommendation To ensure accurate results use a 10 mm path length quartz cuvette and matched pairs from the same manufacturing lot of quartz cuvettes for your zero...

Page 90: ...eference cuvette with a cuvette containing your analyte dissolved in mobile phase solvent and run a sample scan 8 Use the storage review subtract and review and replay functions to analyze the data ob...

Page 91: ...en displays the words Lamp off and the lamp icon appears with an X through it Use the Lamp key Shift 1 to shut the lamp off or turn the lamp on manually display the number of times the lamp has ignite...

Page 92: ...a timed event See Configuring event inputs contact closures on page 64 for more information on programming the lamp through the external contact closure 3 3 10 Shutting down the detector If you need...

Page 93: ...Waters be prepared to provide this information Completed normal operation checklist for the method you are using Nature of the symptom Instrument serial number Flow rate Operating wavelength and pres...

Page 94: ...rly filter and degas solvents to prolong column life reduce pressure fluctuations and decrease baseline noise Warning To prevent injury always observe Good Laboratory Practices when you handle solvent...

Page 95: ...Flushing Removing and cleaning If flushing is ineffective remove and clean the flow cell Replace flow cell components as required Note Always replace the flow cell gaskets when cleaning rebuilding or...

Page 96: ...flow cell inlet tubing Tip For more efficient flushing replace the column with a union or piece of tubing 3 Connect the column inlet tubing to the flow cell inlet 4 If another instrument is downstrea...

Page 97: ...et tubing connections to the flow cell 2 Flush the flow cell for approximately 15 minutes at the recommended flow rate Decreasing system pressure indicates the flow cell is clean 3 If the flow cell re...

Page 98: ...tweezers Torque wrench set to 0 9 N m 128 in oz or 8 in lb with 1 4 inch flat blade screwdriver bit Ethyl alcohol or methanol Flow cell rebuild kit Lint free swab Nitrogen Save all parts after removal...

Page 99: ...phase such as a water acetonitrile mix and either cap the flow ports or dry the flow cell using pure lab gases such as helium nitrogen or air for 5 to 10 minutes 4 4 3 3 Disassembling the flow cell T...

Page 100: ...wab into the cuvette end of the flow cell assembly and remove the split ring and the cuvette lens 3 Using the screwdriver remove the exit lens mount screw 4 Using plastic tweezers pull out the exit wi...

Page 101: ...rogen 3 Using the Flow Cell Rebuild Kit replace any flow cell parts that are scratched burred damaged or do not come clean using nitrogen 4 Rebuild the flow cell by following the procedure in the next...

Page 102: ...xit window into the flow cell body 11 With the tan colored exit window holder facing down use the torque wrench to tighten the exit window screw into the flow cell body to 0 9 N m 128 in oz or 8 in lb...

Page 103: ...e screws on the front plate of the flow cell assembly 2 Gently pull the assembly toward you 3 Insert the new flow cell assembly into the detector 4 Tighten the captive screws 5 Reconnect the inlet out...

Page 104: ...nd lamp such as tungsten The detector performs several self diagnostic tests when powered on or when the Calibrate key is selected One of these tests is the lamp optimization software routine After th...

Page 105: ...ove the lamp 1 Extinguish the lamp using the keypad by pressing Shift Lamp and then pressing Shift Lamp again Tip Using the keypad to extinguish the lamp allows the lamp to cool faster because of the...

Page 106: ...Lamp assembly captive screws 7 Withdraw the lamp assembly from the lamp housing Notice To prevent shattering the glass use care when disposing of the lamp as the lamp gas is under slight negative pres...

Page 107: ...gure 2 If you see any particles or dirt on the new lamp clean the lamp with a gas duster or lens tissue 3 Record the serial number located on a label attached to the lamp connector wire using the proc...

Page 108: ...in line with the alignment pin on the lamp housing Tip There is no additional alignment required Figure 4 8 Aligning the lamp 2 Gently push the lamp forward until it bottoms into position 3 Tighten t...

Page 109: ...display and keyboard diagnostic tests on page 118 If you do not record the new lamp serial number following the procedure in this section the lamp warranty is voided The detector software records and...

Page 110: ...this is a new lamp enter 0 hours are 0 9 Press the HOME key 10 At the OK to store message press Enter to store the serial number and date of installation or press Cancel to cancel the entry 11 When th...

Page 111: ...der The detector requires two 100 to 240 VAC 50 to 60 Hz F 3 15 A 250 V fast blo 5 20 mm IEC fuses Suspect a fuse is open or otherwise defective when the detector fails to power on the fans do not ope...

Page 112: ...With minimum pressure withdraw the spring loaded fuse holder Figure 4 16 Removing fuse holder 4 Remove and discard both fuses 5 Make sure that the new fuses are properly rated and then insert them int...

Page 113: ...it displays the word Error in brackets Error in place of the run time absorbance on the absorbance screen Tip To reduce the likelihood of errors be sure the cuvette holder is empty the flow cell cont...

Page 114: ...tests by pressing DIAG and then selecting 1 Reset instrument If no sticky diagnostic tests are active the wrench icon does not appear on the absorbance screen When you power off the detector sticky d...

Page 115: ...Reset instrument Resets all diagnostic tests to defaults Renders sticky diagnostic tests inactive and removes the wrench icon 2 Sample ref energy Makes it possible to view sample and reference energy...

Page 116: ...Using the input and output diagnostic tests You use the input and output diagnostic tests to display and reset the autozero offsets fix set absorbance fix set the voltage on the 2 V output monitor con...

Page 117: ...y This test sets the voltages on the analog output channels based on the current AU setting This is a sticky diagnostic test 5 2 2 5 Setting a fixed voltage output From the Input Output choice list pr...

Page 118: ...display and keypad diagnostic tests press DIAG and then press 4 Requirement Make sure the detector is powered off and the power cord is disconnected before you replace the lamp To change the lamp 1 F...

Page 119: ...agnostic test 1 From the Lamp display keypad choice list press 3 Test display to run the test Result The display fills from top to bottom and from right to left then returns to the Lamp display keypad...

Page 120: ...e test peaks from the Other diagnostics choice list to turn off this diagnostic test To override the optical filter 1 From the Other diagnostics choice list press 2 Optical filter override to manually...

Page 121: ...rective actions are described in Online Help 5 3 2 Power surges Power surges line spikes and transient energy sources can adversely affect detector operations Be sure that the electrical supply used f...

Page 122: ...d Contact Waters Technical Service In dual wavelength mode time scale is incorrect Sampling rate on controller is 1 pt s In dual wavelength mode you must select a data sampling rate of 1 point per sec...

Page 123: ...tector cell is filled with solvent and free of air bubbles and the current lamp is installed correctly Flushing the flow cell can sometimes correct the problem of apparently low lamp intensity which c...

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Page 125: ...he failure of individuals to comply with any safety precaution when installing repairing or operating any of its instruments or devices The following symbols warn of risks that can arise when you oper...

Page 126: ...a serious health threat Warning Risk of tipping Warning Risk of explosion Warning Risk of high pressure gas release Warning To avoid injury from bursting nonmetallic tubing heed these precautions whe...

Page 127: ...al fluids that you handle are infectious Specific precautions appear in the latest edition of the US National Institutes of Health NIH publication Biosafety in Microbiological and Biomedical Laborator...

Page 128: ...condary containment in the event of spills Requirement Use eye protection when refilling or replacing solvent bottles Requirement Wear clean chemical resistant powder free gloves when handling samples...

Page 129: ...otection lorsque vous vous trouvez proximit de tubes en polym re pressuris s Eteignez toute flamme se trouvant proximit de l instrument Evitez d utiliser des tubes s v rement d form s ou endommag s Ev...

Page 130: ...ici riducendo notevolmente la pressione di rottura dei tubi stessi Advertencia se recomienda precauci n cuando se trabaje con tubos de pol mero sometidos a presi n El usuario deber protegerse siempre...

Page 131: ...inform que si le mat riel est utilis d une fa on non sp cifi e par le fabricant la protection assur e par le mat riel risque d tre d fectueuses Vorsicht Der Benutzer wird darauf aufmerksam gemacht das...

Page 132: ...one per garantire protezione contro gli incendi so8stituire i fusibili con altri dello stesso tipo aventi le caratteristiche indicate sui pannelli adiacenti alla copertura fusibili dello strumento Adv...

Page 133: ...li incendi sostituire i fusibili con altri dello stesso tipo aventi le caratteristiche indicate nel paragrafo Sostituzione dei fusibili del capitolo Procedure di manutenzione Advertencia Para evitar i...

Page 134: ...bols and their associated statements can appear on labels affixed to the packaging in which instruments devices and component parts are shipped Fuse Functional ground Input Output Symbol Description K...

Page 135: ...June 11 2015 715004752 Rev A Page 135 Lower limit of temperature Temperature limitation Symbol Description...

Page 136: ...June 11 2015 715004752 Rev A Page 136...

Page 137: ...s The following table lists the environmental specifications for the 2489 UV Vis detector Table B 1 Physical specifications Attribute Specification Width 34 3 cm 13 5 inches Height 20 8 cm 8 2 inches...

Page 138: ...outlet II Pollution degree c c Pollution Degree 2 A measure of pollution on electrical circuits which may produce a reduction of dielectric strength or surface resistivity Degree 2 refers only to norm...

Page 139: ...e noise b single wavelength wet 8 x 10 6 AU 230 nm 10 points s 30 s segments 1 mL min acetonitrile analytical flow cell Baseline noiseb dual wavelength dry 35 x 10 6 AU 230 nm 280 nm 1 point s 30 s se...

Page 140: ...kPa 69 bar 1000 psi analytical cell Wetted materials 316 stainless steel fused silica PEEK Tefzel a US Patent Numbers US 5 883 721 Table B 6 Waters TaperSlit flow cell specifications Description Volu...

Page 141: ...solvent filters with particulate matter C 1 3 Solvent quality Use HPLC grade solvents for the best possible results Filter solvents through 0 45 m filters before their use Solvents distilled in glass...

Page 142: ...irectly Changes involving two solvents that are not totally miscible for example from chloroform to water require an intermediate solvent such as isopropanol temperature affects solvent miscibility If...

Page 143: ...15 210 4 3 2 propanol 2 35 117 7 15 4 4 Methyl acetate 0 45 56 3 15 17 260 4 5 Methyl ethyl ketone 0 43 80 0 17 330 4 5 Cyclohexanone 2 24 155 7 28 210 4 5 Nitrobenzene 2 03 210 8 14 20 4 6 Benzonitri...

Page 144: ...standard solvents and have M numbers of 0 32 Two liquids with dual M numbers are usually miscible with each other A liquid is classified in the M number system by testing for miscibility with a seque...

Page 145: ...ity Only a finite amount of gas can be dissolved in a given volume of liquid This amount depends on the chemical affinity of the gas for the liquid the temperature of the liquid the pressure applied t...

Page 146: ...ng or by keeping a blanket of helium over the solvent Blanketing inhibits reabsorption of atmospheric gases Sparging can change the composition of mixed solvents C 7 2 Vacuum degassing The in line vac...

Page 147: ...velength selection C 8 1 UV cutoffs for common solvents The table below shows the UV cutoff the wavelength at which the absorbance of the solvent is equal to 1 AU for some common chromatographic solve...

Page 148: ...e UV cutoff nm Mobile phase UV cutoff nm Acetic acid 1 230 Sodium chloride 1 M 207 Ammonium acetate 10 mM 205 Sodium citrate 10 mM 225 Ammonium bicarbonate 10 mM 190 Sodium dodecyl sulfate 190 BRIJ 35...

Page 149: ...avelengths may be necessary to determine the best wavelength for a particular analysis Table C 4 Electronic absorption bands of representative chromophores Chromophore Chemical configuration max nm ma...

Page 150: ...clic 210 230 21 000 Allene C C 3 260 35 000 Allene C C 4 300 52 000 Allene C C 5 330 118 000 Allene C C 2 alicyclic 230 260 3000 8000 Ethylenic Acetylenic C C C C 219 6 500 Ethylenic Amido C C C N 220...

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