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June 11, 2015, 715004754 Rev. A

Page 43

Connecting Signal Cables

This chapter describes procedures for making signal connections between the

detector and other Alliance HPLC system components.

3.1 Component connection overview

The following table summarizes the signal connections used to connect the detector to

the Alliance HPLC system components.

Table 3–1:

Component connector types

Connector type

Component

Ethernet

Chromatography data software, such as

Empower software or MassLynx software,

connected via the Ethernet network.

Tip:

The Ethernet port also supports the

Waters PC-based Autoloader utility for

installing firmware (see the Waters 2414

Refractive Index Detector release notes for

details).

IEEE-488

The following systems are connected via the

IEEE-488 bus:
• Waters data control system, such as

Empower

• Alliance HPLC System with an Alliance

e2695 separations module functioning as

the HPLC system controller

Analog outputs

A/D interface

Event inputs

• System controller (used with the Alliance

HPLC system)

• Waters injector or injector made by

another manufacturer

Ext 1

One optional external column heater

RS-232

For firmware upgrades only

Summary of Contents for 2414

Page 2: ...June 11 2015 715004754 Rev A Page ii...

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

Page 4: ...is used to find out the nature of the potential hazard and any actions which have to be taken Considerations specific to the 2414 Refractive Index Detector Power cord replacement hazard Waters contac...

Page 5: ...into the drain located on the underside of the device Tubing connected to this drain directs the flow to an appropriate waste container Equipment repair or disposal Direct questions regarding repair o...

Page 6: ...Confirms that a manufactured product complies with all applicable European Community directives or Australia EMC compliant Confirms that a manufactured product complies with all applicable United Stat...

Page 7: ...ard curve The concentration range for standards must include the entire range of QC samples typical specimens and atypical specimens Quality control Routinely run three QC samples that represent subno...

Page 8: ...r the internal functioning of the equipment Class B products are suitable for use in both commercial and residential locations and can be directly connected to a low voltage power supply network EC au...

Page 9: ...nd purpose vii Intended use of the 2414 Refractive Index Detector vii Calibrating vii Quality control vii EMC considerations vii Canada spectrum management emissions notice vii ISM Classification ISM...

Page 10: ...40 2 6 4 Connect the valve pack drip tray 41 3 Connecting Signal Cables 43 3 1 Component connection overview 43 3 1 1 Supported IEEE 488 and Ethernet configurations 44 3 2 Ethernet signal cable connec...

Page 11: ...cs failure at startup 64 5 1 3 Using online Help 64 5 1 4 Using the keypad 64 5 1 5 Navigating the user interface 68 5 2 Operating in RIU mode 69 5 2 1 Using the display in RIU mode 69 5 2 2 Navigatin...

Page 12: ...r Messages Diagnostic Functions and Troubleshooting 97 7 1 Error messages 97 7 1 1 Startup error messages 97 7 1 2 Operational error messages 98 7 2 Performing diagnostic functions 98 7 2 1 Accessing...

Page 13: ...and devices 118 A 6 Warnings that address the replacing of fuses 121 A 7 Electrical and handling symbols 123 A 7 1 Electrical symbols 123 A 7 2 Handling symbols 124 B Specifications 127 B 1 Physical...

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Page 15: ...ulk properties of the solvent and the solutes under investigation exploiting the improbability that solvents and solutes would share an identical refractive index 1 2 Operating principles 1 2 1 Optica...

Page 16: ...he medium also affects its RI At a fixed wavelength the relationship between the density of a medium and its RI is generally but not necessarily linear The most important of the factors that affect th...

Page 17: ...a medium depends on these properties Angle at which the light enters the new medium the angle of incidence Refractive indices of the old and new media The angle of a refracted light beam through the...

Page 18: ...1 n2 RI of medium 2 You can use Snell s Law to calculate the RI of a sample solution from the angle of incidence the RI of the solvent and the angle of refraction 1 2 1 3 Using changes in RI for samp...

Page 19: ...try The 2414 RI detector can measure extremely small changes in refractive index to detect the presence of sample The small difference in RI between a reference solution and a sample solution is refer...

Page 20: ...he reference side of the flow cell and the solution in the sample side of the flow cell Of these refractors only the solution in the sample side of the flow cell changes over the course of a run As a...

Page 21: ...constant representing the fixed values By detecting how far the image shifts x the refractometer measures the difference in RI n between the solvent sample solution and the solvent alone The shift in...

Page 22: ...mation 1 3 Detector description The detector is designed for high performance liquid chromatography applications It can operate as a standalone unit with an integrator or with a Waters system controll...

Page 23: ...provides measured refractive index signals to nano RIU sensitivity Purge mode Before performing a measurement the 2414 RI detector can be programmed automatically or by using the front panel keypad t...

Page 24: ...ecycle on keypad panel active Auto Zero on inject on keypad panel active Auto Zero delay from 0 to 30 seconds Chart mark on keypad panel active Auto Zero on keypad panel active 1 3 3 2 410 emulation m...

Page 25: ...configuration for flow allows mobile phase to pass through the sample flow cell and then to waste or to the recycle valve During analysis the solvent sample takes this path 1 Enters through the inlet...

Page 26: ...et tubing port 2 Passes through the sample in tube of the countercurrent heat exchanger 3 Flows through the end cap heat exchanger 4 Flows through the sample side of the flow cell 5 Flows out to the c...

Page 27: ...ystem where you can define a duration 1 3 4 2 Recycle mode You engage the recycle mode via the detector s front panel or remote control interface like a data system or controller A programmatically co...

Page 28: ...lines are coaxial which facilitates heat exchange between incoming and outgoing fluids 1 3 5 2 Flow cell The flow cell consists of two fused quartz hollow prisms Each cell has an inlet and outlet One...

Page 29: ...moving through the sample and reference sides of the flow cell goes out through the pressure relief valve to the waste reservoir The figures on page 26 and page 27 indicate the flow paths during norma...

Page 30: ...sing lens through the aperture and collimating lens to form a beam The light beam passes through the sample and reference sides of the flow cell to the mirror The light beam is reflected back through...

Page 31: ...r dc power supply The power source for the detector the dc power supply provides voltage for the analog and digital circuitry 1 3 7 1 Filter noise The 2414 RI detector uses a Hamming filter and RC fil...

Page 32: ...bance Figure 1 10 Filter time constant comparison 1 3 7 1 2 RC filter A finite impulse response filter which simulates a first order resistor and capacitor filter This filter is available in 410 mode...

Page 33: ...If an event occurs to change Polarity while Polarity 2 is TRUE an appropriate error message appears For instance a front panel key press causes a warning message to appear on the front panel display a...

Page 34: ...diagnostic tests include Central processing unit CPU Serial communication interface SCI Electrically erasable programmable read only memory EEProm RAM Display test pattern Application program checksu...

Page 35: ...ctor ensure that the required components are present none of the shipping containers or unpacked items are damaged 2 2 Unpacking and inspecting The detector s shipping carton contains besides the dete...

Page 36: ...ses Warranties and Support Services Note Ensure that the serial number on the detector s rear panel nameplate or inside the front left hand panel matches the number on its validation certificate 2 3 S...

Page 37: ...re 2 1 Aligning pins with slots 3 Lower the front of the module that you are adding so that its front alignment pin rests in the front alignment slot on the previously added module 4 Repeats steps 1 a...

Page 38: ...to a suitable wall outlet 3 Press the power switch on the front door to power on the detector Result The detector runs a series of startup diagnostic tests while the lamp LED blinks green The lamp LED...

Page 39: ...detector to the detector This tubing assembly has been Warning To avoid the harmful effects of personal contact with solvents including inhalation always wear chemical resistant powder free gloves and...

Page 40: ...nto the outlet fitting of the column or another detector 2 6 2 Connect to waste The detector s flow cell is extremely sensitive to backpressure Be sure to use the correct ID waste tubing supplied pre...

Page 41: ...the tubing into the waste container 2 6 4 Connect the valve pack drip tray A factory installed drip tray lies beneath the three valves of the valve pack Be sure to direct the drip tray tube to a suit...

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Page 43: ...or MassLynx software connected via the Ethernet network Tip The Ethernet port also supports the Waters PC based Autoloader utility for installing firmware see the Waters 2414 Refractive Index Detecto...

Page 44: ...n operate a 2414 RI detector controlled by a data control system in one of these configurations Where all system modules including the detector communicate with the data system via an IEEE 488 bus int...

Page 45: ...operation and for firmware upgrades via the Waters Autoloader utility see the Waters 2414 Refractive Index Detector release notes Requirements You cannot use the detector s Ethernet port while you are...

Page 46: ...connections needed to support these configurations Figure 3 3 Alliance HPLC system with the detector under Empower software control via the IEEE 488 bus interface You use the data control software to...

Page 47: ...om the detector s rear panel connector to the injector as shown in the table below For information on injection trigger signals from a manual injector see Connecting injection trigger signals on page...

Page 48: ...control you can make the following signal connections using the 2414 RI detector s analog out event in connectors Autozero on inject Chart mark on inject Both chart mark and autozero on inject Inject...

Page 49: ...dule and the 2414 RI detector 3 4 1 2 Generating a chart mark on injecting To generate a chart mark on the detector at the start of an injection from the Alliance e2695 separations module make the con...

Page 50: ...Inject Start Pin 9 Chart Mark Pin 2 Inject Start Pin 10 Chart Mark Table 3 6 Detector connections to an Alliance e2695 separations module Alliance e2695 separations module connector B Detector connec...

Page 51: ...2414 RI detector to chromatography data software through a two channel eSAT IN Module make the connections shown in the table and figure below Table 3 7 Analog output connections to the eSAT IN modul...

Page 52: ...each injection Inject start signal from a contact closure signal with each injection Each time the detector receives a signal from a manual injector it performs the corresponding autozero inject star...

Page 53: ...tors Pin 2 Autozero black Table 3 9 Chart mark connections to a manual injector Detector connector A Manual injector connector Pin 9 Chart Mark red Two spade lug terminal connectors both cables can be...

Page 54: ...tor connector A Manual injector 1 Inject Start 2 Inject Start 3 Ground 4 Purge 5 Purge 6 Polarity 7 Polarity 8 Ground 9 Chart Mark 10 Chart Mark 11 Ground 12 Switch 13 Switch Detector connector A Manu...

Page 55: ...thod setting When Polarity Enable is closed connected to an instrument polarity determines peak polarity Polarity open disconnected generates negative polarity Polarity closed connected generates posi...

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Page 57: ...of mixed solvents Outgassing of dissolved gases 4 2 Selecting a solvent An ideal solvent for your analysis has these characteristics Good solubility characteristics for your application Significantly...

Page 58: ...a high quality water purification system If the water system does not provide filtered water filter it through a 0 22 m membrane filter before use 4 2 4 Buffers When you use buffers dissolve salts fir...

Page 59: ...Pentane 1 358 Ethyl bromide 1 424 Acetone 1 359 Methylene chloride 1 424 Ethanol 1 361 Cyclohexane 1 427 Methyl acetate 1 362 Ethylene glycol 1 427 Isopropyl ether 1 368 N N dimethyl formamide DMF 1...

Page 60: ...ncrease in temperature but the solubility of He in benzene increases with an increase in temperature 4 3 1 3 Effects of partial pressure The mass of gas dissolved in a given volume of solvent is propo...

Page 61: ...as the solvent passes through the vacuum chamber At higher flow rates lesser amounts of gas per unit volume of solvent are removed Surface area of the degassing membrane The length of the degassing m...

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Page 63: ...rtup diagnostic tests If the detector passes all tests the initial home RIU or 410 mode screen appears Figure 5 1 Initial home screen RIU mode Figure 5 2 Initial home screen 410 mode Note The 2414 RI...

Page 64: ...etector provides limited context sensitive help When you press Shift HOME from a point in the program associated with an online Help screen the Help screen appears Figure 5 3 Example of an online Help...

Page 65: ...ing to the desired 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 ch...

Page 66: ...or down toward the end Other screens can have special instructions for the use of the s and t keys for example the Display Contrast screen Next Displays a screen with additional options related to the...

Page 67: ...0 9 above Recycle Activate the recycle or diverter valve Recycle is a toggle function The current recycle state is indicated by icon 4 See 0 9 above System Info Displays system information including s...

Page 68: ...hen press Enter Note The s and t keys do not increment or decrement numerical field entries Instead use the numerical keypad Enters a decimal point Also positions the cursor at the last entry in a lis...

Page 69: ...g the HOME key At the first use of the detector the home screen shows the factory set defaults After the first use the home screen shows the settings displayed before the detector was last powered off...

Page 70: ...sterisk that indicates current conditions are not stored as a method Remote IEEE address If the detector is controlled by a data system or other control device over the IEEE 488 bus it displays a remo...

Page 71: ...e screen press Next These are the secondary functions in RIU mode RIU FS Filter type Time constant Voltage offset RIU offset Enable disable keypad entries The settings that you specify in the secondar...

Page 72: ...ult is 500 The RIU Full Scale setting is a parameter used to generate an analog output voltage detector output with the appropriate scaling factor The value expressed in RIU corresponds to the RI valu...

Page 73: ...zero is requested via the front panel You can disable this parameter by pressing any numerical key to deselect this box Enable keypad chart mark Selected by default this parameter results in a chart m...

Page 74: ...the secondary function screens 5 2 4 Programming timed events threshold events and methods Note Programmed methods are available in RIU mode only You can store and retrieve up to 10 methods The detect...

Page 75: ...event list is sorted automatically when you press Next The 2414 RI detector allows programming of the timed events shown in the following table To program a new timed event 1 On the keypad press the...

Page 76: ...o return to the method choice list to store the method 11 Press Reset to reset the run clock to 0 If the 2414 RI detector is configured with another external device the inject start event programmed f...

Page 77: ...itial Threshold value as a timed event 3 Press Enter to advance to the next Switch field or press the s or t key to move among the five fields on the threshold events screen 4 When the Switch field is...

Page 78: ...HOD Shift Temp C 2 Press 3 Retrieve a method Result The last method number stored or retrieved is displayed in the method number box 3 Enter the number of the method you want to retrieve and press Ent...

Page 79: ...ou press Enter the following conditions occur All timed events are deleted All threshold events are disabled All other operating parameters of the method RIU FS Filter Time Constant etc are set to def...

Page 80: ...displaying the Full Scale value used by the Waters 410 and 2410 Detectors and driving the Detector Out signal on the 2414 RI detector output To switch the detector from RIU mode to 410 mode press the...

Page 81: ...ld Icon field name Function Home1 status Home2 entry Sensitivity Shows the current sensitivity setting Home1 status Temp C entry Detector Temperature Shows the current temperature of detector oven Num...

Page 82: ...ctions of the 410 mode home screen From the home screen you can access the following functions either from the home screen or by pressing the Next key Sensitivity The Waters 410 compatible Integrator...

Page 83: ...ta system The offset is added after any polarity has been applied to the output default is 0 mV Voltage offset range is 0 to 2000 mV Autozero on inject The Autozero On Inject Enable enables an autozer...

Page 84: ...software connected to the detector via the Ethernet connector A data control system running Empower software connected to the detector via the IEEE 488 bus see Connecting to a Waters data control sys...

Page 85: ...t run when contact closure changes from On closed to Off open Ignore Do not respond to Inject start input This is the default setting Use the Enter key and the numeric keypad or the s and t keys to se...

Page 86: ...the recycle input to turn on diverter valve High If the contact closure is open Recycle mode is engaged If the contact closure is closed Recycle mode is disengaged Low If the contact closure is closed...

Page 87: ...n adjust the noise level peak height peak direction and the temperatures of the internal oven and column heaters to optimize the performance of the detector 5 6 1 Attenuation and sensitivity guideline...

Page 88: ...1 1 Temperature These screens control the internal detector oven and if configured Waters column heater module These screens are the same for RIU and 410 modes Figure 5 20 Temperature screen 1 Figure...

Page 89: ...el is controlled by the software The signal source for the channel is user programmable When the Detector Oven Temperature channel output type is selected the channel will output a 0 to 2 V signal on...

Page 90: ...efractive index change trace For RIU1 and RIU2 press CE to reset to auto T1 represents the left side of the trace or ending time to be displayed default is 30 T2 represents the right side of the trace...

Page 91: ...ty water and flush the system for 10 minutes at 2 mL min 2 Replace the 100 water mobile phase with a solution of 90 methanol 10 water and flush the system for 10 minutes at 2 mL min 3 Cap the flow por...

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Page 93: ...A contaminated fluid path can cause baseline noise drift inaccurate sample refraction and other operational problems If you suspect the detector s fluid path is contaminated follow the procedure belo...

Page 94: ...Set the detector to purge mode for approximately 5 minutes and operate it for an additional 5 minutes in normal mode 4 Restart the pump or solvent delivery system and then set the flow rate to 5 mL mi...

Page 95: ...e not visible In those cases you must remove the fuse and test it for electrical continuity For information on detecting the cause of a blown fuse refer to Hardware troubleshooting on page 105 To repl...

Page 96: ...d for your requirement 6 Insert the new fuse into the fuse holder 7 Insert the fuse holder into its receptacle in the detector s rear panel and gently push on the fuse until it locks into position War...

Page 97: ...cle power and then contact Waters Technical Service if the error persists see Accessing the diagnostic functions on page 98 Most errors in this category are encountered on startup Messages requiring y...

Page 98: ...it any diagnostic test when it has ended press DIAG to return to the diagnostics choice list or HOME to return to the Home screen 7 2 1 Accessing the diagnostic functions You can use several diagnosti...

Page 99: ...stic tests affects results To clear changes to the voltage output select 1 Reset diagnostics from the diagnostics choice list or cycle power to the detector The following table lists the diagnostics t...

Page 100: ...ze LED screen 7 2 3 Using the input and output diagnostic functions Use the input and output diagnostic functions to perform these tasks Fix that is specify voltage Monitor event out switch and eight...

Page 101: ...closure inputs A solid filled in circle indicates the contact closure is closed On High An open empty circle indicates the contact closure is open Off Low 2 For the outputs SW1 perform these steps a...

Page 102: ...lumn Heater Displays the internal oven and if any the column heater status Simulate peak Specifies the generation of test peaks to calibrate your integrator or data system Valve Monitors the status of...

Page 103: ...of peak repetition 3 Enter a period for repeating the peaks in seconds from 1 to 99 4 Press the Run key Result The generation of the peaks is indicated by the real time display of the change in peak...

Page 104: ...or serial number Problem symptom s RIU FS or measurement range in RIU mode or sensitivity setting in 410 mode Internal oven temperature Flow rate Filter setting Type of column column temperature setti...

Page 105: ...el fuse No power at outlet Confirm that the ac outlet is energized by connecting another electrical unit known to be in working order and determining whether it operates front panel display fails to i...

Page 106: ...many possible causes of baseline noise by identifying the rate at which the baseline is changing and purging the flow cell with the proper solvent Cycling If the baseline is cycling determine the per...

Page 107: ...e in Decontaminating the fluid path on page 93 Flow fluctuations rapid or slow drift Correct pump problems replace pump seals ensure that valves are operating properly Improper ambient environment Ver...

Page 108: ...erm cycling Disconnect other instruments on the electrical circuit use a different wall outlet determine the suitability of the line voltage use power conditioner Radio frequency noise short or long t...

Page 109: ...end to become constant or within range within 4 minutes which can indicate that the column was not equilibrated or that the solvent is not properly degassed Baseline noise random Flow cell can contain...

Page 110: ...ion times Incorrect flow rate Verify flow rate Incorrect solvent composition Change solvent composition Column heater module not on Power on column heater module Column not equilibrated Equilibrate co...

Page 111: ...dening or loss of column efficiency can be the cause If only one peak in a chromatogram is badly shaped the peak component can be interacting with the column through a chemical mechanism that does not...

Page 112: ...Improper connection between detector and recorder Autozero detector baseline Inspect cabling between unit and recorder Solvent and sample have similar refractive indices Select a different solvent Sen...

Page 113: ...efficiency Clean or replace column Change in mobile phase composition Correct mobile phase pH or ionic composition Incorrect flow rate Change flow rate Dirty flow cell Electronic noise Clean the fluid...

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Page 115: ...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 116: ...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 117: ...luids 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 Laboratories...

Page 118: ...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 119: ...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 120: ...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 121: ...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 122: ...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 123: ...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 ince...

Page 124: ...ymbols and their associated statements can appear on labels affixed to the packaging in which instruments devices and component parts are shipped Functional ground Input Output Symbol Description Keep...

Page 125: ...June 11 2015 715004754 Rev A Page 125 Lower limit of temperature Temperature limitation Symbol Description...

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Page 127: ...le lists the environmental specifications for the 2414 RI detector Table B 1 Physical specifications Attribute Specification Width 34 3 cm 13 5 inches Height 20 8 cm 8 2 inches Depth 61 0 cm 24 0 inch...

Page 128: ...tains to instruments that receive their electrical power from a local level such as an electrical wall outlet II Pollution degree 3 3 Pollution Degree 2 A measure of pollution on electrical circuits w...

Page 129: ...0 H2O 3 0 10 9 RIU in 410 2410 emulation mode 1 s FTC RC filter 1 0 mL min 100 water Drift 1 2 0 10 7 RIU hr RIU FS settings RIU mode 1 0 10 6 to 5 0 10 4 RIU Sensitivity settings 410 mode 1 2 4 8 16...

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