Waters 600e User manual

Waters 600E
Multisolvent Delivery System
User’s Guide
34 Maple Stre
et
Milford, MA 0175
7
WAT174-02TP, Revision
3
View More Waters Manuals at
www.Chromatography.co

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 accurate at the time of publication. In no event shall Waters
Corporation be liable for incidental or consequential damages in connection with, or
arising from, the use of this document.
© 1993–2003 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF
AMERICA AND IRELAND. ALL RIGHTS RESERVED. THIS DOCUMENT OR
PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE
WRITTEN PERMISSION OF THE PUBLISHER.
Millennium and Waters are registered trademarks, and Empower, Auto•Blend,
ExpertEase, LAC/E, PIC, PowerLine, Radial-Pak, and WISpare are trademarks of Waters
Corporation.
Hanson Transfer Controller is a copyright of Hanson Research Corporation.
All other trademarks or registered trademarks are the sole property of their respective
owners.

Note: When you use the instrument, follow generally accepted procedures for quality
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.
Note: The Installation Category (Overvoltage Category) for this instrument is Level II. The
Level II Category pertains to equipment that receives its electrical power from a local level,
such as an electrical wall outlet.
STOP Attention: Changes or modifications to this unit not expressly approved by the party
responsible for compliance could void the user’s authority to operate the equipment.
Important : Toute modification sur cette unité n’ayant pas été expressément approuvée
par l’autorité responsable de la conformité à la réglementation peut annuler le droit de
l’utilisateur à exploiter l’équipement.
Achtung: Jedwede Änderungen oder Modifikationen an dem Gerät ohne die
ausdrückliche Genehmigung der für die ordnungsgemäße Funktionstüchtigkeit
verantwortlichen Personen kann zum Entzug der Bedienungsbefugnis des Systems
führen.
Avvertenza: eventuali modifiche o alterazioni apportate a questa unità e non
espressamente approvate da un ente responsabile per la conformità annulleranno
l’autorità dell’utente ad operare l’apparecchiatura.
Atención: cualquier cambio o modificación efectuado en esta unidad que no haya sido
expresamente aprobado por la parte responsable del cumplimiento puede anular la
autorización del usuario para utilizar el equipo.

Caution: Use caution when working with any polymer tubing under pressure:
• Always wear eye protection when near pressurized polymer tubing.
• Extinguish all nearby flames.
• Do not use tubing that has been severely stressed or kinked.
• Do not use nonmetallic tubing with tetrahydrofuran (THF) or concentrated nitric or
sulfuric acids.
• Be aware that methylene chloride and dimethyl sulfoxide cause nonmetallic tubing
to swell, which greatly reduces the rupture pressure of the tubing.
Attention : Manipulez les tubes en polymère sous pression avec precaution:
• Portez systématiquement des lunettes de protection lorsque vous vous trouvez à
proximité de tubes en polymère pressurisés.
• Eteignez toute flamme se trouvant à proximité de l’instrument.
• Evitez d'utiliser des tubes sévèrement déformés ou endommagés.
• Evitez d'utiliser des tubes non métalliques avec du tétrahydrofurane (THF) ou de
l'acide sulfurique ou nitrique concentré.
• Sachez que le chlorure de méthylène et le diméthylesulfoxyde entraînent le
gonflement des tuyaux non métalliques, ce qui réduit considérablement leur
pression de rupture.
Vorsicht: Bei der Arbeit mit Polymerschläuchen unter Druck ist besondere Vorsicht
angebracht:
• In der Nähe von unter Druck stehenden Polymerschläuchen stets Schutzbrille
tragen.
• Alle offenen Flammen in der Nähe löschen.
• Keine Schläuche verwenden, die stark geknickt oder überbeansprucht sind.
• Nichtmetallische Schläuche nicht für Tetrahydrofuran (THF) oder konzentrierte
Salpeter- oder Schwefelsäure verwenden.
• Durch Methylenchlorid und Dimethylsulfoxid können nichtmetallische Schläuche
quellen; dadurch wird der Berstdruck des Schlauches erheblich reduziert.

Attenzione: prestare attenzione durante l’utilizzo dei tubi di polimero pressurizzati:
• Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero
pressurizzati.
• Estinguere ogni fonte di ignizione circostante.
• Non utilizzare tubi soggetti che hanno subito sollecitazioni eccessive o son stati
incurvati.
• Non utilizzare tubi non metallici con tetraidrofurano (THF) o acido solforico o nitrico
concentrato.
• Tenere presente che il cloruro di metilene e il dimetilsolfossido provocano
rigonfiamento nei tubi non metallici, riducendo notevolmente la resistenza alla
rottura dei tubi stessi.
Advertencia: se recomienda precaución cuando se trabaje con tubos de polímero
sometidos a presión:
• El usuario deberá protegerse siempre los ojos cuando trabaje cerca de tubos de
polímero sometidos a presión.
• Si hubiera alguna llama las proximidades.
• No se debe trabajar con tubos que se hayan doblado o sometido a altas
presiones.
• Es necesario utilizar tubos de metal cuando se trabaje con tetrahidrofurano (THF)
o ácidos nítrico o sulfúrico concentrados.
• Hay que tener en cuenta que el cloruro de metileno y el sulfóxido de dimetilo
dilatan los tubos no metálicos, lo que reduce la presión de ruptura de los tubos.

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 informé que si le matériel 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 ordnungsgemäß funktionieren.
Attenzione: 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.
Advertencia: el usuario deberá saber que si el equipo se utiliza de forma distinta a la
especificada por el fabricante, las medidas de protección del equipo podrían ser
insuficientes.

Caution: To protect against fire hazard, replace fuses with those of the same type and
rating.
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.
Attenzione: per una buona protezione contro i rischi di incendio, sostituire i fusibili con
altri dello stesso tipo e amperaggio.
Advertencia: sustituya los fusibles por otros del mismo tipo y características para evitar
el riesgo de incendio.

Caution: To avoid possible electrical shock, disconnect the power cord before servicing
the instrument.
Attention : 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 Vermeidung von Stromschlägen sollte das Gerät vor der Wartung vom
Netz getrennt werden.
Attenzione: per evitare il rischio di scossa elettrica, scollegare il cavo di alimentazione
prima di svolgere la manutenzione dello strumento.
Precaución: para evitar descargas eléctricas, desenchufe el cable de alimentación del
instrumento antes de realizar cualquier reparación.

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
Terminale di conduttore con protezione
Borne del conductor de tierra

Frame or chassis terminal
Borne du cadre ou du châssis
Rahmen- oder Chassisanschluss
Terminale di struttura 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
Attenzione, superficie calda o elevata temperatura
Precaución, superficie caliente o temperatura elevada
Commonly Used Symbols (Continued)

Caution, risk of electric shock (high voltage)
Attention, risque de commotion électrique (haute tension)
Vorsicht, Elektroschockgefahr (Hochspannung)
Attenzione, 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
Attenzione, rischio di puntura con ago
Precaución, riesgo de punción con aguja
Caution, ultraviolet light
Attention, rayonnement ultrviolet
Vorsicht, Ultraviolettes Licht
Attenzione, luce ultravioletta
Precaución, emisiones de luz ultravioleta
Commonly Used Symbols (Continued)
UV

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 disattivata
Alimentación eléctrica desconectada
Commonly Used Symbols (Continued)
1
0

United States – FCC Emissions Notes
This device complies with Part 15 of the FCC Rules. Operation is subject to the following
two conditions: (1) this device may not cause harmful interference, and (2) this device
must accept any interference received, including interference that may cause undesired
operation.
Changes or modifications to this unit not expressly approved by the party responsible for
compliance could void the user’s authority to operate the equipment.
Note: This equipment has been tested and found to comply with the limits for a Class B
digital device, pursuant to Part 15 of the FCC Rules. These limits are designed to provide
reasonable protection against harmful interference in a residential installation. This
equipment generates, uses, and can radiate radio frequency energy and, if not installed
and used in accordance with the instructions, may cause harmful interference to radio
communications. However, there is no guarantee that interference will not occur in a
particular installation. If this equipment does cause harmful interference to radio or
television reception, which can be determined by turning the equipment off and on, the
user is encouraged to try to correct the interference by one or more of the following
measures:
• Reorient or relocate the receiving antenna.
• Increase the separation between the equipment and receiver.
• Connect the equipment into an outlet on a circuit different from that to which the
receiver is connected.
• Consult the dealer or an experienced radio TV technician for help.
Shielded cables must be used with this unit to ensure compliance with the Class B FCC
limits.
Canada – Spectrum Management Emissions Notes
Cet appareil numérique de la classe B est conforme à la norme NMB-003.
This Class B digital apparatus complies with Canadian ICES-003.

600E Pump Information
Intended Use
When you develop methods, follow the “Protocol for the Adoption of Analytical Methods
in the Clinical Chemistry Laboratory,” American Journal of Medical Technology, 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 precautions 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, normal, and above-normal levels of a compound. Ensure that quality-control
sample results are within an acceptable range, and evaluate 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.

Table of Contents xvii
Preface......................................................................................................................... xxv
Chapter 1
Introduction ................................................................................................................. 1
1.1 Waters 600E System Overview......................................................................... 1
1.2 Waters 600E System Modules........................................................................... 3
1.2.1 Waters 600 Controller ......................................................................... 3
1.2.2 Waters 600E Pump .............................................................................. 6
1.3 PowerLine Controller Features ......................................................................... 9
1.4 Gradient Controller Features............................................................................. 9
1.5 Options and Accessories ................................................................................. 10
Chapter 2
Basic Operation ........................................................................................................ 13
2.1 Powering Up the System................................................................................. 13
2.1.1 Powering Up IEEE-488 Components ............................................... 13
2.1.2 Powering Up the Stand-Alone 600E System .................................... 16
2.2 Running Diagnostics ....................................................................................... 16
2.2.1 Powerup Self-Diagnostics ................................................................. 16
2.2.2 Running Extended Diagnostics ......................................................... 17
2.3 Using the Controller Front Panel .................................................................... 17
2.3.1 Keypad .............................................................................................. 18
2.3.2 Function Keys ................................................................................... 19
2.3.3 Screen Keys ...................................................................................... 20
2.4 Comparing Controller Types........................................................................... 21
2.4.1 PowerLine Controller Configuration ................................................ 22
2.4.2 Gradient Controller Configuration .................................................... 24
2.5 Powering Down the System............................................................................ 25
Table of Contents

xviii Table of Contents
Chapter 3
Preparing Your 600E System for Operation .................................................. 27
3.1 Sparging Eluents.............................................................................................. 28
3.2 Priming the Pump............................................................................................ 31
3.3 Daily Routine Operation ................................................................................. 34
3.4 Flushing the System........................................................................................ 35
3.4.1 Flushing the System Fluid Lines ...................................................... 36
3.4.2 Flushing the Rheodyne Manual Injector ........................................... 36
3.4.3 Purging the Autoinjector ................................................................... 37
3.4.4 Reconnecting the Column After Flushing the System ...................... 37
3.5 Equilibrating the System ................................................................................. 38
3.6 Changing Eluents ............................................................................................ 41
3.6.1 Adding Fresh Eluent ......................................................................... 41
3.6.2 Changing Between Incompatible Eluents ......................................... 42
Chapter 4
Setting PowerLine Controller Operating Parameters ............................... 45
4.1 Setting Up the 600 as a PowerLine Controller................................................ 47
4.1.1 Configuring the Controller Type ....................................................... 49
4.1.2 Selecting an Integrator ...................................................................... 51
4.1.3 Selecting the Report Type ................................................................. 54
4.2 Verifying that PowerLine Devices Are Active................................................ 61
4.3 Setting Up the 600E Pump.............................................................................. 63
4.3.1 Accessing the Pump Setup Screen .................................................... 65
4.3.2 Setting Pump Setup Parameters ........................................................ 66
4.3.3 Exiting the Pump Setup Screen ........................................................ 71
4.4 Setting Up the Autoinjector............................................................................. 71
4.4.1 Accessing the Autoinjector Setup Screen ......................................... 73
4.4.2 Setting Autoinjector Setup Parameters ............................................. 74
4.4.3 Exiting the Autoinjector Setup Screen .............................................. 75

Table of Contents xix
4.5 Setting Up Dissolution Control....................................................................... 76
4.6 Setting Up the Detectors ................................................................................. 76
4.6.1 Accessing the Detector Setup Screen ............................................... 79
4.6.2 Setting Detector Setup Parameters ................................................... 80
4.6.3 Exiting the Detector Setup Screen .................................................... 83
Chapter 5
Running PowerLine in Direct Control Mode ................................................ 85
5.1 Setting Run Conditions ................................................................................... 86
5.1.1 Setting Pump Parameters .................................................................. 88
5.1.2 Setting External Event Switches ....................................................... 90
5.1.3 Setting Autoinjector Parameters ....................................................... 91
5.1.4 Setting Detector Parameters .............................................................. 93
5.2 Performing a Direct Control Run.................................................................... 94
5.2.1 Setting the Run Time ........................................................................ 96
5.2.2 Downloading Operating Parameters ................................................. 96
5.2.3 Equilibrating the System ................................................................... 97
5.2.4 Starting a Run ................................................................................... 97
5.2.5 Stopping the Pump ............................................................................ 98
5.2.6 Aborting a Run .................................................................................. 99
5.2.7 Changing Parameters During a Run ................................................. 99
5.3 Running a Method from the Direct Control Screen........................................ 99
Chapter 6
Creating PowerLine Methods ........................................................................... 101
6.1 Creating a Gradient Table.............................................................................. 102
6.1.1 Documenting the Gradient Table .................................................... 103
6.1.2 Programming the Gradient Table .................................................... 105
6.1.3 Entering Parameter Values in the Gradient Table ........................... 108
6.1.4 Editing the Gradient Table .............................................................. 109

xx Table of Contents
6.2 Creating an Event Table................................................................................ 111
6.2.1 Documenting the Event Table ......................................................... 112
6.2.2 Programming the Event Table ........................................................ 115
6.2.3 Entering Parameter Values in the Event Table ................................ 118
6.2.4 Editing the Event Table ................................................................... 119
6.3 Creating a Detector Table.............................................................................. 119
6.3.1 Documenting the Detector Table .................................................... 119
6.3.2 Programming the Detector Table .................................................... 120
6.3.3 Entering Parameter Values in the Detector Table ........................... 123
6.3.4 Editing the Detector Table .............................................................. 124
6.4 Creating a Method Table............................................................................... 124
6.4.1 Method Table Overview ................................................................. 124
6.4.2 Programming the Method Table ..................................................... 125
6.4.3 Entering Parameter Values in the Method Table ............................. 128
6.4.4 Editing the Method Table ................................................................ 130
Chapter 7
Running PowerLine Methods ........................................................................... 133
7.1 Using an Injector........................................................................................... 133
7.1.1 Using the Rheodyne 7725i Manual Injector ................................... 133
7.1.2 Using a PowerLine Autoinjector .................................................... 135
7.1.3 Using a Non-PowerLine Autoinjector ............................................ 135
7.2 Setting Initial Operating Parameters ............................................................. 136
7.3 Starting a Run................................................................................................ 139
7.3.1 Accessing the Operate Methods Screen .......................................... 139
7.3.2 Downloading Operating Parameters ............................................... 141
7.3.3 Running a Method .......................................................................... 142
7.4 Modifying Conditions While Running.......................................................... 143
7.4.1 Changing Parameters ...................................................................... 144
7.4.2 Holding Conditions Constant .......................................................... 144