Waters 432 Operator's Manual

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Waters 432 Conductivity

Detector

Operator’s Guide

34 Maple Stree

Milford, MA 01757

71500043202, Revision A

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NOTICE

The information in this document is subject to change without notice and should not be construed as a commitment by Waters Corporation. Waters Corporation assumes no responsibility for any errors that may appear in this document. This document is believed to be complete and a c cu rate at th e time o f p u blication. In no event shall Water s Corporation be liable for incidental or consequential damages in connection with or arising from the use of this document.

1994–2003 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF AMERICA. ALL RIGHTS RESERVED. THIS DOCUMEBNT OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER.

Alliance, Millennium, and Waters are registered trademarks, and Empower, LAC/E, PowerLine, SAT/IN, Sep-Pak, UltraWISP, and WISP are trademarks of Waters Corporation.

All other trademarks or registered trademarks are the sole property of their respective owners.

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When you use the instrument, follow generally accepted procedures for quality

Note:

control and methods development. If you observe a change in the retention of a particular compound, in the resolution

between two compounds, or in peak shape, immediately determine the reason for the changes. Until you determine the cause of a change, do not rely on the separation results.

The Installation Category (Overvoltage Category) for this instrument is Level II. The

Note:

Level II Category pertains to equipment that receives its electrical power from a local level, such as an electrical wall outlet.

STO

Atención:

responsible for compliance could void the user’s authority to operate the equipment.

Importan t :

par l’autorité responsable de la conformité à la réglementation peut annuler le droit de l’utilisateur à exploiter l’équipement.

Achtun g :

ausdrückliche Genehmigung der für die ordnungsgemäße Fun ktion stüchtigkeit verantwortlichen Personen kann zum Entzug der Bedienungsbefugnis des Systems führen.

Avvertenza:

espressamente approvate da un ente responsabile per la conformità annulleranno l’autorità dell’utente ad operare l’apparecchiatura.

Atención:

expresamente aprobado por la parte responsable del cumplimiento puede anular la autorización del usuario para utilizar el equip o .

Changes or modifications to this unit not expressly approved by the party

Toute modification sur cette unité n’ayant pas été expressément approuvée

Jedwede Änderungen oder Modifikationen an dem Gerät ohne die

eventuali modifiche o alterazioni apportate a questa unità e non

cualquier cambio o modificación efectuado en esta unidad que no haya sido

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Caution:

Use caution when working with any polymer tubing under pressure:

• Always wear ey e protection when near pressurized polymer tubing.

• Extinguish all nearby flames.

• Do not use Tefzel tubing that has been severely stressed or kinked.

• Do not use Tefzel tubing with tetrahydrofuran (THF) or concentrated nitric or sulfuric ac id s.

• Be aware that methylene chlor ide and dimethyl sulfoxide cause Tefzel tubing to swell, which greatly reduces the rupture pressure of the tubing.

Attention :

Soyez très prudent en travaillant avec des tuyaux de polymères sous

pression :

• Portez toujours des lunettes de protection quand vous vous trouvez à proximité de tuyaux de polymères.

• Eteignez toutes les flammes se trouvant à proximité.

• N'utilisez pas de tuyau de Tefzel fortement abîmé ou déformé.

• N'utilisez pas de tuyau de Tefzel avec de l'acide sulfurique ou nitrique, ou du tétrahydrofurane (THF).

• Sachez que le chlorure de méthylène et le sulfoxyde de diméthyle peuvent provoquer le gonflement des tuyaux de Tefzel, diminuant ainsi for tem ent 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 Schutzbrille tragen.

• Alle offenen Flammen in der Nähe löschen.

• Keine Tefzel-Schläuche verwenden, die stark geknickt oder überbeansprucht sind.

• Tefzel-Schläuche nicht für Tetrahydrofuran (THF) oder konzentrierte Salpeter- oder Schwefelsäure verwenden.

• Durch Methylenchlorid und Dimethylsulfoxid können Tefzel-Schläuche quel len; dadurch wird der Berstdruck des Schlauches erheblich reduziert.

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Precauzio ne :

prestare attenzione durante le operazioni con i tubi di polimero sotto

pressione:

• Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero pressurizzati.

• Estinguere ogni fonte di ignizione circostante.

• Non utilizzare tubi Tefzel soggetti a sollecitazioni eccessive o incurvati.

• Non utilizzare tubi Tefzel contenenti tetraidrofurano (THF) o acido solforico o nitrico concentrato.

• Tenere presente che il cloruro di metilene e il dimetilsolfossido provocano rigonfiamento nei tubi Tefzel, che riducono not evolmente il limite di pressione di rottura dei tubi stessi.

Advertencia:

manipular con precaución los tubos de polímero bajo presión:

• Protegerse siempre los ojos en las proximidades de tubos de polímero bajo presión.

• Apagar todas las llamas que estén a proximidad.

• No utilizar tubos Tefzel que hayan sufrido tensiones extremas o hayan sido doblados.

• No utilizar tubos Tefzel con tetrahidrofurano (THF) o ácidos nítrico o sulfúrico concentrados.

• No olvidar que el cloruro de metileno y el óxido de azufre dimetilo dilatan los tubos Tefzel, lo que reduce en gran medida la presión de ruptura de los tubos.

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Caution:

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 inform é que si le mat ériel est utilisé d’une façon non

spécifiée par le fabricant, la protection assurée par le matériel risque d’être défectueuses.

Vorsicht:

Der Benutzer wird darauf aufmerksam gemacht, dass bei unsachgemäßer

Verwenddung des Gerätes unter Umständen nicht ordnungs gem äß funktionieren.

Precau zion e :

l’utente deve essere al corrente del fatto che, se l’apparecchiatura viene usta in un modo specificato dal produttore, la protezione fornita dall’apparecchiatura potrà essere invalidata.

Adver tenc ia:

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.

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Caution:

rating.

To protect against fire hazard, replace fuses with those of the same type and

Attention :

Remplacez toujours les fusibles par d’autres du même type et de la même

puissance afin d’éviter tout risque d’incendie.

Vorsicht:

Zum Schutz gegen Feuergefahr die Sicherungen nur mit Sicherungen des

gleichen Typs und Nennwertes ersetzen.

Precauzio ne :

per una buona protezione contro i rischi di incendio, sostituire i fusibili con

altri dello stesso tipo e amperaggio.

Advertencia:

sustituy a l o s fusibles por o tros del mismo tipo y características para evitar el riesgo de incendio.

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Caution:

To avoid possible electrical shock, disconnect the power cord before servicing

the instrument.

Attent i on :

Afin d’éviter toute possibilité de commotion électrique, débranchez le cordon

d’alimentation de la prise avant d’effectuer la maintenance de l’instrument.

Vorsicht:

Zur V ermeidung von Stromschlägen sollte das Gerät vor der Wartung vom

Netz getrennt werden.

Precauzio ne :

per evitare il rischio di scossa elettrica, scollegare il cavo di alimentazione

prima di svolgere la manutenzione dello strumento.

Precaució n:

para evitar descargas eléctricas, desenchufe el cable de alimentación del

instrumento antes de realizar cualquier reparación.

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Commonly Used Symbols

Direct current Courant continu Gleichstrom Corrente continua Corriente continua

Alternating current Courant alternatif Wechselstrom Corrente alternata Corriente alterna

Protective conductor terminal Borne du conducteur de protection Schutzleiteranschluss Ter m ina le di conduttore con protezione Borne del conductor de tierra

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Commonly Used Symbols (Continued )

Frame or chassis terminal Borne du cadre ou du châssis Rahmen- oder Chassisanschluss Ter m ina le di str uttura o telaio Borne de la estructura o del chasis

Caution or refer to manual Attention ou reportez-vous au guide Vorsicht, oder lesen Sie das Handbuch Prestare attenzione o fare riferimento alla guida Actúe con precaución o consulte la guía

Caution, hot surface or high temperature Attention, surface chaude ou température élevée Vorsicht, heiße Oberfläche oder hohe Temperatur Precauzione, superficie calda o elevata temperatura Precaución, superficie caliente o temperatura elevada

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Commonly Used Symbols (Continued )

Caution, risk of electric shock (high voltage) Attention, risque de commot ion électri que (haut e tension ) Vorsicht, Elektroschockgefahr (Hochspannung) Precauzione, rischio di scossa elettrica (alta tensione) Precaución, peligro de descarga eléctrica (alta tensión)

Caution, risk of needle-stick puncture Attention, risques de perforation de la taille d’une aiguille Vorsicht, Gefahr einer Spritzenpunktierung Precauzione, rischio di puntura con ago Precaución, riesgo de punción con aguja

Caution, ultraviolet light

Attention, rayonnement ultrviolet Vorsicht, Ultr a violettes Licht Precauzione, luce ultravioletta Precaución, emisiones de luz ultravioleta

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Commonly Used Symbols (Continued )

Fuse Fusible Sicherung Fusibile Fusible

Electrical power on Sous tension Netzschalter ein Alimentazione elettrica attivata Alimentación eléctrica conectada

Electrical power off Hors tension Netzschalter aus Alimentazione elettrica disat tivata Alimentación eléctrica desconectada

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432 Conductivity Detector Information

Intended Use

The Waters® 432 Conductivity Detector can be used for in-vitro diagnostic testing to analyze many compounds, including diagnostic indicators and therapeutically monitored compounds. When you de velop methods, follow the “Protocol for the Adoption of Analytical Methods in the Clinical Chemistry Laborator y,” American Journal of Medical Tech nol ogy, 44, 1, pages 30–37 (1978). This protocol covers good operating procedures and techniques necessary to validate system and method performance.

Biological Hazard

When you analyze physiological fluids, take all necessary precaut ions and treat all specimens as potentially infectious. Precautions are outlined in “CDC Guidelines on Specimen Handling,” CDC – NIH Manual, 1984.

Calibration

Follow acceptable methods of calibration with pure standards to calibrate methods. Use a minimum of five standards to generate a standard curve. The concentration range should cover the entire range of quality-control samples, typical specimens, and atypical specimens.

Quality Control

Routinely run three quality-control samples. Quality-control samples should represent subnormal, nor m al, and above-norm al levels of a compound. Ensure that quality-control sample results are within an acceptable range, and ev aluate precision from day to day and run to run. Data collected when quality-control samples are out of range may not be valid. Do not report this data until you ensure that chromatographic system performance is acceptable.

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Preface ....................................................................................... 23

Chapter 1

Introduction .................. ............................. ............................. .......... 26

Chapter 2

Installing the Detector ...................................................................... 30

2.1 Selecting the Installation Site................................................ 30

2.2 Unpacking and Inspection..................................................... 31

2.3 AC Power Connection ........................................................... 32

2.4 I/O Signal Connections......................................................... 35

2.4.1 I/O Signal Desc ri pt ions ....... ..... .... ..... ................... ..... . 35

2.4.2 Powe rLine Controller Connections ............................ 37

2.4.3 Empower and Millennium

Connections .................. 39

2.4.4 Data Module Connections ......................................... 44

2.4.5 Chart Recorder Connections..................................... 45

2.4.6 Chart Marker Input Connections................................ 45

2.4.7 Auto Zero Input Connections..................................... 46

2.4.8 Alliance Separatio ns Mo dule Connec tion s ............... . 46

2.5 Making Fluidic Connections.................................................. 48

2.6 Passivating the System......................................................... 53

2.7 Verifying the Detector............................................................ 54

Table of Contents 15

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Chapter 3

Operating the Detector .................................................................... 56

3.1 Controls and Indicators......................................................... 56

3.2 Startup and Shutdown .......................................................... 61

3.3 Operating Recommendations ............................................... 62

Chapter 4

Performing Ion Analysis ................................................................... 64

4.1 Fundamental Considerations................................................ 64

4.2 Configuring the System......................................................... 68

4.3 Eluents for Ion Analysis......................................................... 69

4.3.1 Preparing Anion Eluent....... ................... ..... ..... .... ...... 70

4.3.2 Preparing Cation Eluent .......... .... ..... ..... ..... ..... .......... 70

4.4 Standards for Ion Analysis .................................................... 70

4.4.1 Preparing Anion Standa rds ......... ..... ................... ..... . 71

4.4.2 Injecting Anion Standards.......................................... 72

4.4.3 Preparing Cation Standard s ............. ..... ................... . 74

4.4.4 Injecting Cati on Stan dar ds........... ..... ..... ..... ..... .......... 76

Chapter 5

Maintenance .................................................................................... 78

5.1 Routine Maintenance............................................................ 78

5.1.1 Replacing the Fuse.................................................... 78

5.1.2 Maintaining the Flow Cell........................................... 79

5.2 Cleaning the Detector Exterior.............................................. 82

5.3 Troubleshooting..................................................................... 82

Table of Contents 16

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Appendix A

Specifications ................................................................................... 87

Appendix B

Spare Parts......... ..... ..... ..... ..... .... ..... ................... ..... ..... ..... ..... .......... 90

Appendix C

Ion Chromatography Methods .......................................................... 91

C.1 General-Purpose Anion Analysis Using Conductivity

and UV Detection ................................................................ 91

C.1.1 Prepa rin g Eluent ...... ..... ..... ..... .... .................... .... ..... . 93

C.1.2 Prepa rin g Stand ard s .......... ..... ................... ..... .... ..... . 93

C.1.3 Preparing a Sample .................................................. 93

C.1.4 Empower Data Processing Method........................... 94

C.1.5 Method Valid atio n .............. ..... .... ..... ..... ..... ............... 95

C.1.6 Method Linea rity.. ..... ..... ..... ................... ..... ..... .... ..... . 95

C.1.7 Quantita tion Prec ision... ..... ..... .... ..... ..... ..... ............... 97

C.1.8 Method Detection Limits............................................ 97

C.1.9 Quantita tion Accu racy... ..... ..... .... ..... ................... ..... . 98

C.1.10 Analyte Recovery.................................................... 99

C.1.11 Example of Use..................................................... 100

C.1.12 Using Direct UV Detection .................................... 100

C.1.13 Preparing Lithium Borate/Gluconate 50X

Stock Concentrate................................................... 102

C.1.14 Preparing Lithium Borate/Gluconate Eluent.......... 102

Table of Contents 17

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C.2 Alkali and Alkaline Earth Cations, Ammonium,

and Amines........................................................................ 103

C.2.1 Prepa rin g Elu ent .................... .... ..... ................... .... 104

C.2.2 Prepa rin g Stand ard s .......... ..... ................... ..... .... .... 104

C.2.3 Preparing a Sample ................................................ 105

C.2.4 Empower Data Processing Method......................... 105

C.2.5 Method Detection Limits.......................................... 106

C.2.6 Examples of Use..................................................... 107

C.2.7 Preparing Stock Reagent........................................ 108

Appendix D

Validation Support .......................................................................... 109

Index ..................................................................................... 111

Table of Contents 18

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List of Figures

1-1 Waters 432 Conductivity Detector.................................................26

1-2 Flow Cell Schematic ......................................................................28

2-1 Rear Panel..................................................................................... 33

2-2 Changing the Voltage Setting........................................................34

2-3 I/O Terminal Strip...........................................................................36

2-4 IEEE-488 Address Switch..............................................................38

2-5 Bus SAT/IN Module (Front Panel).................................................. 40

2-6 Bus SAT/IN to Bus LAC/E Connections.........................................42

2-7 Bus SAT/IN to 432 Detector Connections...................................... 43

2-8 Alliance Separations Module Connections to the 432

Detector Auto-Zero on Inject..........................................................47

2-9 Alliance Separations Module Connections to the 432

Detector Chart Mark on Inject........................................................48

2-10 Fluid Connections.......................................................................... 49

2-11 Cutting Po lymeric Tubing...............................................................50

2-12 Ferrule and Compression Screw Assembly...................................51

2-13 Pulse Dampener............................................................................ 53

3-1 Front Panel ....................................................................................56

4-1 Soda Lime Tube.............................................................................67

4-2 System Configuration for Ion Analysis...........................................68

4-3 Chromatogram of a 7-Anion Standard........................................... 74

4-4 Chromatogram of an 8-Cation Standard........................................77

5-1 Installing Operating Voltage Fuses................................................79

5-2 Flow Cell Assembly....................................................................... 81

C-1 Common Anion Standards ............................................................92

List of Figures 19

Page 20

C-2 Calibration Curves for Chloride, Fluoride, and Bromide ............... 95

C-3 Calibration Curves for Nitrite and Nitrate ...................................... 96

C-4 Calibration Curves for Sulfate and Phosphate ..............................96

C-5 100-mL Injection ...........................................................................97

C-6 Typical Drinking Water, No Dilution Required ............................100

C-7 Direct UV Detection ....................................................................101

C-8 100-ppb Anion Standard .............................................................101

C-9 1-ppm Standard .......................................................................... 103

C-10 25-ppb Cation Standard ..............................................................106

C-11 Typical Drinking Water, No Dilution Required ............................107

C-12 Typical Municipal Wastewater, Diluted 1:50, Overlay

of Duplicate Injections.................................................................. 107

List of Figures 20

Page 21

List of Tables

1-1 Limiting Equivalent Conductance of Ions in Water at 25 °C .........29

2-1 Power Cord Wire Identification .....................................................33

2-2 Nominal Operating Voltage...................................................... 34

2-3 I/O Signal Descriptions ........................................................... 36

2-4 IEEE-488 DIP Switch Setting.................................................. 38

2-5 Bus SAT/IN Cable Connections............................................... 44

2-6 Data Module Signal Cable Connections .................................. 44

2-7 Data Module Chart Mark Cable Connections........................... 45

2-8 Chart Recorder Cable Connections......................................... 45

2-9 Autosampler Chart Mark Cable Connections........................... 46

2-10 Autosampler Auto Zero Cable Connections............................. 46

2-11 Connections for Generating Auto-Zero on Inject...................... 47

2-12 Connections for Generating Chart Mark on Inject.................... 48

3-1 Key Descriptions ...........................................................................58

3-2 Setting the Beep Function....................................................... 60

4-1 Shelf-Life of Standards .................................................................71

4-2 Salts for Anion Standard Concentrates.................................... 72

4-3 Anion Concentrate Dilutions.................................................... 72

4-4 Salts for Cation Standard Concentrates .................................. 75

4-5 Cation Concentrate Dilutions ................................................... 75

5-1 Troubleshooting Guide ..................................................................85

A-1 Operational Specifications ............................................................87

A-2 Mechanical Specifications............................................................ 87

A-4 Electrical Specifications................................................................ 88

A-3 Environmental Specifications........................................................ 88

List of Tables 21

Page 22

A-5 Communications........................................................................... 89

B-1 Spare Parts ...................................................................................90

C-1 Required Instru ment ation ...................................................... .......91

C-1 Analysis Conditions ......................................................................92

C-2 IC Processing Method Using Peak Apex for Retention Time....... 94

C-3 Method Validation......................................................................... 95

C-4 Quantitation Precision................................................................... 97

C-5 Quantitation Accuracy................................................................... 98

C-6 Analyte Recovery.......................................................................... 99

C-1 Required Instru ment ation ...................................................... .....103

C-2 Analysis Conditions.................................................................... 104

C-3 IC Processing Method Using Peak Apex for Retention Time..... 105

List of Tables 22

Page 23

Preface

The Waters 432 Conductivity Detector Operator’s Guide details the procedures for unpacking, installing, operating, maint ain ing, and troubleshooting the 432 Conductivity Detector. It also includes appendixes listing specifications and spare parts and describing validation support.

This guide is intended for use by personnel who need to install, operate, maintain, or troubleshoot the 432 Detector. This guide assum es an understandi ng of the princip les of chromatography.

Organization

This guide contains the following:

Chapter 1 Chapter 2

electrical connections.

Chapter 3

general operating instructions.

Chapter 4

for anion and cation analysis.

Chapter 5

tables to aid in problem diagnosis.

Appendix A

Detector.

Appendix B Appendix C Appendix D

support .

describes the features and method of operation of the 432 Detector.

describes the procedures for installing the 432 Detector and making fluid and

describes the controls and indicators of the 432 Detector, and provides

describes the system configuration, eluents, and standards recommended

describes simple maintenance procedures and provides troubleshooting

describes the operational specifications and requirements of the 432

lists the recommended spare parts for the 432 Detector. describes ion chromatography methods. describes the recommended validation protocols and Waters® validation

Chapter 1 Introduction

Features

The Waters® 432 Conductivity Detector (Figure 1-1) 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 stable operation

• Auto baseline/auto zero

• External recorder/integrator and chart mark connections

• Three time constant selections

• “Leak-detected” alarm signal

aters 432

Conductivity Detector

TP01268

OUT

Figure 1-1 Waters 432 Conductivity Detector

Page 27

Method of Operation

This section discusses the method of operation of the 432 Detector. Additional descriptive information appears in these sections:

• Section 2.4.1,

• Section 3.1, Controls and Indicators

• Appendix A, Specifications

Measurement Technique

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 accuracy .

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” (Figure 1-2 set temperature, and then flows through the cell, directly contacting the electrodes. The 5-electrode design permits measurem ent of conduct ivity 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 .

I/O Signal Descriptions

). Column eluent flows through the heater to attain the

Introduction 27

Page 28

TP01271

Ion Detection Theory

1= Reference Electrodes 2= Detection Electrodes

Figure 1-2 Flow Cell Schematic

3= Guard Electrode

Flow Cell Bloc (heated)

Fluid Outlet

The conductance of a solution of known concentration can be calculated using the following equation:

λC

----------

G = measured conductance of the solution, in Siemens (1 S = ohm

C = concentration in equivalents per 1000 cm K = length/area of cell (the cell constant)

λ = equivalent conductance in S cm

Table 1-1 lists the equivalent conductances of some common ions.1 Concentrations above

−5

to 10−3 N, generally exhibit decreased equivalent conductance due to interionic

effects.

−1

)

3–

equiv

−1

Page 29

Table 1-1 Limiting Equivalent Conductance of Ions in Water at 25 °C

Cations

(CH3)2NH

349.8 OH

38.6 F

50.1 Cl

73.5 Br

77.8 I

61.9 NO3

73.3 ClO

51.8 ClO

53.0 IO

−

Anions

−

198.6

55.4

76.4

78.1

76.8

71.5

64.6

67.4

54.5

—

53.1 Formate 54.6

59.5 Acetate 40.9

63.6 Benzoate 32.4

53.6 SO

52.8 CO

69.7 Fe(CN)

−

80.0

69.3

111.0

69.8

1. Henry H. Bauer et al., eds. “Instrumental Analysis,” Allyn and Bacon, Boston (1978), p. 115. Reprinted with permission from the publisher.

Introduction 29

Page 30

Chapter 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 environmental requirements

• Unpacking and inspecting the 432 Detector and accompanying items

• 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 dampener)

• Passivating the detector and other post-column fluid path componen ts

After you have successfully completed this chapter, familiarize yourself with the information in Section 3.1, detector, perform the startup procedure described in Section 3.2,

Controls and Indicators. When you are ready to operate the

2.1 Selecting the Installation Site

Operating Environment

The 432 Detector operates in any standard laboratory environment that provides suitable electrical power and remains within the following ranges:

• Temperatu re: 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:

Startup and Shutdown.

• 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 contacts

• Appreciable shock or vibration

Selecting the Installation Site 30

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Required Space

The 432 Detector requires bench space that measures approximately:

• 10 inches (25 cm) high

• 14 inches (34 cm) wide

• 24 inches (60 cm) deep

Make sure that air can circulate freely through the ventilation slots on both

STO

Attention:

side panels.

Power Requirements

The 432 Detector requires:

• One properly grounded AC voltage outlet.

• Correct voltage and fuse selections as shown in Table 2-2

2.2 Unpacking and Inspection

Unpacking

The 432 Detector is shipped in one carton that contains the following items:

• Waters 432 Conductivity Detector

• Startup Kit

• Validation certificate

• Waters 432 Conductivity Detector Operator’s Gui de

• Packing list

• Declaration of conformity

If you purchased the 432 Detector as part of an ion/liquid chromatograph system, a

Note:

Waters representative will perform the installation and startup.

To unpack the 432 Detector :

1. Locate the packing list.

2. Unp ack 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

Star tup Kit List.

4. Save the shipping carton for future transport or shipment.

Installing the Detector 31

Page 32

Inspection

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 Appendix B,

2.3 AC Power Connection

Spare Parts.

Caution:

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.

To avoid a potential fire hazard and damage to the 432 Detector, make sure that

Power Cord

The power connector is located on the lower-r ight co rner of the rear panel, as shown in

Figure 2-1 Table 2-1

. If a power pl ug other than the one supplied is needed for your location, consult

and observe the existing applicable regulations.

AC Power Connection 32

Page 33

IEEE DIP Switch Cover

Figure 2-1 Rear Panel

Table 2-1 Power Cord Wire Identification

Wire (USA) Wire (International) Connection

Black Brown Hot White Blue Neutral Green Green/Yellow Ground (Earth)

The 432 Detector can be adapted to operate within two voltage ranges at 50 or 60 Hz.

Table 2-2

describes these voltage ranges and the fuse value that is appropriate to each.

Installing the Detector 33

Page 34

Table 2-2 Nominal Operating V oltage

Nominal Voltage (VAC) Fuse

100/120 T 2A 220/240 T 1A

Required Material

You need a flat-blade screwdriver to perform this procedure.

Procedure

Caution:

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

2. Remove the voltage selection barrel and locate the correct voltage setting

3. Reinstall the voltage selection barrel so the desired voltage setting appears through

To avoid the possibility of electrical shock, turn off the front panel power switch

pry open the power connector cover with a flat-blade screwdriver.

(Figure 2-2

the window when you close the power connector cover (Figure 2-2

Voltage Settings

Figure 2-2 Changing the Voltage Setting

4. Determine if you need to change the fuses (see Table 2-2

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 Section 5.1.1, Replacing Fuses.

5. Reins tall the power connector cover and the power cord.

). All units are supplied

AC Power Connection 34

Page 35

2.4 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®

• Data module

• SAT/IN™ module

• Chart recorder

• Device signalling the Chart Marker input

• Device signalling the Auto Zero input

soft wa r e

To meet the regulatory requirements of immunity from external electrical

STO

Attention:

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 termi nal strips. In addition, ensure you always connect the shield of the cable to chassis ground at one instrument only.

2.4.1 I/O Signal Descriptions

The 432 Detector rear panel has an IEEE-488 connector for communication with a PowerLine controller, and a terminal strip (Figure 2-3 signals are described in Table 2-3.

) for the input/output signals. These

Installing the Detector 35

Page 36

+ –

INT

Int

–

REC

Rec

–

LEAK

Leak

MARKER

Marker In

–

MARKER

Marker Ou

OUT

AUTO ZERO

Auto Zero

–

Figure 2-3 I/O Terminal Strip

Table 2-3 I/O Signal Descriptions

Terminal Pairs Function

Rec (+ and –)

Recorder output – A 10-mV full-scale analog output signal appears on these terminals. The measurement range is deter mined by the product of the Base Range and Sensitivity settings: for example, 500 µS (base range) x 0.005 (sensitivity) = 2.5 µS full sca le.

Int (+ and –)

Integrat or ou tp ut – A 1-V fu ll-scale analo g output signal appears on these terminals. The measurement range is selectable: 10, 50, or 100 µS full scale.

Marker Ou t

Marker output – A 1-second contact closure signal appears on these terminals when either of the following events occurs:

Leak

Auto Zero (+ and –)

• The Char t Mark key on the keypad is pressed

• A contact closure signal occurs between the Marker In terminals Leak Alert output – A contact closure signal appears on these termi-

nals 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 I n (+ and –)

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 terminals.

I/O Signal Connections 36

Page 37

Required Material

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 items:

• 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 instruments.

2.4.2 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.

Required Material

You need a 2.5-mm Allen wrench to connect to the 432 Detector.

Procedure

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 Start up K it) into the bus

connector on the rear panel of the 432 Detector (Figure 2-1 the cable into the bus connector on the PowerLine controller.

3. Remove the DIP switch cov er (Figure 2-1

4. R efer to Table 2-4 to set the DIP switches on the rear panel of the 432 Detector

(Figure 2-4

5. Afte r you set th e DIP swi tc hes, re in stall the DIP switch cover.

To operate the 432 Detector in local mode, press the front panel

Note:

illuminated light above the key will go out.

) to a unique IEEE-488 address between 2 and 29.

) using a 2.5-mm Allen wrench.

) and the other end of

Remote

Installing the Detector 37

key. The

Page 38

1 2 3 4 5

OFF

(Address 8 Shown)

Figure 2-4 IEEE-488 Address Switch

The IEEE-488 address DIP switch employs positive logic to determi ne the addre ss of the 432 Detector from the switch settings. Table 2-4

Table 2-4 IEEE-488 DIP Switch Setting

IEEE-488

Address

2 OFF ON OFF OFF OFF 3 ON ON OFF OFF OFF 4 OFF OFF ON OFF OFF 5 ON OFF ON OFF OFF 6 OFF ON ON OFF OFF 7 ON ON ON OFF OFF 8 OFF OFF OFF ON OFF 9 ON OFF OFF ON OFF 10 OFF ON OFF ON OFF 11 ON ON OFF ON OFF 12 OFF OFF ON ON OFF 13 ON OFF ON ON OFF 14 OFF ON ON ON OFF 15 ON ON ON ON OFF 16 OFF OFF OFF OFF ON 17 ON OFF OFF OFF ON 18 OFF ON OFF OFF ON 19 ON ON OFF OFF ON

1 2 3 4 5

DIP Switch Set t ings

Switch 5Switch 1

shows the settings for valid addresses.

I/O Signal Connections 38

Page 39

Table 2-4 IEEE-488 DIP Switch Setting (Continued)

IEEE-488

Address

20 OFF OFF ON OFF ON 21 ON OFF ON OFF ON 22 OFF ON ON OFF ON 23 ON ON ON OFF ON 24 OFF OFF OFF ON ON 25 ON OFF OFF ON ON 26 OFF ON OFF ON ON 27 ON ON OFF ON ON 28 OFF OFF ON ON ON 29 ON OFF ON ON ON

1 2 3 4 5

DIP Switch Set t ings

PowerLine Operation

Under PowerLine control, the 432 Detector is recognized as a 431 Detector and it retains the functionality of the 431 Detector with the following differences:

• 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, t he selected Balance value is sent to the 432 Detector from the PowerLi ne 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.

2.4.3 Empower and Mille nn ium32 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.

Installing the Detector 39

Page 40

Empower and Millennium32 are menu-driven applications specifically designed by Waters for chromatographers. Use the software to:

• Acquire data

• Process d a ta

• Generate and print reports

• Store information (or data) in a central area and share this information with users who have pr oper se cu rity acce ss

To connect the 432 Detector to an Empower or Millennium

computer, be sure to:

• Connect the Bus Satellite Interface (SAT/IN) module to the Bus Laboratory Acquisition and Control/Environment (LAC/E™) card in the Empower computer,

Millennium

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 m ode when it is connec t ed to an Empower and Mille nnium computer.

Bus S AT/IN Module

The Waters Bus SAT/IN module, shown in Figure 2-5, 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/E

Waters SAT/IN Module

CHANNEL 1 CHANNEL 2

EVENTS

CH1

IN INOUT OUT

+ –

1 2 3 4 5 6 7 8

CH2

+ –

Figure 2-5 Bus SAT/IN Module (Front P anel)

CH1CH

I/O Signal Connections 40

Page 41

To prevent damage to the unit, always disconnect the power cord at either the wall

Note:

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, as shown in Figure 2-6

To connect the Bus SAT/IN module to the Bus LAC/E card:

1. Use the I/O distr ibution cable to connect the I/O distribution box to the 9-pin I/O

distr ibut ion po rt on the Bus L AC/E car d a t the b ack of the Millennium

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

Millennium

installation and configuration guides.

computer.

Installing the Detector 41

Page 42

I/O Distribution

Cable

I/O Distribution Port (9-pin) of Bus LAC/E Card

SAT/IN Module Rear Panel

I/O Distribution Box

BCD

DATA

PWR

Serial Cable

AC to DC Converter

Modified Modular Jack Connections

Connect SAT/IN to Port 1 on the I/O Distribution Box

Figure 2-6 Bus SAT/IN to Bus LAC/E Connections

I/O Signal Connections 42

Page 43

Connecting the Bus SAT/IN Module to the 432 Detector

Black

The Bus SAT/IN module connects to the 432 Detector as shown in Figure 2-7. Refer to the procedure following the figure and Table 2-5

for complete details.

STO

Waters SAT/IN Module

Attention:

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 B u s SAT /IN Mo dule Installation Guide.

Waters 432 Detector

Whi

Red

Black

INT

–

REC

–

LEAK

MARKER IN

–

EVENTS

CH1

CHANNEL 1 CHANNEL 2

IN INOUT OU T

+ –

1 2 3 4 5 6 7 8

CH2

+ –

CH1CH

MARKER OUT

AUTO ZERO

–

TP01264

Figure 2-7 Bus SAT/IN to 432 Detector Connections

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.

2. Con nect 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.

Installing the Detector 43

Page 44

3. Connect the Event In terminals of the channel you chose in the previous step to the

Inject St a rt output signal of the Waters Alliance 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 in Table 2-5. Table 2-5 Bus SAT/IN Cable Connections

solvent delivery system or the

432 Detector I/O

Connector Terminal

Int (+ ) White wire Int (–) Black wire Channel 1 or 2

Bus SAT/IN

Cable

2.4.4 Data Module Connections

This section describes how to connect the analog output signal from the 432 Detector to the Waters 746 Data Module.

Attention:

STO

electrical disturbances that may affect the performan ce 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.

Analog Signal

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 in Table 2-6

Table 2-6 Data Module Signal Cable Connections

Wire

Remember to meet the regulatory requirement s of immunity from external

432 Detector I/O

Connec tor Terminal

Bus SAT/IN

Connector

746

Terminal

Red Int (+) (+)

Black Int (– ) (–)

Shield Ground lug None

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 to gether

I/O Signal Connections 44

Page 45

Use the signal to start a Waters 746 Data Module by connecting a signal cable to the module’s data cable (Table 2-7

Table 2-7 Data Module Chart Mark Cable Connections

432 Detector I/O

Wire

Either wire Marker Out Join to both Remote Start

Other wire Marker Out Green wire

Connector

Terminal

2.4.5 Chart Recorder Connections

To connect the 432 Detector to a chart recorder:

1. Attach the Recorder cable (see Appendix B, output terminals, as indicated in Table 2-8

2. Con nect the cable shield to the ground lug on the 432 Detector rear panel.

3. Con nect the other end of the cable to the 10-mV input terminals on the chart recorder, as indicated in Table 2-8

Table 2-8 Chart Recorder Cable Connections

Wire

Red Rec (+) Pen (+)

Black Rec (–) Pen (–)

432 Detector I/O

Connector Terminal

746

Cable

wires (white and red)

Spare Parts) to the 432 Detector REC

Chart Recorder

Terminal

2.4. 6 Chart Marker Input Connections

The 432 Detector accepts a chart mark (start inject) signal from the following devices:

• 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 Table 2-9

Installing the Detector 45

Page 46

Table 2-9 Autosampler Chart Mark Cable Connections

432 Detector I/O

Connector Terminal

Marker In (+) Either Inject Start ter m inal of a pair Marker In (–) Other Inject Start terminal of the same pair

2.4.7 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 Table 2-10

Table 2-10 Autosampler Auto Zero Cable Connections

432 Detector I/O

Connector Terminal

Autosampler

Terminal

Autosampler

Terminal

Auto Zero (+) Either Inject Start ter m inal of a pair Auto Zero (–) Other Inject Start terminal of the same pair

2.4.8 Alliance Separations Module Connections

Connect the detector to Waters Alliance Separations Modules, when it is not under the control of the Millennium

• Auto-Zero on inject

• Chart mark on inject

• Method start

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 Table 2-11

softwa re, to per fo rm the foll o w i n g tasks:

and illustrated in Figure 2-8.

I/O Signal Connections 46

Page 47

Table 2-11 Connections for Generating Auto-Zero on Inject

–

INT

REC

LEAK

MARKER IN

MARKER OUT

AUTO ZERO

+ –

Alliance Separations Modules

(B Inputs and Outputs)

432 Detector (A Inputs)

Pin 1 Inject Start Auto-Zero (+) Pin 2 Inject Start Auto-Zero (–)

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.

Waters All iance B (Inputs and Outputs)

Inject Start + Inject Start – Ground Stop Flow Stop Flow – Hold Inje c t 1+ Hold Inject 1 Hold Inject 2 + Hold Inje c t 2 Ground Chart Ou t Chart Out

+ –

–

Waters 432 Detector A (Inputs)

1 Inject Start +

2 Inject Start - 3 Ground

4 Lamp On/Off + 5 Lamp On/Off -

6 Chart M a rk +

7 Char t M ark -

8 Ground 9 Auto-Zero +

10 Auto-Zero -

Int

Rec

Leak

Marker In

Marker Ou

Auto Zero

Figure 2-8 Alliance Separations Module Connections to the 432 Detector

Generating Char t Mark on Inject

To generate the chart mark function at the start of an injection, make the connections summarized in Table 2-12

Auto-Zero on Inject

and illustrated in Figure 2-9.

Installing the Detector 47

Page 48

Table 2-12 Connections for Generating Chart Mark on Inject

–

INT

REC

LEAK

MARKER IN

MARKER OUT

AUTO ZERO

+ –

Alliance Separations Modules

(B Inputs and Outputs)

432 Detector (A Inputs)

Pin 1 Inject Start Marker In (+) Pin 2 Inject Start Marker I n (–)

Before you can generate a chart mark from an Alliance Separations Module, you must configure the chart mar k signa l at the front panel. The default chart mark signal is Low.

Waters All iance B (Inputs and Outputs)

Inject Start + Inject Start – Ground Stop Flow Stop Flow – Hold Inje c t 1+ Hold Inject 1 Hold Inject 2 + Hold Inje c t 2 Ground Chart Ou t Chart Out

+ –

–

Waters 432 Detector A (Inputs)

1 Inject Start +

2 Inject Start - 3 Ground

4 Lamp On/Off + 5 Lamp On/Off -

6 Chart M a rk +

7 Char t M ark -

8 Ground 9 Auto-Zero +

10 Auto-Zero -

Int

Rec

Leak

Marker In

Marker Ou

Auto Zero

Figure 2-9 Alliance Separations Module Connections to the 432 Detector

Chart Mark on Inject

2.5 Making Fluidic Connections

Fluid lines to a column and waste container connect to the front of the 432 Detector, as shown in Figure 2-10

. To make these connections:

• Cut the tubing.

• Assemble compression fittings and ferrules.

• Connect the tubing to the detector.

Making Fluidic Connections 48

Page 49

This section will guid e you through each of these pr ocedures.

STO

Attention:

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.

aters 4

ductivity Dete

Out to Waste (18 inches,

0.009-inch I.D.)

0.009-inch ID)

In from Column

In From Column

Out To Waste (18 inches.

Figure 2-10 Fluid Connections

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

To cut the tubing:

1. Meas ure 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. G rasp the tubing on both sides of the scribe mar k with cloth-covered pliers. Gently work the tubing back and forth until it separates.

4. File the ends smooth.

Installing the Detector 49

Page 50

Cutting Polymeric Tubing

Waters chromatography systems are supplied with a tubing cutter (similar to the one in

Figure 2-11

procedure for using the tubing cutter.

Note:

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. Estima te the length of tubing required to connect the componen ts. Allow slack so

2. Insert the tubing into the cutter so that the tubing extending from the metal side is

) to facilitate cutting polymeric tubing. This section presents the recommended

To avoid bandspreading caused by angled cuts, always use a tubing cutter. Angled

that the tubing is not pulled tightly around sharp corn ers.

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.

Figure 2-11 Cutting Polymeric Tubing

3. Pres s down on the razor blade to cut the tubing (Figure 2-11 tubing that extends from the clear side of the cutter.

4. Inspec t the cut for burrs or scratches and for the perpendiculari ty of the cut.

). Discard the excess

Assembling Compression Fittings

To assemble each compression fitting:

1. Slide the com pression screw over the tubing end, followed by the ferrule (Figure 2-12

2. Mount the ferrule with its taper end facing the end of the tubing (Figure 2-12

Making Fluidic Connections 50

Page 51

Compression

Screw

0.009-inch I D Tubing

0.009-inch I.D. Tubing

(0.23 mm)

Figure 2-12 Ferrule and Compression Screw Assembly

Ferrule

Connecting to the 432 Detector

To make connections at the column outlet and detector inlet, and at the detector outlet:

1. Insta ll 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.

If you are using a column with 1/4–28 end fittings and there is a length of

Note:

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.

The 432 Detector and IC-Pak series of columns have very deep ferrules.

Note:

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 t he 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 Detector 51

Page 52

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 dampe ner is not required if you are using a Waters 2695 Separations Module.

To install the pulse dampener betwee n the pump and the injector:

1. Assemble the pulse dampener (Figure 2-13 dampener kit.

2. Con nect the large-ID (0. 020-inch) tubing to the pump outlet using a stainless steel compression screw and ferrule.

3. Con nect 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. Pum p AST M 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.

) using the instructions in the pulse

Making Fluidic Connections 52

Page 53

0.020-inch I.D.

0.009-inch I D Tubing

Restrictor Assembly

0.020-inch I D Tubing

Tubing

0.009- Tubing

nch I.D.

From Pum p

rom Pum p Union To Injector

Low Pressure

Filter Assembly

Filter Assembl

Figure 2-13 Pulse Dampener

2.6 Passivating the System

Union

To Injecto

Restrictor Assembl

STO

Passiv ating the system removes potential contamination from the wetted surfaces of all system components. Perform pas sivation on a new system, and subsequently, whenever you suspect that contamination may have occurred. See Section 5.3, help diagnosing performance problems.

Use this procedure for Waters hardware only. For other equipment, check with the manufacturer before you continue with this procedure.

Attention:

the pump seals before you passivate. Use the new pump seals supplied in the Startup Kit and refer to the replacemen t procedure in the pump manual.

If you are installing the 432 Detector into an existing Waters system, replace

Installing the Detector 53

Troubleshooting, for

Page 54

To passivate the system:

1. Rep lace 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. Con nect the power cord to the 432 Detector and plug the other end into an AC power outlet. Push the 432 Detector power switch to tur n on the instrum ent .

Caution:

when you are using solvents.

4. Prime the pump with 6 N nitric acid (HNO 20 minutes to passivate all the wetted parts of the detector. Press the Clear key to stop the overrange alarm.

5. St op 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).

• Flush the system overnight with 100% methanol at a reduced flow rate. By the

next morning the system will be passivated and ready for use.

If you are using a pump with seal-wash capability, skip step 8.

Note:

8. Use a syr inge 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/mi n. It is not nec essary to turn off the 432 Dete ctor unless it will be idle for an extended period (14 days).

To avoid chemical hazards, alwa ys wear safety glasses and gloves

) and run it at a flow rate of 1.2 mL/min for

For best results, alw ays leave the power on to main t a in c e ll 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.

2.7 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.

Verifying the Detector 54

Page 55

Verify the detector when any of these conditions apply:

• When you replace the flow cell

• To verify accuracy

• When you make adjustments

Calibration Procedure

You need solution of 1 mM potassium chloride (KCl) to calibrate the detector.

Note:

Waters suggests one of its Technical Service Representatives perform this

Note:

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.

4. Pump 1 mM KCl solution through the detector (without a column in place).

5. Verify that the front panel output is 147 µS ± 5 µS.

Installing the Detector 55

Page 56

Chapter 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

3.1 Controls and Indicators

Figure 3-1 illustrates the controls and indicators on the front panel of the 432 Detector.

Waters 432

Conductivity Detector

274 500 0.0005

CONDUCT

( S/cm)

BASE

( S/cm/FS)

SENS

IN OUT

Auto Base

Base

Range

Resp.

Figure 3-1 Front Panel

Remote

Chart

Mark

Clear

EnterShift

Auto Zero

Sens.

Range

Bal.

Temp. Pol.

OFF

Controls and Indicators 56

Page 57

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.

Display

The display shows instrument status and parameter values in two 20-character lines of text. Upon startup, Waters 432 Self Check appears briefly. If any error conditions are detected during start up 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.

Error Messages

A corresponding error message is displayed if one of the following conditions occurs:

• ROM/RAM error (checked during startup only)

Error: ROM/RAM

• Leakage detected

Error: Leak

• Temperature control fail ure

Error: 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.

Keypad

Use the keypad to control the operation of the 432 Detector. Table 3-1 describes the function of each key.

Operating the Detector 57

Page 58

Three keys (Balance, Sensitivity Range, and the numeral 1) perform an alternate

Note:

function when they are preceded by the Shift key.

Table 3-1 Key Descriptions

Key Function

Remote key: Toggles betwee n local and remote operating mode s. In

Remote

Pol.

Base Range

Sens.

Range

remote mode, the light above the key is on and all other front panel controls are disabled.

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 illuminated.

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 (avai lable onl y with 100µS 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.

Bal.

Temp.

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.

Balance key: Manually sets the offset (%) of the signal to the exter nal chart reco rder. (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 illumi nate d when the temperature control is on.

Controls and Indicators 58

Page 59

Table 3-1 Key Descriptions (Continued)

Key Function

Response key: Sets the response time constant of the 432 Detector to

Resp.

Auto Zero

Auto

Base

Shift

optimize signal-to-noise ratio. Use the Up and Down key s 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 eluent.

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 agai n to r e turn to normal mode.

Char t

Mark

Enter

Clear

Shift key then Balance key: Di splays 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 disp lay t o the main screen. Th e Enter key i s 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.

Operating the Detector 59

Page 60

Table 3-1 Key Descriptions (Continued)

Key Function

Up key: Increments the current parameter setting.

Down key: Decrements the current parameter setting.

Beep Function

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 Table 3-2 Table 3-2 Setting the Beep Function

Keystroke Key Function

1 2 3 4

Since the void volume in many separations contains highly conductive counter-ions,

Note:

the error alarm sounds upon each injection. See Table 3-2 alarm.

Enter . (decimal point) Enter 1 0 1 0 Enter

Accesses the beep function parameters.

Tur ns on the beep-on-error function. Tur ns off the beep-on-error function. Tur ns on the beep-on-keystroke function. Tur ns off the beep-on-keystroke function. Saves settings and returns to the main screen.

to silence the beep-on-error

Controls and Indicators 60

Page 61

3.2 Startup and Shutdown

Startup Procedure

Perform the following procedure to start the 432 Detector. Typically, this procedure is done at the beginning of each workday.

This procedure assumes that the flow cell has stabilized at the selected temperature

Note:

(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 f or 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 recomm ended for your particular column or application. Do not sparge eluents, since sparge gasses may contain ionic contaminants.

2. Set the response (time cons tant) to the desired setting by pressing the Response key. A standard setting (0.5 seconds) is preferred for most applications.

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 stabilizes.

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 key.

6. Zero the recorder/integrator by pressing the Auto Zero key or have the Auto Zero terminals of the rear panel I/O termi nal strip connec ted to your manual injector or autosampler.

The 432 Detector is now ready for operation.

Standby Setup

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).

Operating the Detector 61

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Long-Term Storage

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.

3.3 Operating Recommendations

Observe the following recommendations for best detector performance.

Temperature Equilibration

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 .

Base Range

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.

Integrator Output

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.

Recorder Output

The Recorder output is attenuated and the Sensitivity Range should be adjusted to provide the appropriate output level.

Operating Recommendations 62

Page 63

Polarity

Signal polarity depends on eluent conductivity. If necessar y, press the Polarity key to obtain peaks rather than dips.

Eluent Handling

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 63

Page 64

Chapter 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:

• Fundamental considerations

• 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.

4.1 Fundamental Considerations

To obtain full performance from the 432 Detector in chromatographic applications, observe the guidelines presented in this section regarding:

• Water

• Containers

• High-pH eluents

• Sample preparation

Water

Water purity (the absence of conductivity) is the most important consideration in ion analysis.

Using water other than ASTM Type I reagent water will compromise the accuracy of

Note:

your results.

Freshly dra wn 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

Fundamental Considerations 64

Page 65

adsorption cartridges produce ASTM Type I reagent water, and are recommended for ion

chromatography applications. Do not use HPLC-grade water or distilled water.

To avoid damage to the detector flow cell, do not allow the flow cell to

STO

Attention:

dry out.

Containers

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.

Attention:

STO

analyzing for cations.

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 (HNO

2. Rins e with plenty of ASTM Type I reagent water. The containers are ready for

3. For analysis in the ppb range, fill each container completely with ASTM Type I

Avoid glass containers (which tend to leach sodium cations) when you are

Type I reagent water.

analysis in the ppm range.

reagent water and let soak overnight.

) and ASTM

Certain applications that involve ppb lev el analysis may require container considerations beyond the scope of this manual. For further instr uctions on trace metal cleaning of plasticware, see “Selection and Cleaning of Plastic Containers for Storage of Trace Element Samples,” JR Mo ody and RM Lindstrom, An alytical Chemistr y, v. 49, Dec 1977, pp. 2264-67, or contact the Waters Technical Services Depart men t.

Cleaning Syringes

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

High-pH eluents (such as hydroxide eluent) absorb atmospheric CO2, which slowly acidifies the eluent causing baseline drift and retention time changes. To m inimize

Performing Ion Analysis 65

Page 66

carbonate absorption, co nnect a soda lime (Ascarite®) tube (Figure 4-1) 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.

Caution:

when you are handling soda lime.

2. F ill t he tube with soda lim e (As c arite) until it reaches 3/4 inches (2 cm) from the top.

3. Add anot her 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 inlet line.

5. Pass the pump inlet line through the hole. Seal the hole with paraffin film.

6. Cha nge the soda lime in the tube when it is exhausted.

To avoid chemical burns, wear gloves, lab coat, and eye glasses

Fundamental Considerations 66

Page 67

End Fittings

Glass Wool

Soda Lime

Polypropylene Tube

Pump Inlet Lin

Figure 4-1 Soda Lime Tube

Reagent Bott

Sample Preparat ion

Sample preparation is very important in ion analysis. Contact the Waters Technical Services Department, if you need assistance.

Performing Ion Analysis 67

Page 68

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. Howev er, 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 h ydrophobic 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

cartr idge to remove hydrophobic compounds.

4.2 Configuring the System

Figure 4-2 shows a typical system configuration. Refer to Section 2.5, M aking Fluidic Connections, for the procedures to cut tubing and assemble fittings.

Eluent

Reservoir

Waters In-Line Degasser

Pump

Optional

Required for non-Waters pumps or Waters pumps with Breeze software, such

† Required for Waters pumps without SILK or non-W aters pumps

as the HPLC 515

Figure 4-2 System Configuration for Ion Analysis

Dampener

Pulse

Guard Column

Holder

Injector

†

Column

Waters

432

Detector

Configuring the System 68

Waste

TP01269

Page 69

Pulse Dampener

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 “

the Pulse Dampener” on page 52.

4.3 Eluents for Ion Analysis

This section describes how to select, prepare, and use eluents for ion analysis.

Attention:

STO

progressively contaminate a recirculating eluent.

General Guidelines

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.

• 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

Never recirculate eluents. Ions from sample and standard injections

Installing

The Waters Solvent Clarification Kit is recommende d for eluent filtration and preliminary degassing. Durapore Millipore 0.45-µm HATF filters may be used for aqueous eluents containing no organi c modifier. For eluents containing organic modifier, use the Durapore filters.

Attention:

STO

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. Tur n 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,

To avoid contamination when you analyze for cations, minimize the time that

0.22-µm filters can be used for all ion chromatography eluents.

Performing Ion Analysis 69

Page 70

transfer the eluent t o a precleaned plastic container, introducing the least possible amount of bubbles in the process.

The Waters In-line Degasser is recomme nded for continuous online degassing.

4.3.1 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.

Preparing Lithium Borate/Gluconate Concentrate

To prepare sodium borate/gluconate concentrate, refer to SectionC.1.13, Preparing

Lithium Borate/Gluconate 50X Stock Concentrate and Section C.1.14, Preparing Lithium Borate/Gluconate Eluent.

4.3. 2 Preparing Cation Eluent

To prepare 1 L of cation eluent, refer to Section C.2.1, Prepar ing Eluent.

4.4 Standards for Ion Analysis

This section describes how to prepare and inject ion standards.

It is recommended to purchase certified 1000-ppm anion standards instead of

Note:

preparing them manually. Cer tify 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 ppm 1 part per million (ppm) = 1 µg/mL = 1 mg/L = 1000 ppb 1 part per billion (ppb) = 1 ng/mL = 1 µg/L = 1000 ppt 1 par t per tr illion (ppt) = 1 pg/mL = 1 ng/L

Standards for Ion Analysis 70

Page 71

Storing Standards

For accurate quantitative results, do not store standards beyond the approximate periods listed in Table 4-1 significantly shorter than shown here.

Table 4-1 Shelf-Life of Standards

Carbonate, ppm 1 day Chloride, ppm 3 weeks All, ppb 1 day Nitrite and carbonate concentrates 1 week All other anion concentrates 1 to 2 months Cation standards 1 month Cation concentrates 6 months

. Be aware that shelf-life depends on many factors and may be

Standard She lf-Life

Cation standards must be stored in properly prepared plasticware. See “

page 65.

4.4.1 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 s alts shown in Table 4-2 to calculate the amount for a salt not listed:

(Mol. Wt. Salt / Mol. Wt. Cation) x 0.1 = g Salt

Containers” on

or use the following formula

Performing Ion Analysis 71

Page 72

Table 4-2 Salts for Anion Standard Concentrates

Salt (Anion) Weight (mg)

Sodium fluoride (F–) 221.0 Sodium chloride (Cl–) 164.9 Sodium nitrite (NO

–

) 150.0

Potassium bromide (Br–) 148.9 Sodium nitrate (NO Potassium phosphate,

monobasic (HPO Sodium sulfate (SO

–

) 137.1

2–

)

2–

) 147.9

141.8

2. Place each s alt in a separate plastic100-mL volumetric flask and dilute to the mark with ASTM T ype I reagent water . Each concentrate contains 1000 ppm of the anion.

3. Combine the amounts listed in Table 4-3

in a 100-mL volumetric flask with ASTM

Type I reagent water. Table 4-3 Anion Concentrate Dilutions

Anion

Amount

(

Fluoride 100 µL 1 ppm Chloride 200 µL 2 ppm Nitrite 400 µL 4 ppm Bromide 400 µL 4 ppm Nitrate 400 µL 4 ppm Phosphate 600 µL 6 ppm Sulfate 400 µL 4 ppm

4.4.2 Injecting Anion Standards

Required Materials

To inject a standard, obtain the following materials:

• Borate/glucona t e e luent – Refer to Section C.1.13,

Borate/Gluconate 50X Stock Concentrate and Section C.1.14, P reparing Lithium Borate/Gluconate Eluent.

Final Concentration

(ppm)

Preparing Lithium

Standards for Ion Analysis 72

Page 73

• 1-cc disposable plastic syringe – Pharm as eal® Stylex® disposable syringe with a

Luer Slip

• 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.

tip, or equivalent.

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 char t)

• Response = STD (0.5 seconds)

• Temperature = 35 °C

• Polarity = +

2. Equi librate 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 t he syr inge tip into the sample loading port and overfill the 100 µL loop at least three times (that is, 300 µL).

5. Inject the sample.

Startup Procedu re ” on page 61.

Figure 4-3

IC-Pak A (4.6 mm x 5.0 cm) column with borate/glucona te eluent at 1.2 mL /min flow rate. The separation of the standard typically takes 12 to 15 minutes with this setup.

shows a representative chromatogram of the 7-anion standard run on an

Performing Ion Analysis 73

Page 74

Figure 4-3 Chromatogram of a 7-Anion Standard

4.4.3 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 , us e only properly prepared plasticware and do not store standards beyond the recommended shelf-lives listed in Table 4-1

Preparing Cation Standard Concentrates

It is recommended that you use certified 1000-ppm cation standards not prepared

Note:

in acid with this method.

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 s alts shown in Table 4-4 to calculate the amount for a salt not listed.

or use the following formula

Standards for Ion Analysis 74

Page 75

(Mol. Wt. Salt / Mol. Wt. Cation) = g Salt If you choose to use other salts, be sure to avoid any combinations that will for m a

precipitate. Table 4-4 Salts for Cation Standard Concentrates

Salt (Cation) Weight (g)

Lithium hydroxide monohydrate (Li+) 6.0476 Sodium chloride (Na+) 2.5421 Ammonium chloride (NH Potassium chloride (K+) 1.9067 Magnesium nitrate hexahydrate (Mg2+) 10.5 466 Calcium nitrate tetrahydrate (Ca2+) 5.8919 Strontium nitrate tetrahydrate (Sr2+) 3.2377 Barium chloride dihydrate (Ba2+) 1.7786

2. Place each s alt in a separate plastic1-L volumetric flask and dilute to the mark with reagent-grade water. Each concentrate contains 1000 ppm of the cation.

)

2.9640

Preparing an 8-Cation Standard

To prepare 1 liter of 8-cation standard:

1. Add the volume of stock (concentrate) standard listed in Table 4-5 volumetric fl a sk.

Table 4-5 Cation Concentrate Dilutions

Cation Amount (mL)

Lithium 0.25 0.25 Sodium 1. 00 1.00 Ammonium 1.00 1.00 Potassium 3.00 3.00 Magnesium 2.00 2.00 Calcium 3.00 3.00 Strontium 5.00 5.00 Barium 5.00 5.00

2. F ill t he flask to the mar k with ASTM Type I reagent water.

Final Concentration

(ppm)

Performing Ion Analysis 75

to a plastic 1-L

Page 76

4.4.4 Injecting Cation Standards

Required Materials

To inject the standard, obtain the following materials:

• 0.1 mM EDT A/ 3 mM HNO

Eluent.

• 1-cc disposable plastic syringe – Pharm as eal

Luer Slip

• Injector or autosampler with a 100-µL loop – Ion chromatogr aphy com m onl y u se s

a 100-µL injection volume. When you use a fixed loop, overfill a minimum of three times.

tip, or equivalent.

cation eluent – Refer to Section 4.3.2,

Stylex® disposable syringe with a

Preparing Cation

Injecting the Standard

You can substitute the method described in Section C.2,

Note:

Cations, Ammonium, and Amines, for the following procedure.

Use this 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 char t)

• Response = STD (0.5 seconds)

• Temperature = 35 °C

• Polarity = – (negative)

2. Equi librate 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 t he syr inge tip into the sample loading port and overfill the 100-µL loop at least three times (that is, 300 µL).

5. Inject the sample.

Startup Procedu re ” on page 61.

Alkali and Alkaline Earth

Figure 4-4

IC-P a k C M/D column with 0.1 mM EDTA/3 mM HNO separation of the standard typically takes 20 to 25 minutes with this setup.

shows a representative chromatogram of an 8-cation standard run on an

eluent at 1.0 mL/min flow rate. The

Standards for Ion Analysis 76

Page 77

Figure 4-4 Chromatogram of an 8-Cation Standard

Performing Ion Analysis 77

Page 78

Chapter 5 Maintenance

This chapter contains information about maintaining the 432 Detector and troubleshooting charts to help you isolate and correct problems.

Caution:

the power cord before you service the instrument .

To avoid the possibility of electric shock, power off the detector and disconnect

5.1 Routine Maintenance

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 suggest ions to help you maintain the 432 Detector :

• Stock the recommended spare parts listed in Appendix B to reduce downtime.

Contact the Waters Service Departmen t for assistance.

• Record the initial installation date and serial number of your instrument in Appendix B

for easy reference.

• Keep a file of typical chromatograms for comparison when you suspect problems.

5.1.1 Replacing the Fuse

To change the operating voltage fuse:

1. Turn off the front panel power switc h and remove the power cord from its connector on the rear panel of the detector.

Caution:

power switch, and unplug the power cord from the rear panel.

To avoid the possibility of electric shock, turn off the front panel

Routine Maintenance 78

Page 79

2. Pry open the power connector cover with a screwdriver.

3. To change the AC power fuses, pull out each fuse holder as though openi ng a drawer. Spare fuses are included in the System Startup Kit. For ordering information, see A ppendi x B,

4. Table 2-2 on page 34 America or Europe).

5. Insta ll the correct fuse in the holder and slide it back into place (Figure 5-1 arrow on each fuse holder points up when in the correct position.

Figure 5-1 Installing Operating Voltage Fuses

lists the operating voltage fuses (for use in either North

Spare Parts.

). The

6. Close the power connector cov er . Then plug the power cord into its connector on the rear panel of the detector.

5.1.2 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.

Attention:

STO

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 (HNO

3. Flush the system again with ASTM Type I reagent water . Do not reconnect the

To avoid damaging the co lum n, re move it befo re you flush the system. Do not

column until the eluent has returned to about pH 7.

Maintenance 79

Page 80

Refer to Appendix B , Spare Parts, to order a replacement flow c ell. The following tools are required to replace the flow cell:

• Phillips-head screwdr iver

• 5/16-inch open-end wrench

• Knife or flat-blade screwdriver

Caution:

any of the following replacement procedures.

1. Unplug the 432 Detector from the power source, and completely disconnect all

2. Rem ove the 432 Detector cover (four Phillips-head screws, two on each side).

3. Remove the two pins and pin holders that hold the cell block cover in place (see

4. Pull off the cover of the flow cell unit and remove the top layer of insulation.

5. Remove the four Phillips-head screws from the upper plate of the cell block, and

6. Carefully disconnect the inlet and outlet fittings from the flow cell.

7. Remove the two Phillips-head screws from the flow cell mounting bracket.

8. Unp lug the flow cell cable connector from its socket in the cell block.

9. Remove the flow cell assembly.

10. I nsta ll the new flow cell by following steps 2 through 9 in reverse order. Be sure to

To avoid electrical hazards, always unplug the power cord before you perform

electrical cables and fluid connections.

Figure 5-2). Use a knife or flat-blade screwdriver to gently pry the pins and holders

out.

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 block.

orient the upper plate of the cell block properly before you install the four screws.

Routine Maintenance 80

Page 81

Pin holder

Upper plate screws

Cell mounting bracket

Cell block cover

Pin holder

Insulation

Upper plate o cell block

Cell mounting bracket screws

Flow cell

Connector

Cell block

Figure 5-2 Flow Cell Assembly

Maintenance 81

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5.2 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.

5.3 Troubleshooting

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 follo wing information availab le when you call Waters regarding a 432 Detector problem:

• Symptom

• Type of column

• Eluent(s)

•Flow rate

• 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 illumina te and the display remains completely blank when the unit is tur ned on), the fuse may require replacement. Refer to Section 5.1.1,

Replacing the Fuse.

Cleaning the Detector Exterior 82

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Startup Diagnostics

The 432 Detector performs startup diagnostics that check the internal memory (both RAM and ROM), and the associated processing circuitry.

Power Supply

The following factors can adversely affect the operation of the 432 Detector:

• Power surges

• Line spikes

• Transient energy sources

Be sure that the electrical supply used for the 432 Detector is properly grounded and free from any of these conditions.

Error Messages

The error messages displayed by the 432 Detector are listed below along with the recommended action for each:

• Error: ROM/RAM

ROM/RAM error (checked during startup only) Call Waters service.

• Error: Leak

Leakage detected Check flow cell and associated plumbing connections.

• Error: Temp

Temperature control failure Call Waters ser v ic e.

• Error: Over Range

Base over-range condition Set Base Range to the next setting above the background conductivity of the eluent.

• Error: Over Flow

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.

Troubleshooting Procedure

As soon as you realize that a problem may exist:

1. Visually examine the integrity of the electrical and fluid connections as you verify proper system configuration and installation.

Maintenance 83

Page 84

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 leak)?

• Are pressure fluctuations in the same range as during normal operation?

3. Isolate the parameter that varies from normal operation. The parameters to observe include:

• Baseline noise

• Peak retention time

• Peak resolution

• Qualitative/quantitative chromatographic results

• System pressure Evaluate the parameters in the order presented above to rule out simpl e causes of

the problem.

4. Use Table 5-1

to determine corrective actions for the problems that you identify.

Removing Bubbles

Bubbles in the flow cell are evident when the noise is equal to or greater than 2 µS. Use this method to remove bubbles.

1. Discon nec t 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. Rea ttach the tubing from the 432 De tector outlet to a waste receptacle (18-inch length of 0.009-inch ID stainless steel).

7. St art eluent flowing through the system at a flow rate of at least 1 mL/min.

8. W ith the eluent flowing, reattach the detector inlet tubing to the column.

9. Allow a few minutes for temperature reequilibration, then check the noise le vel. If it is not reduced, repeat steps 1 through 4, then proceed to steps 10 through 13.

10. A t tach 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.

Troubleshooting 84

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12. A llow the detector to stand overnight (>12 hours) with temperature on and with methanol in the flow cell.

13. Repeat steps 5 through 9.

Table 5-1 Troubleshooti ng Guide

Symptom Possible Cause Solution

Noisy baseline Pulse dampener not installed See “Installing the Pulse

Dampener” on page 52.

Pulsing pump Check the pump; see the

pump manual.

Bubbles in flow c ell Rem ove bubbles and degas

the solvent.

Voltage fluctuation Use the voltage regulator.

Spikes on baseline Dirty flow cell Cl ean the cell.

Flow cell leak Check flow cell fittings and

tighten. If leak continues, replace the flow cell.

Bubbles in flow c ell Rem ove bubbles and degas

the solvent.

Irregular noise on baseline

Excessive bas eli n e drift

Temperature changes in room Cont rol ambient tempera-

ture, locate drafts, and insulate tubing and column, if necessary.

Cell temperature set lower than ambient

Defective column Replace the column. Unstable temperature control Make sure the temperature

Defective cell heater Call Waters service. Temperature changes in room Cont rol ambient tempera-

Cell temperature set lower than ambient

Set the cell temperature to a minimum of 5 °C above ambient.

control is turned on.

ture, locate drafts, and insulate tubing and column, if necessary.

Set the cell temperature to a minimum of 5 °C above ambient.

Maintenance 85

Page 86

Table 5-1 Troubleshooti ng Guide (Continued)

Symptom Possible Cause Solution

Bubbles in flow c ell Rem ove bubbles and degas

Solvent changeover Wait until baseline stabilizes

Flow cell leak Check flow cell fittings and

Detector cannot be zeroed

Continuous noise at high sensitivity (<1µS)

Solvent changeover Wait until the baseline

Bubbles in flow c ell Rem ove bubbles and degas

Pump crossover noise Install a high-sensitivity noise

the solvent.

(purge autosampler a few times).

tighten. If leak continues, replace the flow cell.

stabilizes.

the solvent.

filter on the pump.

Troubleshooting 86

Page 87

Appendix A Specifications

This appendix includes information on:

• Operational specifications

• Mechanical specifications

• Environmental specifications

• Electr ic a l s pe c if ic ations

• Communications

Table A-1 Operational Specifications

Condition Specification

Drift Less than 0.05 µS/hr/°C (ambient)

Base : 200 µS Sensitiv ity: 0.005 Eluent: 1 mM KCI

Noise Less than 0.005 µS/cm

Base : 200 µS Sensitiv ity: 0.005 Eluent: 1 mM KCI

Temperature control Front-panel selectable: OFF, 30, 35, 40, 45,

50, 55, 60, 65 °C Performa nce: 0.5 °C/hr

Response times Fast: 0. 2 5 sec

Standard: 0.5 sec Slow: 1.0 sec

Table A-2 Me chanical Spec ifications

Condition Specification

Cell volum e 0.6 µL Wetted materials 316 stainless steel, PTFE, and PCTFE

Specifications 87

Page 88

Table A-2 Me chanical Spec ifications (Continued)

Condition Specification

Operating pressure 70 psi maximum Height 9.4 inches (23.8 cm) Length 21 inches (53.3 cm) Width 11.5 inches (29.2 cm) Weight 17.7 pounds (8 kg)

Table A-3 Environmental Specifications

Condition Specification

Operating temperature range 4 to 35 °C

(40 to 95 °F)

Operating humidity 20 to 80% RH, noncondensing

Table A-4 Electrical Specifications

Condition Spec ification

Protection class Over-voltage category Pollution degree Moisture protection

Line voltages (grounded

AC), nominal

Class I II 2 Normal (IPXO) 100/120 VAC

220/240 VAC

Line frequency ranges 50 Hz: ±2 Hz

60 Hz: ±2 Hz 100/120 VAC fuse rating T2 A (20 mm) 220/240 VAC fuse rating T1 A (20 mm) Current (Max) 0.6 A

a. Protection Cla ss I – The insulating schem e used in the instrument to

protect you from electrical shock. Class I identifies a single le vel of insulation between live parts (wires) and e xposed conductive parts (meta l 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.

Specifications 88

Page 89

b. Over Voltage Category II – Pertains to inst ruments that receive th eir

electrical power from a local level such as an electrical wall outlet.

c. Pollution Degree 2

may prod uce a reduct ion of dielec tric st rength or surface resistivit y. Degree 2 refers to normally only nonconductive pollut ion. Occasionally, however, a temporary conductivity caused by condensation must be expected.

d. Moisture Protection – Normal (IPXO) – IPXO means that there is NO

Ingress Pro tecti on agai nst an y t ype of dri pping or sprayed wat er. The X is a placeholder to identify protection against dust, if applicable.

Table A-5 Communications

Signal Specification

Recorder output 0 to 10 m V Integrator output 10, 50, 100 µS/1V FS

Marker output Isolated contact output Controller bus IEEE-488, PowerLine™

–

A measure of pollution on el ectrical circuits, which

Specifications 89

Page 90

Appendix B Spare Parts

The parts listed in Table B-1 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 in Table B-1 to keep in stock for use as needed.

Table B-1 Spare Parts

Item Quantity Part Number

Fuse, Time Delay, 1A, 250V, IEC 2 WAT1 65-11 Fuse, Time Delay, 2A, 5x20 mm T 2 WAT165-14 Fitting kit 1 WAT025604 Pump Seal Replacement kit 2 WAT022934 Union 1 WAT097332

The flow cell (part number 043069) is considered a replacement part. Order the

Note:

flow cell only when it is needed for replacement in the Waters 432 Detector.

Fill in the infor ma tion below for easy reference when you order part s or request serv ice.

Installation Date: _____________

Serial Number: ____________

Spare Parts 90

Page 91

Appendix C Ion Chromato grap hy Methods

This appendix provides information about:

• General-purpose anion analysis using conductivity and UV detection

• Alkali and alkaline earth cations, ammoni um, and amines

C.1 General-Purpose Anion Analysis Using

Conductivity and UV Detection

Table C-1 Required Instrumentation

Instrument Part Number

Alliance, 2695 Separations Module or Breeze (with column heater, seal wash, and degasser)

432 Conductivity Detector 043061 busSAT/IN Mod ule 200415 Empower/Breeze data processing Contact Waters UV Detector (optional) Contact Waters

N/A

Ion Chromatography Methods 91

Page 92

Figure C-1 Common Anion Standards

Table C-1 Analysis Conditions

Condition Value

Column IC-Pak A/HR (part number 026765) Eluent Borate/gluconate containing 12% AcCN Back conductivity 240 ±20 µS Degas Continuous Flow rate 1 mL/min. Backpressure 1200 ±200 psi Temperatu re 30 °C for column heater, 35 °C for detector Injection 100 µL Needle wash 12% AcCN in DI water Detection Di rect conductivity Base range 500 Attenuation 50 µS/Volt unattenuated Polarity Positive

Ion Chromatography Methods 92

Page 93

C.1.1 Preparing Eluent

1. Add 20 m L of lithium borate/gluconate concent rate and 120 mL of HPLC-grade acetonitrile (AcCN) into a 1-liter volumetric flask (see Section C.1.13,

Lithium Borate/Gluconate 50X Stock Concentrate, 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.

C.1.2 Preparing Standards

It is recommended that you use certified 1000-ppm anion standards with this method. If unavailable, see Section C.2.7, 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. After you validate the multi-point calibration curve, a single-point calibration curve within the expected analyte concentration range is appropriate for recalibration.

Preparing

Preparing Stock Reagent, for uncertified standard

C.1.3 Preparing 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 C unretained, but there can be a loss of fluoride recovery.

4. Filter samples containing suspended solids through a 0.45-µm aqueous-compatible disk before injection. Failure to filter solids can increase column backpressure.

For best results, ensure sample pH is between 3 and 11.

Note:

Samples treated with a sample preparation disk in the H cations and neutralize high pH, will yield chromatograms similar to suppressed conductivity chromatograms.

Sep-Pak cartridge. Anions pass through

form, used to remove

Ion Chromatography Methods 93

Page 94

C.1.4 Empower Data Processing Method

Table C-2 IC Processing Method Using Peak Apex for Retention Time

Process Values

Integration Peak Width = 30.0

Minimum Area = 3000 Inhibit Intg. = 0 to 2 min. Threshold = 10 to 25 Minimum Height = 150

Calibration Averaging = None

Update RT = Never Peak Match = Closest Quant By = Peak Area Fit Type = Linear for multi-point calibration,

Linear Through Zero for single-point calibration

Report Analyte Name

Analyte Retention Time Peak Area Amounts

Ion Chromatography Methods 94

Page 95

C.1.5 Method Validation

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 validation design are linear above 50 ppm.

Table C-3 Method Val idation

Individual Youden Pair Standard, in ppm

1 2 3 4 5 6 7 8

Cl 0.7 2.0 3.0 15.0 40.0 20.0 50.0 0.5 Br 2.0 3.0 15.0 40.0 20.0 50.0 0.7 0.5 NO SO NO

Analyte Anion

F 2.0 0.7 0.5 3.0 10.0 7.0 20.0 25.0 PO

3.0 40.0 20.0 15.0 50.0 0.5 2.0 0.7

40.0 50.0 0.5 0.7 2.0 3.0 15.0 20.0

15.0 20.0 40.0 50.0 0.5 0.7 2.0 3.0

50.0 40.0 20.0 0.5 3.0 2.0 0.7 15.0

C.1.6 Method Linearity

Cl r2 = 0.9999

= 0.9986

Br r

= 0.9999

Figure C-2 Calibration Curves for Chloride, Fluoride, and Bromide

Ion Chromatography Methods 95

Page 96

NO2 r2 = 0.9999 NO

r2 = 0.9992

Figure C-3 Calibration Curves for Nitrite and Nitrate

SO4 r2 = 0.9999

r2 = 0.9992

Figure C-4 Calibration Curves for Sulfate and Phosphate

Ion Chromatography Methods 96

Page 97

C.1.7 Quantitation Precision

100-PPB Standards

Quantitation Precision is the percent RSD of analyte peak area at each concentration. Data is based on seven replicate injections of the validation standards.

Table C-4 Quantitation Precision

Analyte F CI NO

0.5 0.95 1.11 3.44 5.17 0.32 12.98 7.62

0.7 0.67 1.64 0.78 1.73 0.75 9.29 3.90 2 0.17 0.18 0.56 1.14 0.91 2.91 1.07 3 0.43 0.17 0.20 0.67 0.19 3.49 0.64 15 0.44 0.05 0.28 0.11 0.16 0.50 0.32 20 0.45 0.04 0.05 0.30 0.06 0.52 0.25 40 0.05 0.04 0.03 0.08 0.37 0.26

ppm Concentration

50 0.13 0.02 0.26 0.03 1.56 0.16

C.1.8 Method Detection Limits

Br NO

Figure C-5 100-µL Injection

Ion Chromatography Methods 97

Page 98

Based on this representative chromatogram using a 100-µL injection , the estimate d detection limits, as ppb, at three times signal to noise (S/N) are as follows:

• Fluoride = 50

• Chloride = 25

• Nitrite = 50

• Bromide = 75

•Nitrate = 75

• Phosphate = 125

• Sulfate = 75

Quantitation below these detection limits is not advised. You can obtain lower detection limits using a 250-µL injection.

C.1.9 Quantitation Accuracy

The Certified Performance Evaluation Standards were diluted 1:100 with DI water. Amounts are based on a multi-point calibration curve prepared from certified standards.

Table C-5 Quantitation Accuracy

Analyte F CI NO

Performance

Evaluation

Standard

True V al ue in ppm

2.69 43.00 1.77 15.37 6.29 37.20

Official Anion

Methods Wet

Chem & IC

IC Using

Alliance IC

Pak A/HR and

B/G Eluent

Measured Mean

Measured Std Dev

Ave IC n=3 2.63

IC/Mean 0.956 1.013 1.090 0.975 1.014 1. 001 IC/True

Value

2.75 43.30 1.77 15.42 6.38 37.00

0.26 3.09 0.07 1.15 0.21 2.24

43.87

±0.05

0.978 1.020 1.090 0.979 1.029 0.995

±0.09

1.93 ±0.01

15.04 ±0.06

Ion Chromatography Methods 98

6.47 ±0.09

37.03 ±0.12

Page 99

C.1.10 Analyte Recovery

The Certified Performance Evaluation Standards were diluted 1:100 with typical drinking water. Amounts are based on a multi-point calibration.

Table C-6 Analyte Recovery

Analyte F CI NO

Milford

Drinking W ater

n=3, as ppm

%RSD

Performance

Evaluation Std

MD W + PES

n=3; as ppm

%RSD

% Recovery

Not detected

2.69 43. 00 1.77 15.37 6.29 37.20

2.46 ±0.04

1.51% 0.11% 0.21% 0.10% 0.92% 0.37%

91.4% 102.5% 102.8% 99.5% 85.1% 102.8%

25.82 ±0.04

0.16 0.92 0.27

69.64 ±0.08

Not detected

1.82 ±0.004

0.23 ±0.002

15.52 ±0.02

Not detected

5.35 ±0.05

8.30

0.02

46.46 ±0.17

Ion Chromatography Methods 99

Page 100

C.1.11 Example of Use

Figure C-6 Typical Dr inking Water, No Dilution Required

C.1.12 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. Figure C-7 the chromatogram of the anion standard that demonstrates this selectivity. Generally, the lower the wa velength 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 negat ives peaks at the retention time of the UV transparent anion, such as F, Cl, PO

, and SO4, if present.

shows

Ion Chromatography Methods 100

Waters 432 Operator's Manual

Specifications and Main Features

Frequently Asked Questions

User Manual

Table of Contents

List of Figures

List of Tables

Preface

Chapter 1 Introduction

Chapter 2 Installing the Detector

2.1 Selecting the Installation Site

2.2 Unpacking and Inspection

2.3 AC Power Connection

2.4 I/O Signal Connections

2.4.1 I/O Signal Descriptions

2.4.2 PowerLine Controller Connections

2.4.3 Empower and Mille nn ium32 Connections

2.4.4 Data Module Connections

2.4.5 Chart Recorder Connections

2.4. 6 Chart Marker Input Connections

2.4.7 Auto Zero Input Connections

2.4.8 Alliance Separations Module Connections

2.5 Making Fluidic Connections

2.6 Passivating the System

2.7 Verifying the Detector

Chapter 3 Operating the Detector

3.1 Controls and Indicators

3.2 Startup and Shutdown

3.3 Operating Recommendations

Chapter 4 Performing Ion Analysis

4.1 Fundamental Considerations

4.2 Configuring the System

4.3 Eluents for Ion Analysis

4.3.1 Preparing Anion Eluent

4.3. 2 Preparing Cation Eluent

4.4 Standards for Ion Analysis

4.4.1 Preparing Anion Standards

4.4.2 Injecting Anion Standards

4.4.3 Preparing Cation Standards

4.4.4 Injecting Cation Standards

Chapter 5 Maintenance

5.1 Routine Maintenance

5.1.1 Replacing the Fuse

5.1.2 Maintaining the Flow Cell

5.2 Cleaning the Detector Exterior

5.3 Troubleshooting

Appendix A Specifications

Appendix B Spare Parts

Appendix C Ion Chromato grap hy Methods

C.1.1 Preparing Eluent

C.1.2 Preparing Standards

C.1.3 Preparing a Sample

C.1.4 Empower Data Processing Method

C.1.5 Method Validation

C.1.6 Method Linearity

C.1.7 Quantitation Precision

C.1.8 Method Detection Limits

C.1.9 Quantitation Accuracy

C.1.10 Analyte Recovery

C.1.11 Example of Use

C.1.12 Using Direct UV Detection