Coulochem III Reference Manual Rev A

NOTICES: This system is covered by a limited warranty. A copy of the warranty is included with this manual. The analyst is required to perform routine maintenance as described herein on a periodic basis to keep the warranty in effect. All information in this manual is subject to change without notice and does not represent a commitment on the part of ESA, Inc. The Coulochem III and various components in the system are covered by the following patents: US: 4,233,031; RE32,920; 4,404,065; 4,497,199; 4,511,659; 4,552,013; 4,753,714; 4,804,455; 4,863,873; 4,976,994; 5,104,639. Canada: 1138043 (1982); 1139841 (1983); 1167277 (1984); 1167526 (1984); 1195383 (1985); 1238362 (1988); 1251515 (1989); 1271811 (1990). Japan: 1536120; 1827931; 2018280; 2059320; 2072937. France: 2422948; 0223532; 0227281; 0033188; 0567564. Germany: P3681691.4; P3686030.1; 3174440; 0567564. Italy: 0223532; 0227281; 0033188. EPO: 0122009. UK: 2012435B; 0223532; 0227281; 0033188; 0567564.

 2004 ESA, Inc. All rights reserved. No part of this manual may be reproduced or transmitted in any form or by any means without the written permission of ESA, Inc. ESA and Coulochem are registered trademarks of ESA, Inc. PEEK is a trademark of Victrex, plc. Kalrez, Mylar and Teflon are registered trademarks of E.I. DuPont deNemours & Co. Inc. Kel-F is a registered trademark of the 3M Company. Printed in the United States of America.

WARRANTY ESA, Inc., warrants that each of the products manufactured and sold by ESA to be free from defects in material and workmanship in normal use and service from the date of delivery to you as the original purchaser for the following periods: ESA Instruments one year; Cells, Sensors and Electrodes - 90 days. This warranty does not cover, and no warranty is provided, for parts which by their nature are normally required to be replaced periodically consistent with normal maintenance, including, without limitation, fuses, tubing, pump piston seals, injector rotors, check valves, filters, and any software included in any product. If any product covered by this warranty is returned to the original shipping point, transportation charges prepaid, within the applicable warranty period set forth above and upon examination ESA determines to ESA’s satisfaction that such product was defective in material or workmanship at the time of delivery to you, ESA will, at ESA’s option, repair or replace the product or defective part thereof or refund the original purchase price of the product. The foregoing not withstanding, ESA will not be responsible for damage to any product resulting from misuse, negligence or accident or resulting from repairs, alterations, or installation made by any person or firm not duly authorized by ESA in writing or for any damage to any cell assembly resulting from the flow being impeded. If any cell assembly is used with control modules or potentiostats other than those manufactured by ESA, this warranty shall be void. ESA shall not be liable for failure to comply with statutes relating to safety and health, including, without limitation, standards promulgated under the Occupational Safety and Health Act (OSHA) of 1970, as amended, and regulations issued pursuant thereto. If, at any time during the period ending ninety (90) days after delivery of any product to you, you report and document any error in any software provided with such product and developed by ESA or any failure of any such software substantially to conform to ESA’s software description that limits or prevents use of the software by you, ESA, at ESA’s option, will use reasonable efforts to correct any such error or failure, will replace such software or will terminate your license to use the software and refund the price of the related product. In connection with any such termination and refund, you will return the related product to ESA forthwith upon request. The warranty will apply only to those portions of the software that were developed by ESA and that incorporated all program corrections and modifications, if any, delivered to you. It will not apply to any error or failure due to machine error or to the misuse by or negligence of any person or entity other than ESA or to any software which is modified by any person or entity other than ESA. With respect to equipment sold to you but not manufactured by ESA, ESA MAKES NO WARRANTY OF ANY KIND, EXPRESS OR IMPLIED, INCLUDING, WITHOUT LIMITATION, ANY WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, but will make available to you, to the extent permitted, the warranties of the manufacturer of the relevant equipment following your timely written request.

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Under no circumstances shall ESA be liable for damage to persons or property. All replacement shall be made FOB factory and without liability to ESA for drayage, freight or labor costs. If you fail to pay when due any portion of the purchase price of any product or any other payment required from you to ESA, whether hereunder or otherwise, all warranties and remedies granted in this Section and all of ESA’s warranties and obligations to service, repair, replace, correct or otherwise remedy defects, errors or failures under any other contract between you and ESA may, at ESA’s option be terminated. THIS WARRANTY IS THE ONLY WARRANTY GIVEN BY ESA WITH RESPECT TO PRODUCTS AND THE SOFTWARE PROVIDED WITH THE PRODUCTS AND IS GIVEN IN LIEU OF ALL OTHER WARRANTIES, EXPRESS OR IMPLIED, INCLUDING, WITHOUT LIMITATION, ANY WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE. YOUR EXCLUSIVE REMEDIES AND ESA’s SOLE LIABILITY FOR ANY NONCONFORMITY OR DEFECT IN THE PRODUCTS AND SUCH SOFTWARE WILL BE THOSE EXPRESSED HEREIN. UNDER NO CIRCUMSTANCES WILL ESA’s LIABILITY ARISING FROM THE PERFORMANCE OR FAILURE TO PERFORM OF ANY PRODUCT OR SOFTWARE, IN CONTRACT, IN TORT (INCLUDING NEGLIGENCE), OR OTHERWISE, EXCEED THE PURCHASE PRICE OF THE PRODUCT AND SOFTWARE. IN NO EVENT WILL ESA BE LIABLE, IN CONTRACT, IN TORT (INCLUDING NEGLIGENCE), OR OTHERWISE FOR SPECIAL, INCIDENTAL, CONSEQUENTIAL OR ANALOGOUS DAMAGES, INCLUDING, WITHOUT LIMITATION, DAMAGES RESULTING FROM LOSS OF USE, LOSS OF PROFITS, LOSS OF BUSINESS OR LOSS OF GOODWILL, EVEN IF ESA HAS BEEN ADVISED OF THE POSSIBILITY OF SUCH DAMAGES. This warranty shall be governed by, and construed and enforced in accordance with the substantive laws of the Commonwealth of Massachusetts. This warranty shall be non-transferable and shall run to the benefit of the original purchaser only.

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WARNINGS AND SAFETY PRECAUTIONS The ESA Coulochem® III is an electrochemical detector system for high performance liquid chromatography. It can be used to determine the level of a large number of electroactive compounds in a broad variety of samples including those of biological origin. The following precautions should be followed to minimize the possibility of personal injury and/or damage to property while using the instrument. 1)

Maintain a Well-Ventilated Laboratory. The mobile phase typically contains a volatile organic solvent. Ensure that the laboratory is well ventilated so that a buildup of vaporized solvent cannot occur. The eluent should be collected in a container that minimizes the escape of waste solvent into the atmosphere.

2)

Avoid Open Flames and Sparks. The mobile phase typically contains a flammable organic solvent. Do not use an open flame in the laboratory and do not install any equipment that can cause sparks in the same room as the instrument.

3)

The Instrument must be Plugged into a Grounded Power Line. Ensure that all parts of the system are properly grounded. It is strongly recommended that all parts of the system are connected to a common ground. Do not attempt to bypass the earth ground connection. A serious shock hazard could result.

4)

Treat all Samples and Mobile Phases as if they are Capable of Containing Hazardous Substances or Transmitting Disease. The sample and/or mobile phase may contain compounds that may present a hazard to the operator of the system. Take all precautions to ensure that the mobile phase does not come into contact with the skin or eyes. A sink and eyewash should be located in the laboratory. In the event of an accident wash the affected part with copious quantities of water and seek medical assistance. If you are analyzing biological/clinical samples, treat them in accordance with the infectious disease control program of your institution.

5)

Ensure that the Unit has been Disconnected from the Line before Removing the Cover. Potentially hazardous currents and voltages may be present inside the unit.

6)

Wear Protective Eyewear. Solvents and other chemicals can damage eyes. Install a sink as close as possible to the module. If any solvents or chemicals splash on the skin or eyes, immediately rinse the affected parts in the sink. Obtain medical help as needed.

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7)

Monitor and Maintain Proper Pressure in Tubing and Devices used in the Coulochem Organizer. The maximum recommended pressure rating of tubing and devices used in the Coulochem III Organizer is 275 bar when using PEEK tubing. The maximum pressure limit used also depends on the specific device(s) used in the organizer. Refer to the device’s manufacturers for their maximum rating.

8)

Use CE Approved HPLC Pump(s) or Solvent Delivery Device(s). The pump(s) used with the Coulochem III Organizers must be CE approved and contain a properly installed and maintained pressure safety device and/or pressure sensor that conforms to the requirement of ISO 4126-1.

9)

Avoid Use of Some Solvents with PEEK Tubing. While PEEK tubing has excellent chemical resistance to most common organic solvents, it is attacked by concentrated nitric acid and sulfuric acid, and tends to swell (and weaken) in solutions with high concentrations of chloroform, dimethylsulfoxide, tetrahydrofuran and similar solvents. If these solvents must be used in high concentrations, the use of stainless steel may be appropriate.

10) Use the Instrument in a Proper Manner. Do not use the detector and its accessories in a manner not specified by ESA. Otherwise the safety protection provided by the equipment may be impaired. 11) Replace the Fuse Only with One of the Same Type and Rating. For continued protection against fire, replace the line fuse only with a fuse of the specified type and rating. 12) Do not Touch Heated Surfaces Inside the Thermal Organizer. If the Thermal Organizer temperature is elevated, do not touch heated components. 13) ESA Electrochemical Cells should not be Maintained Above 45°C. If the temperature of the Thermal Organizer is kept above 45°C (i.e., the separation is effected at an elevated temperature), make sure that the cells are placed outside the organizer as elevated cell temperatures will dramatically shorten cell lifetimes.

WARNING: Before connecting this detector to any MS Unit, the ESA HiVoltage Decoupling Union Kit (P/N 70-5974) must be properly installed (See Appendix E).

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SAFETY/OPERATING SYMBOLS The following symbols appearing on the unit or in the manual are defined as follows: This symbol on the instrument indicates that the user should refer to the operating manual before attempting to connect the power/interface cables and operate the system. This symbol on the instrument states that high voltage may be present when panels/covers are removed. Any adjustment, maintenance, and repair of the opened apparatus under voltage should be avoided as far as possible and, if inevitable, must be carried out only by a skilled person who is aware of the hazard involved. This symbol on the instrument states that elevated temperatures may be present in the system. The user should take care that the internal components are not touched when the temperature is elevated. This symbol on the back of the instrument indicates a ground terminal. The blocked WARNING statement used throughout the manual presents dangers that might result in personal injury. The blocked CAUTION statement used throughout the manual presents hazards on conditions that could cause damage to the instrument or the reporting of erroneous results. The blocked NOTE statement used throughout the manual highlights important information about the detector and its use. Failure to follow these statements may invalidate the warranty.

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IMPORTANT OPERATING CONSIDERATIONS A detailed discussion of the operation of the detector is presented in Chapters 5-7. This section describes some considerations that are especially important for optimizing performance of the system. A) Ensure that In-line Filters are used before each Electrochemical Cell in the System ESA cells with model number 501X (e.g., Model 5010A Standard Analytical Cell) and Model Number 502X (e.g., Model 5020 Guard Cell) contain porous graphite electrodes. These electrodes have a very large surface area to ensure a coulometric response. The graphite electrode can act as an efficient filter for small particles, such as silica fines from the column and particulate matter in the sample. These particles tend to clog the electrode and impede the flow of the mobile phase. To avoid clogging the electrode, an in-line filter containing a filter element MUST precede each electrochemical cell in the system. If the system uses a guard cell and an analytical cell, it will be necessary to use two in-line filters. In addition, it is strongly recommended that the aqueous portion of the mobile phase and all samples are filtered through a 0.22 micron filter. The label illustrated below is placed on ESA cells as a reminder that the use of in-line filters is essential. THE ESA, INC. WARRANTY DOES NOT COVER ELECTRODES WHICH HAVE FAILED DUE TO IMPEDED FLOW.

B) Real-Time Clock Battery Backup The clock circuitry contains a battery that is an integrated part of the system. This battery is potted and sealed inside the chip and is not replaceable. The life of the battery is specified with a minimum lifetime of 10 years. C) Use a Line Voltage Conditioning Device If the input power to the Coulochem III is noisy (i.e., fluctuations in the line voltage from the power source such as spikes, sags, dropouts, brownouts, etc.) interruptions in the use of the instrument may be observed. Interruptions usually cause the unit to “reset”; this will stop the unit, remove the potential from the cells and return the display to the initial screen or “greeting” screen.

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On rare occasions, the display may “lock up” or present nonsense information. If a Coulochem III has a locked up display or shows nonsense information: 1. Turn off the power to the unit. 2. Wait about a minute and then turn the power back on. 3. Continue to use the unit as desired. While the Coulochem III includes an in-line filter, excessive fluctuations and brownouts may create problems. If line voltage fluctuations or brownouts are a common occurrence in your laboratory or if you would simply like to ensure that power fluctuations do not lead to lock ups, loss of analytical methods or default storage methods; the use of a line voltage conditioner or power surge protector with power sag protection is recommended. These devices can usually be obtained from sources that carry computer supplies. ESA recommends that the line voltage conditioning device meets or exceeds the following specifications: a) Response time of 1 ns or less. b) Dropout voltage of about 75% or greater than the stated line voltage (e.g., in the USA the line voltage is 120 V so that a dropout voltage of 85 V or higher would be acceptable). c) Clamping level of 130 V for stated line voltages of 120 V or less and clamping level of 275 V for stated line voltages of 230 V or higher. If you have any questions concerning this, please contact the ESA Service Department or its Representative.

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CAUTIONS This guide contains important information about factors that can effect the accuracy of test results and should be reviewed before the detector is used. This is especially important if the Coulochem® III is to be used in a clinical setting as part of a system and method for in vitro diagnostic testing. Inaccurate results might occur if the Coulochem III detector is not used according to the instructions presented in the Coulochem III (50W) User’s Guide and Reference Manual. The following are particularly important: • • 1

Environment of the Coulochem III detector Analysis of the resulting chromatograms Operating Environment

Erroneous results might occur as the result of the following environmental conditions: • • • • • •

Power Line Fluctuations (surge or sag) Improper Line Voltage Strong Radio Frequency Interference (RFI) Strong Electrostatic Discharge (ESD) Large Temperature Fluctuations Extreme Temperature

The use of an Uninterruptible Power Supply (UPS) system is recommended to prevent the potential deleterious effects of power line fluctuations, interruptions or improper line voltage. This is especially important if the AC power to your facility is unstable and/or susceptible to brown-outs, spikes, surges, sags, dropouts and other interruptions. If a UPS is used, ESA recommends that it meets or exceeds the following specifications: •

Transfer time of 10 milliseconds or less.

•

Dropout voltage of about 75% or greater than the stated line voltage (e.g., in the USA the line voltage is 120 V, so that a dropout voltage of 85 V or higher would be acceptable).

•

Clamping level of 130 V for stated line voltages of 120 V or less and clamping level of 275 V for stated line voltages of 230 V or higher.

•

Capacity of 500 VA or greater (this should allow the detector to run for several hours in the event of a total power failure).

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NOTE: A larger capacity for the UPS device is needed if the entire HPLC system is to be covered by this UPS device. In this situation, the total power consumption of your entire HPLC system must be determined and the desired period of time that the system should operate after the power interruption must be determined to calculate the proper capacity of the UPS system. At the very least, a passive line conditioner (such as a surge protector, line filter, etc.) is recommended for the detector. In addition, we strongly recommend that the Coulochem detector not share a circuit with other electrical equipment that draws significant power from the AC line. This equipment includes items such as fume hoods, refrigerators, ovens, centrifuges, refrigerated cooling systems, vacuum pumps, etc. During installation of the detector or if the detector is moved, make sure that the voltage option of the power input receptacle on the back of the detector is set for the same voltage of the AC circuit to which it will be connected. The likely outcome of various power fluctuations is the introduction of a disturbance or artifact in the chromatogram. These should be readily discernable when the operator reviews the chromatogram and can be dealt with as applicable. Strong RFI may also cause disturbances or artifacts in the chromatogram or with the operation of the detector. The ESA Coulochem III has been designed to meet various certification agencies stringent requirements for susceptibility to RFI. However, it is still possible that a strong emitter of RFI could result in interference with the Coulochem III detector. Therefore, the detector should not be placed close to other instruments or machinery that could emit excessive RFI or magnetic fields such as refrigerators, fume hoods, radio transmitter antennae, NMR instruments, etc. In addition, the Coulochem III has been designed to be immune to electrostatic discharge (ESD). However, strong electrostatic discharges to the instrument could cause a disturbance or artifact to the baseline or could interfere with the operation of the detector. ESA strongly recommends that the detector is properly grounded to a good earth ground. This should result in most ESD events being harmlessly conducted to ground and thus avoiding changes or damage to the sensitive electronics. In addition, we recommend that the operator take steps to reduce the generation of static electricity in the vicinity of the detector. Measures to reduce the chance or severity of ESD include: • • • • •

x

Increasing the relative humidity (especially during the winter) Ensuring that the operator touches a good earth ground (e.g., a metal water pipe) just prior to touching the detector Installing antistatic mats in front of the detector Avoiding the use of carpeting near the detector Low or antistatic lab coats

Coulochem III (50W) Reference Manual

Large temperature fluctuations can potentially lead to baseline disturbances or other interference with the operation of the Coulochem III detector. Similarly, operating the detector at temperatures outside of its intended operating temperature range (10°-35°C or 50°-95°F) - might lead to artifacts or other instrumental interference. Therefore, the temperature of the laboratory in which the Coulochem III is used should be controlled. The laboratory temperature should be within the specified operating temperature range and temperature fluctuations should be kept at a minimum. Avoid placing the instrument in drafts (such as near an air conditioning or heating duct, the heat output of a refrigerator or an open window) and in direct sunlight. On rare occasions, the display and/or the detector may “lock up” or present nonsense if one of the above-described situations occurs. If the Coulochem III has a locked up display or shows nonsense information: 1. 2. 3. 4.

Turn off the power to the unit. Wait about a minute. Turn the power back on. Continue to use the detector as desired.

2

Analysis of Chromatograms

In addition to the environmental considerations described above, a variety of operator actions may lead to an analytical error. Typical examples of this include incorrect preparation of the mobile phase, standards or sample; injection of an incorrect sample volume, incorrect instrumental parameters (e.g., the flow rate or the detector potential) or misidentification of the sample. Most of the operator errors and the errors due to environmental situations can be detected by careful review of each and every chromatogram before the results are reported. The operator should examine each chromatogram for baseline disturbances and artifacts. In addition, the operator should examine each chromatogram of the samples for proper retention time of important peaks, for interfering peaks, expected peak shape and for chromatogram integrity. All chromatograms of controls and standards should be similarly scrutinized. If a questionable result is obtained, the sample should be rerun. It is also advisable in questionable cases to rerun controls (and standards if necessary). If the controls and standards do not provide chromatograms that match previously collected chromatograms for these samples, then the entire HPLC system should be examined for problems. For troubleshooting information, see Chapter 8 of this Manual.

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Table of Contents Warranty ...i Warnings and Safety Precautions ...iii Safety/Operating Symbols... v Important Operating Considerations ...vii Cautions ...ix 1

Introduction 1.1 Overview...1-1 1.2 Contents of the Coulochem III (50W) Reference Manual...1-1 1.3 Contents of the Coulochem III (50W) User’s Guide Manual...1-3

2

Setting up the Detector in the Laboratory 2.1 Introduction...2-1 2.2 Unpacking the Detector ...2-1 2.3 Power Requirements ...2-2 2.4 Locating the Detector in the Laboratory...2-3 2.5 Electrical Connections to Other Devices ...2-4 2.5.1 Description of the Rear Panel of the Control Module...2-4 2.5.2 Connections for Testing the Detector Module ...2-10 2.5.2.1 Detectors with a DC Potentiostat Module ...2-10 2.5.2.2 Detectors with a Pulse/Scan Module ...2-10 2.6 Test Protocol ...2-11 2.6.1 Establishing a Test Protocol ...2-11 2.6.2 Powering Up the Detector and Preliminary Operations...2-12 2.6.3 The DC Mode Test Protocol ...2-13 2.6.3.1 Establishing the DC Mode Test Protocol ...2-13 2.6.3.2 Running the DC Mode Test Protocol ...2-15 2.6.4 The Pulse/Scan Mode Test Protocol ...2-17 2.6.4.1 Establishing the Pulse Mode Test Protocol ...2-17 2.6.4.2 Running the Pulse Mode Test Protocol ...2-19 2.7 ESA Coulochem III Worksheets...2-20 2.7.1 DC Mode ...2-21 2.7.2 Pulse Mode ...2-22

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Table of Contents 3

Installing the Detector in a HPLC System 3.1 Introduction...3-1 3.1.1 General Approach to Installing the Detector into an HPLC System..3-2 3.2 Preliminary Activities ...3-3 3.2.1 Passivation...3-3 3.2.2 Cutting Tubing and Making Fittings ...3-4 3.3 Configuring the HPLC System with an Electrochemical Detector ...3-6 3.3.1 Installing the Solvent Delivery System...3-6 3.3.2 Installing the Injector, Pre-Column, Analytical Column and Post Column In-line Filter...3-8 3.3.3 Attaching Electrochemical Cells to the System ...3-9 3.4 Preparing the System for Operation...3-10 3.4.1 Equilibrating the System ...3-10 3.4.2 Preparing the Analytical Column ...3-10 3.4.3 Equilibrating the Cells...3-10

4

Theory of Operation 4.1 Basics of Electrochemistry ...4-1 4.2 What Potential is Needed for an Electrochemical Reaction ...4-2 4.2.1 Current/Voltage Curves or Hydrodynamic Voltammograms ...4-2 4.2.2 The Nernst Equation...4-4 4.3 The Role of Diffusion in Electrochemical Processes ...4-5 4.4 The Reversibility of Electrochemical Reactions ...4-6 4.5 Coulometric vs. Amperometric Detection ...4-9 4.6 Selectivity and Electrochemical Detection ...4-10 4.7 Faraday's Law ...4-12 4.8 Quantitation using an Electrochemical Detector in the Coulometric Mode ...4-13 4.9 What Types of Compounds are Electroactive ...4-16

5

The Coulometric Cell in Electrochemical Detection 5.1 Overview...5-1 5.2 Design of the Coulometric Cell ...5-1 5.3 Coulometric Detection and Selectivity ...5-3 5.4 Operational Benefits of the Coulometric Detector ...5-3 5.5 Installation, Use and Maintenance of ESA Coulometric Cells...5-6

6

The Amperometric Cell in Electrochemical Detection 6.1 Overview...6-1 6.2 Design of the Amperometric Cell ...6-1 6.3 Installation, Use and Maintenance of Amperometric Cells...6-2 6.4 Cleaning the Target Electrode of the Model 5040/5041 Cell...6-3

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Table of Contents 7

Maintenance Activities and Replacing Components 7.1 Introduction...7-1 7.2 Maintenance...7-2 7.2.1 Overview ...7-2 7.2.2 Daily Maintenance ...7-2 7.2.3 Weekly Maintenance...7-3 7.2.4 Monthly Maintenance...7-3 7.2.5 Quarterly Maintenance ...7-4 7.3 Cell Maintenance ...7-4 7.4 Replacing System Components ...7-5 7.4.1 Filter Elements ...7-5 7.4.2 Battery Backed Real Time Clock...7-7 7.4.3 Changing the Line Fuse...7-7 7.5 Cleaning ...7-8 7.5.1 Cleaning the Coulochem III Detector Unit ...7-8 7.5.2 Cleaning the Coulochem Organizer and Thermal Organizer...7-8 7.6 Establishing a System Log...7-9

8

Troubleshooting 8.1 Introduction...8-1 8.2 Internal Electronic Diagnostic Programs...8-1 8.3 Troubleshooting Guidelines...8-2 8.3.1 Overview ...8-2 8.3.2 Erratic/Noisy Baseline...8-3 8.3.3 High Background Currents ...8-5 8.3.4 Increase in Back Pressure...8-7 8.3.5 Loss of Response ...8-8 8.3.6 Inability to Autozero the Signal ...8-9 8.3.7 Guard Cell Parameters and Screen are not Displayed...8-9 8.3.8 Detector Test Procedure ...8-9 8.3.9 Thermal Organizer Problems ...8-10

9

Programming the Detector for DC Timeline Operation 9.1 The Role of Timeline Programming ...9-1 9.2 Establishing DC+Timeline Conditions...9-2 9.3 Performing a DC+Timeline Operation ...9-7 9.4 Using Timeline to Generate Current Voltage Curves or Hydrodynamic Voltammograms...9-10 9.5 Using Timeline to Change Current Ranges Reproducibly During Each Analytical Chromatographic Run………………...………………………..9-16

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Table of Contents 10

Screen Mode Operation 10.1 Rationale for Screen Mode Operation ...10-1 10.2 Using Screen Mode ...10-1

11

Redox Mode Operation 11.1 Overview of Redox Mode Operation...11-1 11.2 Benefits of Redox Mode Operation ...11-3 11.3 Using Redox Mode ...11-4

12

Pulse Mode Operation 12.1 Overview of Pulse Amperometry ...12-1 12.2 Guidelines for Selecting Parameters in Pulse Mode...12-4 12.3 Generating a Pulse Method...12-5 12.3.1 Pulse Mode Parameters...12-5 12.3.2 Establishing/Editing a Pulse Mode Method...12-6 12.4 Initiating and Performing a Pulse Mode Measurement ...12-8 12.5 Pulse + TimeLine Programming...12-10 12.5.1 Establishing Pulse + Timeline Conditions...12-10 12.5.2 Performing a Pulse + Timeline Operation ...12-15 12.5.3 Establishing a Loop ...12-16 12.6 Using Timeline to Generate Current Voltage Curves or Hydrodynamic Voltammograms for Pulse Mode ...12-17 12.7 Using Timeline to change Charge Ranges Reproducibly during each Analytical Chromatographic Run ...12-24

13

Scan Mode Operation 13.1 Overview of Scan Mode Operation ...13-1 13.2 Cyclic Voltammetry...13-1 13.3 Hydrodynamic Voltammetry ...13-5 13.4 Generating a Scan Method...13-6 13.5 Initiating and Performing a Scan ...13-8

14

Coulochem III Organizer Modules 14.1 Overview...14-1 14.2 Unpacking the Organizer ...14-1 14.3 Installing the Coulochem III Organizer Module...14-3 14.3.1 General Information...14-3 14.3.2 Removing the Organizer Chassis Plate...14-5 14.3.3 Installing the Pulse Damper ...14-5 14.3.4 Installing a Rheodyne Manual Sample Injector Valve ...14-6 14.3.5 Installing the In-line Filter ...14-7 14.3.6 Installing the Column Holder ...14-7 14.3.7 Electrochemical Cells ...14-8 14.3.8 Manual Sample Injector Overflow Line Bracket...14-9 14.3.9 Final Assembly ...14-9

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Table of Contents 14.4 Installing the Coulochem III Thermal Organizer Module ...14-10 14.4.1 General Information...14-10 14.4.2 Removing the Organizer Chassis Plate...14-12 14.4.3 Installing the Pulse Damper ...14-13 14.4.4 Installing a Rheodyne Manual Sample Injector Valve ...14-13 14.4.5 Installing the In-line Filter ...14-14 14.4.6 Installing the Temperature Probe...14-14 14.4.7 Installing the Column Holder ...14-15 14.4.8 Electrochemical Cells ...14-16 14.4.9 Manual Sample Injector Overflow Line Bracket...14-17 14.4.10 Final Assembly ...14-17 14.5 Troubleshooting ...14-19 14.6 Maintenance and Cleaning...14-19 14.6.1 Maintenance on a Daily Basis ...14-20 14.6.2 Maintenance on a Monthly Basis...14-20 14.6.3 Cleaning ...14-20 Appendix A Coulochem III Electrochemical Detector Specifications ...A-1 Appendix B Recommended Supplies and Spare Parts... B-1 Appendix C Maintaining Cell Performance for Coulometric Sensors ...C-1 Appendix D Material Safety Data Sheets (MSDS) ...D-1 Appendix E Mass Spectrometer Decoupling Instructions ... E-1 Glossary ...G-1 Index... I-1

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