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Waters 432 conductivity detector operator's guide
Waters 432 Conductivity
Copyright Waters Corporation 2010
All rights reserved
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for a comprehensive list of warning and caution Operating this instrument
When operating this instrument, follow standard quality-control (QC) procedures and the guidelines presented in this section.
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This guide is intended for personnel who install, operate, and maintain the Waters 432 Conductivity Detector.
Intended use of the Waters 432 Conductivity Detector
Use the Waters® 432 Conductivity Detector, as a standalone module or configured as part of an HPLC system, to determine changes in the thermal conductivity of column eluent as compared with that from a reference flow. The Waters 432 Conductivity Detector is for research use only.
To calibrate LC systems, follow acceptable calibration methods using at least five standards to generate a standard curve. The concentration range for standards should include the entire range of QC samples, typical specimens, and atypical specimens.
When calibrating mass spectrometers, consult the calibration section of the operator's guide for the instrument you are calibrating. In cases where an overview and maintenance guide, not operator's guide, accompanies the instrument, consult the instrument's online Help system for calibration Routinely run three QC samples that represent subnormal, normal, and above-normal levels of a compound. Ensure that QC sample results fall within an acceptable range, and evaluate precision from day to day and run to run. Data collected when QC samples are out of range might not be valid. Do not report these data until you are certain that the instrument performs ISM Classification: ISM Group 1 Class B
This classification has been assigned in accordance with CISPR 11 Industrial 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 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, EC authorized representative
Waters Corporation (Micromass UK Ltd.)Floats RoadWythenshaweManchester M23 9LZUnited Kingdom Table of Contents
Table of Contents connections . 2-13 Table of Contents Table of Contents Table of Contents Table of Contents Table of Contents The Waters® 432 Conductivity Detector is specifically designed to be integrated into chromatographic systems. The following features contribute to its performance in measuring the conductivity of column eluents: Unique 5-electrode flow cell design Heat exchanger and a built-in automatic temperature control system for Auto baseline/auto zero External recorder/integrator and chart mark connections Three time constant selections "Leak-detected" alarm signal Waters 432 conductivity detector
Conductivity Detector Method of operation
This section discusses the method of operation of the 432 Detector. Additional descriptive information appears in these sections: The 432 Detector responds to all ions present in the flow cell, since all ions in solution conduct electricity. This allows the 432 Detector to detect a wide variety of sample ions.
The 432 Detector eliminates the eluent's contribution to conductivity with an electronic technique called baseline suppression. The detector measures the eluent conductivity and assigns it a value of zero. Thus, any sample ions appear as positive or negative measurements, relative to the baseline.
The temperature of an ionic solution affects the conductivity of the ions. Generally, a solution's conductivity rises about 2% for every degree Celsius of temperature increase. The special flow cell heater in the 432 Detector minimizes the effect of ambient temperature fluctuations on measurement Flow cell design
The flow cell in the 432 Detector contains five electrodes connected in a measuring circuit: two reference electrodes, two detection electrodes, and a guard electrode that provides a local electrical "ground". Column eluent flows through the heater to attain the set temperature, and then flows through the cell, directly contacting the electrodes. The 5-electrode design permits measurement of conductivity to be made with a very low current at the detection electrodes. The low current employed eliminates impedance and other problems associated with simpler designs, and results in a stable baseline and an extended range of linearity.
Method of operation Flow cell schematic
1=Reference electrodes 1= Reference Electrodes 2=Detection electrodes 2= Detection Electrodes 3=Guard electrode 3= Guard Electrode Flow cell block ck Ion detection theory
The conductance of a solution of known concentration can be calculated using the following equation: G = measured conductance of the solution, in Siemens (1 S = ohm−1) C = concentration in equivalents per 1000 cm3K = length/area of cell (the cell constant) λ = equivalent conductance in S cm2 equiv−1 The table below lists the equivalent conductances of some common ions.1 Concentrations above 10-5 to 10-3 N, generally exhibit decreased equivalent conductance due to interionic effects.
Limiting equivalent conductance of ions in water at 25 °C
1. Henry H. Bauer et al., eds. "Instrumental Analysis," Allyn and Bacon, Boston (1978), p. 115. Reprinted with permission from the publisher.
Ion detection theory 2 Installing the Detector
This chapter guides you through the following steps in preparing the 432 Detector for operation in a chromatographic system:• Selecting an installation site that satisfies the detector's power and Unpacking and inspecting the 432 Detector and accompanying Connecting the detector to your AC power supply Connecting the detector electrically to the other components of your chromatographic system Connecting the detector inlet to the column and the detector outlet to a waste receptacle (and, if required, installing the pulse Passivating the detector and other post-column fluid path After you have successfully completed this chapter, familiarize yourself are ready to operate the detector, perform the startup procedure described in Contents
Selecting the installation site
The 432 Detector operates in any standard laboratory environment that provides suitable electrical power and remains within the following ranges: Temperature: 5 to 35 °C (40 to 95 °F)
Humidity: 20 to 80%, noncondensing
Install the instrument in a clean area that is free from exposure to: Temperature or humidity extremes, which can be found near direct sunlight, heat registers, and air conditioning vents Strong electromagnetic radiation, such as from large motors or arcing Appreciable shock or vibration The 432 Detector requires bench space that measures approximately: 10 inches (25 cm) high 14 inches (34 cm) wide 24 inches (60 cm) deep Caution: Make sure that air can circulate freely through the ventilation
slots on both side panels.
The 432 Detector requires: One properly grounded AC voltage outlet.
Correct voltage and fuse selections as shown in the Installing the Detector Unpacking and inspection
The 432 Detector is shipped in one carton that contains the following items: Waters 432 Conductivity Detector Validation certificate Waters 432 Conductivity Detector Operator's Guide Declaration of conformity Tip: If you purchased the 432 Detector as part of an ion/liquid chromatograph
system, a Waters representative will perform the installation and startup.
To unpack the 432 Detector:
1. Locate the packing list.
2. Unpack the contents of the shipping carton and check the contents against the packing list to make sure that you received all items.
3. Check the contents of the Startup Kit against the Waters 432 Conductivity Detector Startup Kit List.
4. Save the shipping carton for future transport or shipment.
Inspect all items. If you find any damage or discrepancy, immediately contact the shipping agent and Waters. For more information about the instrument warranty, refer to Waters Licenses, Warranties, and Support.
If the shipment is complete and undamaged, record the installation date and serial number of the 432 Detector in the spaces provided in Unpacking and inspection AC power connection
Warning: To avoid a potential fire hazard and damage to the 432
Detector, make sure that the voltage selector in the power connector is set correctly to match the available AC power source, and that the correct fuses are installed before you apply AC power. Power cord
The power connector is located on the lower-right corner of the rear panel. If a power plug other than the one supplied is needed for your location, consult the existing applicable regulations.
Installing the Detector Rear panel
IEEE DIPswitch cover Power cord wire identification
The 432 Detector can be adapted to operate within two voltage ranges at 50 or 60 Hz. The table below describes these voltage ranges and the fuse value that is appropriate to each.
AC power connection Nominal operating voltage
Nominal Voltage (VAC) Fuse
You need a flat-blade screwdriver to perform this procedure.
Warning: To avoid the possibility of electrical shock, turn off the front
panel power switch and unplug the power cord.
To change the operating voltage setting:
1. Remove the power cord from its connector on the rear panel of the controller and pry open the power connector cover with a flat-blade 2. Remove the voltage selection barrel and locate the correct voltage 3. Reinstall the voltage selection barrel so the desired voltage setting appears through the window when you close the power connector cover.
Installing the Detector Changing the voltage setting
4. Determine if you need to change the fuses (s ). All units are supplied with two 2-A fuses installed for 100/120 volt operation. If you operate the unit on 220/240 volt power, change the fuse as outlined in 5. Reinstall the power connector cover and the power cord.
I/O signal connections
The 432 Detector is usually installed as an integral part of a data collection system. You can control the 432 Detector either locally from the keypad on the front panel or remotely from a PowerLine™ controller, such as the Waters 600S.
This section describes the detector's I/O signals and how they connect to the following devices: PowerLine controller Empower™ or Millennium®32 software SAT/IN ™ module I/O signal connections Device signalling the Chart Marker input Device signalling the Auto Zero input Caution: To meet the regulatory requirements of immunity from
external electrical disturbances that may affect the performance of this instrument, do not use cables longer than 9.8 feet (3 meters) when connecting to the screw-type barrier terminal strips. In addition, ensure you always connect the shield of the cable to chassis ground at one instrument only.
I/O signal descriptions
The 432 Detector rear panel has an IEEE-488 connector for communication with a PowerLine controller, and a terminal strip for the input/output signals. Installing the Detector I/O terminal strip
I/O signal descriptions
Recorder output – A 10-mV full-scale analog output
signal appears on these terminals. The measurement range is determined by the product of the Base Range and Sensitivity settings: for example, 500 μS (base range) x 0.005 (sensitivity) = 2.5 μS full scale.
Integrator output – A 1-V full-scale analog output
signal appears on these terminals. The measurement range is selectable: 10, 50, or 100 μS full scale.
Marker output – A 1-second contact closure signal
appears on these terminals when either of the following events occurs:• The Chart Mark key on the keypad is pressed• A contact closure signal occurs between the Marker In terminals I/O signal connections I/O signal descriptions (Continued)
Leak Alert output – A contact closure signal
appears on these terminals if a leak is detected inside the detector.
Auto Zero input – The voltage at the Recorder and
Integrator output terminals is set to the user-selected balance offset level when a contact closure occurs between these terminals.
Marker input – A chart mark ( 0.5 mV for 3
seconds) is added to the Recorder output signal when a contact closure signal appears between these To connect cables to the I/O terminals, use a small flat-blade screwdriver.
Other rear panel connections and DIP switch
In addition to the I/O terminal strip, the rear panel also contains the following IEEE-488 connector – Communication bus for use with a Waters
PowerLine system controller, such as the Waters 600S.
DIP switch – Sets the IEEE-488 address seen by the system controller.
Ground lugs – Used to connect the 432 Detector to an earth ground
connection and also used as a chassis ground connection to other system PowerLine controller connections
The 432 Detector can be programmed remotely by a PowerLine controller (such as the Waters 600S) via the IEEE-488 data communications bus.
You need a 2.5-mm Allen wrench to connect to the 432 Detector.
Installing the Detector To connect the 432 Detector to a PowerLine controller:
1. Turn off the PowerLine controller and the 432 Detector.
2. Plug one end of the IEEE-488 cable (included in the Startup Kit) into the bus connector on the rear panel of the 432 Detector (see th e other end of the cable into the bus connector on the PowerLine controller.
3. Remove the DIP switch cover (see th using a 2.5-mm Allen wrench.
4. Refer to the to set the DIP switches on the rear panel of the 432 Detector (see the ue IEEE-488 address between 2 and 29.
5. After you set the DIP switches, reinstall the DIP switch cover.
Tip: To operate the 432 Detector in local mode, press the front panel Remote
key. The illuminated light above the key will go out.
IEEE-488 address switch
(Address 8 Shown) I/O signal connections The IEEE-488 address DIP switch employs positive logic to determine the address of the 432 Detector from the switch settings. The table below shows the settings for valid addresses.
IEEE-488 DIP switch setting
DIP switch settings
Installing the Detector IEEE-488 DIP switch setting (Continued)
DIP switch settings
Under PowerLine control, the 432 Detector is recognized as a 431 Detector and it retains the functionality of the 431 Detector with the following The Balance field on the detector setup page of the PowerLine controller affects the Integrator Balance and the Integrator Output only.
When you press the Setup key on the controller, the selected Balance
value is sent to the 432 Detector from the PowerLine controller. However, the 432 Detector output does not change to the selected balance until the detector is autozeroed by a contact closure at the Auto Zero input terminals on the rear panel (remote or local mode) or when you press the Auto Zero key on the front panel (local mode only).
Under PowerLine control, the 432 Detector retains the full functionality of local mode operation, except for the following differences: The Recorder Sensitivity ranges of 0.0002 and 0.0001 are not accessible.
The Integrator Sensitivity ranges are not accessible.
The 432 Detector does not automatically perform an Auto Zero after an Auto Base routine has occurred.
Empower and Millennium32 connections
Empower and Millennium32 software perform data acquisition, processing, and management of chromatographic information. This software requires the detector's analog signal to be converted to a digital form.
I/O signal connections Empower and Millennium32 are menu-driven applications specifically designed by Waters for chromatographers. Use the software to: Generate and print reports Store information (or data) in a central area and share this information with users who have proper security access To connect the 432 Detector to an Empower or Millennium32 computer, be sure Connect the Bus Satellite Interface (SAT/IN) module to the Bus Laboratory Acquisition and Control/Environment (LAC/E™) card in the Empower computer, Millennium32 computer, acquisition client, or LAC/E32.
Connect the 432 Detector to the Bus SAT/IN module (Channel 1 or 2).
Remove the IEEE-488 cable from the rear panel of the 432 Detector, if it is connected.
The 432 Detector is in local mode when it is connected to an Empower and Millennium32 computer.
Bus SAT/IN module
The Waters Bus SAT/IN module translates analog signals into digital form. It then transmits these digital signals to the Bus LAC/E card inside the workstation, acquisition client, or LAC/E32.
Installing the Detector Bus SAT/IN module (front panel)
Waters SAT/IN Module CHANNEL 1 CHANNEL 2 Tip: To prevent damage to the unit, always disconnect the power cord at either
the wall outlet or the power supply before you attach or remove the power connection to the Bus SAT/IN module. The Bus SAT/IN module does not have a power switch.
Connecting the Bus SAT/IN module to the Bus LAC/E card
The Bus SAT/IN module connects to the Bus LAC/E through an I/O distribution box.
To connect the Bus SAT/IN module to the Bus LAC/E card:
1. Use the I/O distribution cable to connect the I/O distribution box to the 9-pin I/O distribution port on the Bus LAC/E card at the back of the Millennium32 computer.
2. Use a serial cable to connect the data terminal on the back of the Bus SAT/IN to a port of the I/O distribution box.
3. Configure the serial port for the Bus SAT/IN module as described in the Empower or Millennium32 installation and configuration guides.
I/O signal connections Bus SAT/IN to Bus LAC/E connections
I/O Distribution Por t (9-pin) of Bus LAC/E Card I/O Distribution I/O Distribution Box I/O Distribution Box AC to DC Converter Installing the Detector Connecting the Bus SAT/IN module to the 432 Detector
The Bus SAT/IN module connects to the 432 Detector as shown below. Refer to the procedure following the figure and the complete details.
Caution: To prevent damage to the unit, do not plug in the power cord of
the Bus SAT/IN module until you perform all of the procedures described in the Waters Bus SAT/IN Module Installation Guide.
Bus SAT/IN to 432 Detector connections
Waters 432 Detector Waters SAT/IN Module CHANNEL 1 CHANNEL 2 To connect the 432 Detector to the Bus SAT/IN module:
1. Connect the white wire of the analog cable (included with the Bus SAT/IN module) to the Int + terminal on the rear panel of the 432 Detector. Connect the black wire to the Int – terminal.
I/O signal connections 2. Connect the other end of the cable to either the Channel 1 or Channel 2 connector on the front panel of the Bus SAT/IN module.
3. Connect the Event In terminals of the channel you chose in the previous step to the Inject Start output signal of the Waters Alliance® solvent delivery system or the Waters 717plus (or equivalent) Autosampler.
4. Remove the IEEE-488 cable from the rear panel of the 432 Detector, if it is connected.
The connections from the 432 Detector to the Bus SAT/IN are summarized Bus SAT/IN cable connections
432 Detector I/O
Bus SAT/IN cable
Bus SAT/IN connector
Data module connections
This section describes how to connect the analog output signal from the 432 Detector to the Waters 746 Data Module.
Caution: Remember to meet the regulatory requirements of immunity
from external electrical disturbances that may affect the performance of this instrument, do not use cables longer than 9.8 feet (3 meters) when connecting to the screw-type barrier terminal strips. In addition, ensure you always connect the shield of the cable to chassis ground.
Installing the Detector To send the analog output signal from the 432 Detector to a Waters data module, connect the signal cable in the 432 Detector Startup Kit as described Data module signal cable connections
432 Detector I/O
Marker out signal
The Marker Out terminals of the 432 Detector provide a contact closure output signal when either of the following events occurs: Chart Mark key is pressed Marker In terminals are shorted together Use the signal to start a Waters 746 Data Module by connecting a signal cable to the module's data cable.
Data module chart mark cable connection
432 Detector I/O
Join to both Remote Start wires (white and Chart recorder connections
To connect the 432 Detector to a chart recorder:
1. Attach the Recorder cable (see he 432 Detector REC output terminals, as indicated in th I/O signal connections 2. Connect the cable shield to the ground lug on the 432 Detector rear 3. Connect the other end of the cable to the 10-mV input terminals on the chart recorder.
Chart recorder cable connections
432 Detector I/O
Chart marker input connections
The 432 Detector accepts a chart mark (start inject) signal from the following Waters 717plus Autosampler Any other device that provides a compatible switch closure Waters 717plus Autosampler
To connect the 432 Detector to a Waters 717/717plus Autosampler, connect a signal cable as indicated in the table below.
Autosampler chart mark cable connections
432 Detector I/O
Marker In (+)
Either Inject Start terminal of a pair Other Inject Start terminal of the same pair Installing the Detector Auto Zero input connections
The voltage at the Recorder and Integrator outputs is set to the user-selected balance offset level when a contact closure occurs between the Auto Zero terminals. This section describes how to connect the 432 Detector to the following devices (so that an auto zero occurs at the injection point): Waters 717plus Autosampler Any other device that provides a compatible switch closure Waters 717plus Autosampler
To connect the 432 Detector to a Waters 717plus Autosampler, connect a signal cable as indicated in the table below.
Autosampler Auto Zero cable connections
432 Detector I/O
Auto Zero (+)
Either Inject Start terminal of a pair Other Inject Start terminal of the same pair Alliance Separations Module connections
Connect the detector to Waters Alliance Separations Modules, when it is not under the control of the Millennium32 software, to perform the following tasks: Auto-Zero on inject Chart mark on inject Generating Auto-Zero on inject
To generate the Auto-Zero function on the 432 Detector at the start of an injection, make the connections summarized in the I/O signal connections Connections for generating Auto-Zero on inject
Alliance Separations Modules (B
432 Detector (A inputs)
inputs and outputs)
Pin 1 Inject Start
Pin 2 Inject Start Before you can generate an Auto-Zero from an Alliance Separations Module, you must configure the Auto-Zero signal at the 432 Detector front panel. The default Auto-Zero signal is Low.
Alliance Separations Module connections to the 432 Detector
Auto-Zero on inject
Waters 432 Detector B (inputs and outputs) 4 Lamp On/Off +5 Lamp On/Off - Hold inject 1 – 8 Ground – Marker in Hold inject 2 – Chart out +Chart out – Generating chart mark on inject
To generate the chart mark function at the start of an injection, make the connections summarized in the table below and illustrated in the Installing the Detector Connections for generating chart mark on inject
Alliance Separations Modules (B
432 Detector (A Inputs)
inputs and outputs)
Pin 1 Inject Start
Pin 2 Inject Start Before you can generate a chart mark from an Alliance Separations Module, you must configure the chart mark signal at the front panel. The default chart mark signal is Low.
Alliance Separations Module connections to the 432 Detector
chart mark on inject
Waters 432 Detector B (inputs and outputs) 4 Lamp On/Off +5 Lamp On/Of Hold inject 1 – Hold inject 2 – Making fluidic connections
Fluid lines to a column and waste container connect to the front of the 432 these connections: Making fluidic connections Cut the tubing.
Assemble compression fittings and ferrules.
Connect the tubing to the detector.
This section will guide you through each of these procedures.
Caution: Conductivity detection is sensitive to flow rate fluctuations. If
you use a non-Waters pump or a Waters pump without the SILK microflow compensation algorithm, you must install the pulse dampener kit supplied in the Startup Kit for optimum performance. Refer to the installation procedure in this section.
Conductivity Detector a ste (18 inches. (18 inches, 0.000-inch ID) Cutting stainless steel tubing
You need the following tools to cut stainless steel tubing: A file with cutting edge Two cloth- or plastic-covered pliers Installing the Detector To cut the tubing:
1. Measure the length of 1/16-inch OD, 0.009-inch ID, stainless steel tubing you need to make the following connections:• Column to the detector inlet Detector outlet to a suitable waste container 2. Use a file with a cutting edge to scribe the circumference of the tubing at the desired length.
3. Grasp the tubing on both sides of the scribe mark with cloth-covered pliers. Gently work the tubing back and forth until it separates.
4. File the ends smooth.
Cutting polymeric tubing
Waters chromatography systems are supplied with a tubing cutter (similar to the one in the ) to facilitate cutting polymeric tubing. This section presents the recommended procedure for using the tubing cutter.
Tip: To avoid bandspreading caused by angled cuts, always use a tubing
cutter. Angled cuts leave unswept dead volumes at the connection junction due to the poor fit of the tubing against the connector or port.
To cut a length of polymeric tubing:
1. Estimate the length of tubing required to connect the components. Allow slack so that the tubing is not pulled tightly around sharp corners.
2. Insert the tubing into the cutter so that the tubing extending from the metal side is the length required. Use the proper hole to have a snug enough fit so that the tubing is not flexed by the blade when you cut it.
Making fluidic connections Cutting polymeric tubing
3. Press down on the razor blade to cut the tubing. Discard the excess tubing that extends from the clear side of the cutter.
4. Inspect the cut for burrs or scratches and for the perpendicularity of the Assembling compression fittings
To assemble each compression fitting:
1. Slide the compression screw over the tubing end, followed by the ferrule.
2. Mount the ferrule with its taper end facing the end of the tubing.
Ferrule and compression screw assembly
Compression screw 0.0009-inch ID tubing 0.009-inch I.D. Tubing Installing the Detector Connecting to the 432 Detector
To make connections at the column outlet and detector inlet, and at the
1. Install a compression screw and then a ferrule on the length of 0.009-inch tubing from the column outlet. Use stainless steel fittings on stainless steel tubing and PEEK fittings on PEEK tubing.
Tip: If you are using a column with 1/4–28 end fittings and there is a
length of tubing with 1/4–28 fittings on each end, use the 1/4–28 to Z-detail adapter (included in the Startup Kit) to connect this tubing to the tubing that leads to the detector inlet.
The Waters IC-Pak C column comes supplied with a length of tubing that has a 1/4–28 fitting on one end (column outlet) and a Waters compression screw and ferrule on the other end (detector inlet).
2. Push the free end of the tubing as far as it will go into the IN fitting on the 432 Detector. While you hold it there, use a 5/16-inch open-end wrench to tighten the compression screw 3/4-turn past finger-tight.
Tip: The 432 Detector and IC-Pak series of columns have very deep
3. Remove the compression screw and tubing from the connection and verify that fluid can flow freely.
4. Reconnect the tubing to the IN fitting, making sure to push the tubing all the way into the fitting.
5. Install a ferrule on an 18-inch length of 0.009-inch tubing and connect it to the OUT connection on the 432 Detector. Use stainless steel fittings on stainless steel tubing and PEEK fittings on PEEK tubing.
6. Place the other end of the tube in a waste container. If you are using any Teflon tubing, attach it after the stainless steel or PEEK tubing.
Installing the pulse dampener
To achieve the best performance from the 432 Detector in a chromatographic system with a non-Waters pump, Breeze™ software, or Waters HPLC 515 Pump, you must install the pulse dampener kit supplied in the Startup Kit. The pulse dampener is not required if you are using a Waters 2695 Separations Module.
Making fluidic connections To install the pulse dampener between the pump and the injector:
1. Assemble the pulse dampener (see the ) using the instructions in the pulse dampener kit.
2. Connect the large-ID (0.020-inch) tubing to the pump outlet using a stainless steel compression screw and ferrule.
3. Connect the small-ID (0.009-inch) tubing to the injector inlet using a stainless steel compression screw and ferrule.
4. Disconnect the tubing from the injector inlet.
5. Pump ASTM Type I reagent water at £ 2 mL/min through the pulse dampener assembly until you see a constant stream exiting from the restrictor assembly outlet line.
6. Reconnect the tubing to the injector inlet.
0.009 inch I.D.
Installing the Detector Passivating the system
Passivating the system removes potential contamination from the wetted surfaces of all system components. Perform passivation on a new system, and subsequently, whenever you suspect that contamination may have occurred. elp diagnosing performance problems.
Use this procedure for Waters hardware only. For other equipment, check with the manufacturer before you continue with this procedure.
Caution: If you are installing the 432 Detector into an existing Waters
system, replace the pump seals before you passivate. Use the new pump seals supplied in the Startup Kit and refer to the replacement procedure in the pump manual.
To passivate the system:
1. Replace the column with a union fitting.
2. If the system is not new, flush it thoroughly with ASTM Type I reagent water to remove any residual solvents or salts.
3. Connect the power cord to the 432 Detector and plug the other end into an AC power outlet. Push the 432 Detector power switch to turn on the Warning: To avoid chemical hazards, always wear safety glasses and
gloves when you are using solvents.
4. Prime the pump with 6 N nitric acid (HNO3) and run it at a flow rate of 1.2 mL/min for 20 minutes to passivate all the wetted parts of the detector. Press the Clear key to stop the overrange alarm.
5. Stop the pump.
6. Remove the inlet line from the nitric acid and place it in ASTM Type I reagent water.
7. Flush the system using one of the following methods: Prime and start the pump, then flush it with ASTM Type I reagent water at 1.2 mL/min until you observe a consistent reading of less than 20 μS (base range set to 50 μS).
Passivating the system Flush the system overnight with 100% methanol at a reduced flow rate. By the next morning the system will be passivated and ready Tip: If you are using a pump with seal-wash capability, skip step 8.
8. Use a syringe to flush the back of the pump seals and pistons by slowly running about 5 mL of water into the top hole in the baseplate of the pump heads. Place a tissue under the baseplates to absorb the water.
9. Set the pump flow rate to 0.0 mL/min. It is not necessary to turn off the 432 Detector unless it will be idle for an extended period (14 days).
For best results, always leave the power on to maintain cell temperature; it takes a minimum of 2 to 3 hours once the detector is turned on to equilibrate the flow cell at the selected operating temperature.
Verifying the detector
This procedure is a guideline for verifying that the detector works correctly within its expected operational range. The detector is calibrated before shipping, and recalibration is not normally required.
Verify the detector when any of these conditions apply: When you replace the flow cell To verify accuracy When you make adjustments Tip: You need solution of 1 mM potassium chloride (KCl) to calibrate the
Tip: Waters suggests one of its Technical Service Representatives perform this
To perform the calibration procedure:
1. Turn on the 432 Detector and set the temperature control to 35 °C. Allow 2 to 3 hours for the temperature in the flow cell to equilibrate.
2. Set the base range to 200 μS.
3. Set the Filter Time Response to Fast.
Installing the Detector 4. Pump 1 mM KCl solution through the detector (without a column in 5. Verify that the front panel output is 147 μS ± 5 μS.
Verifying the detector Installing the Detector 3 Operating the Detector
This chapter contains:• A description of front panel controls and displays Procedures for starting up, shutting down, and long-term storage Recommended operating practices Controls and indicators
The figure below illustrates the controls and indicators on the front panel of the 432 Detector.
Waters 432Conductivity Detector Power switch
The power switch (located in the lower-right corner of the front panel) controls power to the 432 Detector. Upon startup, an initialization routine verifies the data in ROM memory, tests RAM memory function, and checks for any internal leakage or an eluent conductivity over-range condition.
The display shows instrument status and parameter values in two 20-character lines of text. Upon startup, Waters 432 Self Check appears Operating the Detector briefly. If any error conditions are detected during startup or normal operation, the appropriate error message is displayed.
The main screen shows the measured conductivity, as well as the base range and sensitivity settings. When you set an operating parameter, the display shows the selected or entered value.
A corresponding error message is displayed if one of the following conditions ROM/RAM error (checked during startup only)Error: ROM/RAM Leakage detectedError: Leak Temperature control failureError: Temp Over-range (above base range setting)Error: Over Range Overflow (above 10,000 μS)Error: Over Flow Press the Clear key to clear an error alarm and message. For a continuing
error condition, the error message remains after the audio alarm is cleared.
Use the keypad to control the operat describes the function of each key.
Controls and indicators Tip: Three keys (Balance, Sensitivity Range, and the numeral 1) perform an
alternate function when they are preceded by the Shift key.
Remote key: Toggles between local and remote
operating modes. In remote mode, the light above the key is on and all other front panel controls are Polarity key: Toggles the polarity of the signal to the
external chart recorder and integrator. When positive polarity is selected, the light above the key is Base Range key: Sets the base sensitivity range of
the 432 Detector to the appropriate value for the eluent being used. The base sensitivity is set to one of ten steps, from 10 μS (maximum gain) to 10,000 μS, using the Up and Down keys or the numeric keypad.
Sensitivity Range key: Sets the sensitivity range
multiplier of the 432 Detector. The sensitivity range has twelve steps, from 0.0001 (maximum sensitivity) to 1.0 (available only with 100 μS multiplier setting), and is set using the Up and Down keys or the numeric keypad. The 10-mV full-scale recorder response is calculated by multiplying the Base Range by the Sensitivity Range to obtain a value of "x" μS / 10 mV FS. The recorder range is 1 to 0.0001 for the 100 μS setting and 0.1 to 0.0001 for the two lower settings.
Shift key then Sensitivity Range key: Sets the
sensitivity range multiplier of the integrator to 100, 50, or 10 μS using the Up and Down keys or the numeric keypad; the integrator output is 100, 50, or 10 μS/1 V, respectively.
Operating the Detector Key descriptions (Continued)
Balance key: Manually sets the offset (%) of the
signal to the external chart recorder. (Use the numeric keypad or Up and Down keys.) Shift key then Balance key: Manually sets the
offset (%) of the signal to the integrator. (Use the numeric keypad or Up and Down keys.) Shift key after Balance key: Changes the polarity
of the offset. Allowable values are –100 to +100%.
Temperature key: Sets the temperature of the
detection cell. Use the Up and Down keys or the numeric keypad to turn temperature control off (Setting 0) or select one of the following eight settings: 30, 35, 40, 45, 50, 55, 60, or 65 °C. The light above the key is illuminated when the temperature control is on.
Response key: Sets the response time constant of
the 432 Detector to optimize signal-to-noise ratio. Use the Up and Down keys or the numeric keypad to choose Setting 1 (Fast, 0.25 sec) for very narrow peaks, Setting 2 (Standard, 0.5 sec), or Setting 3 (Slow, 1.0 sec) to detect wider peaks. Setting 2 is used for most applications.
Auto Zero key: Automatically zeros the Recorder
and Integrator signals to the specified Recorder Balance and Integrator Balance offsets, respectively.
Auto Base key: Automatically sets the base range of
the 432 Detector to the appropriate value for the eluent being used. This is the next highest setting above the actual background conductivity of the Controls and indicators Key descriptions (Continued)
Shift key: Press the Shift key before, not along with,
other keys to access additional functions and also to change polarity when you set balance offset values. When the Shift key is pressed, an asterisk (*) appears at the right side of the display; press Shift again to
return to normal mode.
Shift key then Balance key: Displays the
integrator balance offset value. When setting the balance offset, press Shift to change polarity.
Shift key then Sensitivity Range key: Displays
integrator range value.
Shift key then 1 key: Displays the current, actual
value of the chart recorder balance offset. Press Enter to return to the main screen.
Chart Mark key: When this key is pressed, a
1-second, 1-mV signal is sent to the Recorder terminals and a 1-second contact closure is sent to the Marker Out terminals.
Enter key: When you manually set offsets,
sensitivity range, or base range, pressing Enter records the displayed value and returns the display to the main screen. The Enter key is also used to access the beep setting function.
Clear key: Erases a value input from the keypad.
The Clear key is also used to clear an error alarm and message. For a continuing error condition, the error message remains after the audio alarm is cleared.
Up key: Increments the current parameter setting.
Operating the Detector Key descriptions (Continued)
Down key: Decrements the current parameter
You can set the 432 Detector to beep when a key is pressed and/or an error condition is detected.
Use the Clear key to stop an error alarm. For a continuing error condition,
the error message remains after the beep is cleared.
To set the beep function, follow the procedure in the table below.
Setting the beep function
Accesses the beep function parameters.
. (decimal point) Turns on the beep-on-error function.
Turns off the beep-on-error function.
Turns on the beep-on-keystroke function.
Turns off the beep-on-keystroke function.
Saves settings and returns to the main Tip: Since the void volume in many separations contains highly conductive
counter-ions, the error alarm sounds upon each injection. to silence the beep-on-error alarm.
Controls and indicators Startup and shutdown
Perform the following procedure to start the 432 Detector. Typically, this procedure is done at the beginning of each workday.
Tip: This procedure assumes that the flow cell has stabilized at the selected
temperature (minimum 2 to 3 hours). Standard practice is to leave the detector powered and with the temperature control on unless the instrument will be unused for several days.
Set the temperature at least 5 °C above the highest ambient temperature expected for the duration of the application.
1. Prime the pump with properly degassed eluent and set the flow rate to 1.2 mL/min or to the flow rate recommended for your particular column or application. Do not sparge eluents, since sparge gasses may contain ionic contaminants.
2. Set the response (time constant) to the desired setting by pressing the Response key. A standard setting (0.5 seconds) is preferred for most 3. Set the base value by pressing the Auto Base key or by manually entering the base range that is the next highest setting above the eluent's background conductivity.
4. Turn on the recorder/integrator and run the system until the baseline 5. Depending on whether you are using a recorder or an integrator, do one of the following actions:• If you are using a 10-mV recorder connected to the Recorder terminals on the rear panel, select the desired sensitivity by pressing the Sensitivity Range key, then the appropriate Up or Down arrow key.
If you are using an integrator connected to the Integrator terminals on the rear panel, select the desired sensitivity by pressing the Shift and Sensitivity Range keys, then the appropriate Up or Down arrow Operating the Detector 6. Zero the recorder/integrator by pressing the Auto Zero key or have the Auto Zero terminals of the rear panel I/O terminal strip connected to your manual injector or autosampler.
The 432 Detector is now ready for operation.
To eliminate the need to allow time for the flow cell temperature to equilibrate, leave the 432 Detector turned on at the end of the workday or workweek. Set the temperature control to the operating temperature and the pump flow rate to 0.01 to 0.1 mL/min (depending on the pump).
If the 432 Detector is to be removed from a system for storage or if the system itself is to be stored for a long time, flush the detector/system with 100% water, then 100% HPLC-grade methanol. Leave the methanol in the system after shutdown. If you are removing the 432 Detector from the system, seal the inlet and outlet bulkheads with dead-end fittings or a loop of tubing.
Observe the following recommendations for best detector performance.
The 432 Detector should be powered up and set at the desired operating temperature for two to three hours before use. Select a temperature at least 5 °C above the highest ambient temperature expected during the duration of the application. The detector is usually set at 35 °C, but it should be set higher if the ambient temperature will exceed 30 °C.
You may choose to leave the 432 Detector powered up overnight at a flow rate of 0.01– 0.1 mL/min (depending on the pump) to minimize the daily reequilibration time.
A drifting baseline is one indication that the temperature of the flow cell is not uniform across the flow cell or stable over time.
The Base Range is normally set at the next setting above the background conductivity of the eluent. For example, if the conductivity of borate/gluconate eluent is 270 μS, set the Base Range to 500 μS.
The Integrator output is not attenuated; signals should be below 1 V. Set the Integrator output to 10 μS/V for small signals or to 50 μS/V when you expect a signal greater than 10 μS. If you are using the 432 Detector with chemical suppression, set the integrator output to 100 μS/V.
The Recorder output is attenuated and the Sensitivity Range should be adjusted to provide the appropriate output level.
Signal polarity depends on eluent conductivity. If necessary, press the Polarity key to obtain peaks rather than dips.
Replace your eluent reservoir filter regularly. When you analyze cations, use an all-plastic eluent reservoir filter. Filter and degas eluents to prolong column life, reduce pressure fluctuations, and decrease baseline noise. When you change eluents, flush the flow cell and associated tubing thoroughly with the new eluent.
Operating the Detector 4 Performing Ion Analysis
This chapter provides essential information about ion analysis techniques. Two representative columns serve as typical examples: the Waters IC-Pak A for anions and the IC Pak C M/D for cations. The following topics are discussed:• Configuring the system Selecting and preparing eluents Preparing and injecting standards Also refer to the manufacturer's manual for the particular column you are using. A recommended source for more information about ion analysis in general is Ion Chromatography: Principles and Applications by Paul R. Haddad and Peter E. Jackson, Elsevier Science Publishing, New York, 1990.
To obtain full performance from the 432 Detector in chromatographic applications, observe the guidelines presented in this section regarding: Sample preparation Water purity (the absence of conductivity) is the most important consideration in ion analysis.
Tip: Using water other than ASTM Type I reagent water will compromise the
accuracy of your results.
Freshly drawn ASTM Type I reagent water, conforming to ASTM specification D1193, with total organic carbon <100 ppb is recommended. Sophisticated laboratory water systems that use a combination of reverse osmosis, mixed bed ion exchange, and carbon adsorption cartridges produce ASTM Type I reagent water, and are recommended for ion chromatography applications. Do not use HPLC-grade water or distilled water.
Caution: To avoid damage to the detector flow cell, do not allow the flow
cell to dry out.
Use plastic to contain all anion and cation samples, cation standards, and cation eluents. When you analyze trace level ions in water, polystyrene containers such as tissue culture flasks are recommended; polypropylene or polymethylpentene containers suit most other applications. Use polystyrene tissue culture flasks for long-term storage.
If your system operates on Breeze software or contains a 2695 Separations Module, use 4-mL polycarbonate vials to hold your samples and standards.
Caution: Avoid glass containers (which tend to leach sodium cations)
when you are analyzing for cations.
Performing Ion Analysis Preparing containers for low-level analysis
To prepare plastic containers for low-level analysis:
1. Soak all containers for 5 hours with a 1:1 solution of nitric acid (HNO3) and ASTM Type I reagent water.
2. Rinse with plenty of ASTM Type I reagent water. The containers are ready for analysis in the ppm range.
3. For analysis in the ppb range, fill each container completely with ASTM Type I reagent water and let soak overnight.
Certain applications that involve ppb level analysis may require container considerations beyond the scope of this manual. For further instructions on trace metal cleaning of plasticware, see "Selection and Cleaning of Plastic Containers for Storage of Trace Element Samples," JR Moody and RM Lindstrom, Analytical Chemistry, v. 49, Dec 1977, pp. 2264-67, or contact the Waters Technical Services Department.
To avoid contamination, always rinse a syringe two to three times with ASTM Type I reagent water before you draw standards or samples for injection.
High-pH eluents (such as hydroxide eluent) absorb atmospheric CO2, which slowly acidifies the eluent causing baseline drift and retention time changes.
To minimize carbonate absorption, connect a soda lime (Ascarite®) tube to
the eluent bottle as follows:
1. Insert a 3/4-inch (2-cm) piece of glass wool in one end of a polyethylene tube with end fittings. Attach the end fitting.
Warning: To avoid chemical burns, wear gloves, lab coat, and eye
glasses when you are handling soda lime.
2. Fill the tube with soda lime (Ascarite) until it reaches 3/4 inches (2 cm) from the top.
3. Add another piece of glass wool to the other end of the tube and attach the end fitting.
4. Drill a hole in the cap of the reagent bottle. The hole should be large enough to accommodate the end fitting. Drill a second hole for the pump 5. Pass the pump inlet line through the hole. Seal the hole with paraffin 6. Change the soda lime in the tube when it is exhausted.
Soda lime tube
Polypropylene ube Performing Ion Analysis Sample preparation is very important in ion analysis. Contact the Waters Technical Services Department, if you need assistance.
As a general rule, to analyze a sample of completely unknown ionic concentration, initially prepare at least a 1:100 dilution and inject 100 μL. For best results, injections should contain a total anion concentration of no more than 300 ppm for the IC-Pak A column or a total cation concentration of no more than 10 ppm per ion for the IC-Pak C M/D column.
The sample volume (usually 100 μL) generally equilibrates to the pH of the eluent upon injection. However, for samples with pH values that differ greatly from that of the eluent (for example, strong acids and bases), bring the sample pH close to that of the eluent before you inject the sample, if possible.
Do not inject concentrated samples directly into the mobile phase. Direct injection may cause precipitation of the salts in the sample. Dissolve (or dilute) samples in an appropriate volume of the mobile phase first. If you must use other solvents, watch for precipitation upon injection into the eluent. Always filter samples before you use them.
Cationic samples that contain organic amines may exhibit hydrophobic interaction between the mobile phase and packing. You may use a water-miscible organic mobile phase, such as acetonitrile, as a modifier to reduce this. Pretreat the sample with a Sep-Pak® C18 cartridge to remove Configuring the system
system configuration. Refer to the procedures to cut tubing and assemble fittings.
Configuring the system System configuration for ion analysis
Waaters In-Line Degasser ters in-line degasser* † Required for Waters pumps without SILK or non-W tRequired for non-Waters pumps or Waters pumps with Breeze software, such as the HPLC 515 If your system uses a non-Waters pump or a Waters pump with Breeze software, such as the HPLC 515, use a pulse dampener (supplied in the Startup Kit) to achieve the best performance from the 432 Detector.
Install the pulse dampener between the pump and the injector, as described in Eluents for ion analysis
This section describes how to select, prepare, and use eluents for ion analysis.
Caution: Never recirculate eluents. Ions from sample and standard
injections progressively contaminate a recirculating eluent.
Observe the following guidelines when you prepare eluents for ion analysis: Use only ASTM Type I reagent water with total organic carbon <100 ppb.
Use the highest purity salts and reagents available.
Performing Ion Analysis A pH meter is recommended for checking the pH of eluents; care should be taken to avoid cross contamination. Adjust the pH with potassium hydroxide (KOH) or lithium hydroxide (LiOH). For eluents such as octane sulfonate, test an aliquot of the eluent with pH paper. Never immerse pH paper directly into a batch of eluent.
Use the following formula to prepare eluents:
Formula Wt of Salt x Molarity = g/L Salt
Eluent filtering and degassing
The Waters Solvent Clarification Kit is recommended for eluent filtration and preliminary degassing. Durapore® 0.22-mm filters can be used for all ion chromatography eluents. Millipore 0.45-mm HATF filters may be used for aqueous eluents containing no organic modifier. For eluents containing organic modifier, use the Durapore filters.
Caution: To avoid contamination when you analyze for cations,
minimize the time that the eluent is in contact with the glass filtration apparatus and transfer the eluent to a suitable pre-cleaned plastic container as soon as possible.
After you install a new filter, pass 20 to 30 mL of eluent through the filter under vacuum. Turn off the vacuum, swirl the eluent throughout the flask and discard. Reattach the flask to the filter apparatus and filter the remaining eluent. As soon as filtration is complete, transfer the eluent to a precleaned plastic container, introducing the least possible amount of bubbles in the process.
The Waters In-line Degasser is recommended for continuous online degassing.
Preparing anion eluent
This section presents the procedure for the preparation of sodium borate/gluconate concentrate and eluent.
Consult the manufacturer's manual for your column (IC-Pak Column and Guard Column Care and Use Manual included with Waters columns) for additional instructions on the selection and preparation of eluents.
A recommended source for more information about ion analysis in general is "Ion Chromatography: Principles and Applications," by Paul R. Haddad and Peter E. Jackson, Elsevier Science Publishing, New York, 1990.
Eluents for ion analysis Preparing lithium borate/gluconate concentrate
To prepare sodium borate/gluconate concentrate, re Preparing cation eluent
To prepare 1 L of cation eluent, refer to.
Standards for ion analysis
This section describes how to prepare and inject ion standards.
Tip: It is recommended to purchase certified 1000-ppm anion standards
instead of preparing them manually. Certify all manual standards against National Institute of Science and Technology traceable standards.
Standard concentrations in this manual are defined in terms of mass. For example, 1 mg of sample per liter of water equals a 1 ppm concentration, since 1 L of water has a nominal mass of 1 kg (0.997 kg at 25 °C).
1 part per thousand = 1 mg/mL = 1 g/L = 1000 ppm1 part per million (ppm) = 1 μg/mL = 1 mg/L = 1000 ppb1 part per billion (ppb) = 1 ng/mL = 1 μg/L = 1000 ppt1 part per trillion (ppt) = 1 pg/mL = 1 ng/L For accurate quantitative results, do not store standards beyond the approximate periods listed in the table below. Be aware that shelf-life depends on many factors and may be significantly shorter than shown here.
Shelf-life of standards
Nitrite and carbonate concentrates Performing Ion Analysis Shelf-life of standards (Continued)
All other anion concentrates Cation concentrates Cation standards must be stored in properly prepared plasticware. See Preparing anion standards
This section presents the procedure for preparing a 7-anion standard. If a simpler standard suffices, follow the procedure, but select only three or four salts, such as sodium chloride, sodium nitrate, and sodium sulfate.
Always use salts of at least reagent-grade purity. If you require quantitative results or you use hygroscopic salts, dry the salts overnight at 80 °C before you make solutions. Store the dried salts in a desiccator.
Preparing a 7-anion standard
To prepare a 7-anion standard:
1. Weigh out the amounts of dry salts shown in the table below or use the following formula to calculate the amount for a salt not listed:(Mol. Wt. Salt / Mol. Wt. Cation) x 0.1 = g Salt Salts for anion standard concentrates
Sodium fluoride (F–) Sodium chloride (Cl–) Sodium nitrite (NO –) Potassium bromide (Br–) Sodium nitrate (NO –) Potassium phosphate, monobasic (HPO 2–) Sodium sulfate (SO 2–) Standards for ion analysis 2. Place each salt in a separate plastic100-mL volumetric flask and dilute to the mark with ASTM Type I reagent water. Each concentrate contains 1000 ppm of the anion.
3. Combine the amounts listed in the table below in a 100-mL volumetric flask with ASTM Type I reagent water.
Anion concentrate dilutions
Injecting anion standards
To inject a standard, obtain the following materials: 1-cc disposable plastic syringe – Pharmaseal® Stylex® disposable
syringe with a Luer Slip® tip, or equivalent.
Autoinjector or manual injector with 100-μL loop – Ion
chromatography commonly uses a 100-μL injection volume. When you use a fixed loop, overfill a minimum of three times.
Performing Ion Analysis Injecting the standard
To inject the standard:
1. Set up the 432 Detector as follows: Base Sensitivity = 500 μS Integrator Sensitivity = 10 μS/V Recorder Sensitivity = 0.01 (strip chart) Response = STD (0.5 seconds) Temperature = 35 °C 2. Equilibrate the 432 Detector as described in 3. Rinse a 1-cc disposable plastic syringe two or three times with ASTM Type I reagent water, then load the standard.
4. Place the syringe tip into the sample loading port and overfill the 100 μL loop at least three times (that is, 300 μL).
5. Inject the sample.
representative chromatogram of the 7-anion standard run on an IC-Pak A (4.6 mm x 5.0 cm) column with borate/gluconate eluent at 1.2 mL/min flow rate. The separation of the standard typically takes 12 to 15 minutes with this Standards for ion analysis Chromatogram of a 7-anion standard
Preparing cation standards
This section presents the procedure for preparing an 8-cation standard. If a simpler standard suffices, follow this procedure selecting only those salts that you want in the standard. For accurate quantitative results, use only properly prepared plasticware and do not store standards beyond the recommended shelf-lives listed in the .
Preparing cation standard concentrates
Tip: It is recommended that you use certified 1000-ppm cation standards not
prepared in acid with this method.
Performing Ion Analysis To prepare concentrated stock solutions for an 8-cation standard (prepare
fewer types of cations, if a simpler standard suffices):
1. Weigh out the amounts of dry sa to calculate the amount for a salt not listed.
(Mol. Wt. Salt / Mol. Wt. Cation) = g SaltIf you choose to use other salts, be sure to avoid any combinations that will form a precipitate.
Salts for cation standard concentrates
Lithium hydroxide monohydrate (Li+) Sodium chloride (Na+) Ammonium chloride (NH + Potassium chloride (K+) Magnesium nitrate hexahydrate (Mg2+) Calcium nitrate tetrahydrate (Ca2+) Strontium nitrate tetrahydrate (Sr2+) Barium chloride dihydrate (Ba2+) 2. Place each salt in a separate plastic1-L volumetric flask and dilute to the mark with reagent-grade water. Each concentrate contains 1000 ppm of the cation.
Standards for ion analysis Preparing an 8-cation standard
To prepare 1 liter of 8-cation standard:
1. Add the volume of stock (concentrate) standard listed in the table below to a plastic 1-L volumetric flask.
Cation concentrate dilutions
Final concentration (ppm)
2. Fill the flask to the mark with ASTM Type I reagent water.
Injecting cation standards
To inject the standard, obtain the following materials: 0.1 mM EDTA/ 3 mM HNO cation eluent – Refer to
1-cc disposable plastic syringe – Pharmaseal® Stylex® disposable
syringe with a Luer Slip® tip, or equivalent.
Injector or autosampler with a 100-μL loop – Ion chromatography
commonly uses a 100-μL injection volume. When you use a fixed loop, overfill a minimum of three times.
Performing Ion Analysis Injecting the standard
Tip: You can substitute the method desc
, for the following procedure.
To inject the standard.
1. Set up the 432 Detector as follows: Base Sensitivity = 2000 μS Integrator Sensitivity = 50 μS/V Recorder Sensitivity = 0.01 (strip chart) Response = STD (0.5 seconds) Temperature = 35 °C Polarity = – (negative) 2. Equilibrate the 432 Detector as described in 3. Rinse a 1-cc disposable plastic syringe two or three times with ASTM Type I reagent water, then load the standard.
4. Place the syringe tip into the sample loading port and overfill the 100-μL loop at least three times (that is, 300 μL).
5. Inject the sample.
representative chromatogram of an 8-cation standard run on an IC-Pak C M/D column with 0.1 mM EDTA/3 mM HNO eluent at 1.0 mL/min flow rate. The separation of the standard typically takes 20 to 25 minutes with Standards for ion analysis Chromatogram of an 8-cation standard
Performing Ion Analysis This chapter contains information about maintaining the 432 Detector and troubleshooting charts to help you isolate and correct problems.
Warning: To avoid the possibility of electric shock, power off the
detector and disconnect the power cord before you service the This section contains information designed to help you maintain the 432 Detector. Routine maintenance for the 432 Detector includes: Replacing the fuse Calibrating the detector Maintaining the flow cell Waters service specialists provide maintenance for the 432 Detector on a corrective, as required, basis. Contact the Waters Technical Services Department if you have questions regarding the repair or performance of your instrument.
Follow these suggestions to help you maintain the 432 Detector: Stock the recommended spare parts listed in Appendix B to reduce downtime. Contact the Waters Service Department for assistance.
Record the initial installation date and serial number of your instrument in for easy reference.
Keep a file of typical chromatograms for comparison when you suspect Replacing the fuse
To change the operating voltage fuse:
1. Turn off the front panel power switch and remove the power cord from its connector on the rear panel of the detector.
Warning: To avoid the possibility of electric shock, turn off the
front panel power switch, and unplug the power cord from the 2. Pry open the power connector cover with a screwdriver.
3. To change the AC power fuses, pull out each fuse holder as though opening a drawer. Spare fuses are included in the System Startup Kit. For ordering information, see operating voltage fuses (for use in either North America or Europe).
5. Install the correct fuse in the holder and slide it back into place. The arrow on each fuse holder points up when in the correct position.
Installing operating voltage fuses
6. Close the power connector cover. Then plug the power cord into its connector on the rear panel of the detector.
Maintaining the flow cell
Maintenance for the 432 Detector consists of ensuring the flow cell is free of foreign material. Foreign material in the flow cell may cause baseline drift, cycling, or noise.
Caution: To avoid damaging the column, remove it before you flush the
system. Do not reconnect the column until the eluent has returned to approximately pH 7.
To clean the cell:
1. Flush the system with ASTM Type I reagent water.
2. Flush the system with 20 mL of 6 N nitric acid (HNO3).
3. Flush the system again with ASTM Type I reagent water. Do not reconnect the column until the eluent has returned to about pH 7.
Routine maintenance Refer to , to order a replacement flow cell. The following tools are required to replace the flow cell: 5/16-inch open-end wrench Knife or flat-blade screwdriver Warning: To avoid electrical hazards, always unplug the power cord
before you perform any of the following replacement procedures.
To replace the flow cell:
1. Unplug the 432 Detector from the power source, and completely disconnect all electrical cables and fluid connections.
2. Remove the 432 Detector cover (four Phillips-head screws, two on each 3. Remove the two pins and pin holders that hold the cell block cover in ). Use a knife or flat-blade screwdriver to gently pry the pins and holders out.
4. Pull off the cover of the flow cell unit and remove the top layer of 5. Remove the four Phillips-head screws from the upper plate of the cell block, and remove the plate. Note the orientation of the plate: a notch is cut into the underside to clear one of the components within the cell 6. Carefully disconnect the inlet and outlet fittings from the flow cell.
7. Remove the two Phillips-head screws from the flow cell mounting 8. Unplug the flow cell cable connector from its socket in the cell block.
9. Remove the flow cell assembly.
10. Install the new flow cell by following steps 2 through 9 in reverse order. Be sure to orient the upper plate of the cell block properly before you install the four screws.
Flow cell assembly
Upper plate screws Cell mounting bracket screws Routine maintenance Cleaning the detector exterior
To clean the outside of the 432 Detector, use only a soft lint-free paper or cloth dampened with mild soap and water.
This section contains troubleshooting charts to help you isolate and correct problems with the 432 Detector.
Keep in mind that the source of apparent detector problems may lie within the chromatography or hardware of your system. The Waters Guide to Successful Operation of Your LC System contains detailed chromatographic troubleshooting tables. (Contact your nearest Waters office for information on how to get a copy.) If you cannot correct a problem, contact the Waters Technical Services Department for assistance.
When you call Waters service
To expedite your request for service, have the following information available when you call Waters regarding a 432 Detector problem: Operating pressure Base Range setting Integrator Sensitivity setting Recorder Sensitivity setting Type of injector (automatic or manual) Type of data integrator Detector does not turn on
If your detector is completely inoperative (for example, the lights do not illuminate and the display remains completely blank when the unit is turned on), the fuse may require replacement. Refer The 432 Detector performs startup diagnostics that check the internal memory (both RAM and ROM), and the associated processing circuitry.
The following factors can adversely affect the operation of the 432 Detector: Transient energy sources Be sure that the electrical supply used for the 432 Detector is properly grounded and free from any of these conditions.
The error messages displayed by the 432 Detector are listed below along with the recommended action for each: ROM/RAM error (checked during startup only) Call Waters service.
Check flow cell and associated plumbing connections.
Temperature control failure Call Waters service.
Error: Over Range Base over-range condition Set Base Range to the next setting above the background conductivity of the eluent.
Conductivity overflow (above 10,000 μS) Dilute eluent to remain within measurable range.
Press the Clear key to clear an error alarm and message. For a continuing
error condition, the error message remains after the audio alarm is cleared.
To troubleshoot a problem:
1. Visually examine the integrity of the electrical and fluid connections as you verify proper system configuration and installation.
2. If the results of previous runs are available, compare the current system operation with the system operation before you identified the problem.
For example, if your system usually runs at a certain pressure with a certain method:• Is the system pressure in the same range, or is it drastically higher (possibly caused by a blocked line) or lower (possibly caused by a Are pressure fluctuations in the same range as during normal 3. Isolate the parameter that varies from normal operation. The parameters to observe include:• Peak retention time Qualitative/quantitative chromatographic results Evaluate the parameters in the order presented above to rule out simple causes of the problem.
4. Refer to the to determine corrective actions for the problems that you identify.
Bubbles in the flow cell are evident when the noise is equal to or greater than 2 μS. Use this method to remove bubbles.
To remove bubbles:
1. Disconnect the tubing from the inlet and outlet of the 432 Detector.
2. Attach a 1-mL tuberculin syringe to a priming syringe cannula which is screwed into the inlet of the detector.
3. Flush four times with 1-mL portions of ASTM Type I reagent water.
4. Flush four times with 1-mL portions of HPLC-grade methanol.
5. Flush four times with 1-mL portions of ASTM Type I reagent water.
6. Reattach the tubing from the 432 Detector outlet to a waste receptacle (18-inch length of 0.009-inch ID stainless steel).
7. Start eluent flowing through the system at a flow rate of at least 1 8. With the eluent flowing, reattach the detector inlet tubing to the 9. Allow a few minutes for temperature reequilibration, then check the noise level. If it is not reduced, repeat steps 1 through 4, then proceed to steps 10 through 13.
10. Attach a dead-end fitting to the 432 Detector outlet.
11. Remove the priming syringe cannula and attach a dead-end fitting to the 432 Detector inlet.
12. Allow the detector to stand overnight (>12 hours) with temperature on and with methanol in the flow cell.
13. Repeat steps 5 through 9.
Pulse dampener not Check the pump; see the pump manual.
Bubbles in flow cell Remove bubbles and degas the solvent.
Troubleshooting guide (Continued)
Voltage fluctuation Spikes on baseline Clean the cell.
Check flow cell fittings and tighten. If leak continues, replace the Bubbles in flow cell Remove bubbles and degas the solvent.
Irregular noise on Temperature changes temperature, locate drafts, and insulate tubing and column, if Cell temperature set Set the cell temperature lower than ambient to a minimum of 5 °C above ambient.
Replace the column.
Excessive baseline drift Unstable temperature temperature control is Defective cell heater Call Waters service.
Temperature changes temperature, locate drafts, and insulate tubing and column, if Cell temperature set Set the cell temperature lower than ambient to a minimum of 5 °C above ambient.
Bubbles in flow cell Remove bubbles and degas the solvent.
Troubleshooting guide (Continued)
Excessive baseline drift Solvent changeover Wait until baseline stabilizes (purge autosampler a few Check flow cell fittings and tighten. If leak continues, replace the Detector cannot be Solvent changeover Wait until the baseline Bubbles in flow cell Remove bubbles and degas the solvent.
Continuous noise at Pump crossover noise high sensitivity (<1μS) high-sensitivity noise filter on the pump.
A Safety Advisories
Waters instruments display hazard symbols designed to alert you to the hidden dangers of operating and maintaining the instruments. Their corresponding user guides also include the hazard symbols, with accompanying text statements describing the hazards and telling you how to avoid them. This appendix presents all the safety symbols and statements that apply to the entire line of Waters products.
Warning symbols alert you to the risk of death, injury, or seriously adverse physiological reactions associated with an instrument's use or misuse. Heed all warnings when you install, repair, and operate Waters instruments. Waters assumes no liability for the failure of those who install, repair, or operate its instruments to comply with any safety precaution.
Task-specific hazard warnings
The following warning symbols alert you to risks that can arise when you operate or maintain an instrument or instrument component. Such risks include burn injuries, electric shocks, ultraviolet radiation exposures, and others. When the following symbols appear in a manual's narratives or procedures, their accompanying text identifies the specific risk and explains how to avoid Warning: (General risk of danger. When this symbol appears on an
instrument, consult the instrument's user documentation for important safety-related information before you use the instrument.) Warning: (Risk of burn injury from contacting hot surfaces.)
Warning: (Risk of electric shock.)
Warning: (Risk of fire.)
Warning: (Risk of sharp-point puncture injury.)
Warning: (Risk of hand crush injury.)
Warning: (Risk of exposure to ultraviolet radiation.)
Warning: (Risk of contacting corrosive substances.)
Warning: (Risk of exposure to a toxic substance.)
Warning: (Risk of personal exposure to laser radiation.)
Safety Advisories Warning: (Risk of exposure to biological agents that can pose a serious
Warning: (Risk of tipping.)
Warning: (Risk of explosion.)
Warning: (Risk of eye injury.)
The following warnings can appear in the user manuals of particular instruments and on labels affixed to them or their component parts.
This warning applies to Waters instruments fitted with nonmetallic tubing.
Warning: Pressurized nonmetallic, or polymer, tubing can burst.
Observe these precautions when working around such tubing:• Wear eye protection.
• Extinguish all nearby flames.
• Do not use tubing that is, or has been, stressed or kinked.
• Do not expose nonmetallic tubing to incompatible compounds like tetrahydrofuran (THF) and nitric or sulfuric acids.
• Be aware that some compounds, like methylene chloride and dimethyl sulfoxide, can cause nonmetallic tubing to swell, which significantly reduces the pressure at which the tubing can rupture.
Mass spectrometer flammable solvents warning
This warning applies to instruments operated with flammable solvents.
Warning: Where significant quantities of flammable solvents are
involved, a continuous flow of nitrogen into the ion source is required to prevent possible ignition in that enclosed space. Ensure that the nitrogen supply pressure never falls below 690 kPa (6.9 bar, 100 psi) during an analysis in which flammable solvents are used. Also ensure a gas-fail connection is connected to the LC system so that the LC solvent flow stops if the nitrogen supply fails.
Mass spectrometer shock hazard
This warning applies to all Waters mass spectrometers.
Warning: To avoid electric shock, do not remove the mass spectrometer's
protective panels. The components they cover are not user-serviceable.
This warning applies to certain instruments when they are in Operate mode.
Warning: High voltages can be present at certain external surfaces of
the mass spectrometer when the instrument is in Operate mode. To avoid non-lethal electric shock, make sure the instrument is in Standby mode before touching areas marked with this high voltage warning Safety Advisories This warning applies to Waters instruments that can be used to process material that might contain biohazards: substances that contain biological agents capable of producing harmful effects in humans.
Warning: Waters instruments and software can be used to analyze or
process potentially infectious human-sourced products, inactivated microorganisms, and other biological materials. To avoid infection with these agents, assume that all biological fluids are infectious, observe Good Laboratory Practices, and consult your organization's biohazard safety representative regarding their proper use and handling. Specific precautions appear in the latest edition of the US National Institutes of Health (NIH) publication, Biosafety in Microbiological and Biomedical Chemical hazard warning
This warning applies to Waters instruments that can process corrosive, toxic, flammable, or other types of hazardous material.
Warning: Waters instruments can be used to analyze or
process potentially hazardous substances. To avoid injury with any of these materials, familiarize yourself with the materials and their hazards, observe Good Laboratory Practices (GLP), and consult your organization's safety representative regarding proper use and handling. Guidelines are provided in the latest edition of the National Research Council's publication, Prudent Practices in the Laboratory: Handling and Disposal of Chemicals. The caution symbol signifies that an instrument's use or misuse can damage the instrument or compromise a sample's integrity. The following symbol and its associated statement are typical of the kind that alert you to the risk of damaging the instrument or sample.
Caution: To avoid damage, do not use abrasives or solvents to clean the
Warnings that apply to all Waters instruments
When operating this device, follow standard quality control procedures and the equipment guidelines in this section.
Attention: Changes or modifications to this unit not expressly approved by the
party responsible for compliance could void the user's authority to operate the Important: Toute modification sur cette unité n'ayant pas été expressément
approuvée par l'autorité responsable de la conformité à la réglementation peut annuler le droit de l'utilisateur à exploiter l'équipement.
Achtung: Jedwede Änderungen oder Modifikationen an dem Gerät ohne die
ausdrückliche Genehmigung der für die ordnungsgemäße Funktionstüchtigkeit verantwortlichen Personen kann zum Entzug der Bedienungsbefugnis des Systems führen.
Avvertenza: qualsiasi modifica o alterazione apportata a questa unità e non
espressamente autorizzata dai responsabili per la conformità fa decadere il diritto all'utilizzo dell'apparecchiatura da parte dell'utente.
Atencion: cualquier cambio o modificación efectuado en esta unidad que no
haya sido expresamente aprobado por la parte responsable del cumplimiento puede anular la autorización del usuario para utilizar el equipo.
주의: 규정 준수를 책임지는 당사자의 명백한 승인 없이 이 장치를 개조 또는 변경할 경우,
이 장치를 운용할 수 있는 사용자 권한의 효력을 상실할 수 있습니다.
Safety Advisories Warning: Use caution when working with any polymer tubing under pressure:
• Always wear eye protection when near pressurized polymer tubing.
• Extinguish all nearby flames.
• Do not use tubing that has been severely stressed or kinked.
• Do not use nonmetallic tubing with tetrahydrofuran (THF) or concentrated
nitric or sulfuric acids.
• Be aware that methylene chloride and dimethyl sulfoxide cause nonmetallic tubing to swell, which greatly reduces the rupture pressure of the tubing.
Attention: Manipulez les tubes en polymère sous pression avec precaution:
• Portez systématiquement des lunettes de protection lorsque vous vous
trouvez à proximité de tubes en polymère pressurisés.
• Eteignez toute flamme se trouvant à proximité de l'instrument.
• Evitez d'utiliser des tubes sévèrement déformés ou endommagés.
• Evitez d'utiliser des tubes non métalliques avec du tétrahydrofurane (THF) ou de l'acide sulfurique ou nitrique concentré.
• Sachez que le chlorure de méthylène et le diméthylesulfoxyde entraînent le gonflement des tuyaux non métalliques, ce qui réduit considérablement leur pression de rupture.
Vorsicht: Bei der Arbeit mit Polymerschläuchen unter Druck ist besondere
Vorsicht angebracht:• In der Nähe von unter Druck stehenden Polymerschläuchen stets • Alle offenen Flammen in der Nähe löschen.
• Keine Schläuche verwenden, die stark geknickt oder überbeansprucht sind.
• Nichtmetallische Schläuche nicht für Tetrahydrofuran (THF) oder konzentrierte Salpeter- oder Schwefelsäure verwenden.
• Durch Methylenchlorid und Dimethylsulfoxid können nichtmetallische Schläuche quellen; dadurch wird der Berstdruck des Schlauches erheblich Warnings that apply to all Waters instruments Attenzione: fare attenzione quando si utilizzano tubi in materiale polimerico
sotto pressione:• Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero • Spegnere tutte le fiamme vive nell'ambiente circostante.
• Non utilizzare tubi eccessivamente logorati o piegati.
• Non utilizzare tubi non metallici con tetraidrofurano (THF) o acido solforico o nitrico concentrati.
• Tenere presente che il cloruro di metilene e il dimetilsolfossido provocano rigonfiamenti nei tubi non metallici, riducendo notevolmente la pressione di rottura dei tubi stessi.
Advertencia: se recomienda precaución cuando se trabaje con tubos de
polímero sometidos a presión:• El usuario deberá protegerse siempre los ojos cuando trabaje cerca de tubos de polímero sometidos a presión.
• Si hubiera alguna llama las proximidades.
• No se debe trabajar con tubos que se hayan doblado o sometido a altas • Es necesario utilizar tubos de metal cuando se trabaje con tetrahidrofurano (THF) o ácidos nítrico o sulfúrico concentrados.
• Hay que tener en cuenta que el cloruro de metileno y el sulfóxido de dimetilo dilatan los tubos no metálicos, lo que reduce la presión de ruptura de los Safety Advisories 경고: 가압 폴리머 튜브로 작업할 경우에는 주의하십시오.
• 가압 폴리머 튜브 근처에서는 항상 보호 안경을 착용하십시오.
• 근처의 화기를 모두 끄십시오.
• 심하게 변형되거나 꼬인 튜브는 사용하지 마십시오.
• 비금속(Nonmetallic) 튜브를 테트라히드로푸란(Tetrahydrofuran: THF) 또는
농축 질산 또는 황산과 함께 사용하지 마십시오.
• 염화 메틸렌(Methylene chloride) 및 디메틸술폭시드(Dimethyl sulfoxide)는 비금속 튜브를 부풀려 튜브의 파열 압력을 크게 감소시킬 수 있으므로 유의하십시오.
Warnings that apply to all Waters instruments Warning: The user shall be made aware that if the equipment is used in a
manner not specified by the manufacturer, the protection provided by the equipment may be impaired.
Attention: L'utilisateur doit être informé que si le matériel est utilisé d'une
façon non spécifiée par le fabricant, la protection assurée par le matériel risque Vorsicht: Der Benutzer wird darauf aufmerksam gemacht, dass bei
unsachgemäßer Verwenddung des Gerätes die eingebauten Sicherheitseinrichtungen unter Umständen nicht ordnungsgemäß Attenzione: si rende noto all'utente che l'eventuale utilizzo
dell'apparecchiatura secondo modalità non previste dal produttore può compromettere la protezione offerta dall'apparecchiatura.
Advertencia: el usuario deberá saber que si el equipo se utiliza de forma
distinta a la especificada por el fabricante, las medidas de protección del equipo podrían ser insuficientes.
경고: 제조업체가 명시하지 않은 방식으로 장비를 사용할 경우 장비가 제공하는 보호 수단이
제대로 작동하지 않을 수 있다는 점을 사용자에게 반드시 인식시켜야 합니다.
A-10 Safety Advisories Warning: To protect against fire, replace fuses with those of the type
and rating printed on panels adjacent to instrument fuse covers.
Attention: pour éviter tout risque d'incendie, remplacez toujours les
fusibles par d'autres du type et de la puissance indiqués sur le panneau à proximité du couvercle de la boite à fusible de l'instrument.
Vorsicht: Zum Schutz gegen Feuer die Sicherungen nur mit
Sicherungen ersetzen, deren Typ und Nennwert auf den Tafeln neben den Sicherungsabdeckungen des Geräts gedruckt sind.
Attenzione: per garantire protezione contro gli incendi, sostituire i
fusibili con altri dello stesso tipo aventi le caratteristiche indicate sui pannelli adiacenti alla copertura fusibili dello strumento.
Advertencia: Para evitar incendios, sustituir los fusibles por aquellos
del tipo y características impresos en los paneles adyacentes a las cubiertas de los fusibles del instrumento.
경고: 화재의 위험을 막으려면 기기 퓨즈 커버에 가까운 패널에 인쇄된 것과 동일한
타입 및 정격의 제품으로 퓨즈를 교체하십시오.
Warnings that apply to all Waters instruments Electrical and handling symbols
These can appear in instrument user manuals and on the instrument's front or rear panels.
Electrical power on Electrical power off Alternating current Protective conductor terminal Frame, or chassis, terminal Recycle symbol: Do not dispose in municipal waste.
A-12 Safety Advisories These handling symbols and their associated text can appear on labels affixed to the outer packaging of Waters instrument and component shipments.
Electrical and handling symbols A-14 Safety Advisories This appendix includes information on: Less than 0.05 μS/hr/°C (ambient)Base: 200 μSSensitivity: 0.005Eluent: 1 mM KCI Less than 0.005 μS/cmBase: 200 μSSensitivity: 0.005Eluent: 1 mM KCI Temperature control Front-panel selectable: OFF, 30, 35, 40, 45, 50, 55, 60, 65 °CPerformance: 0.5 °C/hr Fast: 0.25 secStandard: 0.5 secSlow: 1.0 sec Mechanical specifications (Continued)
316 stainless steel, PTFE, and PCTFE Operating pressure 70 psi maximumHeight 9.4 inches (23.8 cm) 21 inches (53.3 cm) 11.5 inches (29.2 cm) 17.7 pounds (8 kg) Operating temperature range 4 to 35 °C Operating humidity 20 to 80% RH, noncondensing Protection classa Over-voltage categoryb Pollution degreec Moisture protectiond Line voltages (grounded AC), nominal 100/120 VAC Line frequency ranges 50 Hz: ±2 Hz60 Hz: ±2 Hz 100/120 VAC fuse rating 220/240 VAC fuse rating a. Protection Class I – The insulating scheme used in the instrument to protect
you from electrical shock. Class I identifies a single level of insulation between live parts (wires) and exposed conductive parts (metal panels), in which the exposed conductive parts are connected to a grounding system. In turn, this grounding system is connected to the third pin (ground pin) on the electrical power cord plug.
b. Over Voltage Category II – Pertains to instruments that receive their elec-
trical power from a local level such as an electrical wall outlet.
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 to normally only nonconductive pollution. Occasionally, however, a temporary conductivity caused by condensation must be expected.
d. Moisture Protection – Normal (IPXO) – IPXO means that there is NO
Ingress Protection against any type of dripping or sprayed water. The X is a placeholder to identify protection against dust, if applicable.
Integrator output 10, 50, 100 μS/1V FS Isolated contact output IEEE-488, PowerLine™ C Spare Parts
The parts listed in the table below are spare parts recommended for installation by you, the customer. Any parts that are not listed may require installation by a trained service representative. Order a supply of the parts listed below to keep in stock for use as needed.
Fuse, Time Delay, 1A, 250V, IEC Fuse, Time Delay, 2A, 5x20 mm T Pump Seal Replacement kit Tip: The flow cell (part number 043069) is considered a replacement part.
Order the flow cell only when it is needed for replacement in the Waters 432 Detector.
Fill in the information below for easy reference when you order parts or request service.
Installation Date: _
D Ion Chromatography Methods
General anion analysis using conductivity and UV
Alliance, 2695 Separations Module or Breeze (with column heater, seal wash, and degasser)432 Conductivity Detector Empower/Breeze data processing Common anion standards
IC-Pak A/HR (part number 026765) Borate/gluconate containing 12% AcCN Back conductivity Ion Chromatography Methods Analysis conditions (Continued)
30 °C for column heater, 35 °C for detector 12% AcCN in DI water Direct conductivity 50 μS/Volt unattenuated To prepare eluent:
1. Add 20 mL of lithium borate/gluconate concentrate and 120 mL of HPLC-grade acetonitrile (AcCN) into a 1-liter volumetric flask (see , for concentrate preparation).
2. Dilute to volume with DI water.
3. Ensure the natural pH is 8.2 ±0.1.
4. Vacuum degas through a 0.45-μm aqueous and solvent-compatible membrane filter.
5. Store in a glass or plastic container at ambient temperature. Discard after 1 month.
It is recommended that you use certified 1000-ppm anion standards with this uncertified standard preparation.
Prepare at least three mixed analyte standards within the expected range of sample analyte concentration. This method is linear from 0.1 to 100 ppm. General anion analysis using conductivity and UV detection After you validate the multi-point calibration curve, a single-point calibration curve within the expected analyte concentration range is appropriate for Preparing a sample
To prepare a sample:
1. Determine the expected range of analyte concentration and other anionic component in the sample matrix. The major analyte should be less than 100 ppm for best results.
2. Dilute the sample with DI water, if necessary.
3. If the sample contains high amounts of neutral organics or is highly colored, pass the diluted sample through a C18 Sep-Pak cartridge. Anions pass through unretained, but there can be a loss of fluoride 4. Filter samples containing suspended solids through a 0.45-μm aqueous-compatible disk before injection. Failure to filter solids can increase column backpressure.
Tip: For best results, ensure sample pH is between 3 and 11.
Samples treated with a sample preparation disk in the H+ form, used to remove cations and neutralize high pH, will yield chromatograms similar to suppressed conductivity chromatograms.
Ion Chromatography Methods Empower data processing method
IC processing method using peak apex for retention time
Peak Width = 30.0Minimum Area = 3000Inhibit Intg. = 0 to 2 min.
Threshold = 10 to 25Minimum Height = 150 Averaging = NoneUpdate RT = NeverPeak Match = ClosestQuant By = Peak AreaFit Type = Linear for multi-point calibration, Linear Through Zero for single-point calibration Analyte NameAnalyte Retention TimePeak AreaAmounts This validation design is abstracted from ASTM/EPA validation. It has been used to validate all anion analysis methods. Many of the methods using this General anion analysis using conductivity and UV detection validation design are linear above 50 ppm.
Individual youden pair standard, in ppm
Calibration curves for chloride, fluoride, and bromide
Ion Chromatography Methods Calibration curves for nitrite and nitrate
Calibration curves for sulfate and phosphate
General anion analysis using conductivity and UV detection Quantitation Precision is the percent RSD of analyte peak area at each concentration. Data is based on seven replicate injections of the validation Ion Chromatography Methods Method detection limits
Based on this representative chromatogram using a 100-μL injection, the estimated detection limits, as ppb, at three times signal to noise (S/N) are as Quantitation below these detection limits is not advised. You can obtain lower detection limits using a 250-μL injection.
General anion analysis using conductivity and UV detection The Certified Performance Evaluation Standards were diluted 1:100 with DI water. Amounts are based on a multi-point calibration curve prepared from Performance True Value 2.69
Official Anion Mean
Chem & IC
Std DevAve IC n=3 2.63 ±0.06 ±0.09 ±0.12 Alliance IC
0.975 1.014 1.001 Pak A/HR and
0.979 1.029 0.995 The Certified Performance Evaluation Standards were diluted 1:100 with typical drinking water. Amounts are based on a multi-point calibration.
Drinking Water detected ±0.04
n=3, as ppm
D-10 Ion Chromatography Methods Analyte recovery (Continued)
n=3; as ppm
Example of use
Typical drinking water, no dilution required
Using direct UV detection
Many anions are UV active in the range of 205 to 214 nm, such as NO2, Br, and NO3, and the use of direct UV detection provides a degree of detector selectivity. The chromatogram of the anion standard that demonstrates this selectivity. Generally, the lower the wavelength of detection, the greater is the response, as seen with the difference between 205- and 214-nm chromatograms. However, the borate/gluconate eluent has some UV absorption which causes General anion analysis using conductivity and UV detection negatives peaks at the retention time of the UV transparent anion, such as F, Cl, PO4, and SO4, if present.
Direct UV detection
Direct UV detection is five times more responsive for nitrite and nitrate than is conductivity detection and therefore provides lower detection limits. The chromatogram in of a 100-ppb anion standard demonstrates the improved sensitivity.
D-12 Ion Chromatography Methods 100-ppb anion standard
Waters 996 photodiode array detector The borate/gluconate eluent has some UV absorption. Use of eluents that are UV transparent, such as hydroxide and carbonate/bicarbonate, provide lower detection limits.
Preparing lithium borate/gluconate 50X stock concentrate
To prepare lithium borate/gluconate 50X stock concentrate:
1. Using a 60-mL plastic syringe, add approximately 25 g of BioRad AG-50W-X12 strong cation exchange resin in the hydrogen form, or equivalent. Wash the resin with five 20-mL portions of DI water to remove any ionic impurities from the resin. Discard the washings.
2. Dissolve 9.06 g of sodium gluconate in approximately 20-mL of DI water. After dissolution, transfer this solution into the 60-mL syringe with resin. Slowly pass this solution into a 1-liter volumetric flask. Wash the resin with five 20-mL portions of DI water, then add the washing to the volumetric flask. Discard the syringe and resin.
An alternative is to use commercially available 50% gluconic acid. However, it comes as a brown solution that, when diluted, gives a yellow tint to the eluent that can affect long-term performance. You can remove the brown color by passing 5 mL of 50% gluconic acid through a C18 General anion analysis using conductivity and UV detection Sep-Pak cartridge. This requires three Sep-Pak cartridges. Use 13.2 mL of 50% gluconic acid for the eluent concentrate.
3. Adjust the volume in the flask to approximately 500 mL with DI water and use a stirring bar. Add 7.2 g of lithium hydroxide monohydrate and 25.5 g of boric acid. With the aid of a magnetic stirrer, mix until all reagents are dissolved.
4. Add 94 mL of 95% glycerol and mix. Remove the stirring bar and fill to the mark with DI water.
5. Store this lithium borate/gluconate concentrate in a plastic container at ambient temperature for up to 6 months. You can store it at 4 °C for up to 1 year, but warm it to ambient temperature before use.
6. A white "string-like" material observed at the bottom of the container indicates significant microbiological growth. If present, discard and prepare again.
Preparing lithium borate/gluconate eluent
To prepare lithium borate/gluconate eluent:
1. In a 1-liter volumetric flask, add 20 mL of the 50X lithium borate/gluconate concentrate and dissolve in 500 mL of DI water. Add 120 mL of HPLC-grade acetonitrile. Mix and fill to the mark with DI 2. Vacuum degas through a 0.45-μm aqueous / organic membrane.
The background conductivity of the lithium borate/gluconate eluent is between 220 and 270 μS.
D-14 Ion Chromatography Methods Alkali and alkaline earth cations, ammonium, and amines
Alliance, 2695 Separations Module or Breeze (with column heater, seal wash, and degasser)432 Conductivity Detector Empower/Breeze data processing 3 mM HNO3/0.1 mM EDTA Back conductivity Alkali and alkaline earth cations, ammonium, and amines Analysis conditions (Continued)
30 °C for column heater, 35 °C for detector 12% AcCN in DI water Indirect conductivity 100 μS/Volt unattenuated To prepare eluent:
1. Add 0.029 g of EDTA as the free acid into a 1-liter plastic volumetric 2. Dissolve in 500 mL of DI water with a stirring bar for 30 min.
3. Add 30 mL of 100 mM HNO3 (or 189 μL of concentrated HNO3).
4. Dilute to volume with DI water.
5. Vacuum degas through a 0.45-μm aqueous-compatible membrane to remove excess EDTA crystals.
6. Store in a plastic container at ambient temperature. Discard after 1 It is recommended that you use certified 1000-ppm anion standards with this uncertified standard preparation.
Prepare at least three mixed analyte standards, using plastic volumetric flasks, within the expected range of the sample analyte concentration. This D-16 Ion Chromatography Methods method is linear from 0.05 to 20 ppm for lithium, sodium, and ammonium, and 0.05 to 50 ppm for potassium, magnesium, and calcium. Above these concentrations, the response is off scale. After the multi-point calibration curve is validated, a single-point calibration curve within the expected analyte concentration is appropriate for future calibrations.
You can use this method for the analysis of Rb, Cs, Sr, and Ba.
Preparing a sample
To prepare a sample:
1. Determine the expected range of analyte concentration and other anionic component in the sample matrix. Sodium should be less than 20-ppm for best results.
2. Dilute the sample with DI water, if necessary.
3. If the sample contains high amounts of neutral organics or is highly colored, pass the diluted sample through a C18 Sep-Pak cartridge. Cations pass through unretained. There can be residual Na contamination from the cartridge.
4. Filter samples containing suspended solids through a 0.45-μm aqueous-compatible disk before injection. Failure to filter solids can increase column backpressure.
Tip: For best results, ensure sample pH is between 2 and 7 (especially
the alkaline earth cations). Samples with pH less than 10 are appropriate for the alkali cations, ammonium, and amines.
5. For samples with pH less than 2, dilute the sample 1:10 with DI water or treat the sample with an Alltech IC-OH cartridge to remove anions and neutralize pH.
Alkali and alkaline earth cations, ammonium, and amines Empower data processing method
IC processing method using peak apex for retention time
Peak Width = 30.0Minimum Area = 3000Inhibit Intg. = 0 to 2 min.
Threshold = 25 to 40Minimum Height = 500 Averaging = NoneUpdate RT = NeverPeak Match = ClosestQuant By = Peak AreaFit Type = Linear for multi-point calibration, Linear Through Zero for single-point calibration Analyte NameAnalyte Retention TimePeak AreaAmounts D-18 Ion Chromatography Methods Method detection limits
25-ppb cation standard
Based on this representative chromatogram using a 100-μL injection in a 2695 Separations Module, the estimated detection limits, as ppb, at three times signal to noise (S/N) are as follows: You can achieve lower detection limits by using a 250-μL injection.
Alkali and alkaline earth cations, ammonium, and amines Examples of use
Typical drinking water, no dilution required
Typical municipal wastewater, diluted 1:50, overlay of duplicate injections
Tip: Alkyl and alkanol amine analysis standards are between 1 and 5 ppm, 3
mM HNO3/0.1 mM EDTA/3% AcCN.
D-20 Ion Chromatography Methods Preparing stock reagent
Because it is difficult to prepare a stock eluent for this column, it is recommended to prepare fresh working eluent.
To prepare stock reagent:
1. In a 1-liter plastic volumetric flask, add 0.029 g of EDTA (as the free acid, not its salts) in 800 mL of DI water. Place on a magnetic stir plate and stir for 10 minutes.
2. While stirring, add 189 μL of concentrated nitric acid and mix for 5 3. Remove the stirring bar and fill to the mark with DI water.
4. Filter through a 0.45-μm aqueous-compatible membrane filter before use. There can be some remaining white crystals on the filter (EDTA). This does not affect the performance of the eluent. Discard the filter.
Alkali and alkaline earth cations, ammonium, and amines D-22 Ion Chromatography Methods E Validation Support
Validation regulation overview
Federal regulatory codes require that instrumentation and automated systems used for generation, measurement, and assessment of data undergo: Operational qualification (performance verification) following repairs, maintenance, and substantial periods of operation Routine maintenance Federal regulatory codes also require that laboratories maintain: Written Standard Operating Procedures (SOPs) that indicate dates of operational qualification and maintenance Tip: Designate a person to be responsible for maintaining federally required
Waters regulatory compliance support
Waters provides a wide range of documentation and services to assist customers in complying with the following areas of Standard Operating Procedure regulatory requirements: Instrument maintenance Error messages, diagnostics, and tests Instrument performance qualification (calibration) Regulatory requirements regarding proper instruction for preparing, programming, and operating the 432 Detector are satisfied by the Waters 432 Conductivity Detector Operator's Guide.
Validation Support satisfies regulatory requirements for routine instrument maintenance. includes: Calibration adjustment Replacing the flow cell Summary of 432 Detector error messages Troubleshooting tables Additional Waters support
For more information about compliance support products and services, contact Waters Technical Service Department.
Waters regulatory compliance support Validation Support PowerLine controller audience and purpose Containers, selecting and preparing EC Authorized Representative electrical symbols anion, preparing Beep-on-keystroke cation, preparing biohazard warning general guidelines Bubbles, removing nitric acid, preparing Bus LAC/E connections Bus SAT/IN connections equipment guidelines Carbonate absorption, minimizing flammable solvents Chart recorder connections Chart recorder offset chemical hazard warning Fluid connections assembling fittings Compression screw making connections Configuration, system handling symbols Bus SAT/IN to Bus LAC/E Initialization self-test sensitivity range multiplier ISM classification safety advisories Sample preparation Sensitivity Range key Soda lime tube, using Leak Alert output cation, injecting cation, preparing mass spectrometer shock hazard Millennium32 connections Storage, long-term Nitric acid eluent, preparing Noisy baseline, troubleshooting System configuration Time constant, detector Power requirements cutting polymeric PowerLine operation purpose and audience Unpacking the 432 Detector Voltage, operating
Organización Meteorológica Comisión Oceanográfica Intergubernamental de la Grupo de cooperación sobre boyas de acopio de datos Poner freno a los actos de vandalismo perpetrados contra las boyas de acopio de datos Las boyas meteorológicas y oceanográficas, a la deriva o fondeadas, son esenciales para afinar nuestros conocimientos sobre los océanos. Miden datos como la presión atmosférica, la temperatura del aire, el viento o la temperatura del mar, no sólo en superficie sino también a diferentes niveles de profundidad (en algunos casos hasta los 500 metros) e, incluso, las boyas a la deriva miden también las corrientes marinas a partir de su trayectoria. Las medidas y las posiciones se transmiten por satélite en tiempo real a los centros meteorológicos y oceanográficos operativos.