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Waters Column BypassModule
Installation and Maintenance Guide
34 Maple StreetMilford, MA 01757
71500032104, Revision A
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.
© 2002 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.
Integrity, Nova-Pak, and Waters are registered trademarks, and Empower and Thermabeam are registered trademarks of Waters Corporation.
Teflon, Tefzel, and Vespel are registered trademarks of E. I. du Pont de Nemours and Company.
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.
STOPAtención: 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 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 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 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 currentCourant continuGleichstromCorrente continuaCorriente continua
Alternating currentCourant alternatifWechselstromCorrente alternataCorriente alterna
Protective conductor terminalBorne du conducteur de protectionSchutzleiteranschlussTerminale di conduttore con protezioneBorne del conductor de tierra
Frame or chassis terminalBorne du cadre ou du châssisRahmen- oder ChassisanschlussTerminale di struttura o telaioBorne de la estructura o del chasis
Caution or refer to manualAttention ou reportez-vous au guideVorsicht, oder lesen Sie das HandbuchPrestare attenzione o fare riferimento alla guidaActúe con precaución o consulte la guía
Caution, hot surface or high temperatureAttention, surface chaude ou température élevéeVorsicht, heiße Oberfläche oder hohe TemperaturPrecauzione, superficie calda o elevata temperaturaPrecaución, 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)Precauzione, rischio di scossa elettrica (alta tensione)Precaución, peligro de descarga eléctrica (alta tensión)
Caution, risk of needle-stick punctureAttention, risques de perforation de la taille d’une aiguilleVorsicht, Gefahr einer SpritzenpunktierungPrecauzione, rischio di puntura con agoPrecaución, riesgo de punción con aguja
Caution, ultraviolet lightAttention, rayonnement ultrvioletVorsicht, Ultraviolettes LichtPrecauzione, luce ultraviolettaPrecaución, emisiones de luz ultravioleta
Commonly Used Symbols (Continued)
UV
Column Bypass Module Information
Intended Use
The Waters® Column Bypass Module can be used for in-vitro diagnostic testing to analyze many compounds, including diagnostic indicators and therapeutically monitored compounds. 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
Table of Contents 14
Chapter 1 Introduction ...................................................................................... 21
1.1 Valve Control Connections.................................................... 23
1.2 Internal Electrical and Helium Connections .......................... 23
1.3 Making Fluidic Connections .................................................. 24
Chapter 2 Operation ......................................................................................... 27
2.1 Column Bypass Module Fluidics Overview........................... 27
2.2 Injecting Calibrants with the Sample Management System.................................................................................. 33
2.3 Injecting Calibrants Manually ................................................ 34
Chapter 3 Troubleshooting ............................................................................... 36
3.1 Inject/Load Valve Problems................................................... 36
3.1.1 Tightening the Pressure-Adjusting Screw.................. 37
3.1.2 Replacing the Rotor Seal........................................... 40
3.1.3 Tightening the Needle Seal ....................................... 41
3.1.4 Testing and Replacing the Position-Sensing Switch ........................................................................ 42
3.2 Control Signal Problems ....................................................... 43
Appendix A Specifications ................................................................................... 44
Appendix B Spare Parts....................................................................................... 46
Index ....................................................................................... 47
Table of Contents 15
List of Figures 16
1-1 2690/2695 Separations Module with Column Bypass Module ...... 201-2 Column Bypass Module and Column Heater................................. 211-3 Column Bypass Module Valves ..................................................... 211-4 Column Bypass Module Valve Control Connections...................... 221-5 Column Bypass Module Electrical and Helium Connections ......... 231-6 Installing a Column in the Column Heater Module ........................ 241-7 Installing the Waste Vial ................................................................ 25
2-1 Column Bypass Module - Column Mode ....................................... 272-2 Column Bypass Module - Bypass (Inject) Mode............................ 292-3 Column Bypass Module - Bypass (Load) Mode ............................ 302-4 Column Bypass Module - Sample Loop ........................................ 32
3-1 Rheodyne 7725i Injector, Exploded View ...................................... 383-2 Rotor Seal Orientation (Viewed from Stator) ................................. 40
List of Figures
List of Tables 17
3-1 Rheodyne 7725i Injector Troubleshooting................................ 353-2 Control Signal Troubleshooting................................................ 42
A-1 Operational Specifications ............................................................ 43A-2 Physical Specifications............................................................ 43A-3 Environmental Specifications .................................................. 44A-4 Power Specifications ............................................................... 44A-5 Communications Specifications for 6-Port Switching
Valve ...................................................................................... 44
B-1 Spare Parts ................................................................................... 45
List of Tables
Preface
The Waters Column Bypass Module Installation and Maintenance Guide is intended for use by personnel who need to maintain and troubleshoot the Waters® Column Bypass Module. This guide provides information on using, troubleshooting, and maintaining the Waters Column Bypass Module as a component of a Waters LC System.
Organization
This guide contains the following:
Chapter 1 describes the function of the Column Bypass Module.
Chapter 2 describes how to operate the Column Bypass Module.
Chapter 3 provides troubleshooting and maintenance procedures.
Appendix A lists specifications.
Appendix B lists spare parts.
Related Documentation
Waters Licenses, Warranties, and Support: Provides software license and warranty information, describes training and extended support, and tells how Waters handles shipments, damages, claims, and returns.
Online Documentation
Empower LIMS Help: Describes how to use the Empower LIMS Interface to export results and import worklists.
Printed Documentation for Software Options
Waters Integrity System Getting Started Guide: Describes features of the Waters Integrity® System and provides step-by-step tutorials that guide a user through the use of the Empower Mass Spectrometry (MS) option.
Documentation on the Web
Related product information and documentation can be found on the World Wide Web. Our address is http://www.waters.com.
Related Adobe Acrobat Reader Documentation
For detailed information about using Adobe® Acrobat® Reader, see the Adobe Acrobat Reader Online Guide. This guide covers procedures such as viewing, navigating, and printing electronic documentation from Adobe Acrobat Reader.
18
Printing This Electronic Document
Adobe Acrobat Reader lets you easily print pages, page ranges, or the entire document by selecting File > Print. For optimum print quantity, Waters recommends that you specify a PostScript® printer driver for your printer. Ideally, use a printer that supports 600 dpi print resolution.
Documentation Conventions
The following conventions can be used in this guide:
Convention Usage
Purple Purple text indicates user action such as keys to press, menu selec-tions, and commands. For example, “Click Next to go to the next page.”
Italic Italic indicates information that you supply such as variables. It also indicates emphasis and document titles. For example, “Replace file_name with the actual name of your file.”
Courier Courier indicates examples of source code and system output. For example, “The SVRMGR> prompt appears.”
Courier Bold Courier bold indicates characters that you type or keys you press in examples of source code. For example, “At the LSNRCTL> prompt, enter set password oracle to access Oracle.”
Underlined Blue Indicates hypertext cross-references to a specific chapter, section, subsection, or sidehead. Clicking this topic using the hand symbol brings you to this topic within the document. Right-clicking and selecting Go Back from the shortcut menu returns you to the origi-nating topic. For example, “Repairable Column Bypass Module control signal problems involve the cable wiring interface between the 2690/2695 Separations Module and the Column Bypass Module as described in Section 3.2, Control Signal Problems.”
Keys The word key refers to a computer key on the keypad or keyboard. Screen keys refer to the keys on the instrument located immedi-ately below the screen. For example, “The A/B screen key on the 2414 Detector displays the selected channel.”
… Three periods indicate that more of the same type of item can optionally follow. For example, “You can store filename1, filename2, … in each folder.”
> A right arrow between menu options indicates you should choose each option in sequence. For example, “Select File > Exit” means you should select File from the menu bar, then select Exit from the File menu.
19
Notes
Notes call out information that is helpful to the operator. For example:
Note: Record your result before you proceed to the next step.
Attentions
Attentions provide information about preventing damage to the system or equipment. For example:
Cautions
Cautions provide information essential to the safety of the operator. For example:
STOPAttention: To avoid damaging the detector flow cell, do not touch the flow cell window.
Caution: To avoid burns, turn off the lamp at least 30 minutes before removing it for replacement or adjustment.
Caution: To avoid electrical shock and injury, turn off the detector and unplug the power cord before performing maintenance procedures.
Caution: To avoid chemical or electrical hazards, observe safe laboratory practices when operating the system.
20
1
Chapter 1IntroductionIn the current Waters® Integrity® configuration, the Column Bypass Module is contained within the 2690/2695 Column Heater module (Figure 1-1 and Figure 1-2). The Column Bypass Module allows you to easily bypass the column in your chromatographic system. Bypassing the chromatography column is necessary when you inject calibrants or analytes that could damage the column packing, or if you desire Mass Spectral data without the need to develop the chromatography.
The Column Bypass Module contains a 6-port, 2-way, pneumatically operated (switching) valve that directs the fluid path to either include or bypass the column. The Column Bypass Module also has a 6-port, 2-way (inject/load) mechanical valve with a needle port for loading calibrants into a sample loop and injecting them into the fluidic path (Figure 1-3).
The Column Bypass Module connects to the 2690/2695 power supply, sample fluidic lines, and electronic control signals.
Figure 1-1 2690/2695 Separations Module with Column Bypass Module
ColumnBypassModule
ColumnHeater
21
1
Figure 1-2 Column Bypass Module and Column Heater
Figure 1-3 Column Bypass Module Valves
TP01449
COLUMN
LOAD
INJECT
BYPASS
Column Bypass Module
(Slides Out for Access)
Column Heater(Door Opens for Access)
TP01447
Front Panel
Inject/LoadValve
Switching Valve
Introduction 22
1
1.1 Valve Control Connections
Figure 1-4 illustrates the valve control connections between the Column Bypass Module and the 2690/2695 Separations Module. These connectors are located externally on the rear panel of both devices.
Figure 1-4 Column Bypass Module Valve Control Connections
1.2 Internal Electrical and Helium Connections
Figure 1-5 illustrates the internal electrical and helium connections between the Column Bypass Module, and the 2690/2695 Separations Module.
6 Switch 3
1 Switch 1
5 Switch 2
2 Switch 1
3 Ground
4 Switch 2
7 Switch 3
8 Switch 4
9 Switch 4
10 Ground
11 Run Stopped
12 Run Stopped
Pin Signal
1 +Valve In
24 Chassis Ground
–Valve In
Black
White
Clear
White
Black
Clear
Legend
2690/2695 SeparationsModule Rear PanelConnector A
CBM Screw Terminal Connector
Valve Control Connections 23
1
Figure 1-5 Column Bypass Module Electrical and Helium Connections
Note: You need to make a helium connection to the ThermaBeam Mass Detector, as described in the Waters ThermaBeam Mass Detector Installation and Maintenance Guide, Section 3.7, Connecting the Helium Supply.
1.3 Making Fluidic Connections
This section describes how to make fluidic connections between the Column Bypass Module and other components in your LC system.
Note: For details on making connections to the ports on the Column Bypass Module valves, refer to Section 2.1, Column Bypass Module Fluidics Overview.
Installing a Column
Connect either a 2-mm I.D. microbore or 3.9-mm I.D. microbore NovaPak® (standard lengths of 15-cm or 30-cm) column to the Column Holder in the Column Heater module.
Electrical Connector
Helium Connection
Front of Unit
Control Lines toHelium Regulator
Introduction 24
1
To install and connect a column in the Column Heater module:
1. Hold the column so that its outlet is on top (arrows pointing up) and secure the column using the column holder (Figure 1-6).
2. Connect the top of the column to the unconnected end of the factory-installed 0.005-inch I.D. tubing that connects to the From Column port.
3. Connect the bottom of the column to the unconnected end of the factory-installed 0.005-inch I.D. tubing that connects to the To Column port.
Figure 1-6 Installing a Column in the Column Heater Module
Connecting to the LC System
You connect the Column Bypass Module between the sample management system of the 2690/2695 Separations Module and the PDA detector. Use precut lengths of tubing obtained from Waters, or use your own accurately cut lengths of the same diameter tubing.
Note: For details on making connections to the ports on the Column Bypass Module valves, refer to Section 2.1, Column Bypass Module Fluidics Overview.
TP01448
Column Holder
Column
Making Fluidic Connections 25
1
To make the connections:
1. Connect the tubing from the sample management system red line outlet to the port labelled From Sample Management on the Column Bypass Module valves. (See Figure 2-1 through Figure 2-4.)
2. Install a 30-inch length of 0.005-inch I.D. tubing between the To Detector outlet port on the Column Bypass Module valves and the PDA detector Inlet port. (See Figure 2-1 through Figure 2-4.)
Installing the Waste Vial
Position the waste vial under the ends of the two waste tubes and move the vial up and into the clip on the front panel (Figure 1-7).
Note: The Inject/Load valve should always be in the LOAD position when not in use.
• When the inject/load valve is in the Load position, any excess calibrant loaded into the sample loop is expelled from one of these waste tubes into the waste vial.
• When the inject/load valve is in the Inject position, any calibrant previously injected into the sample loop enters the fluid path and goes to the detector. Any flushing agent (or mobile phase) injected into the needle port is expelled through the second waste tube into the waste vial.
Figure 1-7 Installing the Waste Vial
Waste Tubes
Waste Vial
Introduction 26
2
Chapter 2Operation
The Column Bypass Module receives commands from the 2690/2695 Separations Module to direct the fluid path in either of the following ways: through the column (Column mode), or around the column (Bypass mode). Front-panel indicators on the Column Bypass Module indicate the currently selected mode.
• During system calibration, the 2690/2695 Separations Module configures the Column Bypass Module 6-port switching valve to direct the fluid path around the column and into the detector. Calibrants can be either manually or automatically injected and results can be observed.
• During normal operation, the 2690/2695 Separations Module configures the Column Bypass Module 6-port switching valve to direct the fluid path through the column.
When the Column Bypass Module is included as part of the Integrity system, Empower software automatically instructs the Column Bypass Module to bypass the column from the flow path. Once Empower software optimizes the interface temperature, it flushes the system and reestablishes the original conditions.
This chapter describes:
• Column Bypass Module fluidics overview
• Injecting calibrants with the sample management system
• Injecting calibrants with a manual injector
2.1 Column Bypass Module Fluidics Overview
Figure 2-1 through Figure 2-4 are fluidics diagrams of the Column Bypass Module showing the 6-port, 2-way, pneumatically operated switching valve, and the 6-port, 2-way, mechanically operated inject/load valve (see Figure 1-3). The operational modes and fluid paths that are set by these valves are:
• Column mode
• Bypass mode
• Sample loop and needle port
Column Mode
In the Column mode, the fluid path is from the 2690/2695 Separations Module, through the column, and out to the detector. Fluid enters switching valve port 2, leaves through port 3,
Column Bypass Module Fluidics Overview 27
2
then passes through the column. From the column, the fluid enters port 6, exits through port 5, and goes to the detector (Figure 2-1).
Figure 2-1 Column Bypass Module - Column Mode
Bypass Mode
In the Bypass mode (Figure 2-2 and Figure 2-3), the fluid path is from the 2690/2695 Separations Module, through the switching valve, through the inject/load valve, back through the switching valve, and out to the detector, completely bypassing the column. There are two sub-modes of operation associated with the Bypass mode:
• Bypass (inject) mode
• Bypass (load) mode
Bypass (Inject) Mode
In the Bypass (Inject) mode, the inject/load valve is rotated to the Inject position and the switching valve is in the bypass position. Any calibrant in the sample loop goes into the fluid stream and to the detector.
Fluid enters switching valve port 2, exits through port 1, enters inject/load valve port 2, exits through port 1 and then enters the sample loop. The calibrant in the sample loop enters inject/load valve port 4, exits through port 3, enters switching valve port 4, exits through port 5, and enters the detector. Any calibrant previously injected into the sample loop goes into the fluid stream and to the detector. (See Figure 2-2.)
TP01307
1
6
54
3
2
Column
SwitchingValve
Outlet(To Detector)
Inlet From 2690/2695Separations
Module
Operation 28
2
Bypass (Load) Mode
In the Bypass (Load) mode, the inject/load valve is rotated to the Load position and the switching valve is in the bypass position. At this point, the sample loop is isolated from the flow path and can be loaded with calibrant, if desired.
Fluid from the 2690/2695 Separations Module enters switching valve port 2, exits through port 1, enters inject/load valve port 2, exits through port 3, enters switching valve port 4, exits through port 5, and enters the detector. (See Figure 2-3.)
Column Bypass Module Fluidics Overview 29
2
Figure 2-2 Column Bypass Module - Bypass (Inject) Mode
TP01307A
Inject/LoadValve
1 2
3
45
6
To WasteVial
Needle Port(Flush)
Sa
mpl
e L
oop
Column
Outlet(To Detector)
ValveSwitching
Inlet From 2690/2695Separations
Module
2 1
6
54
3
Legend
Flow PathNon-flow Path
Operation 30
2
Figure 2-3 Column Bypass Module - Bypass (Load) Mode
TP01307b
Legend
Flow PathNon-flow Path
Column
SwitchingValve
Outlet(To Detector)
Inject/LoadValve
Sam
ple
Lo
op
To WasteVial
Needle Port(Load)
1 2
3
45
6
2 1
6
54
3
Inlet From 2690/2695Separations
Module
Column Bypass Module Fluidics Overview 31
2
Sample Loop and Needle Port
The sample loop and needle port fluid path is shown in Figure 2-4. The standard size loop for this configuration is 20 µl.
Loading the Sample Loop
When the inject/load valve is in the Load position, the sample loop is isolated from the fluid stream. When you inject calibrant into needle port 4, you fill the sample loop. Excess calibrant leaves the sample loop, enters port 1, exits through vent port 6 (which is now connected to port 1), and enters the front-panel waste vial. The desired amount of calibrant remains in the sample loop.
Injecting the Sample
Any calibrant in the sample loop becomes part of the fluid stream when the switching valve is in the Bypass mode and the inject/load valve is in the Inject position. In this configuration, inject/load valve port 1 is isolated from port 6, thereby removing the injection loop from atmospheric pressure. High-pressure fluid flowing into port 2 flows out of port 1 and forces the calibrant out of the sample loop and into the fluid stream at port 4.
Flushing the Needle Port
In the Inject position, the needle port is connected to port 5. At this time, any flushing agent (or mobile phase) injected into the needle port is expelled directly out of port 5, through a waste tube, and into the waste vial.
Caution: To prevent liquid from squirting back at you when you flush the needle port, slowly inject 1-mL of mobile phase using a Rheodyne (P/N 7125-054) needle port cleaner attached to the syringe barrel.
Operation 32
2
Figure 2-4 Column Bypass Module - Sample Loop
Changing the Sample Loop
When you perform the Operational Qualification for the Integrity System 996/2996 PDA Detector, you need to change the sample loop in the Inject/Loop Valve from the standard 20-µL sample loop to a 1-mL sample loop to perform the wavelength accuracy test.
The sample loop is connected between port 1 and port 4 on the Inject/Load Valve (see Figure 1-3 and Figure 2-4). For more information about the Operational Qualification for the 996/2996 PDA Detector, refer to the Waters HPLC Systems Qualification Workbook for the 996/2996 PDA Detector: Part IV, Operational Qualification.
2.2 Injecting Calibrants with the Sample Management System
The sample management system mode of operation configures the Column Bypass Module to bypass the column while the sample management system automates the delivery of one or more calibration standards directly into the fluid path.
STOPAttention: Ensure that the eluent and sample are properly filtered to prevent additional precipitation. Flush the flow passages and vent lines with water after use of salt solutions. Do not allow salt solutions to sit stagnant in the valve overnight or for long periods of time.
Sam
ple
Lo
op
Inject/LoadValve
To WasteVial
Needle Port(Load)
1 2
3
45
6
Injecting Calibrants with the Sample Management System 33
2
Using a 2690 Separations Module and Empower Software
Use the following procedure to control the Column Bypass Module from a 2690/2695 Separations Module that is operating under Empower software control:
1. Rotate the lever on the Column Bypass Module front panel to the Load position. This position isolates any calibrant in the Column Bypass Module sample loop from the injection stream and reduces band spreading.
2. Place the desired standards and samples into the sample management system.
3. In the Method tab of the Empower software Project window, double-click the appropriate instrument method. The Instrument Method window appears.
4. Access the 2690/2695 Separations Module. When using Empower Pro, double-click the icon for your 2690/2695 Separations Module. When using Empower QuickStart, single-click the icon for your 2690/2695 Separations Module. The 2690/2695 Instrument dialog box appears.
5. Set Switch 4 in the 2690/2695 Instrument dialog box according to your Column Bypass Module control requirements:
• For immediate event control, click Switch 4.
• For time-initiated event control, click the Events icon and specify the time you want the switch S4 event turned on and off.
For details on setting up a 2690/2695 Separations Module, refer to the Waters 2695 Separations Module Operator’s Guide.
6. Save the changes to your instrument method, then exit the window.
7. When using Empower Pro, access the Run Samples window. When using Empower QuickStart, access Sample Queue view. Refer to the Empower Software Getting Started Guide.
8. To initiate a run using Empower Pro, access the Run Samples window. To initiate a run using Empower QuickStart, access the Sample Queue view.
Note: If, during data acquisition, you want to change the Column Bypass Module to Column mode, using Empower Pro, press the Stop Flow key in the Run Samples window. When using Empower QuickStart, press the Stop Flow key in the Control Panel view.
2.3 Injecting Calibrants Manually
The manual injection mode allows you to:
• Manually load a calibrant or sample.
• Inject the calibrant into the fluid path.
• Observe the results of that injection. During this mode, the column is bypassed from the fluid path.
Operation 34
2
Procedure
To inject calibrants with a manual injector:
1. Rotate the lever on the Column Bypass Module front panel to the Load position. This position isolates any calibrant in the sample loop from the fluid path.
2. Inject the desired calibrant (or sample) into the needle port on the Column Bypass Module front panel using a calibrated syringe. To obtain best reproducibility for fixed loop injections, overfill the loop with at least two to three loop volumes of sample.
Note: Five to ten loop volumes will provide even more precision. An excess of sample is needed because mobile phase near the wall of the loop is displaced slowly.
3. Use the 2690/2695 Separations Module under Empower software control, to initiate a timed run, without using the sample management system. A run time of 1 to 2 minutes is recommended.
4. Rotate the lever to the Inject position and observe the results of the calibrant injection.
Note: During the timed run, you can load and inject other calibrants and observe their results.
Injecting Calibrants Manually 35
3
Chapter 3Troubleshooting
This chapter presents step-by-step troubleshooting procedures. The information is divided into the following topics:
• Inject/Load valve problems
• Control signal problems
3.1 Inject/Load Valve Problems
Repairable inject/load valve problems involve the Rheodyne 7725i Injector that is used to make manual injections. Table 3-1 lists potential Rheodyne injector symptoms that may occur during operation, along with possible causes and corrective actions.
Table 3-1 Rheodyne 7725i Injector Troubleshooting
Symptom Possible Cause Corrective Action
Siphoning (appears as leakage)
Tubes from ports 5 and 6 do not have outlet ends at the same level as needle port
Place the outlet ends of both tubes at the same horizontal level as that of the needle port.
Rotor seal leakage (liquid dripping between stator and stator ring, or from needle port or vent tube)
Pressure-adjusting screw loose
Tighten pressure-adjusting screw. Refer to Section 3.1.1, Tightening the Pres-sure-Adjusting Screw.
Scratches on rotor seal Replace rotor seal. Refer to Section 3.1.2, Replacing the Rotor Seal.
Needle seal leakage Syringe needle O.D. smaller than needle seal, reducing sample loading accuracy
Try another syringe needle. Gently push on plastic needle guide to deform Teflon® sleeve to provide a better needle fit. See Section 3.1.3, Tightening the Needle Seal.
Inject/Load Valve Problems 36
3
3.1.1 Tightening the Pressure-Adjusting ScrewRequired Tools
To tighten the pressure-adjusting screw, you need a Phillips-head screwdriver and a 5/64-inch Allen wrench.
Adjustment Procedure
To tighten the pressure-adjusting screw:
1. Turn off the 2690/2695 Separations Module and disconnect the power cord.
2. Slide the Column Bypass Module compartment open to access the pressure-adjusting screw.
3. Locate the pressure-adjusting screw on the shaft of the injector (Figure 3-1).
4. Loosen the two knob set screws.
Increase in system backpressure or band broadening
Plugged valve passages or vent lines due to unfiltered eluent or sample, or buffer solutions crystallized in vent lines
Remove the stator and clean the passages with a wire of 0.005-inch maximum diameter or sonicate in water.If the valve passage remains plugged, disconnect the tubing at the vent valve outlet. If this clears the passage, replace the tubing. If the passage remains blocked, disconnect the compression screw at the vent valve inlet. If this reduces the pressure, replace the vent valve. Refer to Section 3.1.2, Replacing the Rotor Seal.
Gradient does not start Position-sensing switch is not sending the inject signal to 2690/2695 Separations Module
Disconnect the switch from the inject start cable at the connector under the valve. Gently insert a small flat-blade screwdriver into the connector to short-circuit the contacts. If the gradient starts flowing, check the switch as described in Section 3.1.4, Testing and Replacing the Position-Sensing Switch.
Caution: To avoid chemical and electrical hazards, always wear safety glasses, turn off the 2690/2695 Separations Module, and disconnect the power cord before you perform this procedure.
Table 3-1 Rheodyne 7725i Injector Troubleshooting (Continued)
Symptom Possible Cause Corrective Action
Troubleshooting 37
3
5. Let the handle slide down the shaft so that its two tabs fit into the slots on the adjusting screw.
6. Use the knob as a wrench to tighten the pressure-adjusting screw approximately 1/20th of a turn. Use the 20 dial markings on the body and the painted spot on the screw to gauge how far to tighten the screw.
7. If the new setting fails to accomplish leak-free operation, repeat step 6 only once. Avoid excessive tightening, which increases rotor seal wear.
8. Finish by retightening the knob set screws onto the flats of the shaft.
Note: If this procedure fails to stop the leak, replace the rotor seal as described in Section 3.1.2, Replacing the Rotor Seal.
Inject/Load Valve Problems 38
3
Figure 3-1 Rheodyne 7725i Injector, Exploded View
Handle
Knob
Set Screws (2)
Needle Guide
Pressure Adjusting Screw
Body
Spring Washers (4)
Needle Port Tube
Needle Seal
60° Stop Ring
Seal Pins (4)
Bearing Ring
Isolation Seal
Thrust Bearing
Rotor Pin
Position Sensing Switch
Rotor Seal
Stator Ring
Stator Locating Pin
Stator Face Assembly
Stator Locating Hole
Handle Screw
Teflon Sleeve in Rotor Seal
Stator
Stator Screws (3)
Troubleshooting 39
3
3.1.2 Replacing the Rotor Seal
Premature rotor seal failure can be caused by any of the following problems:
• Abrasive particles in the sample or mobile phase that scratch the rotor seal surface.
• A needle tip of the wrong type that chips the ceramic stator face, causing deep scratching of the rotor seal surface.
• Buffer or salt crystallization caused by failure to flush the flow passages and needle port with water after the use of aqueous buffers or salt solutions. The crystals scratch the rotor seal surface, resulting in leakage.
Required Tools
To replace the rotor seal (see Figure 3-1), you need the following tools:
• 9/64-inch Allen wrench
• 5/16-inch open-end wrench
• Flat-blade screwdriver
• Phillips-head screwdriver
Replacement Procedure
To replace the rotor seal:
1. Turn off the 2690/2695 Separations Module and disconnect the power cord.
2. Slide the Column Bypass Module compartment open.
3. Leave the injector attached to the front panel of the Column Bypass Module, and leave the knob on.
4. Disconnect the tubing from the back of the valve.
5. Remove the three stator screws.
6. Pull axially to remove the following items:
• Stator and stator face assembly (remove together). If the stator face assembly is damaged, replace it.
• Stator ring.
7. Use a flat-blade screwdriver to pry the rotor seal off the four seal pins. Leave the isolation seal and bearing ring in place.
Caution: To avoid chemical and electrical hazards, always wear safety glasses, turn off the 2690/2695 Separations Module, and disconnect the power cord before you perform this procedure.
STOPAttention: To avoid damaging the injector after you have used buffer solutions, flush the injector with HPLC-grade water.
Inject/Load Valve Problems 40
3
Troubleshooting 41
Reassembling the Injector
To reassemble the injector:
1. Loosen the pressure-adjusting screw one half-turn.
2. Note the original position of the two red dots.
3. Orient the rotor seal as shown in Figure 3-2, with rotor seal slots facing the stator.
Figure 3-2 Rotor Seal Orientation (Viewed from Stator)
4. Replace the stator ring so that the pin in the 60° stop ring enters the mating hole in the stator ring.
5. Install the stator face assembly on the stator. The three pins on the stator face assembly fit only one way into the mating holes in the stator.
6. Install the stator and stator face assembly on the injector so that the pin in the stator ring enters the mating hole in the stator.
7. Tighten each of the three stator screws a little at a time to keep the stator surface parallel to the stator ring surface until all parts are held firmly in place.
8. Retighten the pressure-adjusting screw until the red dots are aligned as you noted them in step 2.
9. Replace the knob and tighten the two set screws against the two flat areas on the shaft.
3.1.3 Tightening the Needle Seal
The needle seal, a Teflon sleeve in the rotor seal, may not seal correctly around a thin needle. A poor seal reduces accuracy in sample loading.
Removing the Needle
To deform the Teflon sleeve to make a good seal:
1. Remove the needle from the needle port.
2. Push gently on the plastic needle guide with the eraser end of a pencil. Do not squash the Teflon sleeve.
3. Repeat if necessary.
Rotor PinNotch
3
Inject/Load Valve Problems 42
Checking the Needle Seal
To check for a proper seal around the needle:
1. Turn the solvent management system off.
2. Fill the syringe with water.
3. Place the injector in the Load position and slowly discharge the water in the syringe into the injector.
4. Note the low resistance to syringe discharge.
5. Repeat steps 1 and 2 with the injector handle halfway between the Load and Inject positions.
6. Verify that the resistance to discharging the syringe is now much greater than in step 3.
Note: The needle seal holds only a few psi (mPa) of pressure, and does not completely prevent syringe discharge when the injector handle is in the halfway position.
3.1.4 Testing and Replacing the Position-Sensing Switchs
The position-sensing switch is a magnetic reed switch actuated by a magnet in the shaft. The switch sends an Inject Start signal to the controller, signalling the controller to begin a run. If a run does not begin after an injection, the switch may be faulty. (The switch is rated for 100 V at 200 mA.)
Required Tools
To test or replace the position-sensing switch, you need the following tools:
• 9/64-inch Allen wrench
• 5/16-inch open-ended wrench
• Phillips-head screwdriver
Testing the Switch
To test the position-sensing switch:
1. Turn off the 2690/2695 Separations Module and disconnect the power cord.
STOPAttention: To avoid damaging the rotor seal, always use a needle of the correct type and gauge to load the injector.
Caution: To avoid chemical and electrical hazards, always wear safety glasses, turn off the Column Bypass Module, and disconnect the power cord before you perform this procedure.
3
Troubleshooting 43
2. Slide the Column Bypass Module compartment open.
3. Disconnect the switch cable from the inject start cable.
4. Using an ohmmeter with needle probes, insert the probes into the switch cable connector.
With the injector handle in the Load position, the resistance reading should be infinite. With the injector handle in the Inject position, the resistance reading should be less than 1 ohm.
Replacing the Switch
Refer to Figure 3-1 to replace the position-sensing switch:
1. Remove the stator and ring and 60° stop ring.
2. Pull the position-sensing switch out of the stop ring.
3. Insert a new position-sensing switch.
4. Reassemble the injector. Refer to Section 3.1.2, Replacing the Rotor Seal.
3.2 Control Signal Problems
Repairable Column Bypass Module control signal problems involve the cable wiring interface between the 2690/2695 Separations Module and the Column Bypass Module. Other possible causes involve 2690/2695 Separations Module malfunctions.
Table 3-2 lists the possible causes of control signal problems that may occur during operation, along with recommended corrective actions.
Table 3-2 Control Signal Troubleshooting
Possible Cause Corrective Action
2690 Separations Module defective
Troubleshoot the 2690/2695 Separations Module according to the related documentation for your LC system.
Empower soft-ware problems
Verify correct operation of the Empower software in accordance with the related documentation for your LC system.
Poor contact between the Column Bypass Module screw terminal connector and screw terminal socket
1. Remove the screw terminal connector from the terminal strip socket.
2. Examine the contacts and clean as necessary.3. Tighten each of the six connector screws as necessary.4. Insert the connector back into the socket.
Interface cable wiring problems
Check all cable connections to the 2690/2695 Separations Module and secure each connector screw.
A
Appendix ASpecificationsThis appendix includes information on:
• Operational specifications (Table A-1)
• Physical specifications (Table A-2)
• Environmental specifications (Table A-3)
• Power requirements (Table A-4)
• 6-Port Switching Valve communication requirements (Table A-5)
Table A-1 Operational Specifications
Parameter Specification
Pressure, maximum 5000 psi (35 MPa)
Wetted surfaces 316 stainless steel, Vespel® alumina ceramic, PEEK(Note: PEEK is not compatible with nitric acid, sulfuric acid and tetrahydrofuran {THF})
Table A-2 Physical Specifications
Parameter Specification
Height (including column heater)
18.3 in (46.5 cm)
Depth 11.8 in (30 cm)
Width 5.8 in (14.7 cm)
Weight 20 lbs (9.1 kg)
Specifications 44
A
Table A-3 Environmental SpecificationsParameter Specification
Ambient temperature, operating 39 to 104 °F (4 to 40 °C)
Ambient humidity, operating 10 to 90%, noncondensing
Ambient temperature, storage −40 to 158 °F (−40 to 70 °C)
Ambient humidity, storage 0 to 100%, noncondensing
Table A-4 Power Specifications
Parameter Specification
DC voltage 24 Vdc
Power requirements 10 VA
Helium Inlet 80 to 85 psi
Table A-5 Communications Specifications for 6-Port Switching Valve
Parameter Specification
Control Input(+Valve In, – Valve In)
Bypass: <+1.8 Vdc (+1.8 to –30 Vdc max.)Column: >+3.0 Vdc (+3.0 to +30 Vdc max.)
Inject Output Inject: closedLoad: open
Maximum voltage: 10 VdcMaximum current: 10 mA
Specifications 45
Spare Parts 46
B
Appendix BSpare Parts
Table B-1 lists the recommended spare parts for installation by the user. Any parts not listed in the table may require installation by a Waters® service representative.
Table B-1 Spare Parts
Item Quantity Part Number
Cable, Event 1 WAT020321
Cable, Shielded Output 1 WAT057235
Syringe, Rheodyne, 25 µl 1 WAT033381
Assembly, Tubing, 30 cm –– WAT055474
Assembly, Filter Element 2 WAT088084
Needle Guide, 7725i 1 WAT055459
Sample Loop, 1 mL (for 996/2996 PDA Detector Oper-ational Qualification testing)
1 WAT096226
Seal, Rotor, 7725i, Vespel 1 WAT055946
Seal, Rotor, 7725i, PEEKa
a. PEEK is not compatible with nitric acid, sulfuric acid, and tetrahydrofuran (THF).
–– WAT055458
Seal, Rotor, 7730/7716, Vespel
1 WAT0055457
Seal, Rotor, 7730/7716, PEEK –– WAT055406
Stator, 7730/7716 –– WAT055455
I NDEX
Index
AAutoinjection mode 32
CColumn installation 23Connections
LC System 24Conventions, documentation 18
DDocumentation
conventions 18related 17
FFluidic connections
waste vial 25Fluidics description
bypass mode 27column mode 26
IInstallation
column 23waste vial 25
LLC System connections 24
MManual injection mode 33
NNeedle seal tightening 41
OOperation
autoinjection 32manual injection 33overview 26
PPosition sensing switch testing 41Pressure adjusting screw 36
RRelated documentation 17Rheodyne 7725i injector
needle seal 41position sensing switch 41pressure adjusting screw 36rotor seal 39troubleshooting 35, 42
Rotor seal replacement 39
SSpare parts 45Specifications 43
Index 47
INDEX
TTroubleshooting
control signals 42fluid path 35Rheodyne 7725i injector 35, 42
VValve control connections 22
WWaste vial installation 25
I
Index 48
