Waters 2410 Manual

Waters 2410
Differential Refractometer
Operator’s Guide
34 Maple Street
Milford, MA 01757
71500241002, Revision 3

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 guide 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 guide.

2001 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF AMERICA.
ALL RIGHTS RESERVED. THIS DOCUMENT OR PARTS THEREOF MAY NOT BE
REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE
PUBLISHER.
Millennium
®
PIC and Waters are registered trademarks, and busLAC/E and PowerStation are trademarks of
Waters Corporation.
Micromass is a registered trademark and MassLynx is a trademark of Micromass Ltd.
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 take steps to determine the
reason for the changes.Until you determine the cause of a change, do not rely upon the
results of the separations.
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.
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 modefication sur cette unité n’ayant pas été expressément approuvée
par l’autorité résponsable de la conformité à la réglementation peut annuler de droit de
l’utilisateur à exploiter l’équipement.
Achtung : Jedewede 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 realizado a esta unidad que no haya sido
expresamente aprobado por la parte responsable del cumplimiento puede anular la
autorización de la que goza el usario para utizar el equipo.
STOP

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 Tefzel tubing that has been severely stressed or kinked.
• Do not use Tefzel tubing with tetrahydrofuran (THF) or concentrated nitric or
sulfuric acids.
• Be aware that methylene chloride 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 (THT).
• Sachez que le chlorure de méthylène et le sulfoxyde de diméthyle peuvent
provoquer le gonflement des tuyaux de Tefzel, diminuant ainsi fortement 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 quellen;
dadurch wird der Berstdruck des Schlauches erheblich reduziert.

Precauzione : 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 notevolmente il limite di pressione di
rottura dei tubi stessi.
Advertencia: manipular con precaución los tubos de polimero bajo presión:
• Protegerse siempre los ojos a proximidad de tubos de polimero 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 o ácidos nítrico o sulfúrico
concentrados.
• No olvidar que el cloruro de metileno y el óxido de azufre dimetilo inflan los tubos
Tefzel lo que reduce en gran medida 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.
Precauzione: 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.
Precauzione: per una buona protezione contro i rischi di incendio, sostituire i fusibili
con altri dello stesso tipo e amperaggio.
Precaución: sustituya los fusibles por otros del mismo tipo y características para evitar
el riesgo de incendio.

Caution: To avoid possible electrical shock, power off the instrument and disconnect the
power cord before servicing the instrument.
Attention: Afin d’éviter toute possibilité de commotion électrique, mettez hors tension
l’instrument et 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
abgeschaltet und vom Netz getrennt werden.
Precauzione: per evitare il rischio di scossa elettrica, spegnere lo strumento e scollegare
il cavo di alimentazione prima di svolgere la manutenzione dello strumento.
Precaución: para evitar choques eléctricos, apague el instrumento y desenchufe el cable
de alimentación antes de realizar cualquier reparación en el instrumento.

Commonly Used Symbols
Direct current
Courant continu
Gleichstrom
Corrente continua
Corriente continua
Alternating current
Courant alternatif
Wechs el s t r om
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
Precauzione, superficie calda o elevata temperatura
Precaución, superficie caliente o temperatura elevada
Caution, risk of electric shock (high voltage)•Attention,
risque de commotion électrique (haute 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, Ultraviolettes Licht
Precauzione, luce ultravioletta
Precaución, emisiones de luz ultravioleta
Commonly Used Symbols (Continued)

Waters 2410 Differential Refractometer Information
Intended Use
The Waters
®
2410 Differential Refractometer can be used for in-vitro diagnostic testing to analyze
many compounds, including diagnostic indicators and therapeutically monitoredcompounds.When
you develop methods, follow the “Protocol for the Adoption ofAnalytical Methods in the Clinical
Chemistry Laboratory,” American Journal ofMedical Technology, 44, 1, pages 30–37 (1978). This
protocol covers good operatingprocedures 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 xi
Table of Contents
How to Use This Guide
Chapter 1
Waters 2410 Theory of Operation
1.1 Overview ......................................................................................................... 1-1
1.2 Theory of Operation........................................................................................1-2
1.2.1 Optical Refraction .............................................................................1-2
1.2.2 Differential Refractometry ................................................................1-7
1.2.3 Common RI Detection Problems ......................................................1-9
1.3 Principles of Operation.................................................................................. 1-10
1.3.1 Fluidics.............................................................................................1-10
1.3.2 Optics ..............................................................................................1-14
1.3.3 Electronics .......................................................................................1-15
Chapter 2
Installing the 2410 Refractometer
2.1 Introduction.....................................................................................................2-1
2.2 Site Selection and Power Requirements..........................................................2-2
2.3 Unpacking and Inspection...............................................................................2-4
2.4 Making Electrical Power Connections............................................................2-5
2.5 Making Fluidic Connections...........................................................................2-5
2.5.1 Connecting a Column or Second Detector.........................................2-6
2.5.2 Connecting to Waste .........................................................................2-7
2.5.3 Connecting to a Drip Tray .................................................................2-8

xii Table of Contents
Chapter 3
Making Signal Connections
3.1 Component Connection Overview ..................................................................3-1
3.2 Making IEEE-488 Signal Connections ...........................................................3-3
3.2.1 Connecting to a Waters Data System Using the IEEE-488 Bus ........3-3
3.2.2 Connecting to a Waters PowerLine System Controller .....................3-7
3.2.3 Connecting to a Manual Injector .......................................................3-7
3.3 Making Non-IEEE-488 Signal Connections...................................................3-8
3.3.1 Connecting to a Stand-Alone 2690 Separations Module.................3-10
3.3.2 Connecting to the Waters 745/745B/746 Data Module ..................3-13
3.3.3 Connecting to a Chart Recorder ......................................................3-15
3.3.4 Connecting to the Waters 845/860 ExpertEase System ..................3-16
3.3.5 Connecting Injection Trigger Signals .............................................3-16
3.3.6 Polarity Connections .......................................................................3-18
3.4 Connecting the External Column Heaters.....................................................3-19
Chapter 4
Preparing Solvents
4.1 Common Solvent Problems.............................................................................4-1
4.2 Selecting a Solvent..........................................................................................4-2
4.3 Solvent Degassing...........................................................................................4-4
4.3.1 Gas Solubility.....................................................................................4-5
4.3.2 Solvent Degassing Methods ..............................................................4-5
4.3.3 Solvent Degassing Considerations ....................................................4-6
Chapter 5
Using the 2410 Refractometer
5.1 Using the Front Panel......................................................................................5-1
5.1.1 Keypad Functions ..............................................................................5-4
5.2 Selecting Parameter Values .............................................................................5-9

Table of Contents xiii
5.2.1 Sensitivity Guidelines ........................................................................5-9
5.2.2 Scale Factor Guidelines ...................................................................5-10
5.2.3 Time Constant Guidelines ...............................................................5-11
5.2.4 Temperature Guidelines (Ext1 °C, Ext2 °C, Int °C) .......................5-12
5.2.5 Polarity Guidelines ..........................................................................5-13
5.3 Starting Up the 2410 Refractometer..............................................................5-13
5.4 Shutting Down the 2410 Refractometer ........................................................5-15
Chapter 6
Maintenance Procedures
6.1 Cleaning the Fluidic Path.................................................................................6-1
6.2 Replacing Fuses...............................................................................................6-3
Chapter 7
Troubleshooting
7.1 Troubleshooting Overview ..............................................................................7-1
7.2 Chromatography Troubleshooting...................................................................7-2
7.2.1 Abnormal Baseline.............................................................................7-3
7.2.2 Erratic or Incorrect Retention Times .................................................7-7
7.2.3 Poor Peak Resolution ........................................................................7-9
7.2.4 Incorrect Qualitative/Quantitative Results ......................................7-11
7.3 Diagnostics ....................................................................................................7-13
7.3.1 Operating the Startup Diagnostics....................................................7-13
7.3.2 Operating the User-Initiated Diagnostics ........................................7-13
7.4 Hardware Troubleshooting ............................................................................7-16
Chapter Appendix A
Specifications
Chapter Appendix B
Spare Parts/Accessories

xiv Table of Contents
Appendix C
Warranty Information
C.1 Limited Express Warranty...............................................................................C-1
C.2 Shipments, Damages, Claims, and Returns.....................................................C-4
Index

Table of Contents xv
Figures
1-1 Waters 2410 Differential Refractometer..................................................1-1
1-2 Effect of Density on RI............................................................................1-4
1-3 Refraction of Light...................................................................................1-5
1-4 Presence of Sample Changes the Photodiode Signal............................... 1-6
1-5 How Refraction Changes
f .......................................................................1-8
1-6 Waters 2410 Refractometer Fluidics......................................................1-11
1-7 Waters 2410 Refractometer Fluidic Paths..............................................1-13
1-8 Waters 2410 Differential Refractometer Optics Bench
Assembly Light Path.............................................................................. 1-15
2-1 Major Steps in Installing the 2410 Differential Refractometer................2-1
2-2 Dimensions of the 2410 Refractometer ................................................... 2-2
2-3 Waters 2410 Refractometer Rear Panel ...................................................2-4
2-4 Fluidic Connections.................................................................................2-6
2-5 Ferrule and Compression Screw Assembly .............................................2-7
3-1 Waters 2410 Differential Refractometer Rear Panel................................3-2
3-2 Overview of Connecting Components to the
2410 Differential Refractometer.............................................................. 3-3
3-3 Waters Millennium System IEEE-488 Connections................................3-4
3-4 Waters 845/860 System IEEE-488 Connections......................................3-4
3-5 Waters Alliance System IEEE-488 Connections .....................................3-5
3-6 Waters PowerLine System Controller IEEE-488 Connections................3-7
3-7 Waters 2410 Rear Panel Analog-Out/Event-In Connectors..................... 3-9
3-8 Auto Zero Connections Between the 2690 Separations Module
and the 2410 Refractometer...................................................................3-11
3-9 Chart Mark Connections Between the 2690 Separations Module
and the 2410 Refractometer...................................................................3-12
3-10 Chart Mark and Auto Zero Connections Between
the 2690 Separations Module and the 2410 Refractometer................... 3-13

xvi Table of Contents
3-11 Connections to a Waters 745/745B/746 Data Module........................... 3-14
3-12 Analog Output Connections to a Chart Recorder.................................. 3-15
3-13 Analog Output Connections to the Bus SAT/IN Module.......................3-16
3-14 Auto Zero Connectionto a Manual Injector...........................................3-17
3-15 Chart Mark Connections to a Manual Injector ...................................... 3-18
3-16 2410 Refractometer External Column Heater Ports .............................. 3-20
5-1 Display, LED Indicators, and Keypad......................................................5-2
5-2 Effects of Sensitivity Settings................................................................5-10
5-3 Effects of Filter Time Constant Settings................................................5-12
6-1 Removing and Replacing Fuses............................................................... 6-4

Table of Contents xvii
Tables
1-1 Fluidic Line Inner Diameters............................................................ 1-12
2-1 Installation Site Requirements.................................................................2-3
3-1 Component Connection Summary...........................................................3-1
3-2 Waters 2410 Refractometer Inject Start Connections........................ 3-6
3-3 Waters 2410 Connections to a Manual Injector ................................ 3-7
3-4 Waters 2410 Analog-Out/Event-In Connections............................... 3-9
3-5 Auto Zero Connections Between the 2690 Separations Module
and the 2410 Refractometer ........................................................... 3-10
3-6 Chart Mark Connections Between the 2690 Separations Module
and the 2410 Refractometer ........................................................... 3-11
3-7 Chart Mark and Auto Zero Connections Between the
2690 Separations Module and the 2410 Refractometer................... 3-12
3-8 Analog Output Connections to a 745/745B/746 Data Module........ 3-13
3-9 Analog Output Connections to a Chart Recorder............................ 3-15
3-10 Analog Output Connections to the Bus SAT/IN Module................. 3-16
3-11 Auto Zero Connections to a Manual Injector.................................. 3-17
3-12 Chart Mark Connections to a Manual Injector................................ 3-18
3-13 Polarity Options............................................................................. 3-19
4-1 Refractive Indices of Common Solvents..................................................4-3
5-1 Indicator LED Functions..........................................................................5-3
5-2 Keypad Functions............................................................................ 5-4
6-1 Voltage and Fuse Requirements...............................................................6-4
7-1 Abnormal Baseline Troubleshooting ................................................ 7-4
7-2 Retention Time Troubleshooting...................................................... 7-7

xviii Table of Contents
7-3 Resolution Troubleshooting ........................................................... 7-10
7-4 Incorrect Results Troubleshooting.................................................. 7-12
7-5 User Diagnostics............................................................................ 7-14
7-6 Waters 2410 Hardware Troubleshooting......................................... 7-16
A-1 Operational Specifications ...................................................................... A-1
A-2 Integrator Output..............................................................................A-2
A-3 Optical Component Specifications....................................................A-2
A-4 Environmental Specifications...........................................................A-2
A-5 Dimensions ......................................................................................A-3
A-6 Electrical Specifications...................................................................A-3
A-7 Power Source Specification..............................................................A-3
B-1 Recommended Spare Parts...............................................................B-1
C-1 Waters 2410 Warranty Periods.................................................................C-4

xix
How to Use This Guide
Purpose ofThis Guide
The Waters 2410 Differential Refractometer Operator’s Guide describes the features and use of the
Waters
®
2410 Differential Refractometer and provides installation and maintenance procedures.
Audience
This guide is intended for use by anyone interested in installing, using, maintaining, and
troubleshooting the 2410 differential refractometer.
Structure ofThis Guide
The Waters 2410 Differential Refractometer Operator’s Guide is divided into chapters and
appendixes. Each page is marked with a tab and a footer to facilitate access to information within
the chapter or appendix.
The table below describes the material covered in each chapter and appendix.
Chapter 1, Waters 2410
Theory of Operation
Describes the product and the principles of differential
refractometry and 2410 differential refractometer operation.
Chapter 2, Installing the
2410 Refractometer
Describes the 2410 differential refractometer installation
procedures.
Chapter 3, Making
Signal Connections
Describes how to connect other components of your
chromatography system to the 2410 differential refractometer.
Chapter 4, Preparing
Solvents
Discusses the importance of filtering and degassing solvents for
effective operation.
Chapter 5, Using the
2410 Refractometer
Describes how to power on and off and operate the 2410
differential refractometer.
Chapter 6, Maintenance
Procedures
Describes maintenance and parts replacement procedures for the
2410 differential refractometer.
Chapter 7,
Troubleshooting Provides tables describing symptoms, possible causes, and corrective actions for 2410 differential refractometer operational
problems.

xx
Related Documents
The following table lists other documents related to the operation of the Waters 2410 Differential
Refractometer.
Appendix A,
Specifications Provides specifications for the 2410 differential refractometer.
Appendix B, Spare
Parts/Accessories Lists the recommended spare parts for the 2410 differential refractometer.
Appendix C, Warranty
Information
Includes warranty and service information for the 2410 differential
refractometer.
Waters 2690 Separations Module
Operator’s Guide
Describes the procedures for unpacking, installing, using,
maintaining, and troubleshooting the Waters 2690
Separations Module.
Waters 600E Multisolvent
Delivery System User’s Guide
Describes the procedures for installing, using, maintaining,
and troubleshooting the Waters 600E Multisolvent Delivery
System.
Waters Bus SAT/IN Module
Installation Guide
Provides the procedures for installing the Waters Bus
SAT/IN Module.
Millennium Software User’s
Guide, Vol. I and Vol. II
Describes the Millennium Chromatography Manager
software used in both the Millennium 2010 workstation and
the Millennium 2020 client/server system.