SpectraSYSTEM P4000 Gradient Pump User Guide

SpectraSYSTEM P4000 Gradient Pump User Guide
SpectraSYSTEM
P4000 Gradient Pump
User Guide
A0099-97004 Revision A
August 2008
© 2008 Thermo Fisher Scientific Inc. All rights reserved.
SpectraSYSTEM and Develop File are trademarks of Thermo Fisher Scientific Inc. in the United States.
Cheminert is a registered trademark of Valco Instruments, Inc. Kel-F is a registered trademark of the 3M Co.
Luer-LOK is a registered trademark of Becton-Dickinson and Company. Teflon and Tefzel are registered
trademarks of E.I. du Pont de Nemours & Co. Tygon is a registered trademark of Saint-Gobain Performance
Plastics Corporation.
Thermo Fisher Scientific Inc. provides this document to its customers with a product purchase to use in the
product operation. This document is copyright protected and any reproduction of the whole or any part of this
document is strictly prohibited, except with the written authorization of Thermo Fisher Scientific Inc.
The contents of this document are subject to change without notice. All technical information in this
document is for reference purposes only. System configurations and specifications in this document supersede
all previous information received by the purchaser.
Thermo Fisher Scientific Inc. makes no representations that this document is complete, accurate or errorfree and assumes no responsibility and will not be liable for any errors, omissions, damage or loss that might
result from any use of this document, even if the information in the document is followed properly.
This document is not part of any sales contract between Thermo Fisher Scientific Inc. and a purchaser. This
document shall in no way govern or modify any Terms and Conditions of Sale, which Terms and Conditions of
Sale shall govern all conflicting information between the two documents.
Release history: Revision A released August 2008
For Research Use Only. Not regulated for medical or veterinary diagnostic use by U.S. Federal Drug
Administration or other competent authorities.
Regulatory Compliance
Thermo Fisher Scientific performs complete testing and evaluation of its products to ensure full compliance with
applicable domestic and international regulations. When the system is delivered to you, it meets all pertinent
electromagnetic compatibility (EMC) and safety standards as described below.
EMC Directive 2004/108/EC
EMC compliance has been evaluated by TUV Rheinland of North America.
CISPR 11: 1998
EN 61000-4-4: 2004
EN 55011: 1998, A1:1999, A2, 2002
EN 61000-4-5: 2001
EN 61000-3-2: 2000
EN 61000-4-6: 2003
EN 61000-3-3: 1995, A1: 2001
EN 61000-4-11: 2001
EN 61000-4-2: 2001
EN 61326-1: 1997, A1: 1998, A2: 2001, A3: 2003
EN 61000-4-3: 2002
CFR 47: 2007
Low Voltage Safety Compliance
Low voltage safety compliance has been evaluated by TUV Rheinland of North America.
This device complies with Low Voltage Directive 2006/95/EC, harmonized standard EN 61010-1: 2001,
IEC 61010-1: 2002, UL 61010A-1: 2004, and CAN/CSA 22.2 61010-1: 2004.
Changes that you make to your system may void compliance with one or more of these EMC and safety standards.
Changes to your system include replacing a part or adding components, options, or peripherals not specifically
authorized and qualified by Thermo Fisher Scientific. To ensure continued compliance with EMC and safety standards,
replacement parts and additional components, options, and peripherals must be ordered from Thermo Fisher Scientific
or one of its authorized representatives.
FCC Compliance Statement
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.
CAUTION Read and understand the various precautionary notes, signs, and symbols contained inside
this manual pertaining to the safe use and operation of this product before using the device.
Notice on Lifting and Handling of
Thermo Scientific Instruments
For your safety, and in compliance with international regulations, the physical handling of this Thermo Fisher Scientific
instrument requires a team effort to lift and/or move the instrument. This instrument weighs 18 kg (40 lbs) and is too
heavy for one person alone to handle safely.
Notice on the Proper Use of
Thermo Scientific Instruments
In compliance with international regulations: Use of this instrument in a manner not specified by Thermo Fisher
Scientific could impair any protection provided by the instrument.
Notice on the Susceptibility
to Electromagnetic Transmissions
Your instrument is designed to work in a controlled electromagnetic environment. Do not use radio frequency
transmitters, such as mobile phones, in close proximity to the instrument.
For manufacturing location, see the label on the instrument.
WEEE Compliance
This product is required to comply with the European Union’s Waste Electrical & Electronic
Equipment (WEEE) Directive 2002/96/EC. It is marked with the following symbol:
Thermo Fisher Scientific has contracted with one or more recycling or disposal companies in each
European Union (EU) Member State, and these companies should dispose of or recycle this product.
See www.thermo.com/WEEERoHS for further information on Thermo Fisher Scientific’s compliance
with these Directives and the recyclers in your country.
WEEE Konformität
Dieses Produkt muss die EU Waste Electrical & Electronic Equipment (WEEE) Richtlinie 2002/96/EC
erfüllen. Das Produkt ist durch folgendes Symbol gekennzeichnet:
Thermo Fisher Scientific hat Vereinbarungen mit Verwertungs-/Entsorgungsfirmen in allen EUMitgliedsstaaten getroffen, damit dieses Produkt durch diese Firmen wiederverwertet oder entsorgt
werden kann. Mehr Information über die Einhaltung dieser Anweisungen durch Thermo Fisher
Scientific, über die Verwerter, und weitere Hinweise, die nützlich sind, um die Produkte zu
identifizieren, die unter diese RoHS Anweisung fallen, finden sie unter www.thermo.com/
WEEERoHS.
Conformité DEEE
Ce produit doit être conforme à la directive européenne (2002/96/EC) des Déchets d'Equipements
Electriques et Electroniques (DEEE). Il est marqué par le symbole suivant:
Thermo Fisher Scientific s'est associé avec une ou plusieurs compagnies de recyclage dans chaque état
membre de l’union européenne et ce produit devrait être collecté ou recyclé par celles-ci. Davantage
d'informations sur la conformité de Thermo Fisher Scientific à ces directives, les recycleurs dans votre
pays et les informations sur les produits Thermo Fisher Scientific qui peuvent aider la détection des
substances sujettes à la directive RoHS sont disponibles sur www.thermo.com/WEEERoHS.
CAUTION Symbol
CAUTION
VORSICHT
ATTENTION
PRECAUCION
AVVERTENZA
Electric Shock: This instrument uses
high voltages that can cause personal
injury. Before servicing, shut down the
instrument and disconnect the instrument
from line power. Keep the top cover on
while operating the instrument. Do not
remove protective covers from PCBs.
Elektroschock: In diesem Gerät werden
Hochspannungen verwendet, die
Verletzungen verursachen können. Vor
Wartungsarbeiten muß das Gerät
abgeschaltet und vom Netz getrennt
werden. Betreiben Sie Wartungsarbeiten
nicht mit abgenommenem Deckel. Nehmen
Sie die Schutzabdeckung von Leiterplatten
nicht ab.
Choc électrique: L’instrument utilise des
tensions capables d’infliger des blessures
corprelles. L’instrument doit être arrêté et
débranché de la source de courant avant
tout intervention. Ne pas utiliser
l’instrument sans son couvercle. Ne pas
elensver les étuis protecteurs des cartes de
circuits imprimés.
Descarga eléctrica: Este instrumento
utiliza altas tensiones, capaces de
producir lesiones personales. Antes de
dar servicio de mantenimiento al
instrumento, éste debera apagarse y
desconectarse de la línea de alimentacion
eléctrica. No opere el instrumento sin sus
cubiertas exteriores quitadas. No remueva
las cubiertas protectoras de las tarjetas
de circuito impreso.
Shock da folgorazione. L’apparecchio è
alimentato da corrente ad alta tensione
che puo provocare lesioni fisiche. Prima di
effettuare qualsiasi intervento di
manutenzione occorre spegnere ed isolare
l’apparecchio dalla linea elettrica. Non
attivare lo strumento senza lo schermo
superiore. Non togliere i coperchi a
protezione dalle schede di circuito
stampato (PCB).
Chemical: This instrument might contain
hazardous chemicals. Wear gloves when
handling toxic, carcinogenic, mutagenic,
or corrosive or irritant chemicals. Use
approved containers and proper
procedures to dispose waste oil.
Chemikalien: Dieses Gerät kann
gefährliche Chemikalien enthalten. Tragen
Sie Schutzhandschuhe beim Umgang mit
toxischen, karzinogenen, mutagenen oder
ätzenden/reizenden Chemikalien.
Entsorgen Sie verbrauchtes Öl
entsprechend den Vorschriften in den
vorgeschriebenen Behältern.
Chimique: Des produits chemiques
dangereux peuven se trouver dans
l’instrument. Proted dos gants pour
manipuler tous produits chemiques
toxiques, cancérigènes, mutagènes, ou
corrosifs/irritants. Utiliser des récipients
et des procédures homologuées pour se
débarrasser des déchets d’huile.
Química: El instrumento puede contener
productos quimicos peligrosos. Utilice
guantes al manejar productos quimicos
tóxicos, carcinogenos, mutagenos o
corrosivos/irritantes. Utilice recipientes y
procedimientos aprobados para
deshacerse del aceite usado.
Prodotti chimici. Possibile presenza di
sostanze chimiche pericolose
nell’apparecchio. Indossare dei guanti per
maneggiare prodotti chimici tossici,
cancerogeni, mutageni, o
corrosivi/irritanti. Utilizzare contenitori
aprovo e seguire la procedura indicata per
lo smaltimento dei residui di olio.
Heat: Before servicing the instrument,
allow any heated components to cool.
Hitze: Warten Sie erhitzte Komponenten
erst nachdem diese sich abgekühlt haben.
Haute Temperature: Permettre aux
composants chauffés de refroidir avant
tout intervention.
Altas temperaturas: Permita que lop
componentes se enfríen, ante de efectuar
servicio de mantenimiento.
Calore. Attendere che i componenti
riscaldati si raffreddino prima di
effetturare l’intervento di manutenzione.
Fire: Use care when operating the system
in the presence of flammable gases.
Feuer: Beachten Sie die einschlägigen
VorsichtsmaBnahmen, wenn Sie das
System in Gegenwart von entzündbaren
Gasen betreiben.
Incendie: Agir avec précaution lors de
l’utilisation du système en présence de
gaz inflammables.
Fuego: Tenga cuidado al operar el
sistema en presencia de gases
inflamables.
Incendio. Adottare le dovute precauzioni
quando si usa il sistema in presenza di gas
infiammabili.
Eye Hazard: Eye damage could occur
from splattered chemicals or flying
particles. Wear safety glasses when
handling chemicals or servicing the
instrument.
Verletzungsgefahr der Augen:
Verspritzte Chemikalien oder kleine
Partikel können Augenverletzungen
verursachen. Tragen Sie beim Umgang mit
Chemikalien oder bei der Wartung des
Gerätes eine Schutzbrille.
Danger pour les yeux: Dex projections
chimiques, liquides, ou solides peuvent
être dangereuses pour les yeux. Porter des
lunettes de protection lors de toute
manipulationde produit chimique ou pour
toute intervention sur l’instrument.
Peligro par los ojos: Las salicaduras de
productos químicos o particulas que
salten bruscamente pueden causar
lesiones en los ojos. Utilice anteojos
protectores al mnipular productos
químicos o al darle servicio de
mantenimiento al instrumento.
Pericolo per la vista. Gli schizzi di
prodotti chimici o delle particelle presenti
nell’aria potrebbero causare danni alla
vista. Indossare occhiali protettivi quando
si maneggiano prodotti chimici o si
effettuano interventi di manutenzione
sull’apparecchio.
General Hazard: A hazard is present that
is not included in the above categories.
Also, this symbol appears on the
instrument to refer the user to instructions
in this manual.
Allgemeine Gefahr: Es besteht eine
weitere Gefahr, die nicht in den
vorstehenden Kategorien beschrieben ist.
Dieses Symbol wird im Handbuch
auBerdem dazu verwendet, um den
Benutzer auf Anweisungen hinzuweisen.
Danger général: Indique la présence
d;un risque n’appartenant pas aux
catégories citées plus haut. Ce symbole
figure également sur l’instrument pour
renvoyer l’utilisateur aux instructions du
présent manuel.
Peligro general: Significa que existe un
peligro no incluido en las categorias
anteriores. Este simbolo también se utiliza
en el instrumento par referir al usuario a
las instrucciones contenidas en este
manual.
Pericolo generico. Pericolo non
compreso tra le precedenti categorie.
Questo simbolo è utilizzato inoltre
sull’apparecchio per segnalare all’utente
di consultare le istruzioni descritte nel
presente manuale.
When the safety of a procedure is
questionable, contact your local Technical
Support organization for Thermo Fisher
Scientific San Jose Products.
Wenn Sie sich über die Sicherheit eines
Verfahrens im unklaren sind, setzen Sie
sich, bevor Sie fortfahren, mit Ihrer
lokalen technischen
Unterstützungsorganisation für Thermo
Fisher Scientific San Jose Produkte in
Verbindung.
Si la sûreté d’un procédure est incertaine,
avant de continuer, contacter le plus
proche Service Clientèle pour les produits
de Thermo Fisher Scientific San Jose.
Cuando la certidumbre acerca de un
procedimiento sea dudosa, antes de
proseguir, pongase en contacto con la
Oficina de Asistencia Tecnica local para
los productos de Thermo Fisher Scientific
San Jose.
Quando e in dubbio la misura di sicurezza
per una procedura, prima di continuare, si
prega di mettersi in contatto con il
Servizio di Assistenza Tecnica locale per i
prodotti di Thermo Fisher Scientific San
Jose.
CAUTION Symbol
CAUTION
Electric Shock: This instrument uses
high voltages that can cause personal
injury. Before servicing, shut down the
instrument and disconnect the instrument
from line power. Keep the top cover on
while operating the instrument. Do not
remove protective covers from PCBs.
Chemical: This instrument might contain
hazardous chemicals. Wear gloves when
handling toxic, carcinogenic, mutagenic,
or corrosive or irritant chemicals. Use
approved containers and proper
procedures to dispose waste oil.
Heat: Before servicing the instrument,
allow any heated components to cool.
Fire: Use care when operating the system
in the presence of flammable gases.
Eye Hazard: Eye damage could occur
from splattered chemicals or flying
particles. Wear safety glasses when
handling chemicals or servicing the
instrument.
General Hazard: A hazard is present that
is not included in the above categories.
Also, this symbol appears on the
instrument to refer the user to instructions
in this manual.
When the safety of a procedure is
questionable, contact your local Technical
Support organization for Thermo Fisher
Scientific San Jose Products.
C
Contents
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi
About This Guide. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .xi
Safety and Special Notices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .xi
Manual Conventions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xii
Good Laboratory Practices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .xiii
Contacting Us . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xv
Chapter 1 Installation and Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1
Start-up Checklist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Unpacking. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
LC System Connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Performance Verification. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Specifications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Chapter 2 A Quick Example. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .25
Learning Your Way Around . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Instrument Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Practice Examples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
Chapter 3 Basic Operations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .35
Theory of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
SpectraSYSTEM 4000 Pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Instrument Startup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
Some Routine Operations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
The File(s) Menu. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Purging Solvent Lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
Running the Pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
The Commands Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
Status . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
Monitoring Pump Performance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
Shutting Down at the End of the Day . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
Chapter 4 Advanced Operations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .65
The Options Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
The Queue Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
Thermo Scientific
SpectraSYSTEM P4000 Gradient Pump User Guide
ix
Contents
The Develop File. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
Creating a Develop File Program. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
Develop File Examples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
Running the Develop File . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98
Preview Record . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106
The Tests Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108
Chapter 5 Required Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .127
Maintenance Schedule. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
Maintenance Log. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
Extending the Maintenance Period . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132
Maintenance Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
Maintenance Tips . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153
Repair Instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
Appendix A Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .157
Theory of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
Troubleshooting Your Pump. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
Appendix B Glossary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .167
Appendix C Kits and Parts Lists . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .175
Accessory Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175
Standard Maintenance Kit. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 176
Standard LC Fittings Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 176
Piston Flush Seal Kit (10 mL) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177
Piston Seal Kit (30 mL) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177
Solvent Inlet Tube Kit. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177
Solvent Tube Extension Kit. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177
Manual Injection Valve Bracket Kit. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178
Narrow-bore Upgrade Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 181
Index
x
SpectraSYSTEM P4000 Gradient Pump User Guide
Thermo Scientific
P
Preface
About This Guide
This guide describes how to install and maintain the SpectraSYSTEM P4000 gradient pump,
as well as how to control the gradient pump from the front panel keypad.
Safety and Special Notices
Caution!
A caution alerts you to situations that could result in personal injury. It also tells you how to
avoid them.
High Voltage!
A high voltage caution alerts you to the presence of high voltage and to the potential injury
that could occur from electrical shock were you to come in contact with a specific
instrument area or component. It also tells you how to avoid contact with the high-voltage
areas in your instrument.
Hot Surface!
A hot surface caution alerts you to potential injury that could occur from coming in contact
with a heated surface or area on or in an instrument. It also tells you how to avoid contact
with the heated surfaces in your instrument.
Note
Notes alert you to the correct operating or maintenance procedures needed to prevent
equipment or data damage. They also alert you to important exceptions, side effects, or
unexpected occurrences that may result from certain action(s).
Hint
Hints call out general rules or shortcuts. They specify ways to obtain the best performance
and results from your instrument.
Thermo Scientific
SpectraSYSTEM P4000 Gradient Pump User Guide
xi
Preface
Manual Conventions
This manual uses several conventions. Among them are menu displays, text conventions
(brackets, slashes, and so on), and standard words.
Displays
We will depict the two-line display as shown below. Note that in menu illustrations, the
triangular cursor location is indicated by a caret (>).
>FILES
QUEUE
COMMANDS
TESTS
OPTIONS
A two-line menu display
Frequently the two lines shown on the display are only part of a longer menu which you
would see by pressing the down-arrow key. In this manual, menus having more than two lines
are represented as follows:
Edit File
1
File Name
-------------------------------------------------------->Solvent Program
Options
Timed Events
A menu longer than two lines
Text Conventions
Three typographic conventions are used to differentiate between keys, menus, and fields.
Brackets
Brackets, [ ], indicate instrument keys, For example: Press the [MENU] key.
Slashes
Slashes, / /, are text conventions used around menu choices. For example: From the Main
Menu, select /FILES/, /Edit/.
Capitalization
Capitalization is used to make field and menu names appear just as they do on the display.
Generally the first letters of field names are capitalized. For example: In /Solvent Program/ go
to the Flow field.
xii
SpectraSYSTEM P4000 Gradient Pump User Guide
Thermo Scientific
Preface
Standard Words
We have also standardized the meanings of two words: “select” and “enter.”
Select
The word “select” is used when you need to choose from among available options. For
example, to select a particular menu choice, you would move the cursor to the appropriate
choice and press [ENTER]. To “select” a field entry, move the cursor to the appropriate field
and use the [+] and [–] keys to scroll to the desired choice.
Enter
The word “enter” is used when you need to specify individual alphanumeric digits. To “enter”
a particular value, move the cursor to the field and use the [+] and [–] keys to increment or
decrement each digit in the field until the desired value or letter appears.
Good Laboratory Practices
To obtain optimal performance from your LC system and to prevent personal injury or injury
to the environment, do the following:
• Keep good records
• Read the manufacturers’ Material Safety Data Sheets for the chemicals being used in your
laboratory
• Remove particulate matter from your samples before you inject them into the liquid
chromatograph
• Use HPLC grade solvents
• Connect the drainage tubes from the pump, autosampler, and detector to an appropriate
waste receptacle. Dispose of solvents as specified by local regulations
Keep Good Records
To help identify and isolate problems with either your equipment or your methodology, keep
good records of all system conditions (for example,% RSDs on retention times and peak areas,
peak shape and resolution). At a minimum, keep a chromatogram of a typical sample and
standard mixture, well documented with system conditions, for future reference. Careful
comparison of retention times, peak shapes, peak sensitivity, and baseline noise can provide
valuable clues to identifying and solving future problems.
Thermo Scientific
SpectraSYSTEM P4000 Gradient Pump User Guide
xiii
Preface
Chemical Toxicity
Although the large volume of toxic and flammable solvents used and stored in laboratories can
be quite dangerous, do not ignore the potential hazards posed by your samples. Take special
care to read and follow all precautions that ensure proper ventilation, storage, handling, and
disposal of both solvents and samples. Become familiar with the toxicity data and potential
hazards associated with all chemicals by referring to the manufacturers’ Material Safety Data
Sheets (MSDS).
Sample Preparation
Always consider the solubility of your sample in the solvent/mobile phase. Sample
precipitation can plug the column, tubing or flowcell causing flow restriction. This
obstruction can result in irreparable damage to the system. To avoid damage caused by
particulate matter, filter samples through 0.45 or 0.2 micron (or less) filters.
Solvent Requirements
Many chemical manufacturers provide a line of high-purity or HPLC-grade reagents that are
free of chemical impurities. Routine filtration of all solvents or eluents through a 0.45 or
0.2 micron (or less) fluorocarbon filter before placing them in the solvent reservoir
significantly prolongs the life and effectiveness of the inlet filters, check valves and seals,
injector, and column. Typically, HPLC-grade solvents do not require filtration.
Choose a mobile phase that is compatible with the sample and column you have selected for
your separation. Remember that some solvents are corrosive to stainless steel.
Solvent Disposal
Make sure you have a solvent waste container or other kind of drain system available at or
below the benchtop level. Most solvents have special disposal requirements and should not be
disposed of directly down a drain. Follow all governmental regulations when disposing of any
chemical.
High-pressure Systems and Leaks
LC systems operate at high pressures. Because liquids are not highly compressible they do not
store much energy. Accordingly, there is little immediate danger from the high pressures in an
LC system. However, if a leak occurs, correct it as soon as possible. Always wear eye and skin
protection when operating or maintaining an LC system. Always shut down the system and
return it to atmospheric pressure before attempting any maintenance.
xiv
SpectraSYSTEM P4000 Gradient Pump User Guide
Thermo Scientific
Preface
Contacting Us
There are several ways to contact Thermo Fisher Scientific for the information you need.
Y To contact Technical Support
Phone
Fax
E-mail
Knowledge base
800-532-4752
561-688-8736
[email protected]
www.thermokb.com
Find software updates and utilities to download at mssupport.thermo.com.
Y To contact Customer Service for ordering information
Phone
Fax
E-mail
Web site
800-532-4752
561-688-8731
[email protected]
www.thermo.com/ms
Y To copy manuals from the Internet
Go to mssupport.thermo.com and click Customer Manuals in the left margin of the
window.
Y To suggest changes to documentation or to Help
• Fill out a reader survey online at www.thermo.com/lcms-techpubs.
• Send an e-mail message to the Technical Publications Editor at
[email protected]
Thermo Scientific
SpectraSYSTEM P4000 Gradient Pump User Guide
xv
1
Installation and
Specifications
Introduction
This chapter contains information necessary to install your Thermo
Scientific, SpectraSYSTEM P4000 pump. The step-by-step
instructions describe how to set the voltage, how to connect tubing,
and how to prime and purge the pump. Use the checklist on the next
page to complete pump installation. Also, be sure you read the Safety
Information at the front of this manual before proceeding with any
installation. If you have any questions or need further assistance,
refer to the Preface for the product support or technical assistance
numbers.
Thermo Scientific
1
Start-up Checklist
This list is a brief summary of tasks that should be completed to
install your pump. Complete installation information is contained in
this chapter.
Inspect your instrument
Check for parts shortages
Set the voltage
Place the pump
Connect the power cord
Check initial response to power-on
Hardwire to eight-pin port, using external
function connector, making electrical connection
to other SpectraSYSTEM instruments
Install kits or accessories
Prepare and connect solvents
Connect inlet lines
Prime with solvent
Purge solvent lines
Connect to system
This pump was installed by:
______________________________________ ______________
(Name)
(Date)
2
Thermo Scientific
Unpacking
INSPECT YOUR
INSTRUMENT
Your pump was shipped in a special container designed to provide
excellent protection from routine wear and tear encountered in transit.
After unpacking, inspect your pump and its accessories for missing
parts and/or physical damage. If damage is found, notify both the
carrier and your sales representative. DO NOT return any goods
without prior authorization from Thermo Fisher Scientific.
The contents of your ship kit is as follows:
1
1
1
1
1
Pump
Accessory Kit (A4070-010)
Tubing Kit (See Appendix C)
Declaration of Conformity
SpectraSYSTEM documentation CD
OPTIONS
AVAILABLE
A variety of options, kits, and accessories is available for your pump.
Refer to Appendix C for a description and parts list for each. If you
purchased an inert/biocompatible pump, the correct tubing and liquid
ends were installed at the factory before shipment. For a list of all
available accessories, upgrades, and kits, contact your Thermo Fisher
Scientific sales representative. Note that all upgrades require
installation by Thermo Fisher Scientific.
NOTE: The pump features a bypass valve pre-installed as standard
equipment.
Thermo Scientific
3
Figure 1.1 The SpectraSYSTEM P4000 pump
4
Thermo Scientific
Installation
LIFTING AND
CARRYING THE
PUMP
The correct way to carry the pump is to use the two hand holds, one
located underneath the front of the pump, and the other at the top of
the back, near the power switch. Grasp the pump well underneath the
front when lifting and carrying.
SETTING THE
VOLTAGE
All pumps are configured at shipment for 230 VAC (50/60 Hz)
operation. Depending upon the country of use, you might need to
change the voltage setting.
NOTE: Check the position of the voltage select barrel located on the
rear of the instrument. If the indicated voltage setting is not
consistent with your area, DO NOT CONNECT THE POWER CORD!
Figure 1.2 shows the pump rear panel.
Thermo Scientific
5
Lift Point
I
ON
O
OFF
Communication
Port (COMM)
Power Switch
TERMINAL CONFIGURATION
8-Pin Port
Communication
Port to SCM400
(DEGAS)
Fan
Power Input
Voltage Selector/
Fuse Cover
230Vac
PU-Z003E/DT
Power Cord
Connection
Figure 1.2 Rear panel
6
Thermo Scientific
The voltage setting can be easily modified as follows:
1. Remove the tape label covering the power entry receptacle.
2. Ensure that the power cord is not connected to the pump.
3. Use a small, flat blade screwdriver to pry open the power
selector/fuse cover to expose the voltage selector barrel. You
will probably hear the top edge of the cover snap as it is pried
open (Figure 1.3).
PRY OPEN HERE
REMOVE, TURN
& REPLACE
TO CHANGE VOLTAGE
115Vac
230Vac
PU-Z006E/FM
230Vac
230Vac
Figure 1.3 Power selector/fuse cover, closed, with "voltage window."
The opened power selector/fuse cover, with barrel selector removed.
4. Remove the plastic selector barrel by pulling it straight out.
5. Rotate the barrel until you see the desired voltage (either
115 VAC or 230 VAC) and insert the barrel back into the
housing with the desired voltage visible.
6. Firmly snap the housing cover back in place. Be sure that the
selected voltage is visible in the voltage window.
HINT: Use two thumbs to push up on the top half of the cover as you
push in. The voltage selected will be visible through the window.
Thermo Scientific
7
PLACING THE
PUMP
The pump weighs approximately 38 pounds (18 kg) and requires at
least 6 inches (16 cm) of bench width and at least 19 inches (48 cm)
of bench depth. If used with a manual injector bracket, the pump
requires 9 inches (23 cm) of bench width. The pump needs a space at
least 15 inches (38 cm) high.
Place the pump on a level surface. Leave 2 inches (6 - 7 cm) behind
the instrument for good air flow and access to electrical connections.
Keep the pump away from heating and cooling ducts, and avoid
exposing the pump to direct sunlight. The pump should be placed to
the far left of your LC system if it is used with a SpectraSYSTEM
autosampler or detector.
CONNECTING THE
POWER CORD
Attach the AC power cord (Figure 1.3). Plug the power connector
into an appropriately grounded power outlet.
NOTE: For safe operation and optimum performance, the pump must
be connected to a properly grounded power receptacle.
CHECKING INITIAL
RESPONSE TO
POWER ON
Turn the power on by pressing the power switch (Figure 1.2). With
the pump's front panel facing you, the power switch is located in the
back, on the upper right-hand side. The fan starts and the display
shown in Figure 1.4 appears for one second.
Version
(X.YY.ZZ)
Figure 1.4 Brief power-up message
If this message does not appear, double-check the electrical
connections and try turning on the pump once more, watching the
screen closely. If the message still does not appear, contact your
Thermo Fisher Scientific representative.
Next, the display shows the Status Screen, similar to Figure 1.5.
Status
Flow
PSI
MaxP
STOP
1.00
154
3000
Figure 1.5 4000 Status Screen
8
Thermo Scientific
HARDWIRING
EXTERNAL EVENTS
Pin 6 on the eight-pin port on the back of the pump (Table 1.1) allows
you to control another device, such as a column switching valve or
fraction collector. If you plan to control such a device or instrument
using the pump's Timed Events feature, insert the external function
connector into the eight-pin port.
The pins are labeled both on the port and on the external function
connector. Ensure that the pin numbers match whenever plugging
into the connector to the port.
Hardwire your device using the 4-connection cable. Loosen pin 6's
small screw, insert the wire, then tighten the screw.
You must also insert the external function connector if you use a
SpectraSYSTEM autosampler. More information about making
hardwire connections to an autosampler is found on page 17, and in
the SpectraSYSTEM Autosampler User Guide.
Pin assignments for the eight-pin port are shown in Table 1.1:
Table 1.1 Eight-pin port assignments
Pin #
1
2
3
4
5
6
7
8
Description
READY (Output)
+5 VDC 100 mA MAX
GROUND
PRESSURE 0.1 V/1000 PSI
STOP (Input)
TIMED EVENT (Output)
RUN (Input)
INJ HOLD (Output)
All outputs (pin 1, pin 6, and pin 8) are open-collector type, capable
of sinking up to 30 mA at a maximum of 30 VDC.
INSTALLING KITS
OR ACCESSORIES
Refer to Appendix C for complete instructions for installing the
Manual Injection Valve Bracket (column holder).
PREPARING AND
CONNECTING
SOLVENTS
If you did not purchase a solvent degassing apparatus or solvent
bottles from Thermo Fisher Scientific, skip this section. Continue
with Connecting Inlet Lines on the next page.
Thermo Scientific
9
Solvent Bottles
Prepare your solvent bottles by following the steps below.
1. Rinse the bottles with LC-grade solvent to remove any dust.
2. Fill the bottles with appropriate LC-grade solvents.
3. The bottle caps are pre-assembled to include an inlet line and
filter. Ensure that the filters are tightly assembled to their
fittings, and the filter fittings are firmly attached to the inlet
lines. Place the solvent filter/inlet line into each bottle, making
sure that the inlet filter rests on the bottom of the bottle. Cap
the bottle.
4. Attach the appropriate A, B, C, or D label to each solvent bottle
cap to identify it.
5. Run vent lines from each bottle to an appropriate exhaust
apparatus.
Degassing
There are two recommended methods for degassing solvents for use
with your pump: vacuum degassing and helium degassing.
NOTE: Solvent degassing is required when proportioning (blending)
solvent using your SpectraSYSTEM gradient pump. Degassing is not
required for isocratic, premixed pump operation, but is recommended
because of improved detector performance.
If you purchased a Thermo Scientific degasser, set up your degas
apparatus as described in the degasser kit and continue the pump
installation when you have a supply of degassed solvent available.
CONNECTING
INLET LINES
Refer to Figure 1.6 to make the plumbing connections.
10
Thermo Scientific
RUN
STATUS
MENU
STOP
ENTER
PURGE
SpectraSYSTEM
P4000
Crossover Tube
Pressure
Transducer
Transducer
Tube
Bypass Valve
Knob
Outlet
Liquid
End
Bypass
Valve
Transducer
Check
Valve
Bypass Valve Tube
Inlet
Liquid
End
Solvent
Inlets
Outlet Port
(to autosampler or
manual injection valve)
Inlet Check Valve
Solvent Inlet Tube
Proportioning
Valve
Drip Tray
PU-Z002E/FM
Waste Tube
Lift Point
Figure 1.6 Front panel, cover removed
Thermo Scientific
11
BYPASS VALVE
The bypass valve is shown in Figure 1.7. Do not attach a line to the
waste/prime port (the middle port) now, but have the solvent waste
tube (provided in the accessory kit) ready. You will attach the tube to
the waste/prime port after priming the pump.
PRIMING AND
PURGING
Each pump is shipped with methanol in the pump heads and
connecting tubing. If the first solvent you'll use is not miscible with
methanol, the pump must first be primed with an intermediary
solvent. Once primed, you should purge the pump to remove any air
bubbles.
HINT: It is best to prime the pump initially with methanol to fully wet all
internal surfaces. Priming with 100% water can often result in trapped air
due to the high surface tension of the water. Trapped air affects flow
stability.
Priming the Pump
To prime the pump with your solvent and simultaneously flush the
methanol out, you will need the 20 mL priming syringe and Luer
adapter found in the accessory kit. You will also need a solvent waste
container.
1. Remove the waste line from the waste/prime port of the bypass
valve, if connected.
2. Install the Luer adapter to the waste/prime port (Figure 1.7).
Tighten to finger-tight, then use a wrench to tighten
approximately 1/4-turn. (This allows the priming syringe to be
attached and detached conveniently.)
12
Thermo Scientific
Open
Closed
Bracket
mounting
screws
Bypass Valve
Waste/Prime port
Luer adapter
Bypass outlet
tube
Bypass inlet
tube
Figure 1.7 Bypass valve with Luer adapter connected
3. Make sure that the 20 mL Luer-tip priming syringe is fully
depressed. Connect the syringe to the adapter in the
waste/prime port (Figure 1.8) twisting the syringe slightly to
make a leak-free connection.
4. Position a solvent waste container nearby to collect the syringe
discharge, since two or three syringe volumes may be needed
to prime the pump. Make sure that all tube connections are airtight.
Figure 1.8 Bypass valve with syringe attached
Thermo Scientific
13
5. Open the bypass valve by turning the knob fully
counterclockwise.
NOTE: Solvents flow through the pump when the pump is purging or
when a file is initialized (loaded as a run file).
6. Turn on power to the pump (if it not already on) and press
[PURGE]. The cursor should appear under the word Purge on
the display. Press [+] until the word ALL appears on the
P4000.
NOTE: Do not purge ALL or BOTH unless all solvent lines are
connected, and contain solvent. If this is not the case, select the
appropriate solvent (A, B, C, D), instead.
NOTE: Purging starts whenever the cursor is moved out of the
Time field using the [ENTER] key.
Purge
Flow
Time
ALL
1.00
0.0
Figure 1.9 Purge Menu (P4000)
NOTE: The Flow field may be labeled Pres instead of Flow,
depending on the purge mode that is set. The purge mode can be
changed from the [MENU], /OPTIONS/, /More/, Purge Mode field.
7. For analytical applications: Move the cursor to the Flow field.
Set the flow rate to 10 mL/min. If the purge mode is pressure,
set the pressure to 1000 psi (69 bars, 7 MPa).
For narrow-bore applications: Move the cursor to the Flow
field. If the purge mode is pressure, set the pressure to
1000 psi (69 bars, 7 MPa).
8. Move the cursor to the Time field, then press [ENTER]. You
will hear the pump's motor start.
9. Slowly pull the syringe plunger back, thus creating a small
vacuum in the solvent lines and drawing the solvent from the
solvent bottles (or the vacuum degasser), into the pump heads.
Do not draw back so far as to remove the plunger from the
syringe barrel!
If more than one draw is necessary to prime the pump, (i.e., the
syringe fills with air before solvent enters the pump heads),
press [STOP], close the bypass valve (turn clockwise), remove
the syringe, and depress the plunger. Reconnect the syringe,
open the bypass valve, press [PURGE], enter a flow rate or
time and press [ENTER] and finish drawing the solvents into
the pump as described at the beginning of this step.
If you notice a leak in one of the fittings, or need to stop the
solvent flow, press [STOP]. This will immediately stop flow
through the pump.
14
Thermo Scientific
10. When solvent steadily appears in the syringe and no air bubbles
are present, press [STOP], then close the bypass valve.
11. Gently remove the syringe and empty it into the solvent waste
container.
12. Remove the Luer adapter from the valve port and store it, along
with the priming syringe, for later use.
13. Connect the solvent waste line to the waste/prime port of the
valve and tighten to finger-tight. Route the waste tubing to an
appropriate solvent waste container.
Thermo Scientific
15
Purging the Pump
With the pump primed, you now must purge the lines containing your
chosen solvents.
1. Ensure that the solvent inlet filters inside each solvent bottle
are in a vertical position so that air within the filter will not be
trapped.
2. Fully open the bypass valve.
3. We suggest that you initially purge the lines with a volume of
10 mL.
Follow the steps below to begin the purge operation:
a) Press [PURGE] to reach the Purge Menu. If desired,
change the purge mode by pressing [MENU] and selecting
/OPTIONS/, /More/, Purge Mode. (The words in the top
line of the display will change, depending on your purge
mode preference.)
b) Select "ALL" or the solvent of choice in the Purge field,
then press [ENTER]. (See Figure 1.10).
c) If the purge mode is Flow, do the following:
For analytical applications: If the purge mode is Flow,
enter a flow of 10.00 mL/min. [If pressure, set to 1000 psi
(69 bars, 7 MPa).]
For narrow-bore applications: If the purge mode is Flow,
enter a flow of 5.00 mL/min. [If pressure, set to 1000 psi
(69 bars, 7 MPa).]
d) Press [ENTER] and enter a time of 1.00 minute. Ten
milliliters should be enough volume to remove any trapped
air and ensure that the pump and tubing are cleansed of any
contaminants.
e) Press [ENTER]. The pump's motor will start. Purging will
automatically stop after one minute and will initialize the
pump. After the pump has stopped, be sure to close the
purge valve.
Purge
Flow
Time
ALL
10.00
1.00
Figure 1.10 Purge Menu with flow and time values entered
If you choose to purge without setting a time in the Time field, you
can stop the purge by pressing [STOP]. This will immediately stop
flow through the pump.
4. After you completed the purge, and the pump has stopped,
close the bypass valve.
16
Thermo Scientific
LC System Connections
Once the pump is purged, you can plumb it to the rest of your
chromatographic system. Figure 1.7 and Figure 1.8 illustrate the
bypass valve, showing the pump's outlet. Using a pre-cut piece of
stainless steel tubing, connect the outlet to your autosampler or
manual injection apparatus.
If you are using a manual injector valve from Thermo Fisher
Scientific, Appendix C for complete information for installing the
injector valve bracket.
If you are using a SpectraSYSTEM autosampler, the pump can send a
ready signal to the autosampler through pin 1, and can receive a stop
signal from the autosampler through pin 5. In addition, the ground
contact (pin 3) must also be connected to the autosampler's ground
contact. For complete information on how to make these
connections, refer to the Chapter 1 of the SpectraSYSTEM
Autosamplers User Guide. The table below summarizes the hardwire
connections necessary between a SpectraSYSTEM autosampler and
pump.
Table 1.2 Pump connections to SpectraSYSTEM autosampler
Pump
READY (Output)
GROUND
STOP (Input)
RUN (Input)
INJ HOLD (Output)
Autosampler
Pin 1
Pin 3
Pin 5
Pin 7
Pin 8
Thermo Scientific
Pin 5 PUMP READY
Pin 1 GROUND
Pin 3 PUMP STOP
Pin 4 GRAD START
Pin 7 INJ HOLD
17
Performance Verification
After installing your SpectraSYSTEM pump, for best results, run a
performance test to verify that the instrument is working properly.
Common tests for pumps include flow accuracy and precision,
gradient linearity, and compositional accuracy. Procedures for these
performance tests are described below.
FLOW ACCURACY
There are many ways to test pump flow accuracy: graduated cylinder
vs. time, calibrated flowmeter, or gravimetric vs. time. The procedure
below describes how to measure the flow accuracy using a
gravimetric procedure. In general you will set the pump to a flow
rate, collect eluent for a specified time in a weighed flask, and
determine the flow rate and accuracy. The actual flow rate can also
be used to calibrate your pump. This test will take approximately
20 minutes.
Test Setup
Column or flow restrictor: To deliver 1000 psi backpressure
Flow rate: Any flow rate to be tested for accuracy
Mobile phase: MeOH or other appropriate mobile phase
Experimental
Using the following steps to calculate the flow rate accuracy:
1. Prepare the desired mobile phase for the accuracy test.
2. Filter and degas the mobile phase.
3. Purge the pump and the column or flow restrictor being used.
4. Set the pump at the desired flow rate and start.
5. Weigh a clean flask capable of holding approximately
10 minutes of volume at the set flow rate. Include top and
any other items attached to the flask.
HINT: A volumetric flask is ideal because it helps to minimize
evaporation. Also, wrap tube and top of flask with aluminum foil to
help eliminate evaporation.
6. Record this weight.
7. Place the flask under the flow stream from the pump and start
timing.
HINT: Use a stopwatch for the timing portion of the test. The more
accurate the stopwatch the better the results.
18
Thermo Scientific
8. Collect the pump's eluent for 20 minutes.
9. Reweigh the flask and record.
10. Calculate the actual flow rate:
[(Weightfull - Weightempty) / Density] / Time = Flow Rate
11. Set a flow rate of 0.5 mL/min using MeOH as the mobile
phase.
Flow Rate = [(22.8577 g - 14.8858 g) / 0.7894 g/mL] / 19.9687 min
Flow Rate = [(7.9719 g) / 0.7894 g/mL] / 19.9687 min
Flow Rate = 10.0987 mL / 19.9687 min
Flow Rate = 0.506 mL / min
12. Calculate the flow accuracy of the pump:
Flow Accuracy = 100 × ⎢(FRset - FRactual) / FRset⎟
Flow Accuracy = 100 × 0.006 mL / min
Example
Using the data from step 10 gives the following results:
Flow Accuracy = 100 × ⎢(0.5 mL/min - 0.506 mL/min)/0.5 mL/min ⎢
Flow Accuracy = 100 × (0.006 mL / 0.5 mL/min)
Flow Accuracy = 1.2%
HINT: For flow rates above 0.5 mL/min, it is best to install an in-line
100 μL mixer replacing the 3 μL mixer.
Specification
Typical flow accuracy results are less than 1.5% (absolute) using this
gravimetric procedure in the 100 μL/min to 2 mL/min flow rate
range.
The factory specification is less than 1% (absolute) using a calibrated
flowmeter and methanol as the mobile phase.
FLOW PRECISION
Flow precision checks the reproducibility of the flow rate. This
procedure is simply running the accuracy test 7 or more times and
calculating a percent relative standard deviation. This procedure will
require at least a couple of hours.
Thermo Scientific
19
Test Setup
Set up the flow precision test above to perform 7 or more replicate
runs.
Column or flow restrictor: To deliver 1000 psi back pressure
Flow rate: Any flow rate that accuracy is to be tested
Mobile phase: MeOH or other appropriate mobile phase
HINT: For flow rates above 0.5 mL/min, it is best to install a 100 μL mixer
in-line replacing the 3 μL mixer.
Experimental
1. Repeat the flow accuracy test above for 7 or more replicate
runs.
2. Determine the flow rate for each replicate.
3. Determine the average flow rate for the replicate runs.
4. Determine the standard deviation for the replicate runs.
5. Determine the relative standard deviation for the replicate runs.
Using the following example values:
Average = 0.502 mL/min
Standard Deviation = 0.004
Relative Standard Deviation (RSD) = 100 × SD/Average
Yields:
RSD = 100 × (0.004/0.502 mL/min)
RSD = 100 × 0.00797
RSD = 0.797%
Specification
Typical results for flow precision are less than 1% (absolute).
However, results can vary based on temperature, mobile phase, and
flow rate.
GRADIENT
PERFORMANCE
Gradient performance is measured by testing gradient linearity and
step/compositional accuracy. This procedure consists of running a
pump method, which establishes a baseline, runs a linear gradient,
and then runs 20% steps. This procedure can be run on a single pair
of pump solenoid/switching valves or both pairs for quaternary
gradient pumps. Each pair of valves will take approximately
50 minutes.
20
Thermo Scientific
Depending data system, you will need a way to record the absorbance
levels throughout the gradient run. With a computer data system or
strip chart recorder this is automatically done for you. However, with
an integrator this may require some manual recording of these levels.
HINT: The levels of importance are 0% spiked mobile phase, 100% spiked
mobile phase, and each of the 20% steps.
Test Setup
Flow Restrictor:Deliver 1,000 to 2,000 psi
Mobile phases:
A and/or C: Methanol
B and/or D: Spiked Methanol with 7 ppm
Ethyl Paraben
Flow rate:
0.5 mL/min (or other desired flow rate)
Detection:
UV at 254 nm
Experimental
1. Prepare mobile phases, filter through a 0.45-micron filter, and
degas.
2. Purge pump lines with the appropriate mobile phase.
3. Purge entire system with methanol.
4. Setup the following gradient method for the pump:
NOTE: This procedure is for a quaternary pump. For a binary pump
simply eliminate the gradient profile for C & D.
5. Run the gradient profile of step 4.
6. Record the 0%, 100%, and each 20% absorbance level of the
spiked methanol.
6. Subtract the 0% level from all other recorded levels.
7. Determine 1% absorbance level of spiked methanol from the
recorded 100% level.
8. Record each of the 20% absorbance levels of spiked methanol.
HINT: Record data after the particular step has stabilized.
Specification
The factory specification is that the linear gradient falls within ±1%.
Each step must fall within ±1% of it’s respective theoretical value.
Thermo Scientific
21
Figure 1.11 Gradient Profile
22
Thermo Scientific
Specifications
P4000:
Gradient pump using a dual in-line and floating piston design,
bayonet-mounted liquid ends, and patented ceramic check valves.
Physical:
14.5” (37 cm) × 6” (15 cm) × 18.5 (47 cm) (H × W × D)
38.6 lb. (18 kg)
Wetted Surfaces:
316 stainless steel, analytical and narrow-bore
PEEK, semi-prep
other surfaces for all pumps: Teflon®, Tefzel®, sapphire,
polyethylene
Delay Volume:
< 800 μL, analytical
< 500 μL, narrow-bore (P4000)
Flow Rate Range:
0.01 - 10.00 mL/min, analytical or narrow-bore pumps
0.01 - 30.00 mL/min, semi-prep PEEK pumps
Flow Accuracy:
< 1% at 1.0 and 4.0 mL/min, analytical or semi-prep
< 1% at 0.2 and 1.0 mL/min, narrow-bore
Flow Precision:
Typically < 0.2% at 0.5 mL/min or greater
Gradient Linearity:
< 1.0% at 1.0 mL/min from 5 - 95% composition, analytical
< 1.0% at 0.5 mL/min from 5 - 95% composition, narrow-bore
Compositional Accuracy:
< 1.0% at 1.0 mL/min from 5 - 95% , composition analytical
< 1.0% at 0.5 mL/min from 5 - 95% composition, narrow-bore
Compositional Precision:
Typically < 1.0% at 1.0 mL/min analytical
Typically < 0.2% at 0.3 mL/min, narrow-bore
Pressure Range:
42 MPa or 420 bar or 6000 psi, analytical or narrow-bore
28 MPa or 280 bar or 4000 psi, semi-prep
Pressure Pulsation:
Typically < 1.0% at 1 mL/min
Method Files:
9 method + 1 Shutdown + 1 Develop, P4000
Communications:
Remote Inputs: Ready, Run, Stop
Timed Events
Analog Pressure output
RS-232 (P4000)
Environmental
10-40 °C
5-95% RH noncondensing
Power:
115/230 VAC, 50/60 Hz
T4A 200VA
Safety/EMC Compliance:
CSA, TÜV, FCC, CE Mark, EMC and Low Voltage Directives
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2
A Quick Example
Introduction
This chapter provides you with the concepts you'll need for using
your gradient pump. It also introduces you to the instrument's pump's
screens and menus. In this chapter you will set up a few typical
options, purge your solvent lines, and run a flow stability test.
If you already feel comfortable with how to move through menus and
displays, just scan this chapter and proceed to Chapter 3. If you want
more practice with the pump, follow the instructions in this chapter
closely. Since the object of this chapter is to become familiar with
the keypad and menus, we won't provide detailed explanations of the
examples shown. More information can be found in one of the
succeeding chapters.
If you haven't installed your pump, be sure that you read the Safety
Information section and follow the procedure in Chapter 1.
Throughout our explanations, we encourage you to explore the
general architecture of the instrument's menus and screens. Use the
menu trees in the front pocket of the manual as your guide.
Learning Your Way Around
AS EASY AS 1-2-3!
It's easy to learn your way around a SpectraSYSTEM pump. Just
remember these three easy rules:
1. The ([∧], [∨], [<], [>]) move the cursor in the direction printed
on the key.
HINT: Press [MENU] to jump quickly to the top of the menu
structure.
2. The shape of the cursor determines how you make a selection:
•
If a triangular cursor appears, press [ENTER].
•
If a blinking square cursor appears, press the [+] or [-] keys
to scroll up or down through preset choices, or to increase
or decrease alphanumeric entries
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3. There are four ways to accept (and automatically save) an
entry. Just move the cursor out of the field by any of the
following methods:
•
Pressing [ENTER]
•
Using the arrow keys
•
Pressing [MENU]
•
Pressing [STATUS]
NOTE: You won't be able to leave a menu if errors are present or if you
haven't filled in all the necessary entries.
Several visual clues help you move through the pump's menus and
enter values.
1. Top-level menu choices are displayed in all-capital letters;
lower-level menu choices are displayed in upper- and lowercase letters.
2. A field's square cursor changes to an underscore cursor when
you're scrolling through preset choices or entering numerical
values and characters.
3. A solid down-arrow (T) on the right side of some displays
indicates that the current menu continues on additional screens.
To access additional menu lines, press the down-arrow key,
[∨].
4. The last line of a longer menu is frequently a blank line
(without a solid down-arrow).
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Instrument Control
Take a look at the keypad and two-line display located on the front
panel (Figure 2.1). This is the command center from which you'll
access menus and control the instrument's operations. A brief
explanation of the keys and the main menus and screens follows.
RUN
STATUS
MENU
STOP
ENTER
SpectraSYSTEM
P4000
PU-Z004/DT
PURGE
Figure 2.1 The P4000 pump's keypad.
The keypad of each SpectraSYSTEM instrument consists of twelve
keys. Four keys directly control the instrument's operation: [RUN],
[STOP], [STATUS], and, on the pump, a blank key called [PURGE].
The remaining keys ([MENU], [ENTER], [∧], [∨], [<], [>], [+], [-])
either access commands or are used to set parameters and move
around the display. The function of each is explained below.
[RUN]
Generally, pressing [RUN] starts a run or sets up the conditions
specified for the beginning of a run.
The specific [RUN] operation depends on the instrument's state:
1. If the pump's state is STOP, pressing [RUN] automatically
prepares the last file loaded to be run and sets the pump to the
conditions specified for the start of the run (t0 conditions).
2. If the pump's state is READY, (that is, the initial conditions
already exist), pressing [RUN] starts the run.
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[STOP]
Pressing stop halts an operation in progress. (Specifically, the
[STOP] key aborts a run in progress by stopping solvent flow through
the pump.)
[STATUS]
Pressing [STATUS] displays the Status Screen (Figure 2.3). From
the status screen you can monitor the run in progress. You can also
access the Status Menu. See page 29 for more information.
[PURGE]
The unlabeled key is the only variable key in the whole
SpectraSYSTEM family. On the pump, the blank key is the
[PURGE] key. The key's name appears on the nameplate below the
key.
The [PURGE] key brings the PURGE Screen to the display. Purge
parameters can be changed, and the purge operation started from this
display. Refer to Purging Solvent Lines in Chapter 1 and to Priming
and Purging the Pump in Appendix A for complete information.
[MENU]
Pressing [MENU] displays the Main Menu (Figure 2.2). Each main
menu item is explained in detail in the rest of this manual. For FILES
and COMMANDS see Chapter 3, for QUEUE and OPTIONS and
TESTS, see Chapter 4.
[ENTER]
Pressing [ENTER] accepts a selected choice or menu entry. The
[ENTER] key also advances the cursor to a new field, either on the
same line of the display or in the line below.
ARROWS
Pressing any arrow key (up, down, left, or right) moves the cursor in
the direction indicated on the key. If the cursor is on the first or the
last line of a menu, the up- and down-arrow keys move you "up" or
"down" in the menu structure.
[+] and [-]
Pressing the [+] and [-] keys scrolls you through a field's available
choices or changes the value of alphanumeric entries. Holding down
either key will continuously scroll the list of choices forward or
backward until you release the key.
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In fields that require numeric entries, the value of each digit is
increased or decreased by one unit each time you press the [+] or [-]
key. In fields that accept either numeric or alphabetic entries, such as
the File Name field, the [+] and [-] key scroll through the alphabet
from A to Z, then through the numbers 0 to 9, and finally to a slash,
hyphen and blank space.
In other fields, the [+] key advances you through a preset list of
choices while the [-] key takes you back through the list.
MENUS
AND SCREENS
Your pump has two kinds of displays: menus and screens. Menus
require you to make selections or enter specific values. Screens
display information that cannot be edited. The Menu Tree in the front
pocket illustrates the structure and content of the pump's menus and
screens.
Main Menu
The Main Menu (Figure 2.2) is the top level of the menu structure. It
gives you access to five other menus: FILES, QUEUE, TESTS,
COMMANDS, and OPTIONS. To see the Main Menu, press the
[MENU] key at any time.
>FILES
·QUEUE
·COMMANDS
·TESTS
·OPTIONS
Figure 2.2 Main Menu (P4000)
From the Files Menu you can edit, load, copy, or delete files. The
Commands Menu lets you hold, continue, or reset the pump. In the
Options Menu, you can set up or change your instrument's
configuration. From the Queue Menu you can edit or change the
order and number of files in the queue. Refer to Chapters 3 and 4,
and Appendices B and C for more information on any of the
instrument's menus.
Status Screen
The Status Screen (Figure 2.3) appears whenever you turn on the
instrument or press the [STATUS] key. The P4000's Status Screen
displays the pump's state, flow, current pressure, and the maximum
pressure setting. Two additional lines show the current solvent
composition and the elapsed time of a run. Below the Status Screen
is the Status Menu, described next.
Status
Flow
PSI
READY
1.00
0
MaxP
3000T
Figure 2.3 The first two lines of the P4000 Status Screen
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Status Menu
Just below the Status Screen is the Status Menu. To access the Status
Menu, press the down-arrow key from the Status Screen. The Status
Menu lets you review and edit run parameters during a run. Chapter
3 discusses the Status Menu in more detail.
MESSAGES
There are three different kinds of messages that can appear on the
pump's display: user messages, confirmation messages, and error
messages.
User messages
User messages (Figure 2.4) tell you about an existing instrument
condition or ask for further action. Some of these will only appear on
the display for three seconds. An example of a message requiring
further action is shown in Figure 2.4.
To install or remove
liquid ends, press ENTER
Figure 2.4 Example of a user message
Confirmation messages
Confirmation messages (Figure 2.5) indicated on the display by
asterisks, appear for one second after an operation has been carried
out successfully.
* *
File Loaded
* *
Figure 2.5 An example of a confirmation message
Error messages
Error messages (Figure 2.6), indicated on the display by exclamation
points, are displayed whenever an undesirable condition exists that
prevents the instrument from carrying out an operation. Error
messages remain on the display until you press a key.
!!
MAX PRESSURE
!!
EXCEEDED
Figure 2.6 An example of an error message
Practice Examples
This section will take you step-by-step through four operations:
1. Relabeling two solvents so that the display shows H2O and
MeOH instead of A and B (P4000 only).
2. Changing pressure units.
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3. Purging these two solvent lines.
4. Running a flow stability test.
These examples assume that the pump is properly installed and that
the bypass valve's outlet is routed to a solvent waste container. [The
solvents you actually use need not be water and methanol (MeOH),
but the solvents connected to lines A and B must be miscible. If they
are not, skip the example in Purging Lines A and B; it calls for
mixing the two solvents which may damage the liquid ends.]
LABELING
SOLVENTS
The pump display can be changed to suit your own needs. The
solvent labels in the P4000 are a good example.
To change solvent labels:
1. Press [MENU].
2. Use the right-arrow key [>] to move the cursor to /OPTIONS/.
Press [ENTER] to access the Options Menu.
3. The cursor is next to Solvent Selection. Press [ENTER] to
select /Solvent Selection. A display similar to the one in Figure
2.7 should appear:
Select Solvents
[
A
]
[
B
]
[
C
]
[
D
]
Figure 2.7 Solvent Selection Menu
4. The cursor should be in the [ A ] field. Press [+] until H2O
appears. If you accidentally go past it, press [-] to go back.
5. Press [ENTER]. The cursor should now be in the [ B ] field.
6. Again, press [+] until MeOH appears. The display should now
look like the one below:
Select Solvents
[ H2O ]
[ MeOH ]
[
C
]
[
D
]
Figure 2.8 Labels assigned to solvents A and B
You have just relabeled the two solvents A and B. Wherever a
display would ordinarily show A and B, it will now show H2O and
MeOH instead. C and D will remain unaffected. You will notice this
as you continue with the examples.
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CHANGING
PRESSURE UNITS
To change pressure units on your display:
1. Press [MENU] and move the cursor to /OPTIONS/. Press
[ENTER].
2. From the Options Menu, press [∨]. Select /More/, and press
[ENTER]. The display now looks like Figure 2.9.
Pressure Units
PSIν
Purge Mode
Flow
Figure 2.9 The More Menu
3. The cursor should be in the Pressure Units field. Press [+] until
the field shows MPa. You have just changed the display so
that all pressures will be shown in megapascals. If you prefer
other units, press [+] until you see another preset choice that
suits you.
4. Move the cursor down to the Purge Mode field. Use [+] to see
the Pressure selection (pressure). Press [+] once more, to
change the purge mode selection back to Flow.
5. You may exit the More Menu in several ways. This time, press
[STATUS] to exit the menu.
6. Use [∨] to look at the Status Menu, which shows the run file.
PURGING LINES
A AND B
You might already be familiar with the purge operation from
Chapter 1. If so, skip this example and go on to Running a Flow
Stability Test page 34.
In this example you will purge two solvent lines simultaneously,
using equal compositions.
1. Open the bypass valve on the pump. Make sure that the bypass
valve outlet is routed to a solvent waste container.
2. Press [STATUS]. The Status Screen should show that the
pump is stopped.
3. Notice that by pressing [∨] you can see the current solvent
compositions, and the H2O and MeOH labels you set earlier.
Your Status Screen and Status Menu should be similar to
Figure 2.10.
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Status
Flow
MPa
MaxP
STOP
1.50
1
21
-------------------------------------------------------Stat
H2O
100.0
0.0
MeOH
C
D
0.0
0.0
0.0
(the pressure units displayed may be different depending on your selection in the example
above.)
Figure 2.10 Status Screen and Status Menu (P4000)
4. Leave the Status Screen and go to the Purge Menu by pressing
[PURGE] (the blank key).
5. The cursor should be in the Purge field (Figure 2.11). Press [+]
until the word BLEND appears.
Purge
Flow
Time
H 2O
1.50
0.0
1
Figure 2.11 Purge Menu (P4000)
6. Press the down-arrow key. The display will show four solvents
(Figure 2.12).
H2O
MeOH
C
D
100
0
0
0
Figure 2.12 Blend solvent compositions (P4000)
7. Enter 50 (percent) for the composition of the first solvent using
the [-] key. Notice that the composition of the second solvent
automatically changes so that the total remains 100%. The
display now looks similar to Figure 2.13.
H2O
50
MeOH
50
C
D
0
0
Figure 2.13 Equal compositions of two solvents (P4000)
8. Press [∧]. You will return to the top of the Purge Menu.
9. To begin the purge cycle, move the cursor to the Time field by
pressing [ENTER] as many times as necessary. Enter 2.0
minutes in the Time field, then press [ENTER] again.
The pump will purge the A and B solvents for two minutes at the flow
rate shown in the Flow field. You can stop the purge operation at any
time by pressing [STOP]. After the purge is complete, the pump will
automatically initialize the run file.
Remember to close the bypass valve immediately following the purge
so that solvent flow returns to the LC.
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RUNNING A FLOW
STABILITY TEST
The flow stability test is a common test of the pump's performance.
The pump must be in a READY, EQUIL, or RUN state for this test to
be initiated, and the flow rate must be greater than 0 mL/min. If you
need to change the flow rate, edit the flow setting from the Status
Menu, then proceed with the steps below.
1. When the Status Screen shows READY, press [MENU] and
select /TESTS/.
2. Select /Diagnostics/ from the Tests Menu.
3. The cursor should be on /Flow Stability/. Press [ENTER].
After a short time, the flow stability rating followed by a numerical
value will appear. STABLE flow corresponds to a reading between 0
and 25, ACCEPTABLE is between 26 and 90, and UNSTABLE is a
value greater than 90.
NOTE: The Flow Stability test may show unstable flow during gradient
compositional changes or during column equilibration.
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3
Basic Operations
Introduction
The type of chromatographic analyses you do will determine how you
choose to use your pump. The P4000 is a gradient pump used for
methods development, and characterized by automatically varying
solvent compositions during a run. This chapter contains a pump
theory of operation, some recommended LC pump practices, and
describes how the pump is used to perform basic operations: editing,
loading and running a file, purging the pump, and viewing the pump's
status.
Theory of Operation
The pump is typically the second of six components in an LC system
(solvent degasser is the first). A pump delivers a steady flow of one
or more solvents to a sample-injection instrument (generally an
autosampler). This solvent flow continues through the column and on
to a detector. From the detector, a signal is passed to an integrator, a
recorder or another kind of data system capable of collecting the data
and allowing the data of the injected sample to be analyzed.
A gradient pump works by first pulling a filtered and degassed
solvent into a proportioning valve. The P4000 pump has four
proportioning valves. Solvents are measured by percentages,
specified by you, and mixed inside the pump. The solvents then
travel to the pump head where a piston meters the flow of the mixture
to an outlet tube. The pump's outlet tubing then connects the solvent
stream to an automatic or manual injector.
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SpectraSYSTEM P4000 Pump
The SpectraSYSTEM P4000 pump has been designed for ease of use
and unsurpassed performance. It can be used as a stand-alone pump
or as a module in a totally automated LC system.
The P4000 pump provides low-pressure quaternary mixing for
accurate proportioning of binary, ternary, or quaternary gradient
mobile-phase compositions and solvent switching when used for
isocratic applications. The P4000 pump contains as many as
210 lines in as many as 9 method files (40 lines per file maximum).
Table 3.1 File Characteristics for P4000 pump
File Characteristics
Time Lines/File
Total Number Files
Total Time Lines
P4000
40
9 + Shutdown + Develop
210 lines
The SpectraSYSTEM P4000 pump is engineered for reliability and
ease of maintenance. Easy maintenance helps to ensure that your
chromatography results are accurate and remain accurate. A built-in,
patented Maintenance Log (Chapter 5) allows you to follow the life
span and use of seals, pistons, and check valves. If service is ever
required, the resident diagnostics and modular design of the pump
will keep downtime to a minimum. The simplicity and durability of
the pump means that a minimum of spare parts need to be kept on
hand.
OPTIONS
Narrow-bore
LC refers to the use of narrow-ID, 2.0-3.0 mm columns for LC
separations. To optimize instruments for narrow-bore LC, standard
LC hardware must be modified to reduce extra-column volume and
gradient delay. Specific hardware modifications include minimizing
the pump’s dead volume, the detector flowcell’s volume, and the
volume contained in any interconnecting tubing and fittings. The
lower system volume of narrow-bore LC increases sample
concentration, which results in greatly improved sensitivity.
Decreased solvent consumption reduces operating costs as compared
to standard (4.0-4.6 mm ID) LC separation techniques.
NOTE: For narrow-bore applications we recommend you use a 20 μL
sample loop in Thermo Scientific autosamplers.
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Narrow-bore Hardware Modifications
Pump: For the SpectraSYSTEM gradient pump, the following
hardware changes have been made to create a compatible pump for
narrow-bore applications.
•
The solvent inlet line (from the gradient valve to the inlet check
valve) is changed from 0.060-inch to 0.030-inch ID tubing and
shortened from 12 cm to 10 cm.
•
All interconnecting tubing has been changed from 0.020-inch
ID to 0.010-inch ID.
•
The outlet liquid end is changed from the standard to the lowvolume design.
Autosampler: For the SpectraSYSTEM narrow-bore autosampler
the pump-to-autosampler tubing (~ 6 inches long) and pre-heat tubing
(~ 24 inches long) has been changed from 0.020-inch ID to
0.007-inch ID.
Operational Considerations
Injection Mode: We recommend that you consider the following in
setting up your methods and LC instrumentation for most narrowbore applications. Due to the manner in which the pump firmware
stores certain values and references the hardware, the minimum
operating pressure for these pumps is 200 psi.
We recommend that you use the PushLoop® mode for narrow-bore
applications; however, depending on your sample volume, other
injection modes may be more beneficial (see below) in some cases.
The following algorithms define the amount of sample needed for
your injection volume:
PushLoop
Pull Loop
Full Loop
(Injection Volume + 15 μL)
(Injection Volume + 1.1 μL)
(Injection Volume x 1.33) + 70 μL
In PushLoop mode, the smallest setable sample volume is 0.1 μL.
Allowed volume increments are also 0.1 μL. You should not inject
more than 10 μL without changing the standard 20 μL loop. The
autosampler is supplied with a 20 μL loop.
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Reducing Extra Column Volume: Minimizing extra column
volume maximizes analytical efficiency. Band-broadening can occur
in several ways. With narrow-bore columns, it is vital that good
plumbing connections are made. Otherwise, a dead volume created
by a bad fitting can result in much lower than expected efficiency and
peak resolution. Use zero dead volume (ZDV) fittings only with the
proper nuts and ferrules. Cut tubing with the proper tools. If
possible, electropolish metal tubing after cutting. For polymer tubing
(for example PEEK), use a "guillotine" cutter to ensure straight, rightangle cuts.
Instrument Startup
Be sure you have installed the pump according to Chapter 1 and have
completed the Start-up Checklist.
When you turn on the pump, the Status display appears. This allows
you to check the instrument settings before entering your parameters.
As described in Chapter 2, it is possible to edit a file under the Status
Menu. If you have turned on the pump before, and have used this
editing capability, the Status Menu will contain the parameters last
saved before the pump was turned on.
At powerup, there are no messages to alert you that a file is being
loaded, but you can check the file number and name by pressing the
[STATUS] key. Then use the down-arrow key to scroll to the file
listing. (Both the [STATUS] key and the creation of files are
discussed later in this chapter.)
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Some Routine Operations
Ordinarily, you will probably perform these operations with your
pump every day:
•
Edit a file (or create a new file) and/or load a file to run
•
Run your samples
•
Purge the solvent lines
•
Check the pump's status
•
Monitor pump performance
•
Shut down the pump at the end of the day
The rest of this chapter is devoted to explaining these basic
operations. If you need to, refer to the keypad "rules" in Chapter 1.
You may also want to refer to the basic menu structure on the quick
reference card in the front pocket of your manual.
The File(s) Menu
In this section we describe how to set up the files that control solvent
composition, pressures, flow rates, and run times. Each of the file
operations accessed from the Files Menu is briefly defined, then
described in detail.
To access the File(s) Menu, press [MENU] and select /FILES/. The
Files Menu is shown in Figure 3.1.
>Edit
·Copy
·Load
·Delete
Figure 3.1 The Files Menu (P4000)
Edit
Select /Edit/ if you want to change the name and/or parameters in an
existing file, or create a new file. Select the file by file number or
letter (file S is the Shutdown file, file D is the Develop File™).
Load
Select /Load/ to load the file you want the pump to use when in
operation. Make a file selection by file number. We refer to a loaded
file as the run file.
Copy
Select /Copy/ to copy the contents of one file into another. Select
both files by file number.
Delete
Select /Delete/ to delete the contents of a file and return all file
parameters to their default values. An information message allows
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you to cancel the delete operation before the file is deleted. Once
deleted, the file cannot be retrieved.
EDITING A FILE
Each file consists of four parts: File Name, Solvent Program, Options,
and Timed Events. Selecting /Edit/ gives you access to these areas
(Figure 3.2).
Edit File
1
File Name
1
----------------------------------------------------->Solvent Program
·Options
·Timed Events
Figure 3.2 The Edit Menu
To use the Edit Menu, select the file number in the Edit File field
using the [+]/[-] keys. The P4000 pump has file numbers 1 through 9.
The P4000 pump has a Shutdown file (file number S), discussed on
page 51. In addition, the P4000 has a Develop file (D) which is
explained in detail in Chapter 4.
If a particular file cannot be accessed, the File Protect feature for that
file has been turned on, preventing changes or deletions. When the
File Protect feature has been turned on, a message appears on the
display, indicating that the file cannot be edited. (File Protection is
discussed in OPTIONS, More Menu in Chapter 4.)
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File Name
You can enter or change the File Name using the [+]/[-] keys. The
File Name field is eight characters long. Any name you enter is
automatically remembered by the pump, so you do not need to
formally "save" the name. All of the parameters discussed below are
automatically remembered by the pump when you edit a file from the
Files Menu.
NOTE: When changing the parameters of a run file from the Status Menu,
you must select /Save File/ in order for the pump to remember your changes
for future runs. The pump will, however, use your changes until the next
time the file is re-initialized, regardless of whether the file has been saved.
Editing a run file from the Status Menu is discussed in more detail later.
The Solvent
Program Menu
The Solvent Program Menu contains time lines used to construct
gradient or isocratic solvent parameters. Time lines consist of a time,
a percentage composition (in whole and half percents in the P4000),
and a flow rate (in mL/min). When /Solvent Program/ is selected, the
display shows (Figure 3.3):
Time
0.0
A
B
C
D
Flow
100
0.0
0.0
0.0
1.00
Figure 3.3 A P4000 solvent program
NOTE: If you used the Solvent Selection Menu (P4000) to label solvents as
described in Chapter 2, the display will show the assigned labels in place of
A, B, C, and D. This is true for all displays showing solvents.
If you are not going to use one of the solvents set up for your pump,
you can change the display so that it shows only three solvents.
Complete information for changing the solvents displayed is found
under Display Solvents on page 49.
General Rules for
Entering Time Lines
A file should contain a minimum of two time lines. If a file contains
one time line, the pump will not be able to maintain a RUN state,
although it will show READY.
The solvent compositions and flow for the zero (0.0) time line are
always editable. The Time field 0.0 itself is not editable; it remains as
time zero. To add new times you must create new time lines.
To create a new time line, use the down-arrow key to go to a blank
line, or press [ENTER] until a blank line appears, then press the [+]
key. The new line, automatically incremented one minute past the
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previous line, will be displayed. The cursor will appear in the Time
field of the new line. The Time field in any new line is editable.
HINT: Don't add all your time lines to the file at once. Enter values for
solvent composition and flow in the first time line, then add a new time line.
All of the solvent composition and flow rate values will be copied to the new
line automatically. In fact, a new line is always identical to the one just
before it, with the exception of the Time field as discussed earlier. Solvent
composition values can be incremented easily then, from the previous
values.
The solvent compositions in each time line must always add to 100.
As soon as one percentage is changed, the pump automatically
changes the remaining solvents to keep the total equal to 100.
Solvent compositions can be set to whole (X.0) or half (X.5) percents
in the P4000.
You can enter times in any order you choose. The pump will
automatically rearrange the time lines on the display so that they are
in chronological order as soon as you move the cursor off the Time
field.
Gradients occur between adjacent pairs of time lines. The pump
continually adjusts the solvent composition to achieve the
composition shown in the next time line by the time specified.
One of twenty-one preset gradient curves can be selected from the
Options Menu (P4000). Creating step gradients is explained on
page 48.
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Thermo Scientific
Changing Solvent Composition — A P4000 Example
Start with the display shown in Figure 3.4.
Time
0.0
A
B
C
D
Flow
100
0.0
0.0
0.0
1.00
----------------------------------------------------1.0
2.0
100
0.0
0.0
0.0
1.00
100
0.0
0.0
0.0
1.00
Figure 3.4 Example P4000 solvent program
Then decrease solvent A by 20, solvent B automatically increases so
that the total is 100 (Figure 3.5).
1.0
100
0.0
0.0
0.0
1.00
2.0
80.0
20.0
0.0
0.0
1.00
Figure 3.5 Example P4000, continued
Then, if you decrease solvent B by 15, the difference is applied to
solvent C (Figure 3.6), and so on.
1.0
100
0.0
0.0
0.0
1.00
2.0
80.0
5.0
15.0
0.0
1.00
Figure 3.6 Example P4000, continued
If all solvent fields have been filled in, a change made to one field
will cause the adjacent field, to the right, to reflect the difference. If
that next solvent goes to 0, the solvent after it is affected, and so on.
In this example the pump would pump 100% solvent A until
1.0 minute into the run. After one minute the pump changes the
composition such that at 2.0 minutes into the run solvent A would be
at 80%, and solvents B and C at 5% and 15%, respectively.
The Options Menu
Under Files, Edit
The Options Menu consists of maximum and minimum pressure
levels, an equilibration time, and, in the P4000, a gradient curve type
and option to display certain solvents. You are not required to set any
values in this menu, although you should always set a maximum
pressure level, since the pump will warn you whenever operating
pressures exceed this level.
You can access the Options Menu from /FILES/, /Edit/ (Figure 3.7) or
by pressing [ENTER] or [∨] on the blank line at the bottom of the
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43
Solvent Program, then selecting /Options/. The Options Menu is
shown in Figure 3.7.
Maximum Pressure
3000
Minimum Pressure
0
----------------------------------------------------Equilibration Time
0.0
Gradient Curve
Linear
Display Solvents
A
B
C
D
Figure 3.7 The Files, Options Menu (P4000)
Maximum, Minimum Pressures
Change the pressure levels as desired. The maximum value is
6000 psi, the minimum is 0 psi. (The allowed values depend on the
pressure units, selected in /OPTIONS/, /More/, Pressure Units.) The
maximum pressure value must be greater than the minimum pressure.
The pump will not allow you to set maximum and minimum pressures
that are inconsistent. If you are unable to edit one value, try to edit
the other.
NOTE: Pressure units (psi, bar, or MPa) are selected from the Main Menu,
/OPTIONS/, /More/, described in Chapter 4.
During operation, the pump continuously monitors the actual
operating pressure and displays this on the Status Screen. If the
actual pressure ever exceeds the maximum pressure level, or falls
below the minimum pressure level set here, an error message will
alert you to this circumstance. For more information about using this
error feature, refer to Chapter 4, Options, Error Recovery.
Equilibration Time
If desired, enter a value for the equilibration time. The equilibration
time is the time that the pump will maintain the conditions (solvent
composition and flow) specified on the first line of a run file (the zero
time line), before showing READY on the Status Screen.
Gradient Curve
The P4000 pump has the ability to automatically run a Gradient
Curve between time lines. There are twenty-one choices in this field:
Linear, Convex 1 to Convex 10, and Concave 1 to Concave 10.
These shapes are shown in Figure 3.8. The curve is applied to all
solvents participating in the gradient.
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Thermo Scientific
Convex 10
Convex 1
Linear
(COMPOSITION)
FINAL %
Concave 1
Concave 10
INITIAL %
|
|
(TIME)
BEGIN
END
Figure 3.8 Gradient Curves in the P4000
The selected curve operates between adjacent pairs of time lines of
the Solvent Program. Gradient curves are usually used by specifying
only two time lines (a zero time line and one other time line). The
pump automatically runs the chosen gradient curve between these two
time lines.
Concave gradients begin with a small and end with a high rate of
change. Convex gradients begin with a high rate of change and end
with a small rate of change. Linear gradients have a constant rate of
change between solvent compositions.
For example, if the Solvent Program contained the time lines shown
in Figure 3.9, and the gradient curve were Convex 3, then, over
12 minutes, solvent A would follow the shape of Convex 3, starting at
a composition of 20%, and ending with a composition of 100%.
Solvent C would follow a complimentary shape, starting at a
composition of 80%, and ending with a composition of 0% (Figure
3.10).
0.0
12.0
20.0
0.0
80.0
0.0
1.00
100
0.0
0.0
0.0
1.00
Figure 3.9 A two-solvent program
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45
PU-Z012E/DG
100 -
-
A
-
% Solvent
70 -
50 -
30 -
-
C
-
-
-
-
0
12
Time (in Minutes)
Figure 3.10 Solvents A and C, using a Convex 3 gradient curve.
When three solvents are involved, as in the Solvent Program shown in
Figure 3.11, Solvent A follows the designated curve from 0% to
100% over 12 minutes. Solvents B and C follow a similar curve
(remember, it is the rate of change that defines the curve shape), from
70% to 0%, and 30% to 0%, respectively, over the same 12 minutes
(Figure 3.12).
0.0
12.0
0.0
70.0
30.0
0.0
1.00
100
0.0
0.0
0.0
1.00
Figure 3.11 A three-solvent program
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PU-Z010E/DG
100 -
-
A
% Solvent
70 -
-
B
30 -
-
C
-
0
12
Time (in Minutes)
Figure 3.12 Solvents A, B, and C using a Convex 3 gradient curve
The most common gradient is linear. When Linear is selected in the
Gradient Curve field, solvent composition follows a straight line,
ramping from one solvent percentage to the next, between time lines.
Figure 3.13 shows the composition profile of the solvent program
shown in Figure 3.11.
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47
PU-Z011E/DG
100 -
-
% Solvent
70 -
A
-
30 -
B
-
C
-
-
-
-
0
12
Time (in Minutes)
Figure 3.13 Solvents A, B, and C using a Linear gradient curve
Creating Step Gradients
Creating step gradients is easy. Figure 3.14 is an example showing a
simple step gradient solvent program.
Time
0.0
A
B
C
D
Flow
100
0.0
0.0
0.0
1.00
----------------------------------------------------5.0
100
0.0
0.0
0.0
1.00
5.1
80.0
20.0
0.0
0.0
1.00
10.0
80.0
20.0
0.0
0.0
1.00
10.1
60.0
40.0
0.0
0.0
1.00
15.0
60.0
40.0
0.0
0.0
1.00
15.1
30.0
70.0
0.0
0.0
1.00
20.0
30.0
70.0
0.0
0.0
1.00
Figure 3.14 A two-solvent step gradient solvent program (P4000)
Each "step" is created by specifying a very short amount of time
(0.1 minutes) in which the gradient is allowed to operate. In the
example, the pump establishes the conditions in the Time = 0.0 min
time line, and maintains this until the Time = 5.0 min time line
(because both time lines have identical compositions). The first step
takes place between 5.0 and 5.1 minutes. The pump establishes the
conditions in the Time = 5.1 minutes time line and maintains them
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Thermo Scientific
until 10.0 minutes, when the next step occurs. The last step occurs at
15.0 minutes.
HINT: System baseline may take time to stabilize.
Display Solvents
In the Display Solvents field you can change the number of solvents
displayed in the file. You should be certain that the composition for
any undisplayed solvent is zero in all time lines before you try to turn
off its display. The pump automatically checks the solvent
composition field for each solvent and prevents you from turning off
the display for any one whose composition is not zero.
Use the [+] / [-] key to see and select the set of solvents that suits your
needs. A minimum of three solvents must be displayed at all times.
Automatic solvent proportioning rules apply only to the solvents that
are displayed.
HINT: It is simpler to program a three-solvent display since the automatic
solvent proportioning rules make binary program creation straightforward
(e.g. if you select Display Solvents ABC and want to program only A and B,
then you only need to select values in the A field. If you want to program B
and C, then you only need to select values in the B field. If you want to
program A and C, then select values in the C field.)
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49
Timed Events Menu
You can access the Timed Events Menu from /FILES/, /Edit/, or by
pressing [ENTER] or [∨] on the bottom line of the Options Menu,
then selecting /Timed Events/. The Timed Events Menu is shown in
Figure 3.15.
Time
Event
0.00
Off
Figure 3.15 The Timed Events Menu
A timed events output is available at the external events port. This
output can be used to control an external device such as a column
switching valve. The output can be turned on or off up to six times
per run.
The time on the first line of the menu remains 0.0. Additional timed
events are added much like time lines in the Solvent Program. Use
the down-arrow key to reach a blank line, then press [+]. As many as
five additional lines can be added. The Time field in any added line
is editable. Enter times and turn the output Off or On in any of the
six Event fields using the [+] / [-] keys.
For example, Figure 3.16 shows a timed events example.
Time
Event
0.00
Off
------------------------------------------------------6.50
On
9.00
Off
Figure 3.16 Timed events example
In this example, a timed event output (signal) would be sent (turn on)
at 6.5 minutes into the pump's run and become inactive (turn off) two
and a half minutes later, at 9.00 minutes.
To delete a line, move the cursor to the time field, then press and hold
[-]. The time field will eventually become blank, and the line will
disappear. If the time has more than one non-zero digit (for example
6.50), then decrement each digit, starting with the left-most digit.
Release the [-] key after each field becomes blank and decrement the
next digit.
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THE SHUTDOWN
FILE
The Shutdown file (file number S) is an easy way to assist you with
the proper maintenance of the pump. Since your pump and column
should never be allowed to sit idle with salts or corrosive materials in
them (including water), the Shutdown file lets you automatically flush
the pump and the column at the completion of a series of samples.
Whenever the pump detects that it has been in a READY state,
without a run being initiated, for a specified period of time, it
automatically loads, initializes, and runs the Shutdown file.
NOTE: The pump's clock is reset anytime the [RUN] or [STOP] key is
pressed, or anytime the /Reset/ command is issued.
Some instances when you might want to use a shutdown file are:
•
To keep solvent at a very low flow rate flowing through the
LC
•
To clean the column and keep the pump running
•
To clean the column, then stop solvent flow through the
LC
The Shutdown file's Solvent Program, Options, and Timed Events
Menus are exactly the same as those for other files with one
exception: in the last line of the Options Menu, the display shows a
Time from READY field instead of the Equilibration Time field
(Figure 3.17). Gradient curves are not available in the Shutdown file.
Minimum Pressure
Time from READY
0
Off
Figure 3.17 Last lines of the Options Menu of the Shutdown file
The Shutdown file will be automatically loaded and run if the current
run file has not been started, either manually or remotely, in the time
specified in the Time from READY field. There are ten preset times
(in minutes) that can be selected in this field: 5, 10, 20, 30, 45, 60, 90,
120, 240, or 480. The Shutdown feature can be turned off completely
by selecting "Off." The Time from READY timer starts as soon as
the Status Screen shows READY.
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51
If the Shutdown file is loaded automatically, the pump will do one of
the following when it reaches the last time line of the Shutdown file:
•
If the flow rate is greater than zero (> 0) the pump will
maintain (Hold) the last time line's solvent composition and
flow rate indefinitely.
•
If the flow rate is zero (= 0) then the pump stops, and STOP is
shown on the Status Screen. In addition, the pump remembers
the run file that had been used just before the Shutdown file
was automatically invoked, and restores that file to the run file
position.
If the Shutdown file is loaded manually (/FILES/, /Load/), then it
remains the run file until another file is loaded.
NOTE: If the pump will be operated unattended for an extended
period of time, ensure that the solvent reservoir and waste containers
have sufficient capacity.
If you use an autosampler, specify the time interval to be longer than
the cycle time on the autosampler (for example., [1.5 × cycle time], or
[cycle time + 20 minutes]). If you perform manual injections, set the
time interval to the maximum time likely between injections.
Depending on your own circumstances, you may wish to turn the
Time from READY to "Off."
LOADING A FILE
When you select /Load/ the display shown in Figure 3.18 appears.
>Load File
1:(filename)
Figure 3.18 The Load display
Select a file by number, then press [ENTER]. A message will
confirm that the file was loaded. A loaded file is referred to as the
"run file."
As soon as a file is loaded, the pump will initialize the file, i.e., bring
the pump to the conditions specified on the zero (0.0) time line of the
file. The Status Screen shows INIT until these conditions are
achieved. If an equilibration has been specified, the pump will then
show EQUIL until the equilibration time has been reached.
After initialization and equilibration, the Status Screen then shows
that the pump is READY. The pump's clock will not start unless a
run is triggered either manually by pressing [RUN], from a properly
connected (hardwired) autosampler, or from a data system.
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COPYING A FILE
When you select /Copy/ the display shows (Figure 3.19):
Copy File 1:(filename)
to File 2:(filename)
Figure 3.19 The Copy display
Select both files by number. Press [ENTER] after both files are
specified only if you are sure you want to copy the parameters from
the top file into the other file. The Copy command overwrites the
selected file. Once overwritten, previous values cannot be retrieved.
If you do not want to copy a file, leave the display by pressing
[MENU] or [STATUS], or the up-arrow key. Press [ENTER] to
proceed with the copy operation, and a message confirms its
completion.
If you try to copy to a protected file, a message will appear indicating
that the file is protected. You will then be returned to the Copy
display.
DELETING A FILE
When you select /DELETE/ the display shows (Figure 3.20):
>Delete File 1:(filename)
Figure 3.20 The Delete display
Select a file by number, then press [ENTER]. A message is
displayed, asking you to confirm the selection. Press [ENTER] only
if you are sure you want to delete the file. Once deleted, a file cannot
be retrieved. If you do not want to delete the file, leave the display by
pressing [MENU] or [STATUS], or the up-arrow key. A
confirmation message concludes a successful file deletion.
When you delete a file you are actually returning all parameters in the
file to their default values.
If you try to delete a protected file, a message will appear indicating
that the file is protected. You will then be returned to the Delete
display.
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53
Purging Solvent Lines
If none of the solvent lines has solvent in it, refer to Priming the
Pump in Chapter 1.
Air will slowly diffuse through the thin-wall Teflon inlet tubing, and
into the solvent. If the pump flow has been turned off or if any of the
solvent lines from the solvent reservoirs to the proportioning valve
have not been used in the past several hours, those lines should be
purged with degassed solvent before use.
The purge operation can be activated when the pump is in any state.
NOTE: Open the bypass valve prior to purging, or else ensure that
your chromatographic column can withstand the purge parameters
you set before performing any purge.
THE PURGE MENU
Pressing the [PURGE] key brings the Purge Menu to the display
(Figure 3.21).
NOTE: DO NOT move the cursor out of the Time field using the
[ENTER] key until your LC is ready to start a purge.
Purge
A
Flow
Time
1.00
0.0
-------------------------------------------------------(If BLEND is selected under Purge for P4000)
A
100
B
C
0
D
0
0
Figure 3.21 The Purge Menu
Purging can be accomplished in one of three purge modes: Flow,
Pressure, or Both. In Flow mode, purging is regulated by the flow
rate from the pump. In Pressure mode purging is accomplished at a
specific fluid pressure. In Both mode purging is accomplished using
both flow and pressure parameters. Internal limits are designed to
protect your LC system: in flow mode, the pump will accept rates of
0.1 to 10.0 mL/min. The maximum pressure the pump allows in flow
mode is the maximum pressure value from the current run file; in
pressure mode, the maximum flow the pump will reach in attaining
the set pressure is 6 mL/min.
NOTE: Purging in Pressure or Both mode with the bypass valve open may
not allow sufficient pressure to be generated in the system. The pump will
operate at maximum flow, but the target pressure may not be achieved.
Ensure that your analytical column can withstand the purge pressure (or use
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Thermo Scientific
a flow restrictor or old column), and do not open the bypass valve.
The purge mode can be changed from /OPTIONS/, /More/, Purge
Mode, described in Chapter 4. The top line of the Purge Menu will
show either Flow or Pres, depending on the purge mode you select.
Purge (field)
There are six choices available in the Purge field for the P4000 (A, B,
C, D, All, Blend). When Blend is selected, one additional line can be
accessed by pressing the down-arrow key (not the [ENTER] key!).
These are used to set solvent compositions. You should enter values
for the solvent composition in these fields before you initiate a purge.
Initially, the compositions will be copied automatically from the first
line of the Run File. These solvent fields follow the same
proportioning rules as the fields in the Solvent Program; they must
total 100. Unlike the solvent program however, all solvents are
automatically displayed, regardless of the selections made from
Display Solvents.
Flow or Pres
The flow rate is taken from the last line of the run file. Use this field
to select a flow rate. If the purge mode is Pres (pressure) instead of
Flow, the pressure is automatically set to one-half (50%) of the
maximum pressure level set in the run file.
NOTE: To change the purge mode you must go to /OPTIONS/, /More/,
Purge Mode. Refer to Chapter 4 for complete information.
Time
The Time field is used to set the length of time you want the pump to
purge. If the field remains 0.0, purging, once started, will continue
until stopped.
NOTE: When you check the purge operation from the Status Screen, the
P4000 displays PURGE. The time remaining to complete the purge is
shown on line 4.
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55
Starting a Purge
To initiate a purge cycle, move the cursor out of the Time field by
using the [ENTER] key.
If the time stays set to 0.0, purging will continue until you press
[STOP], or a file is initialized. If a time is entered, the pump will
purge until the time set. After completing a purge, the pump
automatically initializes the run file.
When running in a timed purge mode, the Status Screen will indicate
the time remaining to purge (that is, counts backwards toward 0.0). If
the [PURGE] key is pressed once more, the Purge Menu is again
displayed and the time countdown continues, uninterrupted, unless
the cursor is moved out of the Time field by pressing [ENTER], at
which point the timer will be reset and purging will restart.
The pump will retain the selections made in the PURGE display as
long as the power to the pump is on.
If power is turned off, the pressure parameter must be re-entered.
However, the flow rate setting is retained in NOVRAM.
Stopping a Purge
There are three ways to stop a purge cycle:
56
•
Press the [STOP] key
•
Load (initialize) a file by selecting a file using /FILES/, /Load/
•
Allow the pump to complete the purge (if a time has been
specified), at which point the pump will automatically initialize
the last run file.
Thermo Scientific
Running the Pump
To perform a run:
•
purge the solvent lines if necessary,
•
establish a READY state (may require the pump to INIT,
and/or EQUIL)
•
inject the sample, and
•
press [RUN].
If you are performing a manual injection, fill the injection loop, check
that the Status Screen shows READY, inject the sample, and press
[RUN] in succession. If an autosampler is hardwired to the pump,
simply initiate the autosampler run sequence.
NOTE: Generally, when operated manually, if the pump is stopped (i.e., the
Status Screen shows STOP), the run file can be initialized by pressing
[RUN], and then started by pressing [RUN] again when the Status Screen
shows READY.
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57
ESTABLISHING
READY
The READY state means that the pump has reached the conditions
specified on the first line of the solvent program and is ready to start a
run.
Initializing a file
To achieve the READY state, initialize a run file, by any of the
following methods:
•
Load a file by selecting /FILES/, /Load/, a file number,
then pressing [ENTER]. This simultaneously loads the run
file and initializes it.
•
If the pump is stopped (Status Screen shows STOP), press
[RUN]. This initializes the run file, without starting the
run.
•
If the pump is stopped or in run, reset the run file by
selecting /COMMANDS/, /Reset/. This initializes the run
file without starting the run.
Wait for the pump to reach zero time line conditions. If an
equilibration time was specified in the file, the Status Screen will
show EQUIL for this period of time.
PRESSING [RUN]
As soon as the pump shows READY, begin the run by pressing the
[RUN] key. This starts the pump's clock.
NOTE: Even though the pump may be ready, your column and the
rest of your LC system may not be! Take into account your own LC
application and ensure that your column is at chemical equilibrium
and that the other instruments in your system are ready before your
proceed with any injection.
As soon as the [RUN] key is pressed, the pump begins to operate
based on the time lines in the Solvent Program of the run file, and the
Status Screen is displayed. The Status field on the P4000 Status
Screen shows RUN, indicating that the pump is operating from the
parameters in the run file. Line 4 shows the time into the run. Status
is fully described on page 61.
STOPPING THE
PUMP
There are a number of ways to stop the pump, depending on what you
wish to do next. More information regarding the Commands Menu,
referred to below, is found on page 60.
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Thermo Scientific
Using a Hold
Command
If you want to stop the pump's clock momentarily (but not stop
solvent flow), and plan to resume the run where it was stopped, press
[MENU], and select /COMMANDS/, /Hold/. To resume, select
/COMMANDS/, /Continue/.
By Resetting the
Pump's Clock
If you want to restart the current run, press [MENU] and select
/COMMANDS/, /Reset/. This stops the pump's clock and returns to
the zero time line, automatically initializing the file. Restart the run
by pressing [RUN] after the pump shows READY.
By Pressing [STOP]
If you want to completely stop the pump, press [STOP]. This aborts
the run and stops solvent flow through the pump. If you want to
resume with the same file, you must initialize the run file by pressing
[RUN], waiting for the pump to show [READY], then pressing
[RUN] again.
WHILE THE PUMP
IS RUNNING
There are several messages which can appear in the Status field.
These are discussed in detail in the Status section below.
While the pump is running you may do several things without
disturbing pump operation:
•
edit files (/FILES/, /Edit/).
•
check some pump performance parameters.
•
edit the run file from the Status Menu. (This has an effect
on the current run - see page 63 for more information.)
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59
The Commands Menu
The Commands Menu is reached by pressing [MENU] and selecting
/COMMANDS/.
When /COMMANDS/ is selected the display shows (Figure 3.22):
>Reset
·Hold
-------------------------------------------------------·Repeat
Figure 3.22 The Commands Menu
RESET
Reset is used when the pump is in RUN and you want to abort the run
without stopping the pump's flow. Reset reinitializes the file (i.e.,
resets the timer to zero). The result is that the pump reestablishes the
conditions of the zero time line, and returns the pump to a READY
state. Pressing [RUN] restarts the run.
HOLD/CONTINUE
Hold is used to stop and hold the pump's clock. It causes the pump to
maintain the operating conditions used at the moment the Hold
command was issued, including flow rate. These conditions are
maintained indefinitely unless:
•
A Continue command is used from the Commands Menu,
at which point the timer continues from the point at which
it was held.
•
The [STOP] key is pressed.
•
The Reset command is selected.
Whenever a Hold command is issued, the word Continue will replace
the word Hold in the Commands Menu.
To select any of the commands on the Commands Menu, move the
cursor to the desired line, and press [ENTER] to issue the command.
The display returns to the Status Screen.
REPEAT
The Repeat command initiates two different events, depending on
what the pump is doing when the command is issued:
1) If a queue is running, the /Repeat/ repeats the run current in
progress. (To skip to the next run, select /Reset/.)
2) If a queue is not running, then /Repeat/ is identical to /Reset/.)
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Thermo Scientific
Status
The Status Screen appears whenever the pump is powered on, a file is
initialized, or the [STATUS] key is pressed. The Status Screen,
consisting of four lines in the P4000, shows the pump's current
operating values. Below the Status Screen is the Status Menu, where
you can view and, if necessary, edit parameters of the run file.
STATUS
MESSAGES
The Status field of the Status Screen can show any of the following
messages:
Table 3.2 Status Messages
(time)
•
The time into the run (or time remaining if a timed
purge).
EQUIL
•
The pump is equilibrating the LC by maintaining the
conditions on the first line of the run file for the
equilibration time specified in the file.
HOLD
•
/COMMANDS/, /Hold/ has been selected. The pump is
maintaining the conditions that existed when the Hold
command was issued, including the flow rate. To
continue, select /COMMANDS/, /Continue/.
INIT
•
The pump is initializing a file.
NRDY
•
(Not Ready) The pump is waiting for a signal from
another instrument.
PURGE
•
The pump is purging.
Q (time)
•
The file listed in Order 1 of the queue is running. The
time into the current run is shown.
QEQUIL
•
The pump is equilibrating the LC based on a file listed in
the queue.
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Table 3.2 Status Messages, continued
QREADY
•
A queue has been loaded and the run specified in Order
1 can be started.
Q RUN
•
Appears briefly when a queue is run.
QSTOP
•
A run listed in the queue has been stopped.
READY
•
The pump has achieved the conditions on the first line of
the run file, and the equilibration time has elapsed (if
set). A run can be started. READY, shown in Status.
RUN
•
The pump is running the file.
STOP
•
All mobile-phase flow through the pump is stopped.
SYNC
•
This is a remote communications message that appears
briefly whenever a run is started. At lower flow rates, it
may be seen for longer periods of time.
P4000
Status
Flow
PSI
MaxP
READY
1.00
0
3000
------------------------------------------------------Stat
0.0
A
100.
B
C
D
0.0
0.0
0.0
File 1:EXAMPLE
Time
A
B
C
D
Flow
0.0
100
0.0
0.0
0.0
1.00
1.0
100
0.0
0.0
0.0
1.00
2.0
80.0
5.0
15.0
0.0
1.00
Maximum Pressure
Minimum Pressure
Equilibration Time
Gradient Curve
3000
0
0.0
Linear
>Save File
(appears only if the run file is changed)
Figure 3.23 Example P4000 Status Screen and Status Menu
The first and second lines of the P4000's Status Screen show the state,
flow, pressure, and maximum pressure setting.
The third and fourth lines of the Status Screen show the pump's time
and current solvent composition.
Solvent compositions are displayed to tenths of a percent (0.1).
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EDITING A RUN
FILE
The remaining lines, which comprise the Status Menu, show the file
name, solvent program, and options of the run file. If the solvent
program and options do not appear, the Status Lock feature has been
turned on. (Status Lock is described in Chapter 4, under the
OPTIONS, More Menu.)
All time lines of the run file's solvent program may be edited while
the pump is running, as may the maximum and minimum pressures,
and the equilibration time. Any changes take effect as soon as the
cursor leaves each field. However, the changes are saved only when
the /Save File/ command is selected below Gradient Curve (P4000).
HINT: Any time the parameters of the Run File are changed, the /Save File/
command will appear at the bottom of the Status Menu. Note that you
cannot save changes using the /Save File/ command if file protection for that
file has been turned "On".
When changing the parameters of a run file from the Status Menu,
you must select /Save File/ in order for the pump to remember your
changes. The pump will, however, use your changes until the next
time the file is initialized by selecting /FILES/, /Load/. Use the same
methods described on page 41 and on page 43 to edit the parameters
of a run file. New lines may be added to the run file. Timed events
may only be edited using /FILES/, /Edit/, /Timed Events/, and will
not take effect until that file is loaded (becomes the run file).
Monitoring Pump Performance
The pump has the capability to automatically monitor its own
performance and warn you if a flow problem exists. The way the
pump responds to error conditions is set in the Error Recovery menu
of OPTIONS, selected from the Main Menu. These options (not to be
confused with File Options), are described in detail in Chapter 4.
Certain flow and pressure conditions are monitored continuously. For
example, if a time line with a zero flow rate has been encountered, the
pump responds with the appropriate error message shown below. As
mentioned earlier, you can select the pump's response to certain
conditions.
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! !
ZERO FLOW RATE
! !
! !
MAX PRESSURE EXCEEDED
! !
Figure 3.24 Example error messages resulting from flow problems
"ZERO FLOW RATE" indicates that a time line with a zero flow rate
has been encountered. "MAX PRESSURE EXCEEDED" is a
condition whose pump response is user-selectable in /OPTIONS/,
/Error Recovery/.
The pump can also initiate a Flow Stability Test. This test is run by
selecting /TESTS/, /Diagnostics/, /Flow Stability/. The results are
continuously displayed until another key is pressed. The results
consist of 2 parts. The first is a summary of the performance
evaluation (STABLE, ACCEPTABLE, or UNSTABLE) and the
second is a number that indicates a position in each range. Further
explanation of this test can be found in Appendix A.
Shutting Down at the End of the Day
Some shut down suggestions when you conclude your work with the
pump for the day:
64
•
Do not leave buffers in the pump or in your LC; purge the
pump (50/50 MeOH/water is a good solvent) if it has just
concluded a run using buffered solutions. (Don't leave 100%
H2O in your LC.)
•
Leave the column full of a solvent recommended by the
column manufacturer.
•
Make use of the Shutdown file. The pump will automatically
maintain the conditions specified on the last line of the file.
This is particularly useful if you prefer to have a small but
continuous flow of solvent through your LC system while it is
idle.
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4
Advanced Operations
Introduction
This chapter focuses on two menu items: Options, used to set some
important features, and Queue, used to build a list of files to run. This
chapter also describes what the P4000 pump's Develop File is and
how to use it. The majority of this chapter discusses the Develop
File.
The Options Menu
The Options Menu (accessed from the Main Menu, not from
/FILES/), contains seldom-changed features such as the pump's
response to certain electrical and flow conditions, user-selected
display and operational preferences, and file protection.
·FILES
·QUEUE
·COMMANDS
·TESTS
>OPTIONS
Figure 4.1 The Main Menu with /OPTIONS/ selected
The P4000 Options Menu is shown in Figure 4.2.
>Solvent Selection
·Error Recovery
-------------------------------------------------------·More
Figure 4.2 The P4000 Options Menu
ERROR RECOVERY
MENU
The pump continuously checks pressure so that problems can be
indicated on the display immediately. The pump can also sense a
power failure or power interruption. The Error Recovery Menu
(Figure 4.3) is used to preset the pump's response to detecting error
conditions in any of these three operating parameters.
AC Power Fail
Stop
@Maximum Pres
Stop
Figure 4.3 The Error Recovery Menu
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65
Each field in the Error Recovery Menu can be set to one of three
selections:
Selection
Pump's Response
Stop
The pump stops immediately if the condition is
encountered
Continue
The pump continues as if the condition had not
occurred
Shutdown
The pump immediately stops, then loads and
runs the Shutdown file.
AC Power Fail
The pump may sense a power interruption at any time. Select Stop,
Continue, or Shutdown in the AC Power Fail field to stop, continue,
or shut down the pump as soon as power is restored.
NOTE: If the power switch is turned off while the motor is running, the
pump considers this a power failure and will respond accordingly as soon
as power is restored.
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@Maximum Pres
In a file's Options Menu, you can change the maximum pressure level
(the default is 6000 psi). If the pump's operating pressure ever
exceeds this value, the pump will operate based on your selection in
the @Maximum Pres field. Select Stop, Continue, or Shutdown to set
the pump's response to sensing operation at maximum pressure.
MORE MENU
To access other options, select /More/ (Figure 4.4).
·Error Recovery
>More
Figure 4.4 Selecting /More/
The More Menu (Figure 4.5) contains additional, miscellaneous, user
preferences, such as the units the pressure is displayed in and how
quickly field choices scroll when the [+] and [-] keys are pressed and
held.
Pressure Units
PSI
Purge Mode
Flow
------------------------------------------------------Delay Volume
0.0
Cursor Speed
Medium
Status Lock
Off
Ready Output Active
File Name
Hi
Protect
1:(filename)
Off
2:(filename)
Off
3:(filename)
Off
4:(filename)
Off
5:(filename)
Off
6:(filename)
Off
7:(filename)
Off
8:(filename)
Off
9:(filename)
Off
Figure 4.5 The More Menu
Pressure Units
Select either PSI, BAR, or MPa as your preferred units. All menus
and screens that show pressure units will reflect the selection.
Purge Mode
Select either Flow, Pressure, or Both in the Purge Mode field. Your
selection is reflected on the Purge Menu and is used as the primary
purge parameter. Select Flow if you wish to purge based on a flow
rate, Pressure if you want to purge based on an operating pressure, or
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67
Both if you wish both flow and pressure parameters to govern
purging.
Purging in pressure mode requires a certain amount of back pressure
in the system. Ensure that your analytical column can withstand the
purge pressure (or use a flow restrictor or old column), and do not
open the bypass valve.
Delay Volume
The gradient delay volume is the volume of mobile phase that the
pump will pump before allowing a SpectraSYSTEM autosampler to
proceed with an injection. This prevents an injection from occurring
before the gradient has reached the column. Determine the delay
volume of your system, and enter this value (in mL) in the Delay
Volume field. (The pump must be properly hardwired to the
autosampler for the delay volume to be recognized by the
autosampler. The pump sends a signal to the autosampler's Inject
Hold input while the delay volume is being pumped.)
Cursor Speed
Cursor Speed is used to change how quickly choices scroll on the
display when the [+] and [-] keys are pressed and held, and how
quickly a menu scrolls (up and down) when the arrow keys are
pressed and held. Select Fast, Medium, or Slow.
Status Lock
Status Lock prevents a run file from being edited from the Status
Menu. When Status Lock is On, the Status Menu only shows the run
file name (and number). The rest of the run file cannot be accessed.
Status Lock is different from File Protection (below). A protected file
cannot be saved from the Status Menu (using the /Save File/
command), although it can be viewed (from the Status Menu), nor can
it be edited from /FILES/. Status Lock, on the other hand, prevents a
run file from being seen (and hence edited) from the Status Menu.
The file remains editable from /FILES/.
The Status Screen is unaffected by Status Lock; it can always be
viewed.
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Ready Output Active
The Ready output, located on the back of the pump, continuously
sends an electrical signal to any device hardwired to it. Use the
Ready Output Active field to choose whether the signal is either a 5V
signal (Hi) or a 0V signal (Lo) whenever the pump is in a READY
state. If the pump is not READY, the other signal is output.
Whenever the pump's Ready Output is hardwired to a
SpectraSYSTEM autosampler to coordinate injections, it should be
set to provide "Hi" voltage in the READY state.
File Protection
Each numbered file can be safeguarded against accidental or
unauthorized changes by turning on the file protection feature. When
File Protection for a specific file is turned On, that file cannot be
edited, deleted, or copied to. Initially, all files are editable (file
protection is Off.) Use the [+] or [-] key to select Off or On in the
Protect field.
SOLVENT
SELECTION MENU
The Solvent Selection Menu displays four solvent labels (A, B, C,
and D), each of which can be changed to one of thirteen preset
solvent names. Matching a label to the actual solvents that are
connected to the four inlet lines is a convenient way to avoid
confusion. When you label solvents, each of the pump's displays that
would normally show lettered solvent labels A, B, C, and D will
instead show the name you set. The labels available are: H2O,
MeOH, ACN, Phos (Phosphate), Acet (acetate), TFA, Buff (Buffer),
Acid, Base, THF, IPA, MeCl, and HEX.
To change a label, select /OPTIONS/, /Solvent Selection/. The
display in Figure 4.6 appears.
Select Solvents
[
A
]
[
B
]
[
C
]
[
D
]
D
]
Figure 4.6 Solvent Names display
Use the [+]/[-] keys to select a label in any of the four fields.
Figure 4.7 shows an example.
Select Solvents
[ H2O ]
[ MeOH ]
[ IPA ]
[
Figure 4.7 Solvent labels for A, B, and C chosen
NOTE: If your solvent isn't in the list of available choices, retain the default
letter as the solvent label.
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The Queue Menu
The Queue Menu is used to edit, load or delete a chronological list of
files the pump will run, and the number of times each file is run. By
linking several files together by means of a queue you can match
specific pump files with injections in your sequence. Any regular file
(numbers 1 - 9 in the P4000) can be put into the queue. (The
Shutdown and Develop files are not queue-able.)
You can create a queue with as many as ten lines. Access the Queue
Menu by pressing [MENU] and selecting /QUEUE/ (Figure 4.8).
·FILES
>QUEUE
·COMMANDS
·TESTS
·OPTIONS
Figure 4.8 The Main Menu, showing /QUEUE/ selected
When you select /QUEUE/ the display shown in Figure 4.9 appears.
>Edit
·Load
·Delete
Figure 4.9 The Queue Menu
This section describes how the queue works, and how to edit, load,
delete, and run a queue. It also explains how a running queue can be
paused, stopped, or edited.
HOW THE QUEUE
WORKS
The pump looks at the first line of the queue to determine which file
to run. It then runs that file as many times as specified, with each run
being initiated by a manual or remote RUN command.
To use the queue:
1. Edit the Queue.
2. Load the Queue.
3. Initiate the [RUN] manually or remotely each time a new
injection/run needs to be started.
NOTE: Set the autosampler cycle time equal to the pump run time.
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Thermo Scientific
The pump will run based on the files listed in the queue. For the
example queue shown in Figure 4.10, the pump would run File #4 ten
times, then File #2 five times, and then File #1 twenty times, for a
total of 35 runs. Note that the file number is not the same as the file's
Order (chronological position) in the queue.
Order
1
File:Name
4:(filename)
#Runs
10
-----------------------------------------------------2
2:(filename)
5
3
1:(filename)
20
Figure 4.10 An Example queue
Editing, loading, and deleting a queue are explained on the following
pages.
EDITING THE
QUEUE
To view, build, or change the queue, select /Edit/. A display similar
to Figure 4.11 appears.
Order
File:Name
#Runs
1
#:(filename)
(1-999)
------------------------------------------------------
Figure 4.11 The Queue's Edit Menu
Order
The field is not editable. As you add more lines to the queue, this
field automatically displays the numerical order of lines in the queue.
File:Name
The File:Name field is used to select the name of each file to be run.
Use the [+] / [-] keys to select one file for each line.
#Runs
The field is used to enter the number of times you want a particular
file to be run before the next file (Order 2) is loaded.
Adding Lines to the
Queue
Once the File:Name and #Runs fields for Order 1 are filled in, you
can add an additional line to the queue by pressing the down-arrow
key or [ENTER]. The cursor will move to the Order 2 line. For each
line that you add, select a file name and enter a value for the number
of times you want the file to run. You may add as many as nine lines
(for a total of 10 lines).
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71
Order
File:Name
#Runs
1
#:(filename)
(1-999)
-----------------------------------------------------2
2:EXAMPLE
5
Figure 4.12 Adding lines to the queue
HINT: You can repeat a file name several times in the queue.
Deleting Lines
from the Queue
To delete lines from the queue, put the cursor in the File:Name field,
and press [-] until the File:Name field is blank. The remaining entries
will be re-sorted as soon as the cursor is moved off the line.
LOADING THE
QUEUE
The Queue Menu's Load command simultaneously loads the queue
into the pump and begins running the queue by initializing the file in
Order 1. When /Load/ is selected, the display prompts you to confirm
the operation (Figure 4.13). Press [ENTER] if you wish to proceed.
(If you do not want to load the queue, exit the display by pressing
[MENU], or [STATUS], or the up-arrow key.) A confirmation
message will appear, completing the load operation.
>Load Queue
Figure 4.13 Loading the queue
If you load a queue while another file is running, the queue will
immediately take over the pump's operation. The Order 1 file
becomes the run file and is initialized.
Once the queue is loaded, you are returned to the Status Screen. The
Status Screen will show QREADY, as soon as the pump is ready to
begin a run using the file specified in Order 1.
DELETING THE
QUEUE
The Queue Menu's command is used to erase the entire queue.
When /Delete/ is selected, the display prompts you to confirm the
operation (Figure 4.14). Press [ENTER] only if you wish to delete
the entire queue. A confirmation message completes the operation.
If you do not want to delete the queue, press the up-arrow key, or
[STATUS] or [MENU].
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>Delete Queue
Figure 4.14 Deleting the queue
If you only want to delete certain lines of the queue, use the Edit
Menu. Refer to the paragraph Deleting Lines from the Queue on page
72, for more information.
You can delete the queue at any time, regardless of whether or not it
is running. If you delete a running queue, the current run is
completed. The file that had been in Order 1 remains as the run file.
NOTE: Deleting a queue has no effect on the files themselves, it simply
erases the list of files.
The contents of the queue is lost whenever the pump is switched off
or a power failure occurs. The file that had been in Order 1 will be
the run file when power is restored.
RUNNING A QUEUE
To run a queue, simply load it by selecting /QUEUE/, /Load/. When
the pump's Status Screen shows QREADY, you can begin running the
first file in the queue by pressing [RUN] or by having another LC
instrument trigger the run. The pump will continue to run the file in
Order 1 each time a new run is started, until it has been run the
number of times specified in the #Runs field. The pump then loads
the file designated in Order 2 and uses that file the number of times
specified in that line, and so on, until the entire queue has been run.
CHECKING A
QUEUE'S
PROGRESS
You can track the progress of a running queue from the Queue Menu.
To view the progress of a running queue:
1. Press [MENU].
2. Select /QUEUE/. Note that when a Queue is loaded, the Queue
Menu (Figure 4.15) changes. The /Load/ selection is replaced
by /Pause/.
·Edit
>Pause
·Delete
Figure 4.15 The Queue Menu when a queue is loaded (running)
3. Select /Edit/ to view the running queue. The display will look
similar to Figure 4.10.
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While the queue is running, you can see the #Runs field automatically
decrease by one with each run (injection). When the last run is made
for a file, the queue is automatically re-sorted. The information for
Order 2 is moved up to Order 1, and the information for Order 3 is
moved up to Order 2. This process continues until the queue becomes
empty, is paused, or is deleted.
You can also see the progress of the current run in the queue from
Status. When a queue is running in the P4000, a Status Screen similar
to Figure 4.16 will be displayed. Note that the Status Screen shows Q
RUN on line 2.
Status
Flow
PSI
Q RUN
1.00
1250
MaxP
3000
------------------------------------------------------Stat
6.4
A
B
C
D
80.0
10.0
10.0
0.0
Figure 4.16 A P4000 Status Screen when a queue is running
As always, the Status Menu shows the run file. The run file can be
edited from the Status Menu (if Status Lock is Off), as normal.
EDITING A
RUNNING QUEUE
You can edit a running queue in order to add, delete, or edit lines
(File, #Runs). All lines of a running queue except the Order 1 line are
editable. Refer to the procedure outlined in the Edit section on page
71 to edit the queue. If you need to make a change to the Order 1 line
of a running queue, you must first pause the queue as described in
Using a Pause Command on page 75. Note that the pump will always
finish the current run before pausing.
EDITING A FILE IN
THE QUEUE
You can edit any file in the queue that has not yet been run by
selecting /FILES/, /Edit/. Since the pump only loads the file in Order
1 once, any changes made to the file specified in Order 1 do not take
effect while the queue is running. If the same file is specified later in
the queue, then the changes will be recognized, since the edited file is
loaded at a later time. To edit the file shown in Order 1, you can
either edit the run file from the Status Menu, or pause the queue (see
page 75) then edit the file. If you pause the queue (and the #Runs for
Order 1 is greater than 1), the edited file will be loaded as soon as the
queue is re-loaded.
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Thermo Scientific
LOADING OTHER
FILES
When a queue is running, you may not load any other file from the
Files Menu without first pausing or deleting the queue. If you try to
load a file while a queue is running, the information message shown
in Figure 4.17 appears. You are then returned to the Files Menu. As
described on page 75 you can load another file into the pump by first
pausing the queue. You can load another file into the queue by
editing the queue.
** Queue Loaded **
Cannot Load File
Figure 4.17 File load error message when the queue is loaded
STOPPING A
QUEUE
There are several ways to stop a queue, depending on what you wish
to do next.
NOTE: You do not need to stop the queue in order to edit it.
Using a Hold
Command
If you want to stop the pump's clock momentarily and plan to resume
the run in the queue, press [MENU], and select /COMMANDS/,
/Hold/, to hold the pump at the current compositions. The pump will
hold until a Continue command is issued.
To resume, select /COMMANDS/, /Continue/.
Using a Pause
Command
If you want to finish the current run, but then pause the queue so that
the pump can run another file, or so that you can edit the Order 1 line
of the queue, press [MENU] and select /QUEUE/, /Pause/. The
/Pause/ selection is only present if the queue is running. Remember,
you can always edit the queue itself to move a particular file into the
queue, but you cannot edit the first line of the queue if the queue is
running. Whenever the queue is paused, the letter Q will disappear
from the Status Screen.
Use /Pause/ if you need to interrupt the running of a queue for the
purpose of relegating the pump to another task.
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75
To resume running the queue, re-load the queue by selecting
/QUEUE/, /Load/. When the Status Screen shows QREADY, press
[RUN] to start the queue.
By Pressing [STOP]
You can stop the current run in the queue by pressing the [STOP]
key. The pump will immediately stop, and the clock will be reset to
zero. You may restart the same run by initializing the run file by
pressing [RUN]. (The run file will be the last file in Order 1 remember, the queue automatically re-sorts the queue after all the
runs of a file have been performed.) When the pump shows
QREADY, restart the run as you normally would. The pump
continues its operation based on the queue.
By Resetting the
Pump's Clock
Another way to reset the pump's clock and to restart the current run is
to press [MENU] and select /COMMANDS/, /Reset/. When the
Status Screen shows QREADY, restart the run by pressing [RUN].
By Aborting a
Queue
You may abort the queue by deleting it. To do this select /QUEUE/,
/Delete/. The current run of a deleted queue will be completed and
the file in Order 1 will remain the run file.
The Develop File
INTRODUCTION
The Develop File is a flexible and powerful tool that can help you
automate methods development. Available only on the P4000, this
file is used to "program" a comprehensive set of fixed and changing
parameters that includes solvent composition, gradient curves, and
run time. By letting the pump systematically increment these
parameters, and by studying the resulting chromatogram(s), you can
identify the optimum set of conditions for obtaining your best
chromatogram or for focusing on a single peak. We recommend that
only chromatographers experienced in methods development use the
Develop File.
The Develop File sets up the beginning and ending solvent
composition parameters, and specifies a solvent interval (isocratic
mode) or curve (gradient mode) by which those conditions will
change from run to run. Essentially, you "program" the Develop File
to perform the first run at one set of conditions and to automatically
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Thermo Scientific
increment those conditions through successive runs. All solvent
proportioning and switching is done based on the "program" you set
up in the file.
For example, in isocratic mode, using two solvents, you might specify
that solvent A start at 20% and end at 100% (the pump automatically
adjusts solvent B's composition to begin at 80% and end at 0%), with
the time interval set at 10%. The pump would then set up nine runs,
with each isocratic composition being different from the previous
one. Table 4.1 illustrates the resulting solvent composition for each
run in an isocratic, two-Solvent Develop file.
Table 4.1 Solvent Composition For Each Run In An Isocratic,
Two-Solvent Develop File
Run #
1
2
3
4
5
6
7
8
9
Solvent A%
20
30
40
50
60
70
80
90
100
Solvent B%
80
70
60
50
40
30
20
10
0
The flow rate, run time, and equilibration time you specify remain
constant from run to run.
You won't build a file whose menu, line by line, looks at all like
Table 4.1, though. Instead, you will simply specify all the starting
and ending parameters and any interval, along with other typical file
parameters such as flow rate and run time. From these, the pump will
automatically deliver solvents at the proper proportions.
The example in Table 4.1 is very basic. We will present many
examples in this chapter to help you understand exactly what can be
accomplished with the Develop File.
You should become familiar with an isocratic develop file before you
read about creating a gradient develop file. The examples in the first
half of this section are designed to provide you with a foundation of
isocratic program editing and then continue to the more advanced
editing features of gradient programs.
At the end of this chapter are form sheets for you to copy and use in
planning your own Develop File. They list all of the fields shown in
each display. There are six different sheets: three for isocratic
development (2, 3, or 4 solvents) and three for gradient development
(2, 3, or 4 solvents).
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Creating a Develop File Program
To edit the Develop File, select [MENU], /FILES/, /Edit/ and choose
file D which is called DEVELOP. This filename is permanent and
cannot be changed (Figure 4.18).
Edit File
File Name
D
DEVELOP
----------------------------------------------------->Setup
·Options
·Timed Events
·Preview
Figure 4.18 The Edit Menu of a Develop File
The menu selections are similar to those found in regular file editing,
except that Solvent Program is replaced by Setup, and a Preview
selection is added (Figure 4.18). The Setup Menu should be edited
first, as the Options Menu is affected by selections made in the Setup
Menu.
Program Modes
The Develop File operates in either of two program modes: isocratic
or gradient. The fields displayed in the Setup Menu will vary based
on the mode you select on the first line of the Setup Menu, and on the
number of solvents you specify on the second line.
SETUP
When you select /Setup/, a display similar to the one shown in
Figure 4.19 appears.
Program
Number of Solvents
Isocratic
2
Figure 4.19 The first lines of a Develop File's Setup Menu
Program
Number of Solvents
Select either Gradient or Isocratic in the Program field. Then choose
the number of solvents you want to use in the Number of Solvents
field. You must always use at least two solvents.
NOTE: Verify the program mode and the number of solvents you
intend to use before going any further into Setup or going into the
Options Menu. Although there are some common fields between
Setup Menus, not all entries you make will be retained if either the
program mode or the number of solvents is later changed. You
cannot retrieve your previous menu settings by returning the
Program/Number of Solvents settings to their earlier values.
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Thermo Scientific
Common Fields in
the Setup Menu
The Setup Menu in gradient (grad) mode is similar to the Setup Menu
in isocratic (iso) mode, but it is not identical. There are minor
differences between the Options Menu in both modes, too. In several
instances menus contain fields with identical names, in other
instances, field names are different, but imply a very similar function.
The fields of an isocratic, two-solvent program (Figure 4.20) are
described below. These fields provide the foundation for all other
isocratic and gradient programs. Other fields, specific to certain
Program/Number of Solvent combinations, are explained below each
menu in the Other Isocratic Programs (page 80) and Gradient
Programs (page 84) sections.
Iso / Two Solvents
The two-solvent isocratic Setup Menu is shown in Figure 4.20. All of
the fields in this Setup Menu are explained below.
Program
Isocratic
Number of Solvents
2
-------------------------------------------------------Solvent 1
A
Solvent 2
B
Start
%
(Solvent 1)
0
End
%
(Solvent 1)
100
% Interval
Flow Rate
Run Time
20
1.0
10.0
Figure 4.20 The Develop File's Setup Menu showing some common
fields
Solvent 1
Select one of the four solvents to be designated as your first solvent.
Solvent 2
Select one of the four solvents to be designated as your second
solvent.
NOTE: The pump does not allow you to select the same solvent for Solvent
1 and Solvent 2.
Start %
End %
Enter values for the beginning and ending percentages of the first
solvent. This would be solvent A in the menu shown in Figure 4.20
because A was selected to be Solvent 1.
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79
% Interval
Set an interval by which the first solvent will change (increase or
decrease, depending on the start and end percentages) for every
successive run. The pump will automatically set up each successive
run to increase or decrease the first solvent by this percent. The pump
automatically adjusts the second solvent accordingly.
Flow Rate
Run Time
Enter values as desired.
OTHER ISOCRATIC
PROGRAMS
This section describes fields that are specific to three- and foursolvent isocratic Setup Menus.
Iso / Three Solvents
When using three solvents in an isocratic program, the Setup Menu
(Figure 4.21) asks for more solvent information than when only a
two-solvent program is used.
Program
Isocratic
Number of Solvents
3
-------------------------------------------------------D
Solvent Not Used
Solvent 1
A
Start
%
A
0
End
%
A
100
% Interval
Solvents 2,3
Relative Start %
Relative End %
20
B
C
50
50
0
100
20
Relative % Interval
Flow Rate
Run Time
1.0
10.0
Figure 4.21 The three-solvent isocratic Setup Menu
Solvent Not Used
Select the solvent that you will not use.
NOTE: The pump does not allow you to select the same solvent for Solvent
Not Used and Solvent 1.
Solvents 2,3
The Solvents 2,3 field is filled in automatically, based on the solvents
remaining after selections have been made in the Solvent Not Used
field and the Solvent 1 field. This line simply labels to Solvents 2
and 3 for you.
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Thermo Scientific
RelativeStart %
There are two fields on the RelativeStart % line. These correspond to
Solvents 2 and 3. The relative start percentages for the two solvents
always add to 100. You enter a value for Solvent 2. Solvent 3 is
entered automatically. The values in these fields are relative to
Solvent 1's composition.
For example, if the Start % of Solvent 1 were 60%, then the
remaining 40% would be comprised of a combination of solvents 2
and 3. If you want the relative ratio of Solvents 2 and 3 to be 50 / 50,
you would enter a RelativeStart % of 50 for Solvent 2. Then, the
actual proportions delivered by the pump would be:
Solvent 1
Solvent 2
Solvent 3
60%
20%
20%
(40% × 0.5 = 20)
(40% × 0.5 = 20)
The total of all solvents is 100%.
If solvents 2 and 3 were not entered as 50 / 50, but 20 / 80 instead,
then the actual proportions delivered by the pump would be:
Solvent 1
Solvent 2
Solvent 3
60%
8%
32%
(40% × 0.2 = 8)
(40% × 0.8 = 32)
You do not need to make these calculations yourself to see what the
pump will actually do. After you complete your program in the Setup
Menu, you can preview the composition of the resulting runs by
selecting /Preview/ as described in conjunction with several examples
beginning on page 87.
Relative End %
Enter a value for the relative end % of Solvent 2. Solvent 3 is entered
automatically.
Relative % Interval
Enter a value for the relative percent increment from run to run of
Solvents 2 and 3. If you do not wish to increment Solvents 2 and 3,
make sure the Relative End % is the same as the RelativeStart %.
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81
More Information About
Relative Percentages
When you set up a program to increment the relative percents of
solvents 2 and 3, you are creating a "sub-loop" in the program. This
"sub-loop" is created because Solvent 1, itself, has been set up to
increment from run to run.
The set of runs that the Develop File creates in these situations goes
like this:
•
In the first run, the proportions of solvents 1, 2, and 3 are taken
directly from the Setup Menu.
•
In the next runs, Solvent 1 will maintain its initial composition,
while Solvents 2 and 3 increment. Solvent 1 will maintain its
initial proportion for as many runs as are necessary, while
Solvents 2 and 3 are systematically incremented.
•
Only when the "sub-loop" is complete (i.e., when Solvents 2
and 3 reach their specified Relative End % values), will
Solvent 1 change by its increment value.
•
Then the sub-loop of Solvents 2 and 3 begins again, as
Solvent 1 remains fixed at its new composition.
Iso / Four Solvents
The isocratic four-solvent case is similar to the isocratic three-solvent
case. However, when four solvents are specified (Figure 4.22), one of
the four solvent compositions must remain fixed. The entries made in
the Setup Menu determine how Solvents 1, 2, and 3 vary to comprise
the percentage difference (100% - Fixed Solvent %) for each run.
Program
Isocratic
Number of Solvents
4
------------------------------------------------------D
Fixed Solvent
20
Fixed Solvent %
Solvent 1
A
Start
%
A
0
End
%
A
100
% Interval
Solvents 2,3
RelativeStart %
Relative End %
20
B
C
100
0
0
100
Relative % Interval
Flow Rate
Run Time
20
1.0
10.0
Figure 4.22 The four-solvent isocratic Setup Menu
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Thermo Scientific
Fixed Solvent
Select the solvent whose composition will remain fixed.
Fixed Solvent %
Enter a value for the fixed solvent composition.
OPTIONS MENU
(ISOCRATIC)
The isocratic Options Menu (Figure 4.23) is used to set all of the
same parameters as that of normal files, with three additions. Note
that the Options Menu in isocratic mode is different than in gradient
mode. The Options Menu for gradient programs is described on
page 86.
Access the Options Menu by selecting /Options/ from the Edit Menu.
The Options Menu is the same for any isocratic program, regardless
of the number of solvents specified in the Setup Menu.
Equilibration Time
#Runs/Interval
0.0
1
-------------------------------------------------------#Develop Cycles
Maximum Pressure
Minimum Pressure
Total # of Runs
1
3000
0
(Not editable, filled in automatically)
Figure 4.23 An isocratic program's Options Menu
Equilibration Time
Enter a value for the equilibration time. It is very important to set an
equilibration time in the Develop File so that the column can
equilibrate prior to the beginning of the next run.
#Runs/Interval (Develop File)
Enter the number of times you want each run repeated, before the
solvents' compositions are incremented. (An interval, in this case,
refers to one set of solvent compositions.)
#Develop Cycles (Develop File)
Select the number of times (9 maximum) the entire program will be
executed. Usually this is only once (1), but you might enter another
number if you want to run the entire program again using a different
column, or, if you have a column oven, at a different column
temperature.
Maximum Pressure
Minimum Pressure
Enter values for maximum and minimum pressure levels, if desired.
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83
Total # of Runs
The Total # of Runs field is filled in automatically, based on the
program entered in the Setup Menu, and the values specified in the
#Runs/Interval and the #Develop Cycles fields. The pump can
contain as many as 999 runs in the Develop File.
GRADIENT
PROGRAMS
The gradient program mode is used to program the pump to perform a
series of varying gradient runs. Unlike the isocratic mode where you
increment Solvent 1 by a percentage, in the gradient mode you
specify the total number of gradient curves you want performed.
During each successive run, the pump follows a different gradient
shape (for example., Linear, Convex 2, then Concave 2). Note that
this is not the same as specifying the curve itself (that is., Concave 7
or Convex 4), as you do in regular files.
You vary the relative percentages of Solvents 2 and 3 just as you do
in the isocratic program mode, except that, when the pump performs
the gradient, the other solvents also follow gradient shapes.
This section describes the fields that are specific to two-, three-, or
four-solvent gradient Setup Menus. The fields that are common to all
Setup Menus are described on page 79.
Grad / Two Solvents
Program
Gradient
Number of Solvents
2
--------------------------------------------------------Solvent 1
A
Solvent 2
B
Start
%
A
0
End
%
A
100
1
#Curves
Flow Rate
Run Time
1.0
10.0
Figure 4.24 The two-solvent gradient Setup Menu
#Curves (Develop File)
Select the total number of gradient shapes you want the pump to run.
If, for example, you select 1, as in Figure 4.24, then the curve would
be "Linear" - a straight line from 0% to 100% over the course of the
run. If, instead, you select 3, then three runs would be performed:
Linear, Concave 2, and Convex 2. shows the curves that correspond
to each numerical selection. Refer to Chapter 3 to see the shape of
each curve.
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Thermo Scientific
Table 4.2 The Number Of Curves Selected And The
Corresponding Gradient Shapes Performed
#Curves Shapes, Performed
1
Line
3
Line, Cvx2, Ccv2
5
Line, Cvx2, Ccv2, Cvx3, Ccv3
7
Line, Cvx2, Ccv2, Cvx3, Ccv3, Cvx5, Ccv5
9
Line, Cvx2, Ccv2, Cvx3, Ccv3, Cvx5, Ccv5, Cvx7, Ccv7
11
Line, Cvx2, Ccv2, Cvx3, Ccv3, Cvx5, Ccv5, Cvx7, Ccv7, Cvx10, Ccv10
(Line = linear, Ccv = concave, Cvx = convex)
Grad / Three
Solvents
During a three-solvent gradient run, the ratio of solvents 2 and 3
remains constant. As the gradient is generated, it is Solvent 1's ratio
to Solvent 2 and its ratio to Solvent 3 that changes. This is commonly
called a solvent strength gradient. In successive runs, Solvents 2 and
3 take on a new ratio, which is maintained as the gradient is
generated.
Program
Gradient
Number of Solvents
3
--------------------------------------------------------Solvent Not Used
D
Solvent 1
A
Start
%
A
80
End
%
A
20
Solvents 2,3
RelativeStart %
Relative End %
B
C
100
0
0
100
Relative % Interval
20
#Curves
Flow Rate
Run Time
1
1.0
10.0
Figure 4.25 The three-solvent gradient Setup Menu
All the fields in this menu have been explained in the previous pages.
Grad / Four
Solvents
When four solvents are specified (Figure 4.26), one of the four
solvents' compositions must remain fixed. The entries made in the
Setup Menu determine the ratios between Solvents 1, 2, and 3 that
will be maintained as each gradient curve is performed.
Thermo Scientific
85
Program
Gradient
Number of Solvents
4
-------------------------------------------------------Fixed Solvent
D
Fixed Solvent %
0
Solvent 1
A
Start %
End
%
A
0
A
100
Solvents 2,3
RelativeStart %
Relative End %
B
C
100
0
0
100
Relative % Interval
20
#Curves
Flow Rate
Run Time
1
1.0
10.0
Figure 4.26 The four-solvent gradient Setup Menu
All fields in this menu have also been explained previously.
OPTIONS MENU
(GRADIENT)
The gradient Options Menu (Figure 4.27) is used to set all of the same
parameters as those found in an isocratic Options Menu, with two
differences, #Runs/Curve and Run Time Increment. To access the
Options Menu select /Options/ from the Files Menu. The Options
Menu is the same for all gradient programs, regardless of the Number
of Solvents specified in the Setup Menu. (Note the Options Menu for
an isocratic program is different from a gradient program. Refer to
page 83 for the isocratic Options Menu.)
Equilibration Time
#Runs/Curve
0.0
1
-------------------------------------------------------#Develop Cycles
Run Time Increment
Maximum Pressure
Minimum Pressure
Total # of Runs
1
0.0
3000
0
(not editable, filled in automatically)
Figure 4.27 A gradient program's Options Menu
#Runs/Curve (Devlop File)
Select the number of times you want each run of a gradient shape
performed, before the solvent parameters are incremented.
Run Time Increment
Enter a value (in minutes) to increment the run time of multiple
develop cycles. The Run Time Increment only operates if there is
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Thermo Scientific
more than one develop cycle (#Develop Cycles). After the pump
completes an entire cycle of runs, it increments the Run Time by the
time entered here.
For example, if you specify five develop cycles, a run time of 10.0
minutes, and a Run Time Increment of 5.0 minutes. The resulting run
times for each develop cycle will be those shown in Table 4.3.
Table 4.3 An example showing the use of Run Time Increment
and five develop cycles
Cycle
Run Time
Each run in Cycle 1
Each run in Cycle 2
Each run in Cycle 3
Each run in Cycle 4
Each run in Cycle 5
10.0 minutes
15.0 minutes
20.0 minutes
25.0 minutes
30.0 minutes
Develop File Examples
This section describes the Preview Screen and presents seven
different examples of isocratic and gradient programs. The Setup
Menu for each example program is presented, along with a preview of
each run, and a graph showing solvent compositions from run to run.
The last section of this chapter contains form sheets for planning and
recording your own Develop File, as well as a Preview Record on
which you can record the composition of each run shown in the
Preview Screen.
PREVIEW SCREEN
Once you have built a program using the Setup and Options Menus,
you can preview the programmed run compositions by selecting
/Preview/ (Figure 4.28).
Edit File
File Name
D
DEVELOP
-----------------------------------------------------·Setup
·Options
·Timed Events
>Preview
Figure 4.28 The Develop File's Edit Menu, with Preview selected
The Preview Screen differs, depending on the program mode. An
isocratic Preview Screen is shown in Figure 4.29; a gradient Preview
Screen is shown in Figure 4.30.
Thermo Scientific
87
Isocratic
80.0
Cyc:1
20.0
IntRun#1
0.0
0.0
#
1
Figure 4.29 An isocratic Preview Screen
Gradient
0.0
Cyc:1
100
0.0
CrvRun#1
0.0
#
1
---------------------------------------------------100.0
0.0
0.0
0.0
Line
Figure 4.30 A gradient Preview Screen
Selecting Runs
There is only one active cursor location (field) in the Preview Screen:
the far-right field, underneath the word IntRun# (isocratic) or
CrvRun# (gradient). Use the [+] / [-] keys in this field to select a run
number to preview. The solvent compositions for each run will be
displayed to the left, in the solvent composition fields, as you select
run numbers. Note that the figures showing Preview Screens in the
rest of this chapter list subsequent runs under a dashed line.
(Normally, the dashed line indicates that you press [∨] to view
subsequent lines.)
The gradient Preview Screen contains one additional line for each run
number, accessed by using the down-arrow key. This line shows the
ending solvent compositions and, in the far-right corner, indicates the
gradient curve shape for that run. Refer to Chapter 3 for a description
of gradient curve shapes. "Ccv" denotes concave, "Cvx" denotes
convex, and "Line" denotes linear.
Program Mode
The program mode is shown in the left-hand corner of the first line.
Cycle
The number corresponding to the develop cycle for the selected run is
shown in the middle of the top line.
IntRun# , CrvRun#
The number immediately following the word "IntRun#" and
"CrvRun#" at the top, right-hand corner, changes automatically if
more than one run per interval or one run per curve has been selected.
You will see this number change as you increment and decrement the
run number.
The Preview Screen shows all runs for a given program even when
the Develop File is running. The preview screen only changes when
the Develop File is edited.
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Thermo Scientific
EXAMPLE 1
ISO / TWO
SOLVENTS
This example shows a two-solvent isocratic program (Figure 4.31).
Solvent A starts at 20% and will increment 20% for each run until it
reaches the last run at 80%. The total number of runs is four (4).
Thermo Scientific
89
Setup
This example assumes that the #Runs/Interval = 1 and the #Develop
Cycles = 1.
Program
Isocratic
Number of Solvents
2
-------------------------------------------------------Solvent 1
A
Solvent 2
B
Start
%
A
20
End
%
A
80
% Interval
20
Flow Rate
1.0
Run Time
10.0
Figure 4.31 The Setup Menu for Example 1
Preview
The Preview Screen for this program is shown in Figure 4.32.
Isocratic
20.0
Cyc:1
80.0
IntRun#1
0.0
0.0
#
1
-------------------------------------------------------40.0
60.0
0.0
0.0
#
2
60.0
40.0
0.0
0.0
#
3
80.0
20.0
0.0
0.0
#
4
Figure 4.32 The Preview Screen for Example 1
The graph below shows how solvents A and B change each time a
new run begins. Note that the dashed line in the isocratic graphs
indicate that the pump changes solvent composition between the end
of one run and the beginning of the next. Solvent composition does
not change during the run in an isocratic program.
90
Thermo Scientific
Figure 4.33 The two-solvent composition path for Example 1
Thermo Scientific
91
EXAMPLE 2
GRAD / TWO
SOLVENTS
This example shows a simple gradient program using two solvents
(Figure 4.34). Solvent A starts at 80% and ends at 20%. One curve
shape, linear, is performed over the 10.0 minute run time. The total
number of runs in this program is one (1).
Setup
This example assumes that the #Runs/Curve = 1, the #Develop Cycles
= 1, and the #Curves = 1.
Program
Gradient
Number of Solvents
2
--------------------------------------------------------Solvent 1
A
Solvent 2
B
Start
%
A
80
End
%
A
20
#Curves
1
Flow Rate
1.0
Run Time
10.0
Figure 4.34 The Setup Menu for Example 2
Preview
Gradient
80.0
Cyc:1
20.0
0.0
CrvRun#1
0.0
# 1
-------------------------------------------------------20.0
80.0
0.0
0.0
Line
Figure 4.35 The Preview Screen for Example 2
Since only one curve is specified, the Develop File in this example is
identical to a two time-line Solvent Program, which specifies a linear
gradient curve.
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Thermo Scientific
0
100
10
90
1
80
30
70
40
60
50
50
60
40
70
30
80
20
90
10
100
0
PERCENT SOLVENT B
PERCENT SOLVENT A
20
Figure 4.36 The two-solvent gradient composition path for Example 2
Notice that the solid line in the gradient graphs indicates that the
pump changes solvent composition during the run.
Thermo Scientific
93
EXAMPLE 3
GRAD / TWO SOLVENTS WITH
MULTIPLE CURVES
Example 3 (Figure 4.37) is similar to Example 2, except that five
curves are specified. The total number of runs in this program is
five (5).
Setup
This example assumes the #Runs/Curve = 1, the #Develop Cycles =
1, and the #Curves = 5.
Program
Gradient
Number of Solvents
2
--------------------------------------------------------Solvent 1
A
Solvent 2
B
Start
%
A
100
End
%
A
0
#Curves
5
Flow Rate
1.0
Run Time
10.0
Figure 4.37 The Setup Menu for Example 3
Preview
Gradient
Cyc:1
CrvRun#1
100.0
0.0
0.0
0.0
# 1
0.0
100.0
0.0
0.0
Line
-------------------------------------------------------100.
0.0
0.0
0.0
# 2
0.0
100.
0.0
0.0
Cvx2
-------------------------------------------------------100.
0.0
0.0
0.0
# 3
0.0
100.
0.0
0.0
Ccv2
-------------------------------------------------------100.
0.0
0.0
0.0
# 4
0.0
100.
0.0
0.0
Cvx3
-------------------------------------------------------100.
0.0
0.0
0.0
# 5
0.0
100.
0.0
0.0
Ccv3
Figure 4.38 The Preview Screen for Example 3
If, in the Options Menu, we had specified the #Runs/Curve = 2, then a
total of ten runs would have been seen in the Preview Screen instead
of five.
The graph in Figure 4.39 illustrates the gradient curves performed in
this example.
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Thermo Scientific
100
0
10
90
4
80
20
2
70
60
40
50
1
50
40
60
3
70
30
20
80
5
90
100
PERCENT SOLVENT B
PERCENT SOLVENT A
30
5
10
0
TIME
Figure 4.39 The two-solvent gradient curve composition path for Example 3
Thermo Scientific
95
EXAMPLE 4
GRAD / TWO
SOLVENTS, WITH
RUN TIME
INCREMENT
Example 4 (Figure 4.40) shows how a run time increment can be used
in the gradient program mode. The Setup Menu is similar to Example
4, but the Options Menu in this example specifies a Run Time
Increment of 10.0 minutes, and four (4) Develop Cycles. The total
number of runs created by this program is four (4).
Setup
This example also assumes the #Runs/Curve = 1, and the
#Curves = 1.
Program
Gradient
Number of Solvents
2
--------------------------------------------------------Solvent 1
A
Solvent 2
B
Start
%
A
80
End
%
A
20
#Curves
1
Flow Rate
Run Time
1.0
10.0
Figure 4.40 The Setup Menu for Example 4
Options
Equilibration Time
#Runs/Curve
0.0
1
-------------------------------------------------------#Develop Cycles
4
Run Time Increment
10.0
Maximum Pressure
3000
Minimum Pressure
0
Total # of Runs
4
Figure 4.41 The Options Menu for Example 4
96
Thermo Scientific
Preview
Gradient
Cyc:1
CrvRun#1
80.0
20.0
0.0
0.0
# 1
20.0
80.0
0.0
0.0
Line
(10.0 minute run time)
-------------------------------------------------------80.0
20.0
0.0
0.0
# 2
20.0
80.0
0.0
0.0
Line
(20.0 minute run time)
-------------------------------------------------------80.0
20.0
0.0
0.0
# 3
20.0
80.0
0.0
0.0
Line
(30.0 minute run time)
-------------------------------------------------------80.0
20.0
0.0
0.0
# 4
20.0
80.0
0.0
0.0
Line
(40.0 minute run time)
Figure 4.42 The Preview Screen for Example 4
Figure 4.43 The composition paths for a two-solvent gradient with run time
increment for Example 4
Thermo Scientific
97
Running the Develop File
LOADING THE
DEVELOP FILE
Load the Develop File just as you would any other file (/FILES/,
/Load/). If the total number of runs programmed by the Develop File
exceeds 999, a message will indicate that this maximum has been
reached, preventing the file from being loaded successfully. If the
message appears, edit the Develop File to reduce the total number of
runs. The following fields affect the total number of runs:
% Interval
Relative % Interval
#Curves
#Develop Cycles
#Runs/Interval or #Runs/Curve
(Setup Menu)
(Setup Menu)
(Setup Menu)
(Options Menu)
(Options Menu)
After you make changes you can double-check the total number of
runs from the Options Menu.
STATUS
The Status Screen for an example Develop File is shown in
Figure 4.44. Note that all four solvents are automatically displayed,
regardless of the Number of Solvents specified in the Setup Menu.
Status
Flow
PSI
READY
1.00
1250
MaxP
6000T
--------------------------------------------------------Stat
0.0
File
Time
A
20.0
B
80.0
C
D
0.0
0.0
D:DEVELOP
C
Flow
0.0
20.0
A
80.0
0.0
1.00
10.0
70.0
30.0
0.0
1.00
Maximum Pressure
Minimum Pressure
Equilibration Time
Gradient Curve
#Run/Total#Runs
B
6000
0
0.0
Convex 3
04/10
Figure 4.44 An example Status of a gradient Develop File
98
Thermo Scientific
Status Menu
The Status Menu shows one new line, #Run/Total#Runs. This line
displays the current run number and the total number of runs
programmed in the file. The Gradient Curve field shows the curve
shape of the current run.
The same Status Menu editing capabilities of a run file also apply
when a Develop File is loaded, except that the run file may not
contain more than two time-lines.
COMMANDS
All commands function as they would for any other file. However,
the pump sees the Develop File as one file, not as a series of runs.
This is important if you reset the file, using /COMMANDS/, /Reset/.
When the reset command is issued, the pump will return to the initial
conditions of Run #1. The entire program then restarts as soon as the
pump receives a RUN command.
Develop File Review
The examples presented in the preceding pages give you a good idea
of the flexibility and power of the Develop File. You can combine
many of the concepts presented in these examples into your own
Develop File, depending on your needs.
For instance, in a gradient program you might combine the run time
increment with three or more curves, instead of one. The pump
would perform all the curve shapes, then increment the run time and
run all the shapes again.
In an isocratic program you might specify two develop cycles, instead
of one, and run the second cycle with a different column. You could
do the same thing with a gradient program.
The Develop File can greatly assist you in automating methods
development. The form sheets supplied in the following pages can
help you plan and document the conditions necessary for optimizing
your chromatography.
Thermo Scientific
99
Develop File Record
Setup
Program
Number of Solvents
Date:
Isocratic
2
Solvent 1
________
Solvent 2
________
Start %
(Solvent 1)
(0 - 100)
________
End
%
(Solvent 1) (0 - 100)
________
% Interval
(1 - 100)
________
Flow Rate
Run Time
Options
(0.0 - 650)
________
Equilibration Time
(0.00 - 99.9)
________
#Runs/Interval
(1 - 9)
________
#Develop Cycles
(1 - 9)
________
Maximum Pressure
(1 - 6000 psi)
________
Minimum Pressure
(0 - 5999 psi)
________
Total # of Runs
(fill in from display)
________
NOTES:
100
________
Thermo Scientific
PERCENT SOLVENT A
100
10
90
20
80
30
70
40
60
50
50
60
40
70
30
80
20
90
10
100
0
Thermo Scientific
PERCENT SOLVENT B
0
101
Develop File Record
Setup
Program
Number of Solvents
Date:
Gradient
2
Solvent 1
________
Solvent 2
________
Start %
End%
(Solvent 1)
(Solvent 1)
#Curve
(0 - 100)
________
(0 - 100)
________
(1,3,5,7,9,11)
________
Flow Rate
Run Time
Options
Equilibration Time
#Runs/Curve
(0.0 - 650)
(0.00 - 99.9)
(1 - 9)
________
________
________
#Develop Cycles
(1 - 9)
________
Run Time Increment
(0.0 - 650)
________
Maximum Pressure
(1 - 6000 psi)
________
Minimum Pressure
(0 - 5999 psi)
________
Total # of Runs
(fill in from display)
________
NOTES:
102
________
Thermo Scientific
PERCENT SOLVENT A
100
10
90
20
80
30
70
40
60
50
50
60
40
70
30
80
20
90
10
100
0
Thermo Scientific
PERCENT SOLVENT B
0
103
Develop File Record
Date:
Setup
Program
Number of Solvents
Gradient
4
Fixed Solvent
Fixed Solvent %
________
________
(0 - 100)
Solvent 1
Start %
End
________
(0 - 100)
________
(Solvent 1) (0 - 100)
________
(Solvent 1)
%
Solvents 2,3
(
)
(
RelativeStart %
(0 - 100)
________
________
Relative End %
(0 - 100)
________
________
Relative % Interval
#Curves
(1 - 100)
(1,3,5,7,9,11)
Flow Rate
)
________
________
________
Run Time
(0.0 - 650)
________
Equilibration Time
(0.00 - 99.9)
________
#Runs/Curve
(1 - 9)
________
Options
#Develop Cycles
Run Time Increment
(0.0 - 650)
________
Maximum Pressure
(1 - 6000 psi)
________
Minimum Pressure
(0 - 5999 psi)
________
Total # of Runs
(fill in from display)
NOTES:
104
________
(1 - 9)
Thermo Scientific
________
Thermo Scientific
105
Preview Record
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Thermo Scientific
107
The Tests Menu
The Tests Menu allows you to access the pump’s built-in diagnostics,
part of the pump’s advanced features. This section assumes that the
source of the problem is known to be the SpectraSYSTEM pump. If
you are not certain that the pump is the source of trouble, refer to
General LC System Troubleshooting, on page 159.
To access the Tests Menu, select /TESTS/ from the Main Menu
(Figure 4.45):
·FILES
·QUEUE
·COMMANDS
>TESTS
·OPTIONS
Figure 4.45 Main Menu with /TESTS/ selected
The Tests Menu (Figure 4.46) consists of five items. Tests are
divided into three specific menus, for convenience: diagnostic tests,
calibration tests, and service tests. The Maintenance Log is described
fully in Chapter 5.
>Software Version
·Diagnostics
-------------------------------------------------------·Maintenance Log
·Calibration
·Service
Figure 4.46 Tests Menu
ABOUT RUNNING
TESTS
Tests are internal computer programs that exercise the pump's
hardware and circuitry and verify operation. If any abnormal
behavior is found it is reported as a message or an electronic circuit
board failure code. In most cases the test isolates the problem to the
failed module or component.
108
Thermo Scientific
Active and Passive
Tests
There are active and passive tests. Passive tests can be initiated at any
time as they do not affect either file memory or pump performance.
Passive tests are usually initiated by pressing [ENTER]. Active tests
require that the pump be idle before being initiated, since the pump's
valves and motor maybe engaged during the test. Usually, active tests
are initiated by pressing [RUN], and are stopped by pressing [STOP].
Some tests stop by themselves. An active test should not be
performed while the pump is in operation, as it will interfere with
pump operation.
NOTE: Pressing [STOP] during a passive test can interrupt pump
operation.
Initiating Tests
To initiate a test, move the cursor to the test's name and press
[ENTER]. Always follow the instructions displayed on the pump
when you initiate a test. In some cases the message, "Pump must be
stopped to run test" may be displayed if you attempt to run an active
test while the pump is in RUN. Alternately, if the pump is stopped
when you initiate a test you may see the message, "The Pump Must
Be Running to Perform This Test." In most cases, the pump will
initiate an active test if the pump is in EQUIL or READY.
Usually you will press [ENTER] to initiate a passive test, or [RUN] to
initiate an active test. Some tests display instructions. In these cases,
follow the instructions in the message to proceed.
Test Results
After each test is run a message appears advising you of the results of
the test. In most cases, if trouble is found, the message indicates the
failure or failed component. Specific test menu descriptions begin
below.
Flow Stability and
Hardware Series
Test Routines
Under most circumstances the Flow Stability and Hardware Series
tests will provide a thorough evaluation of the condition of your
pump. We recommended that these two tests be used first if the
performance of the pump is in question. The Flow Stability test is
described on page 110 and the Hardware Series Tests is described on
page 121.
Thermo Scientific
109
SOFTWARE
VERSION
(PASSIVE)
Selecting /Software Version/ displays the version of software
contained in the pump. The particular version of software resident in
your pump will vary depending upon the date of manufacture or upon
the date of software upgrade.
THE DIAGNOSTICS
MENU
The Diagnostics Menu (Figure 4.47) contains three items commonly
used to evaluate the pump and an additional selection to allow the
pressure transducer's output to be zeroed.
>Flow Stability
·Measured Parameters
-------------------------------------------------------·Check Valve Test
·Zero Pressure
Figure 4.47 The Diagnostics Menu
Flow Stability
Your pump is constantly monitoring its flow stability while pumping.
An internal software program allows the pump to determine when
flow stability has been adversely affected by leaking check valves,
out-gassing solvents, or other abnormal conditions.
The Flow Stability test (a passive test) evaluates how even the solvent
flow is through the pump. The pump must be pumping solvent for
this test to be run. The test can be initiated when the Status Screen
shows any one of these states: EQUIL, or READY (all pumps). In
addition, time, QEQUIL, or QREADY can be displayed on the P4000
pump. Completion of the test however, is dependent on the pump
being in a READY state. When the pump is not in a ready state the
Flow Stability screen will read, "Not Ready". When flow stability is
being evaluated, the screen will read, "Test in Progress. Please Wait."
The pump's cam must go through at least 10 pump cycles (cam
revolutions) to accurately assess stability. The value displayed is
affected by the compressibility of the solvent being pumped and the
compliancy of the hardware (tubing, column, and so on).
Therefore, the results are reported in two ways: a) an overall
judgment of the pump performance which appears in the upper lefthand corner of the display, and b) a number indicating where, within
the range, the result lies. This number is shown in the upper righthand corner of the display. Three flow stability readings are possible:
110
Thermo Scientific
Stable
Acceptable
Unstable
( 0- 25)
(25 - 90)
( > 90)
Unless a very volatile or compressible solvent is being pumped, for
example hexane, a number near the higher end of the range (60-90)
probably indicates that the system is not ideal, and the results of
further troubleshooting might improve the flow stability.
NOTE: The Flow Stability Test may show unstable flow during column
equilibration.
If the results of the flow stability test are abnormal for your LC
application, follow these steps to locate the problem:
1. Test the integrity of the inlet and transducer check valves by
running a Check Valve test. (See page 112.)
2. Verify that the mobile phase solvents are adequately degassed.
3. Refer to the recommendations of General LC System
Troubleshooting Techniques in Appendix A, page 159.
The flow stability assessment will remain displayed until you stop the
test by pressing any one of the following keys: [ENTER], [∧],
[MENU], or [STATUS].
Measured Parameters
Measured Parameters (a passive test) shows the measured flow rate
(Figure 4.48). The number in parentheses indicates the flow rate
setting in the run file. The calculated flow is based on the flow
calculated during the last 360° of motor rotation.
Calc Flow
(2.00)
1.95
Figure 4.48 An example of the Measured Parameters Menu
The measured parameters will remain on the display until the test is
stopped by pressing any one of the following keys: [ENTER], [∧],
[MENU], or [STATUS].
Thermo Scientific
111
Check Valve Test
The Check Valve test (an active test) can help you further diagnose
the source of flow stability problems. If the results indicate a
defective check valve, the test should be repeated to ensure the results
were not due to an isolated transient condition, such as a single air
bubble.
This test affects flow accuracy while the test is running. Do not run
the test during an analysis. The pump can be in EQUIL, or READY
and the flow rate must be 2.5 mL/min or less (for standard liquid
ends). If the test is activated and the current flow rate is greater than
2.5 mL/min, a message will be displayed, prompting you to reset the
flow rate. In general, the flow rate must be set to one-quarter or less
of the maximum flow rate of the liquid ends.
HINT: Defective check valves tend to perform better at higher column
pressures. Lowering the column pressure by decreasing the flow rate
increases the test's sensitivity of marginally operating check valves.
NOTE: The inlet check valve is located at the base of the inlet liquid end.
The transducer check valve is located at the base of the pressure transducer.
To activate the test select /Check Valve Test/. Follow the displayed
instructions. A message will inform you of the 8 - 10 pump cycle
delay before test completion. During this time, the condition of the
inlet and transducer check valves of the pump are monitored. When
the monitoring period is complete, pumping returns to normal
constant flow control and a message is displayed, showing the results.
Messages are listed in Table 4.1.
NOTE: Changing mobile phase concentration may cause the test to report
a good check valve as defective. Stabilize composition before running the
test.
Pressing [RUN] after the results are displayed will rerun the test.
Stop the test by pressing [ENTER] or [∧].
112
Thermo Scientific
Table 4.4 Check Valve Test Results
Both check valves good
Both check valves are performing well.
Transducer check valve
is defective
The transducer check valve should be replaced. See Chapter 5, Required
Maintenance for instructions.
Inlet check valve may
be defective
The inlet check valve may be defective. An air bubble lodged in the check
valve or piston seal or a slight leak in an inlet fitting may cause this message
to be displayed. Verify that solvents are adequately degassed and that
fittings are tight. Purge the pump and rerun the test to verify the message.
If this same message is displayed, replace the inlet check valve. See Chapter
5, Required Maintenance for instructions.
Bubbles or leaks likely.
Check degas
The check valves are not the cause of flow problems. Verify that solvents
are adequately degassed and that fittings are tight. Observe the inlet tubing
while purging the pump. If air bubbles are seen, increase the helium flow
rate (if helium degassing), or tighten the leaking fitting. Tighten bottle caps.
Ensure solvent supply is vented.
NOTE: Pulse dampeners should not be used with SpectraSYSTEM pumps.
The flow is dynamically controlled and will be adversely affected by
compliant loads.
Test aborted, Pump not
referenced in 10 Cycles
The pump is not able to establish a reference column pressure within
10 pump cycles. The pump has serious flow problems. Verify that the
solvents used are miscible in all concentrations encountered. If possible for
your column, increase the column pressure by raising the flow rate. The
check valves require more than 100 psi column pressure to operate properly.
Defective check valves will usually operate well enough at higher pressures
to allow the test to run.
Test aborted
By Operator
The test was stopped before the pump could count 8 cycles.
Thermo Scientific
113
Zero Pressure
The pump allows the pressure transducer's output to be zeroed
automatically without the need for adjustment of potentiometers. The
pump should be stopped before selecting /Zero Pressure/. The display
will show:
Release system pressure,
then press ENTER key.
Figure 4.49 Release system pressure prior to zeroing the pressure
transducer's output
Release the system pressure by opening the bypass valve or removing
the column from the system to ensure that the transducer is actually
sensing zero system pressure. Otherwise, a message showing
"Unable to Zero" will appear. Follow the instructions to complete
zeroing the pressure. To abort the test, press [ENTER], or [∧], or
[STOP]. This will return the zero setting to its previous value.
THE MAINTENANCE
LOG
The Maintenance Log is fully described in Chapter 5, Required
Maintenance.
THE CALIBRATION
MENU
The Calibration Menu (Figure 4.50) contains a Flow Calibration
"test". The flow Calibration can be run in one of three modes.
·Flow Calibration
Figure 4.50 The Calibration Menu
Flow Calibration
The Flow Calibration Menu performs some internal calculations
based on user-measured values. This is a passive test but it uses the
[RUN] key. Unless run incorrectly, this test will not interfere with
pump operation. Select /Flow Calibration/ to access the flow
calibration menus.
The test is run in one of three modes. Select either Meter, Fixed
Volume or Fixed Time from the Flow Calibration Menu (Figure 4.51).
Depending on your selection, the display will allow you to enter values
that enable the test to be completed. These are described in
Figure 4.51.
114
Thermo Scientific
·Fixed Time
·Meter
·Fixed Volume
Figure 4.51 Flow Calibration Menu
Table 4.5 Flow Calibration Modes
Calibration Mode
Value to Enter
Fixed Time
Measured Volume (mL)
Fixed Volume
Measured Time (min)
Meter
Measured Flow
Fixed Time: Measured Vol (mL)
This calibration mode assumes that you have collected and measured
a specific volume pumped during a fixed time period. Selecting
Fixed Time displays the Fixed Time Menu (Figure 4.52).
·Calculated Vol.
·10.0 mL
·Measured Vol.
9.5 mL
Figure 4.52 The Fixed Time Menu
(NOTE: The Measured Vol. value was artificially input to
demonstrate the menu function.)
1. Enter the theoretical volume for the fixed time period in the
Calculated Vol. field.
2. Enter the measured volume for the fixed time period in the
Measured Vol. field.
3. Press [RUN] to initiate the test. During the test, the pump
determines a new flow correction factor to compensate for the
inaccuracy in the flow rate. The Flow Correction Menu
displays the results of the test. Figure 4.53 shows example
results.
OLD
Flow Correction
100.00%
Use
NEW
101.00%
Figure 4.53 The Flow Correction Menu with example fixed time
calibration results
Thermo Scientific
115
OLD: The previous flow rate correction factor default or from
a previous flow calibration test.
NEW: The new flow rate correction factor based on the
Measured Vol. value.
Flow Correction: Use the [+]/[-] keys and press [ENTER] to
select one of three choices:
Use:
Use NEW flow correction factor. The factor remains in
effect until you turn off the pump or initiated the
NOVRAM.
Save: Save NEW flow correction factor to NOVRAM. This
factor remains in effect until you save a different value
over it, or until you reinitialize the NOVRAM
(TESTS/SERVICE/Test 271). Test 271 replaces this
value with the default flow correction factor.
Scrap: Discard the NEW (previous) flow calibration factor and
keep the current value.
Press [RUN] to complete the test.
Fixed Volume: Measured Time (min)
This calibration mode assumes that you have externally timed the
period in which a specific volume has been pumped.
Selecting Fixed Volume displays the Fixed Volume Menu
(Figure 4.54).
·Calculated Time.
·10.00 min
·Measured Time
9.50 min
Figure 4.54 The Fixed Volume Menu
(NOTE: The Measured Time value was artificially input to demonstrate the
menu function.)
1. Enter the theoretical time period for the fixed volume in the
Calculated time field.
2. Enter the measured time for the fixed volume in the Measured
Time field.
3. Press [RUN] to initiate the test. During the test the pump
determines a new flow correction factor to compensate for the
flow rate inaccuracy.
OLD
100.00%
Flow Correction
Use
NEW
101.00%
Figure 4.55 The Flow Correction Menu with example fixed volume
calibration results
116
Thermo Scientific
OLD: The previous flow rate correction factor default or from
a previous flow calibration test.
NEW: The new flow rate correction factor based on the
Measured Vol. value.
Flow Correction: Use the [+]/[-] keys and press [ENTER] to
select Use, Save, or Scrap as described above.
4. Use the [+]/[-] keys to choose Use, Save, or Scrap.
5. Press [RUN] to complete the test.
Meter: Measured Flow (mL/min)
This calibration mode assumes that you have externally measured the
precise flow rate that the pump is operating at while set at a specific
flow rate.
Selecting /Meter/ displays the Meter Menu (Figure 4.56).
·Selected Flow.
·1.00 Mn
·Measured Flow
1.00 Mn
Figure 4.56 The Meter Menu
(NOTE: The Measured Flow value was artificially input to demonstrate the
menu function.)
1. Enter the set flow in the Selected Flow field.
2. Enter the measured flow in the Measured Flow field.
3. Press [RUN] to initiate the test. During the test the pump
determines a new flow correction factor to compensate for the
inaccuracy in the flow. The Flow Correction Menu displays
the results of the test. Figure 4.57 shows example results.
OLD
Flow Correction
100.00%
Use
NEW
101.00%
Figure 4.57 The Flow Correction Menu with example fixed volume
calibration results
4. Use the [+]/[-] keys to choose Use, Save or Scrap as described
for the Fixed Time Menu.
5. Press [RUN] to complete the test.
To exit the flow calibration menus without entering any values, press
[∧] until you return to the Calibration Menu.
NOTE: Do not press [STOP] unless you have already entered a new
value in one of the flow calibration menus. If the pump is in RUN,
doing so will interfere with your analysis. Use [∧] to exit the flow
calibration menus instead.
Thermo Scientific
117
THE SERVICE
MENU
The Service Menu (Figure 4.58) contains several service-related tests,
including the Hardware Series test.
>Current History
·Lifetime History
------------------------------------------------------·ROM Test
(200)
·RAM Test
(201)
·Cycle Step Count
(205)
·External Inputs
(206)
·Display Test
(208)
·Transducer Range
(209)
·Motor Step/Valve
(211)
·Hardware Series
(220)
·Initialize NOVRAM
(271)
Figure 4.58 The Service Menu
The numbers in parentheses refer to a similar test found in earlier
SpectraSYSTEM pumps. They are included for the convenience of
users and service personnel familiar with this previously-used
numbering scheme.
Current History
By selecting /Current History/ you access a chronological list of
operating state changes. The negative number on the far left indicates
the time (in minutes) between the time the Current History Menu was
accessed and the state change occurred. More specific information
about reading the Current History Menu is found in the
SpectraSYSTEM Pumps Field Repair Manual.
To exit the Current History, press [ENTER].
Lifetime History
By selecting /Lifetime History/ you access a log of five measured
items relating to the entire time the pump has been in operation. An
example Lifetime History Menu is shown in Figure 4.59.
The top line shows 1) the total time that the pump's motor has been
running in hours (Hr) and 2) the total number of strokes in thousands
(kSt) taken by the cam; the bottom line shows 3) the number of times
the pump has been powered-on (on), 4) the number of times the pump
has been powered-down (off), and 5) the number of times an error
occurred when NOVRAM was written to upon power-down (bad).
118
Thermo Scientific
Press any one of the following keys to exit the Lifetime History
screen: [ENTER], [∧], [MENU], or [STATUS].
1.2 Hr
24on
2.3kSt
23off
0bad
Figure 4.59 The Lifetime History Menu
ROM Test
(Passive)
The ROM test (200) verifies the integrity of the ROM (Read Only
Memory) in your pump. The ROM is where all of the built-in
programs for the pump operation are stored. If faults are found in any
part of ROM, a message indicating that the test has failed will be
displayed. Press [STOP] to stop this test. Do not to press [STOP]
more than once or else pump operation will be interfered with.
If a failure is indicated, contact Thermo Fisher Scientific.
RAM Test
(Active)
The RAM test (201) verifies the integrity of the RAM (Random
Access Memory) in your pump. The RAM is where your pump files
are stored and where temporary calculations are performed. The
pump must be stopped (STOP) for this test to be implemented. If any
faults are found with RAM, the message shown in Figure 4.60 is
displayed.
RAM TEST failed
Figure 4.60 RAM Test failure message
Contact Thermo Fisher Scientific if this test indicates a failure.
Cycle Step
Count (Passive)
Normally, 12,800 motor drive pulses are required for one revolution
of the pump motor, as detected by the cam sensor. The Cycle Step
Count test (205) displays a count of the number of pulses required for
the last complete motor revolution.
The Cycle Step Count test is a dynamic measurement of the number
of steps counted, the lag amount (the number of steps the count has
shifted since the last revolution), and the number of seconds required
for the last revolution.
Thermo Scientific
119
Generally, the number of steps should be 12,800 ± 64. The lag value
varies due to the load on the pump. This number should be steady, or
fluctuate no more than ±120 steps.
Start the test by selecting /Cycle Step Count/. To stop the test, press
any one of the following keys: [ENTER], [∧], [MENU], or
[STATUS].
NOTE: The Cycle Step Count test requires that a full cam revolution has
occurred.
External Inputs
(Passive)
The External Inputs test (206) allows you to conveniently monitor the
status of two of the external input lines, STOP and RUN. The STOP
line causes the pump to stop pumping when momentarily grounded.
A momentary ground at the RUN input line causes the run time clock
to begin. Use this test if you are having difficulty interfacing your
pump to a controlling device, such as a SpectraSYSTEM
autosampler.
To run the test, select /External Inputs/. The display continuously
shows the current state of the STOP and RUN inputs (updates every
0.5 second). "Lo" means the input is grounded (active) and "Hi"
means the input is "high" (inactive).
To stop the test, press either [ENTER], [∧], [MENU], or [STATUS].
Display Test
(Passive)
The Display test (208) exercises the pump's display. When initiated,
the display shows staggered alphanumeric characters that scroll from
left to right. Pressing [STOP] freezes the display; pressing [RUN]
resumes movement.
This test is also a keyboard test. Pressing the cursor keys will cause
the alphanumeric display to scroll in that direction.
Other keys can be tested by first pressing [ENTER] to access the key
test. The display will verify other keys such as [STATUS] or
[MENU], as soon as each is pressed.
To return to the scrolling alphanumeric characters, press [RUN],
[RUN]. To stop the test, and return to the Service Test Menu, press
[STOP], [STOP].
Contact your representative if the display appears unusual.
120
Thermo Scientific
Transducer Range
(Active)
Transducer Range is not truly a test. It contains a field where you
must enter the calibration value (in mV) for a replaced pressure
transducer. The range is located on the replacement transducer wire.
Your pump features advanced circuit designs which allow the
pressure transducer range adjustment to be set by entering a value
from the keyboard. No adjustment of potentiometers is necessary.
Your pump comes from the factory preset to the proper range. The
value is stored in a NOVRAM. Do not change the transducer
calibration setting unless the pressure transducer or System PCB are
replaced. The calibration number is recorded on a tag attached to the
transducer cable. The System PCB and pressure transducer are not
user-serviceable parts. A qualified service representative must
perform any repair or replacement.
Motor Step/Valve
(Active)
The Motor Step/Valve test (211) exercises the pump motor and the
switching valve on the pump. When activated, the pump motor is
continuously stepped and each switching valve is sequentially opened
and closed at a rate of 1 valve per 0.512 seconds. This test is useful
for detecting an intermittently failing switching valve or pump motor.
Select /Motor Step Valve/ to initiate the test. Follow the instructions.
Each open valve is shown dynamically on the display. The test will
continue until one of the following keys is pressed: [ENTER], [∧],
[STOP], [MENU], or [STATUS].
Hardware Series
(Active)
The Hardware Series test (220) is an extensive evaluation of the
System Printed Circuit Board (PCB), switching valve, pump motor,
and pressure transducer. The System PCB contains all of the circuitry
for the operation of the pump, except for the display functions. Once
activated, the test exercises and diagnoses the condition of various
circuits. The pump must be idle (not pumping) before activating the
test. This test will not affect pump files.
NOTE: The external events connector (if present) must be removed from the
rear of the pump before initiating the Hardware Series test. Otherwise,
"Board Failure: Code 8" may occur.
Thermo Scientific
121
To activate the test, select /Hardware Series/. Follow the instructions
given on the display. Typically, the display will show:
Release system pressure
Then press ENTER.
Figure 4.61 Initial Hardware Series test message
Open the column bypass valve or otherwise remove column pressure
from the transducer since the pump will operate during the test and an
excessively high column pressure might be generated if not bypassed.
Press [ENTER] to continue the test.
Once the test is activated the display will appear as in Figure 4.62.
Test in Progress
Figure 4.62 Hardware Series test message, after pressing [ENTER]
The pump's components are tested in the following order.
1. Pressure transducer and circuitry
2. Input/output ports
3. Solvent switching valve and circuitry (P4000 only)
4. Motor drive circuitry
5. Cam marker and circuitry
6. Motor revolution and sine/cosine circuitry
As each portion of the test is completed a message is displayed. If all
components and circuitry are within specifications, the messages
shown in Figure 4.63 are displayed during the test.
If a failure is detected during the test, the failure message is
displayed. It remains displayed until [ENTER] is pressed (the test
resumes).
To stop the test, press [STOP], or press [∧] to return to the Service
Menu.
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Thermo Scientific
TRANSDUCER TEST PASSED
------------------------------------------------------PORT TEST PASSED
------------------------------------------------------VALVE TEST PASSED ** (P4000 only)
------------------------------------------------------MOTOR DRIVE TEST PASSED
------------------------------------------------------CAM MARKER TEST PASSED
------------------------------------------------------REVOLUTION TEST PASSED
------------------------------------------------------Hardware series tests completed - PASSED
Figure 4.63 No problems found during the Hardware Series test.
If problems are found during the test, a message suggesting the most
likely failure is displayed, although in some cases other failures are
possible. The messages shown in Table 4.6 indicate a possible
System PCB failure.
Thermo Scientific
123
Table 4.6 Errors Indicating A Possible System PCBA Failure
BOARD FAILURE: CODE XXX
If the failure is identified as a component on the system printed circuit
board or if the test cannot determine the failed component, a message is
reported where XXX is a 1-, 2- or 3- digit number.
8
Remove the external events connector from the rear of the pump. This
test exercises the input lines and may be affected by attached cabling.
1-100
Pertains to failures of the system printed circuit board. Contact your
service representative.
101, 102, 103
Indicates the proportioning/switching valve's electrical system (P4000)
has failed. Refer to Chapter 5, under Solvent Proportioning/Switching
Valve Replacement.
104, 105
Possible broken or loose pump motor cable wire.
124
Too many motor steps were needed to complete a cam revolution. You
may have a loose motor coupler or faulty System PCB. Contact your local
sales/service representative for service information.
125
Too few motor steps were needed to complete a cam revolution. You may
have a faulty cam sensor or System PCB. Contact your local sales/service
representative for service information.
TRANSDUCER UNPLUGGED
The pressure transducer was not detected. The connector going to the
pressure transducer should be checked. Access to this connector requires
the removal of the pump's outer protective cover. Because there are safety
issues involved in its removal, this should only be performed by a
qualified service technician. The location of this connector as well as the
proper procedure for removing the outer cover are outlined in the Field
Repair Manual.
CANNOT ZERO TRANSDUCER
The transducer circuitry is not able to compensate for the zero offset of
the transducer. Make sure that the system is at zero column pressure
(column bypassed) before starting the test. If so, replace the transducer.
Contact your local sales/service representative for assistance.
CHECK FUSE F1
(P4000) Fuse F1 is used to protect the valve drive circuitry from internally
shorted valves which may short internal circuitry on the system PCB.
Checking the status of F1 requires the removal of the outer cover of the
pump. Because of safety issues involved in the removal of this cover a
trained service technician should perform this evaluation. Contact your
local sales/service representative for assistance.
CAM MARKER NOT FOUND
This message indicates that the sensor that detects cam revolutions is not
operational. Either the motor coupling is loose or the cam sensor is
defective. This requires tightening of the motor-to cam coupler if loose,
or replacement of the cam sensor. Both of these actions require the
removal of the top cover. Because of safety issues involved in the
removal of this cover a trained service technician should perform this
evaluation.
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Thermo Scientific
Table 4.7 Errors Indicating A Possible System PCBA Failure, continued
VALVE CABLE UNPLUGGED
(P4000) The test has detected that the solvent proportioning/switching
valve cable is disconnected from the System PCB. There are two
locations where the valve is connected. The first location is underneath
the proportioning valve assembly. To access this location the
proportioning valve must first be removed. This is achieved by
unscrewing the finger-tight captive fasteners which hold the assembly in
place. Once this is accomplished the cable connector is visible. Press
together the connector attached to the cable coming from the
proportioning valve assembly to the mating connector located nearby in
the chassis of the pump. If this fails to remedy the problem then the
second cable connection location should be checked. Correction of the
problem at this second location requires the removal of the top cover.
Because of safety issues involved in the removal of this cover a trained
service technician should perform corrective action.
CAM SENSOR FAILURE
The cam sensor cable is disconnected or defective and needs to be
replaced. Correction of this problem requires the removal of the top
cover. Because of safety issues involved in the removal of this cover a
trained service technician should perform corrective action. Contact your
local sales/service representative for service information.
NO CURRENT TO MOTOR
The test has detected no current flow through the pump motor. Either the
motor cable is unplugged on the System PCB or the entire drive circuitry
is defective. Correction of this problem requires the removal of the top
cover. Because of safety issues involved in the removal of this cover a
trained service technician should perform corrective action. Contact your
local sales/service representative for information on obtaining a
replacement.
Initialize NOVRAM
(Active)
The pump must be stopped for /Initialize NOVRAM/ to be
completed.
NOTE: Initialize NOVRAM (271) has a profound effect upon the
pump's non-volatile RAM. Do not initialize the NOVRAM unless you
fully understand all consequences associated with this action.
By initializing the NOVRAM all files are reset to their default values,
with the exception of the run file. All user-preferences set in
/OPTIONS/ are also returned to their default values. In addition, any
changes that had been made to the Liquid End Type and Flow
Calibration parameters are erased, and these selections are returned to
their default values. In general, any field value or selection that is
normally retained when the pump is turned off and then on will be
reset to its default value when the NOVRAM is initialized.
NOTE: The pressure transducer's range value is not erased when the
NOVRAM is initialized.
HINT: To retain a single file while initializing the NOVRAM: load it (so
that it becomes the run file), stop the pump, initialize NOVRAM, make a
change to a value within the run file from Status, then select /Save File/ at
the bottom of the Status Menu.
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5
Required Maintenance
Introduction
When properly maintained, your Thermo Scientific SpectraSYSTEM
pump will provide years of trouble-free operation. It is important that
your pump receive routine preventive maintenance to ensure
reliability and optimum performance. Properly performed routine,
preventive maintenance also helps keep your warranty valid. Your
pump is designed to encourage proper maintenance by making
maintenance parts easy to access, replace and record.
This chapter describes the Maintenance Log Menu and how to use it.
Some hints to help you extend the maintenance period of your pump
are also included. Easy-to-follow, step-by-step required maintenance
procedures are also contained in this chapter so that you can keep
your pump in optimum working condition. A few maintenance tips
for parts of the pump not directly involved with solvent flow follow
the maintenance procedures. The last section contains replacement
procedures for two user-serviceable parts: fuses and the solvent
switching valve.
NOTE: Maintenance of the pump is the responsibility of the user. Routine
maintenance is not provided under warranty. However, planned
maintenance contracts are generally available. Please contact your local
representative if you are interested in purchasing a planned maintenance
contract.
THE BENEFITS OF
PROPER
MAINTENANCE
As with most things, there is tremendous benefit in doing things right
the first time. For example, an unusually fast seal failure may
indicate either incorrect installation or a scratched piston. A
scratched piston may be caused by improper installation of the seal or
piston, by allowing the pump to sit idle with a buffered eluant in it, or
by failing to filter your eluants.
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127
Maintenance Schedule
Table 5.1 Gradient Pump Preventative Maintenance Schedule
Frequency
Procedure
Performed By
Daily
Check waste reservoir.
Empty as required.
User
Check solvent reservoir.
Replenish as required.
User
Replace piston seals.
User
Replace backflush seal.
User
Annually
Check pistons and Kel-F seals.
Sonicate any parts as required.
Maintenance Log
The Maintenance Log provides a convenient way for you to record
maintenance performed on the liquid ends and set intervals for
periodic maintenance. When a maintenance interval has been
exceeded the pump will automatically display a message indicating
that maintenance may need to be performed.
MAINTENANCE
LOG MENU
The Maintenance Log is accessed by selecting /TESTS/ from the
Main Menu, then selecting /Maintenance Log/ (Figure 5.1 and
Figure 5.2).
·FILES
·QUEUE
·COMMANDS
>TESTS
·OPTIONS
Figure 5.1 Main Menu with TESTS selected
·Software Version
·Diagnostics
--------------------------------------------------->Maintenance Log
·Calibration
·Service
Figure 5.2 Tests Menu with Maintenance Log selected
The Maintenance Log Menu (Figure 5.3) consists of a table, used to
record dates and volumes, followed by one field used to enter a value
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Thermo Scientific
relating to flow, and two additional menu items. All of these are
described in this section.
ITEM
Seal1
DATE
DUE
VOL
8 AUG95
200
201
-------------------------------------------------------Seal2
8 AUG95
200
201
Piston1
18AUG95
600
400
Piston2
18AUG95
600
400
Inlet
8 AUG95
600
201
X-ducer
8 AUG95
600
201
·Maintenance Position
·Liquid End Type
Figure 5.3 Maintenance Log Menu
The Maintenance
Table
The top half of the Maintenance Log Menu is a table (Figure 5.3).
ITEM, DATE, DUE, and VOL
The ITEM field remains fixed. "Seal 1" and "Seal 2" should be
paired with "Piston 1" and "Piston 2", respectively, to identify the
inlet and outlet liquid ends. "Inlet" refers to the inlet check valve,
while "X-ducer" refers to the transducer check valve.
Enter the date (day/month/year) in the DATE field for the last time
maintenance was performed on each item.
The pump keeps "liters pumped" counters, in the VOL (volume) field,
for each major maintenance item (the pump seals, pistons, and check
valves). You may set a DUE volume in liters for each item. When
the DUE volume is exceeded by the volume of liters pumped (VOL),
the reminder "MAINTENANCE DUE - SEE PUMP LOG" is
displayed. This message will appear each time a file is initialized.
You may choose to use this feature to set regular intervals for
maintenance, such as seal changes, pump/column cleaning, or simply
to serve as a reminder to verify that the system is operating properly.
The interval remains set until either the date has been updated or the
DUE value has been increased.
In the example shown in Figure 5.3, a fairly complete maintenance
was done on 8 Aug 95, when both seals and check valves were
replaced.
Setting Intervals
The volume of mobile phase that you can expect to pump before the
pump requires maintenance is very dependent upon the eluant being
pumped and your adherence to good chromatographic practices. To
obtain the maximum lifetime and best performance from your pump,
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129
read Extending the Maintenance Period on page 132. Pump pistons
and check valves have been known to last for years. Even the seals
themselves can last more than a year for some applications.
An initial guideline for setting up your Maintenance Log for the first
time is to set both seal DUE counters to 200 liters, and the check
valve and piston DUE counters to 600 liters. Setting a value of zero
(0) for any DUE interval inactivates the Maintenance Log for that
specific item. To inactivate the entire Log, a zero (0) must be entered
for all DUE intervals. Your specific maintenance interval can be
determined by observing pump performance over time.
When a Maintenance Message is Displayed
Whenever the interval has been exceeded, and the message
"MAINTENANCE DUE - SEE PUMP LOG" is displayed, you
should either verify that the pump needs maintenance or that the
pump is operating properly. If a maintenance interval is exceeded
and you find that the pump does not require maintenance, increase the
DUE interval by another 50 liters from the previous setting. Once you
have established an expected interval for your system, use that
interval for routine preventive care.
If you find that the interval before component failure is either
unacceptable or variable, then the source of the problem must be
identified. Read this chapter and Appendix A. Poor chromatographic
practices are by far the most common source of problems. Specific
procedures for inspecting and changing parts begin on page 133.
Maintenance
Position
Selecting /Maintenance Position/ prepares the pump for liquid end
removal or replacement. The display shown in Figure 5.4 appears:
To install or remove
liquid ends press ENTER
Figure 5.4 Maintenance position message
The maintenance position puts the pump's cam into a position to
facilitate liquid end removal.
Liquid End Type
The Liquid End Type Menu allows you to select the proper liquid end
for a specific application. Do not change the flow range unless you
are installing liquid ends with capacities different from those
purchased with the pump.
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Thermo Scientific
Liquid End Type
Flow Range
Normal
0 - 10 mL/Min
Figure 5.5 Liquid End Type Menu
Selecting Normal, Bio, or SemiPrp
If you select Normal, Bio, or SemiPrp (semi prep) in the Liquid End
Type field, the Flow Range field changes automatically to
corresponds to the preset ranges for these liquid ends. (Normal = 0 10 mL/min, SemiPrp = 0 - 30 mL/min,
Bio = 0 - 10 mL/min, and Other.
Selecting Other
If you select Other in the Liquid End Type field, the Flow Range field
becomes active, allowing you to enter your own flow range.
After changing the liquid end type on the display, press [ENTER]. A
message (Figure 5.6) will prompt you to write down the old values in
the Maintenance Log which correspond to the liquid ends you
presumably just removed. It is important to keep records for each set
of liquid ends you use. If you reinstall the "old" liquid ends, you will
need to reenter the dates and statistics for the "old" ends into the
Maintenance Log table. You are also reminded to enter new values
into the log's VOL field, which correspond to the newly installed
liquid ends. Normally the VOL field is not edited, but when new
liquid ends are installed, ensure that the VOL fields for the
appropriate maintenance items are reset to 0.
Write down old values
ENTER new values in log
Figure 5.6 Reminder to keep proper records when liquid ends are
changed
Flow Correction
An additional menu appears whenever you press an arrow key,
[ENTER], or [+]/ [-] from the display shown in Figure 5.7. This
menu allows you to set a Flow Correction, if desired. OLD and NEW
values are displayed. For no Flow Correction, enter 100.00%.
NOTE: The Flow Correction menu is the same as the menu displayed when
a flow calibration (/TESTS/, /Calibration/, /Flow Calibration/) has been
initiated, except that it does not include the Use, Save, or Scrap option.
OLD
Flow Correction
100.00%
NEW
100.00%
Figure 5.7 Flow Correction Menu
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131
Flow correction is a value, in percent, which adjusts the actual
volume that the pump delivers. As you use the pump, you may feel
that although the pump is set at a specific flow rate, for example
2 mL/min, the pump actually delivers slightly more or less than this
volume per minute. This can be due to a variety of maintenance- or
LC-related reasons (seals, valves, and so on).
If desired, manually enter a flow correction value. This value can be
entered automatically, based on the result of the flow calibration test,
initiated from /TESTS/, /Calibration/, /Flow Calibration/. This test
[which requires you to enter an accurately measured operation value
(time, volume or flow rate)] is fully explained in Chapter 4.
If no correction to the flow is desired, enter a value of 100% in this
field. Values from 90% to 110% are valid. For example, if you pump
for one minute at 1 mL/min and collect 0.95 mL, then the pump is
actually delivering 5% less solvent than expected. To compensate,
enter 105.26% in the Flow Correction field. The pump's Status
Screen will still display a flow rate of 1.0 mL/min, but the pump will
actually deliver 105.26% of what it normally delivers at 1.0 mL/min.
Press [ENTER] to save the value, or simply leave the menu by
pressing an arrow key.
Extending the Maintenance Period
As mentioned earlier, the volume of mobile phase you can expect to
pump before maintenance is due is very much dependent on the way
that the pump is being used. Following these guidelines helps you
extend the life and improve the performance of your pump.
132
•
Use high quality, spectro-grade or HPLC-grade solvents.
These solvents do not usually need to be filtered before use.
•
Filter water and prepared solvents through at least a 0.45micron filter before placing them in the solvent reservoirs to
remove particulate matter and organic contamination.
•
Avoid pH extremes.
•
Verify that the solvents used are miscible in all proportions.
This is very important for a buffered mobile phase.
Precipitation of salts quickly damages maintenance parts.
•
Never leave the pump filled with buffered solvent when not
pumping. Either lower the flow to 0.1 mL/min. or thoroughly
flush the pump. Flush with at least 25 mL of pure filtered
water.
•
The pump should be filled with methanol if it is to be left idle
for more than two days. This avoids the possible growth of
organisms in aqueous solvent systems.
•
Never use hydrochloric acids solutions.
Thermo Scientific
•
Avoid metal ions that can cause corrosion due to
electrochemical processes. Typical metal ions to avoid:
manganese, chromium, nickel, copper, iron, molybdenum.
Maintenance Procedures
SAFETY
PRECAUTIONS
Observe the following safety precautions whenever performing
periodic maintenance.
Caution!
A caution alerts you to situations that could result in personal injury.
It also tells you how to avoid them.
High Voltage!
This icon alerts you to the presence of high voltage and to the
potential injury that could occur from electrical shock were you to
come in contact with a specific instrument area or component. It also
tells you how to avoid contact with the high-voltage areas in your
instrument.
Hot Surface!
This icon alerts you to potential injury that could occur from coming
in contact with a heated surface or area on or in an instrument. It also
tells you how to avoid contact with the heated surfaces in your
instrument.
This section includes procedures for:
•
Complete liquid end maintenance (includes disassembly and
assembly)
•
Check valve replacement
•
Passivating stainless steel parts
Also included are maintenance tips for pump parts that are not
involved with pump flow.
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133
TOOLS
The following tools are useful to have on-hand as you perform
maintenance procedures.
•
Tweezers
•
Open-end wrenches (1/4-inch, 5/16-inch, 1/2-inch)
•
Loupe or magnifying glass
•
Allen wrench (Hex head) 9/64-inch
PREPARATION
Prepare the pump for maintenance before performing any
maintenance procedure.
To prepare the pump for maintenance, flush the pump with 25 mL of
methanol. If an incompatible solvent is resident in the pump, flush
with appropriate intermediate solvents before flushing with methanol.
For example, if chloroform is being used as the mobile phase solvent,
an intermediate flush of 25 mL methylene chloride would be
appropriate before flushing with methanol.
LIQUID END
MAINTENANCE
Complete liquid end maintenance includes procedures for seal and
piston maintenance:
•
Removal
•
Disassembly
— inspection for contamination
— cleaning
— piston inspection
— cleaning/replacing parts if necessary
•
Assembly
•
Installation
For thorough cleaning, piston replacement, or total liquid end
reconditioning, the liquid ends must be removed.
Having a second set of reconditioned liquid ends on hand for quick
replacement will save additional time and allow maintenance to be
performed at your convenience. Contact your local Thermo Fisher
Scientific representative if you are interested in obtaining spare
components. Part numbers are included in Appendix C.
NOTE: Keep the liquid end components as clean as possible.
Contamination decreases seal life significantly.
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Thermo Scientific
Preparation
1. Flush your pump with 25 mL prior to disassembling your
liquid ends. If methanol is not compatible with the mobile
phase in your pump, flush the system with 25 mL of an
intermediate solvent before flushing with methanol.
2. Remove the front cover, exposing the liquid ends (Figure 5.8).
RUN
STATUS
MENU
STOP
ENTER
PURGE
SpectraSYSTEM
P4000
Crossover Tube
Pressure
Transducer
Transducer
Tube
Bypass Valve
Knob
Outlet
Liquid
End
Bypass
Valve
Transducer
Check
Valve
Bypass Valve Tube
Inlet
Liquid
End
Solvent
Inlets
Outlet Port
(to autosampler or
manual injection valve)
Inlet Check Valve
Solvent Inlet Tube
Proportioning
Valve
Drip Tray
Lift Point
PU-Z002E/FM
Waste Tube
Figure 5.8 Pump with front cover removed
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135
Liquid End Removal
To remove the liquid ends from the pump:
PU-Z016E/DT
1. Remove all tubing attached to the pump heads. Turn the nuts
counter-clockwise to remove (Figure 5.9).
Crossover Tube
Outlet
Liquid
End
Pressure
Transducer
Bracket
BypassValve
Tube
Pressure
Transducer
Inlet
Liquid
End
Transducer
Tube
Solvent Inlet
Tube
Inlet
Check
Valve
Figure 5.9 Liquid ends and tubing
2. Position the pump cam to enable the liquid ends to be removed.
To do this, press [MENU], and select /TESTS/, /Maintenance
Log/. Then move the cursor to /Maintenance Position/ and
press [ENTER]. The display shown in Figure 5.10 appears.
To install or remove
liquid ends, press ENTER
Figure 5.10 Maintenance position message
Press [ENTER] to continue. The pump motor rotates for a few
seconds and then is electrically locked into position. While the
motor rotates the display appears as in Figure 5.11. When the
cam is in the maintenance position, the display in Figure 5.12
appears.
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Thermo Scientific
Install or remove liquid
ends when motor stops
Figure 5.11 The display when the motor moves cam to maintenance
position
X-ducer
(date)
(due)
(vol)
· Maintenance Position
Figure 5.12 The display after the motor moves the
cam to the maintenance position
The pump is now in its maintenance position. It will hold this
position (if power is maintained) until a file is initialized or a
purge is started.
3. Remove the inlet check valve (Figure 5.13) from the inlet
pump head.
NOTE: It is not necessary to remove the check valve to replace a piston,
however, it is easier to remove at this time if total liquid end reconditioning
is to be performed.
PU-Z009E/DT
Inlet
check
valve
Figure 5.13 Inlet check valve
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137
4. Push in the outlet (upper) liquid end and rotate it (90 degrees
counter clockwise) until it releases from the pump module.
Remove the liquid end and set it aside.
5. Push in the inlet (lower) liquid end and rotate it (90 degrees
clockwise) until it releases from the pump module. Remove it
and set it aside.
Liquid End
Disassembly
To disassemble the liquid ends:
1. Separate the pump head from the piston holder housing by
removing the two 9/64-inch hex cap screws. Turn the screws
counter-clockwise.
2. Examine the Kel-F® seal in the cylinder bore. If the seal is
damaged (scratched, warped or torn) it must be removed. Use
tweezers to remove it by pulling gently on the seal's inner
circumference. (Be careful not to scratch the cylinder surface!)
3. Examine the pump head for contamination. Flush the pump
head with methanol or place it into an ultrasonic bath.
PU-Z050E/DT
4. Remove the seal holder from the piston holder housing by
grasping both ends of the exposed tube, and pulling gently
(Figure 5.14).
Seal Holder
Piston Seal
(Spring Side Up)
Pump Head
Cylinder
Figure 5.14 Removing the seal holder from the piston holder housing
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Thermo Scientific
5. Carefully remove the piston seals from the seal holders using
the seal removal tool supplied in the accessory kit. Insert the
tool and wiggle it in a circular manner to remove the seal. (See
Figure 5.15). Flush the holders with methanol if contamination
is present.
Seal Removal
Tool
Piston Seal
(Spring Side Up)
PU-Z025E/DT
Seal Holder
Figure 5.15 Using the seal removal tool
6. While retaining the piston holder, remove the 9/64-inch
retaining cap screw (Figure 5.15). This allows the piston
holder to be removed from the piston holder housing. Separate
the holder, piston, spring and housing.
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139
CAUTION! Wear safety glasses! The piston components are spring
loaded and may shoot out! (Figure 5.16).
PU-Z130E/ST
Gently press the piston holder while
you remove the retaining cap screw.
Then slowly let the piston holder out
of the piston holder housing.
Figure 5.16 Retaining the piston holder
7. Examine all parts for wear, corrosion or contamination. Clean
all deposits. Look for wear marks on the stainless steel shaft of
the piston and corresponding wear marks on the inner aspects
of the spring which indicate bending or bowing. If these wear
marks are present, replace the spring (Figure 5.17).
NOTE: It is normal for the piston holder to produce a small amount of
wear particles.
8. Examine the piston carefully under a low-power microscope or
magnifying glass for fine scratches, ridges, or scoring which
can reduce seal life (Figure 5.18). Some apparent scratches are
actually deposits that can be cleaned. The piston can be
cleaned by wiping it gently with a laboratory towlette or cotton
swab that has been immersed in methanol. Replace the piston
if scratched or pitted. The new piston should also be cleaned
prior to installation.
9. Thoroughly flush all components with methanol.
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Thermo Scientific
Bushing
Backup Seal
Bushing
Cap Screws (2x)
Inlet
Check Valve
Pump Head
Kel-F Seal
Piston Seal
Standard Version
Seal Holder
Piston Flush Seal
Retaining
Cap Screw
Lock
Washer
Piston Holder
Housing
Piston
Spring
PU-Z164E/ST
Piston Holder
Figure 5.17 Liquid end components
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141
PU-Z024E/DT
Look for Scratches
in this Portion
of the Piston
Figure 5.18 Piston scratches (under magnification)
Liquid End
Assembly
To replace the piston seal and reassemble the liquid end:
1. Place the seal holder on end on a clean, flat surface. Place the
seal holder into the pump head with the seal down (spring
should face the inside of the pump head.) Install new piston
seals by setting them in position on the seal holder (spring side
up) and gently pressing them into place with the pump head
(Figure 5.19).
NOTE: It is possible to install the seal in the wrong end of the seal holder.
If installed in the wrong end, the seal will not be flush with the top of the
holder. The opposite end of the seal holder is deeper, to accommodate the
piston flush seal. Install the piston seal only in the end closest to the tubes.
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Thermo Scientific
Pump Head
Cylinder
Seal Holder
PU-Z021E/DT
Piston Seal
(Spring Side Up)
Piston Flush Seal
Figure 5.19 Seal installation
2. A piston flush seal (part of a Piston Flush Seal Kit and
normally used with buffers) may be located at the opposite end
of the seal holder. If you use a piston flush seal it should be
replaced once a year. The piston flush seal is not subject to the
higher pressures seen by the piston seal, so maintenance of this
part is only occasionally necessary.
NOTE: The piston guide bushings do not need replacement. Retain
them for new seal replacement.
To replace the piston flush seal:
a. Use the seal removal tool to remove the piston flush seal.
b. Insert a new piston flush seal into the seal holder (spring
side down). The piston flush seal is thicker than a piston
seal. The seal holder's cavity on the piston flush seal side
is deeper to accommodate the larger size.
c. Ensure that the piston flush seal is flush with the edge of
the seal holder. Use the large end of the seal removal tool
to push the seal into the holder.
3. Place the seal holder into the piston holder housing spring side
up (Figure 5.20).
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143
Inlet Check
Valve
Cap
Screws
Figure 5.20 Seal holder alignment
4. If the Kel-F seal is being replaced, put the new seal in the
pump head cavity now.
5. Install the pump head onto the housing using the two 9/64-inch
Allen head screws.
a) For the inlet liquid end, the pump head must be connected
to the piston holder housing as shown in Figure 5.21 (Inlet
check valve down, retaining cap screw to the left).
b) For the outlet liquid end, the pump head must be oriented
as shown in Figure 5.22.
Evenly tighten the screws to forty inch-pounds (tight).
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Thermo Scientific
PU-Z020E/DT
Piston Holder
Housing
PU-Z029E/DT
Piston Holder
Piston Holder
Housing
Pump
Head
Retaining
Cap Screw
Cap Screws
Inlet Check
Valve
PU-Z049E/ST
Figure 5.21 Installing the retaining cap screw
(inlet liquid end)
Piston Holder
Piston Holder
Housing
Pump Head
Retaining
Cap Screw
Cap Screws
Bypass Valve
Tube Connection
(Hidden from View))
Figure 5.22 Installing the retaining cap screw
(outlet liquid end)
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145
6. Install the piston into the piston spring and then place them
both into the piston holder housing (Figure 5.23). Do not press
the piston through the seal at this time.
7. Compress the piston holder into the holder housing and install
the retaining cap screw with its washer as shown in Figure 5.21
and Figure 5.22, so that the screw enters the slot in the piston
holder. This action pushes the piston through the seal. Tighten
the screw until snug.
NOTE: Make sure the retaining cap screw is oriented on the left-hand side
of the piston holder housing for both the inlet and outlet liquid ends.
Piston Holder
Housing
E-Ring
Piston
Figure 5.23 Piston installation
Liquid End
Installation
To install the liquid end assemblies into the pump, the pump must be
in the maintenance position. If the pump has not been turned off
since the liquid ends were removed, the pump motor should still be in
its maintenance position. If not, press [MENU], /TESTS/,
/Maintenance Log/ then move the cursor to /Maintenance Position/
and press [ENTER]. The display shows:
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Thermo Scientific
PU-Z022E/S
Piston Spring
To install or remove
liquid ends, press ENTER
Figure 5.24 Maintenance position message
Press [ENTER] to continue. The pump motor rotates for a few
seconds and then is electrically locked into position. While the motor
rotates the display shows:
Install or remove liquid
ends when motor stops
Figure 5.25 Pump rotating cam to maintenance position
The pump is now in its maintenance position. The display will appear
as shown in Figure 5.26. The liquid ends can be installed.
X-ducer
(date)
(due)
(vol)
· Maintenance Position
Figure 5.26 The display after the motor moves
cam to the maintenance position
1. Replace the inlet liquid end first. This liquid end contains
tapped holes for the inlet check valve and the transducer tube.
Install it by pressing in and turning it approximately 90 degrees
counter-clockwise, until it locks into position. Be sure that the
check valve (or the check valve hole, if the valve is not
installed) is pointed down, and the transducer tube hole is
pointed to the upper left-hand side (10 o'clock) (Figure 5.27).
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147
PU-Z028E/DG
Inlet
Liquid
End
D
B
A
C
Figure 5.27 Inlet liquid end
2. Replace the outlet (upper) liquid end. This liquid end contains
tapped holes for the crossover tube and the bypass valve tube.
This liquid end is installed by pressing in and turning it
approximately 90 degrees clockwise, until it locks into
position. Be sure that the crossover tube hole is pointed up,
and the bypass valve tube is pointed to the lower right-hand
side (4 o'clock).
3. Replace the check valve and tubing. (Do not over-tighten
fittings.) Generally, a 1/16-turn beyond finger-tight is
sufficient to make a leak-free connection.
HINT: If the transducer check valve has been removed you can
distinguish the two check valves: the inlet check valve has a wider
fitting opening than the transducer check valve (connected to the
pressure transducer).
4. After replacing the liquid ends, reset the VOL values in the
Maintenance Log for all replaced components to zero (0).
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POST-INSTALLATION
SEAL CONDITIONING
Follow the procedure below each time you replace liquid end seals.
1. Open the column bypass valve and purge with methanol to
clear air from the solvent lines and liquid ends.
2. Purge the pump through an old column or flow restrictor at
4000 psi for a minimum of 20 minutes. Reduce the flow to
1 mL/min and continue pumping for 15 minutes.
3. Check for solvent leaks. Do not return the pump to every-day
service unless you are sure that no leaks are present.
CHECK VALVE
MAINTENANCE
If the pump has notified you that it is time to replace check valves or
if check valve replacement was recommended in Appendix A,
Troubleshooting, then follow these steps. Check valve maintenance
consists of:
•
•
Inlet check valve removal and installation
Transducer check valve removal and installation
NOTE: The factory-supplied replacement check valves are
manufactured in a clean-room environment and capped to protect
them from contamination. It is very important to maintain a clean
environment when installing them.
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149
Inlet Check Valve
(bottom position)
To remove the existing inlet check valve and install a new one:
1. Remove the solvent inlet tube (Figure 5.27) from the check
valve.
2. Remove the defective check valve by rotating the valve
counter-clockwise with a 1/2-inch open-end wrench.
3. Install the new check valve by rotating clockwise until the
valve is snug against the liquid end cylinder. Reconnect the
inlet tubing. Tighten to finger-tight.
Transducer
Check Valve
To remove the existing transducer check valve (connected to the
Pressure Transducer) and install a new one:
PU-Z015E/DG
1. Remove the transducer tubing from the transducer check valve
and the inlet pump head (Figure 5.28).
Pressure
Transducer
Transducer
Tube
Transducer
Check Valve
Figure 5.28 Transducer check valve
2. Remove the defective check valve by rotating it counter
clockwise with a 1/2-inch open-end wrench.
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Thermo Scientific
3. Install the new check valve by rotating it clockwise until snug,
and tighten with a 1/2-inch open-end wrench. Replace the
connecting tubing. Tighten fittings only enough to stop leaks.
Generally, this is 1/16-turn beyond finger-tight.
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Maintenance Tips
This section contains useful maintenance tips for pump parts not
directly related to solvent flow.
DRIP TRAY
A removable, white plastic solvent drip tray is located underneath the
inlet bracket of your pump.
To remove the tray squeeze the top, front-edge of both sides of the
tray together and carefully pull the tray out. You may need to wiggle
the tray as you pull. If you see solvents in the tray, be particularly
careful not to spill them as the tray is removed.
PU-Z017E/DG
HINT: It may be easiest to use the index finger of each hand to push the
sides together.
Drip Tray
Solvent Value
Mounting Plate
Figure 5.29 Drip tray installed
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PASSIVATION OF
STAINLESS STEEL
COMPONENTS
All the major type 316 stainless steel components used in the
SpectraSYSTEM pumps are passivated prior to assembly to ensure
the removal of porous particles from the surface and to coat the
surface with a layer of chromium oxide, which is highly resistant to
corrosion. All stainless steel replacement parts purchased from
Thermo Fisher Scientific are also passivated.
However, stainless steel components are subject to corrosion from
strong acid solutions (in particular, materials containing halides),
organic acids, and sometimes even water. Resistance to corrosion of
the stainless steel components can be enhanced by using the
following procedures.
CAUTION—Chemical Hazard! Take care when passivating with
strong acids. Wear protective eye covering and protective clothing.
NOTE: Before installing any new parts not supplied from the Factory such
as stainless steel tubing, the parts should first be passivated using the
methods below.
NOTE: DO NOT expose a column to the passivation mixture.
Remove the column before pumping if it is necessary to pump
passivation solvents through the pump. It is preferable, however, to
remove the components from the pump and then passivate them apart
from the system.
1. When the surface area to be passivated is thoroughly clean, it is
passivated by wetting the surface with a 20% nitric acid
solution in deionized water for about 10 minutes at room
temperature.
2. After passivation, thoroughly clean the parts to remove any
residual nitric acid. Wash with deionized water until the
system is neutral to pH paper. Follow up with another wash
using 50-50 water/methanol followed by methanol. When
thoroughly clean, blow dry using nitrogen. (Do not use the
laboratory air system or air from a compressor that may contain
an oily residue.)
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Repair Instructions
If troubleshooting has pointed to a blown fuse in the power entry
module, or to the need to replace the solvent switching valve (P4000),
use the procedures below to make repairs.
POWER ENTRY MODULE
FUSE REPLACEMENT
Instrument power is supplied by two 4.0-amp fuses housed in the fuse
compartment of the power entry module, above the power cord
receptacle.
To replace the fuses:
1. Ensure that the power cord is not connected to the pump.
2. Use a small, flat blade screwdriver to pry open the power
selector/fuse cover. You will probably hear the top edge of the
cover snap as it is pried open.
3. Pull out the fuse holder and discard the bad fuse. Place the
new fuse into the holder with the metal end visible.
4. Snap the fuse holder back into place.
NOTE: If the power selector barrel accidentally comes out, be sure
to replace it so that the correct voltage for your area shows through
the voltage window.
5. Firmly snap the housing cover back in place. Be sure that the
correct voltage is visible in the voltage window.
HINT: Use two thumbs to push up on the top half of the cover as you
push in.
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SOLVENT SWITCHING
VALVE REPLACEMENT
(P4000 ONLY)
To remove the solvent switching valve:
1. Turn off power.
2. Remove front cover.
3. Remove the solvent inlet tube fitting from the inlet check
valve.
4. Loosen the two knurled screws on the front-left and front-right
sides of the inlet bracket.
5. Lift and pull the bracket forward 3 - 4 inches.
6. Disconnect the inlet tubes from the solvent switching valves at
locations A and B.
7. Remove the solvent inlet line from the center port of the valve.
8. Disconnect the solvent switching valve cable at the rear of the
solvent valve cavity, and remove the solvent valve/bracket
fully from the pump.
To install a new solvent switching valve:
1. Connect the replacement valve's cable to the connector at the
rear of the solvent valve cavity.
2. Connect the solvent inlet line to the center port of the
replacement valve. Tighten snugly but only finger-tight.
3. Re-connect the inlet tubes to the valve ports, making sure the
identification of each inlet tube matches the identification of
each valve port location.
4. Install the valve and bracket into the solvent valve cavity.
Tighten the knurled screws.
5. Attach the solvent inlet line to the inlet check valve. Tighten
snugly but only finger-tight.
6. Prime the pump using the technique described in Chapter 1.
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A
Troubleshooting
Introduction
Your SpectraSYSTEM pump is designed to operate trouble-free for
many years when properly maintained. Most pump problems can be
avoided by simple, periodic maintenance, as described in Chapter 5.
However, in the event that an error message is displayed or if a
mechanical or electrical failure is suspected, the problem can be
easily diagnosed. Further, if the diagnosis indicates that a problem
exists with non user-serviceable parts inside the pump, a qualified
Thermo Fisher Scientific service representative can quickly and easily
replace most malfunctioning parts.
This appendix contains information on:
•
Theory of operation
•
General LC system troubleshooting
•
Pump-specific Troubleshooting
•
Error Messages
•
Display Messages
A quick-reference hardware troubleshooting guide is included at the
end of this appendix that includes tips for diagnosing and remedying
hardware problems. This guide can save you time in diagnosing
problems when the symptoms are known.
Theory of Operation
A gradient pump works by first pulling a solvent into a proportioning
valve. The P4000 pump has four proportioning valves. Solvents are
measured by percentages, as specified by the user, and mixed inside
the pump. The solvents then travel to the pump head, where a piston
regulates the flow of the mixture to an outlet tube. The mixed solvent
is routed through the pressure transducer, into a second pump head,
then through a bypass valve (closed during normal operation), and
finally out to the LC system. The pump's outlet tubing is then
generally connected to an automatic injector, such as an autosampler.
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157
Troubleshooting Your Pump
SAFETY
PRECAUTIONS
Observe the following safety precautions whenever troubleshooting
hardware difficulties.
Caution!
A caution alerts you to situations that could result in personal injury.
It also tells you how to avoid them.
High Voltage!
This icon alerts you to the presence of high voltage and to the
potential injury that could occur from electrical shock were you to
come in contact with a specific instrument area or component. It also
tells you how to avoid contact with the high-voltage areas in your
instrument.
Hot Surface!
This icon alerts you to potential injury that could occur from coming
in contact with a heated surface or area on or in an instrument. It also
tells you how to avoid contact with the heated surfaces in your
instrument.
TROUBLESHOOTING
TIPS
Eliminate all Other possible sources of trouble
Before you spend any time trying to diagnose a suspected pump
problem you should verify that the pump is the only source of
difficulty. Systematically eliminate all other instruments in your LC
system as the source of trouble. If you are not sure which component
of your chromatography system is responsible for poor system
performance the General LC System Troubleshooting section
beginning on page 159 of this appendix provides useful suggestions.
When You're Sure Its the Pump
Once you have isolated the pump as the only remaining source of
difficulty, a variety of self-tests are built into the pump to help you
determine if your pump is operating correctly.
The next section describes the tests found in the /TESTS/ menu item.
Instructions for the built-in hardware and electronics diagnostics are
contained within the description of the Tests Menu.
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Thermo Scientific
Table A.1 General Troubleshooting Table
Symptom
Cause/Remedy
1. No flow.
a) Check mobile phase connections.
b) Check for leaks.
c) Check pump troubleshooting guide.
2. High back pressure.
a) Check flow rate and system/column
specifications.
b) Check for tubing or column blockage.
c) Check pump troubleshooting guide.
3. Unstable baseline or
drift.
a) System/column not equilibrated; allow more
time.
b) Check detector troubleshooting guide.
c) Check pump troubleshooting guide.
4. Baseline noise.
a) Check for air bubbles in system, degas
solvents.
b) Check for system/solvent contamination.
c) Check pump troubleshooting guide.
5. No peaks.
a) Check detector and data system connections.
b) Check autosampler troubleshooting guide.
c) Check sample retention with
chromatographic conditions.
6. Contaminating/ghost
peaks.
a) Clean system and column.
b) Check autosampler troubleshooting guide.
c) Check pump troubleshooting guide.
7. Poor peak shape.
a) Check system for leaks.
b) Check fittings and tubing lengths.
c) Check column performance.
d) Check autosampler troubleshooting guide.
e) Check pump troubleshooting guide.
f) Check detector troubleshooting guide.
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Table A.1 General Troubleshooting Table, continued
Symptom
8. Poor retention time
reproducibility.
Cause/Remedy
a) Check system for leaks and bubbles.
b) System/column not equilibrated, allow more
time.
c) Check column performance.
d) Check pump troubleshooting guide.
e) Check autosampler troubleshooting guide.
f) Check data system troubleshooting guide.
a) Check column performance.
9. Poor peak area
reproducibility.
b) Check autosampler troubleshooting guide.
c) Check data system troubleshooting guide.
10. Non-integrated or too
many peaks.
a) Check integrator or data system
troubleshooting guide.
11. No instrument of device
control.
a) Check cable connections.
b) Check system configuration.
c) Check individual instrument troubleshooting
guide.
d) Check integrator or data system
troubleshooting guide.
For more detailed chromatographic troubleshooting, refer to any
HPLC troubleshooting reference book or call your local sales or
service representative.
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Thermo Scientific
Table A.2 Pump-Specific Hardware Problems
Symptom
1. No response when power
is switched on.
Possible Cause
Diagnostic/Remedy
a) Power cord not firmly
installed.
a) Re-seat cord.
b) Power cord defective.
b) None. Replace cord.
c) Power Entry Module Fuse
blown.
c) None. Refer to page 154 for fuse
replacement.
a) Display contrast needs
adjustment.
a) Press [STATUS] then press and hold
the right-arrow key. Press the [+] or
[-] key to adjust the display contrast.
Press [STATUS] again.
b) Internal fuse blown.
b) None. Contact Thermo Fisher
Scientific representative for service.
3. No display.
a) System PCB defective.
a) None. Contact Thermo Fisher
Scientific representative for service.
4. Only fan runs with power
on.
a) Internal fuse blown.
a) None. Contact Thermo Fisher
Scientific representative for service.
b) Display cable
loose/unplugged.
b) None. Contact Thermo Fisher
Scientific representative for service.
5. No response to keypad
entry. Display OK.
a) Keypad defective or
System PCB defective.
a) None. Contact Thermo Fisher
Scientific representative for service.
6. Random display.
a) Display defective.
a) Display Test (page 120). Contact
Thermo Fisher Scientific
representative for service.
7. Pump motor will not run.
a) Max pressure or flow rate set
to zero.
a) Press [STATUS]. Set Maximum
Pressure to a value > 100 psi.
b) Motor defective.
b) Hardware Series Test (page 121).
Contact Thermo Fisher Scientific
representative for service.
c) Motor cable unplugged.
c) Hardware Series Test (page 121).
Contact Thermo Fisher Scientific
representative for service.
2. No display. Fan and
pump run OK.
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Table A.2 Pump-Specific Hardware Problems, continued
Symptom
Possible Cause
Diagnostic/Remedy
d) Drive circuit.
d) Hardware Series Test (page 121).
Contact Thermo Fisher Scientific
representative for service.
e) Internal fuse blown.
e) Hardware Series Test (page 121).
Contact Thermo Fisher Scientific
representative for service.
f) Target pressure low.
f) None. Press [PURGE] and then
reinitialize file.
8. Oil found on inlet
bracket.
a) Main bearing overlubricated.
a) None. Small amount of oil is
normal.
9. Improper composition.
a) Proportioning valve cable
loose.
a) Hardware Series Test (page 121).
Reconnect cable.
b) Proportioning valve
defective.
b) Hardware Series Test (page 121).
Replace solvent proportioning;
switching valve.
c) Proportioning valve drive
defective.
c) Hardware Series Test (page 121).
Contact Thermo Fisher Scientific
representative for service.
d) Cam marker failure.
d) Hardware Series Test (page 121).
Contact Thermo Fisher Scientific
representative for service.
a) Internal fuse defective.
a) Hardware Series Test (page 121).
Contact Thermo Fisher Scientific
representative for service.
b) Proportioning valve drive
failure.
b) Hardware Series Test (page 121).
Contact Thermo Fisher Scientific
representative for service.
10. Proportioning valve does
not click open.
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Table A.2 Pump-Specific Hardware Problems, continued
Symptom
11. Pump will not go
READY (possible
unstable flow)
Possible Cause
Diagnostic/Remedy
a) Check valve failure.
a) Flow Stability Test, page 34.
Check Valve Test page 112.
Replace check valve - see Appendix
C.
b) Immiscible solvents.
b) Flow Stability Test, page 34.
Check Valve Test, page 112.
Change solvent system.
c) Unstable load (column).
c) Flow Stability Test, page 34.
Check Valve Test, page 112. Pump
must see stable, non-compressible
load.
d) Insufficient degas.
d) Flow Stability Test, page 34.
Check Valve Test, page 112.
Increase helium rate (if helium
degas) or decrease flow rate (if
vacuum degas). Use vent line and
good bottle cap seal.
e) Circuitry failure.
e) Hardware Series Test (page 121).
Contact Thermo Fisher Scientific
representative for service.
f) Partially clogged frit or
filter on high pressure side
of pump.
f) Hardware Series Test (page 121).
Replace filter or frit.
a) Pressure transducer failure.
a) None. Contact Thermo Fisher
Scientific representative for service.
b) Circuitry failure.
b) Hardware Series Test (page 121).
Contact Thermo Fisher Scientific
representative for service.
13. Pump goes from RUN
immediately to READY
(will not maintain RUN
state).
a) Run file has only one time
line.
a) View run file. Add time line and
reload file.
14. Pump will not start or
stop remotely.
a) Incorrect wiring.
a) External Inputs Test 120. Correct
wiring.
12. Sudden shift in pressure
display with no flow.
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Table A.3 Pump Operation Error Messages
BELOW MINIMUM
The column pressure has fallen below the file's Minimum Pressure
PRESSURE;
setting. Check for mobile phase leaks.
CHECKSUM ERROR
The program memory may have been corrupted. Make a note of the
BAD PROGRAM;
circumstances which preceded the message and contact Thermo Fisher
Scientific.
CODE ERROR
The program code has errors. Make a note of the software version and
PROGRAM LOST;
the circumstances which preceded the message. Contact Thermo Fisher
Scientific.
CODE ERROR
The program code has errors. Make a note of the software version and
STACK UNDERFLOW;
the circumstances which preceded the message. Contact Thermo Fisher
Scientific.
CODE ERROR
The program code has errors. Make a note of the software version and
STACK OVERFLOW;
the circumstances which preceded the message. Contact Thermo Fisher
Scientific.
CODE ERROR
You may have experience low line voltage ("brown-out") or there may
FALSE POWER FAIL;
be hardware problems. Note the circumstances which preceded the
message and contact Thermo Fisher Scientific.
EXCEEDS FLOW RANGE
A flow rate was entered in the pump file which exceeded the flow rate
capabilities of the pump. The maximum flow rate for standard
SpectraSYSTEM pumps is 10 mL/min. If higher flow rates are needed,
inert/biocompatible liquid ends are available to extend the flow rate to
30 mL/min. Contact your local sales representative for information.
MAX PRESSURE
The column pressure of the system has exceeded the MaxP (maximum
EXCEEDED;
pressure) value entered into the pump file. The file's Maximum
Pressure value may need to be increased. The default value is 3000 psi
for
10 mL/min liquid ends (the maximum is 6000 psi). If your operating
column pressure is increasing, check for column plugging.
MOTOR STALLED
The motor is unable to maintain the combination of requested flow rate
and needed pressure for operation. Reduce flow rate or check for flow
restriction or plugged column frit.
OVER MAXIMUM
The pump has overheated. This may be due to blocked ventilation slots
TEMPERATURE;
or to a hardware malfunction. If necessary, clean the air filter.
POWER FAILURE
A power failure has occurred or the pump was switched off with the
CONTINUE;
motor running. The pump has automatically resumed operation.
("Continue" was selected in /OPTIONS/, /Error Recovery/, AC Power
Fail.)
POWER FAILURE
A power failure has occurred or the pump was switched off with the
STOP;
motor running. The pump has automatically stopped. ("Stop" was
selected in /OPTIONS/, /Error Recovery/, AC Power Fail.)
POWER FAILURE
A power failure has occurred or the pump was switched off with the
SHUTDOWN;
motor running. The pump has automatically loaded and run the
shutdown file. ("Shutdown" was selected in /OPTIONS/, /Error
Recovery/, AC Power Fail.)
ZERO FLOW RATE
A time line (other than Time = 0.0 min) with a zero flow rate was
encountered. To remedy, enter a valid flow rate in the first line of the
pump file. Rates between .01 and 10 mL/min are valid for standard
SpectraSYSTEM Pumps. The optional inert/biocompatible liquid ends
extend the maximum flow rate to 30 mL/min.
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Thermo Scientific
Table A.4 Display Messages
Cannot Load File:
(P4000 only) The five messages below indicate that Develop
File parameters are inconsistent.
Solvent1 = Fixed Solvent
Solv 1 = Solv not used
Start%1 + Fixed% > 100
End % 1 + Fixed % > 100
Solvent 1 = Solvent 2
#Runs > 999
Too many runs are set up. Edit the develop file parameters to
reduce the total number of runs.
Maintenance Due
See Log
A volume milestone has been reached. Consult the
Maintenance Log for component by pressing the [MENU] key
and selecting /TESTS/, /Maintenance Log/. For more
information, refer to Chapter 5.
Memory Full
File Not Copied
There is not enough memory available to copy the parameters
of one file into another. Free memory by deleting an old or
unused file, or by reducing the number of time lines in a file.
Try to copy the file again.
Memory Nearly Full
Data May Not Be Saved
There may not be enough memory available. Double-check the
file to ensure that no parameters or settings were lost. Free
memory by deleting an old or unused file, or by reducing the
number of time lines in a file. Try to save the file again.
No Queue Available
You cannot load a queue if none has been set up first.
Not Enough Room
File Not Saved
The run file changes cannot be saved to the file. Free memory
by deleting an old or unused file, or by reducing the number of
time lines in a file. Try to save the run file once more from the
Status Menu.
Protected File
Cannot Be Copied To
You cannot modify a protected file.
Protected File
Cannot Be Deleted
You cannot modify a protected file.
Protected File
Cannot Be Edited
You cannot modify a protected file.
Queue Loaded
Cannot Load File
When a queue is loaded you cannot load any other file without
first pausing the queue.
Run In Progress
No Testing Allowed
The test cannot be initiated because the pump is in RUN or is in
HOLD.
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Table A.5 Solvent Compatibility for Pump Wetted Surfaces
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Thermo Scientific
B
Glossary
A
autosampler
an instrument designed to automatically inject samples into the sample flow
path with a high degree of precision and reproducibility; sometimes called
an injector
B
baseline
the reference line at the bottom of a chromatogram from which
measurements are made; a baseline represents the chromatogram that would
be drawn if only the mobile phase (with no sample) were run through the
column
binary
capable of mixing or switching between two solvents
biocompatible
describes components that are inert when used with biological samples;
biocompatible components are usually made from titanium, PEEK, Teflon,
quartz, or sapphire
blend
a purge option in the P4000 pump allowing all solvents to be purged
simultaneously
buffer
a substance that can neutralize both acids and bases
C
channel
the path along which something (solvent or information) flows
chromatogram
a plot depicting the separated components in a sample (absorbance units
versus time); each component is shown as a separate peak whose
concentration can be determined by studying the area under the peak
chromatograph
the basic set of instruments needed to perform chromatography: a pump,
injector (manual or automatic), a column, and a detector; various recording
and data handling instruments are common additions
chromatography
a means of separating and analyzing mixtures of chemical substances
column
the packed tube through which a sample is passed for separation; the sample
separates according to the way in which it adheres to the column's packing
material
component
an "ingredient" in a chemical mixture, also the individual parts of a liquid
end assembly
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167
conditioning
the process of preparing the surface of the column wall and introducing the
buffer pH conditions into the column before a run
configuration
the way instruments are interconnected to form a system
cursor
a moving or blinking symbol on the display which indicates where
information is entered
D
default
a value or choice built into a system; if no specific choice is made,
instruments will run (or data analyzed) using the default settings
degassing
removal of dissolved gas (i.e. oxygen) from the solvent to prevent bubbles
from forming in the pump; degassing can be done by vacuum or by sparging
detector
the instrument used to detect the presence of a chemical compound
develop file
in the P4000 pump a file which allows a comprehensive set of parameters to
be changed, automatically and sequentially
diagnostics
ways of detecting and isolating instrument or software problems
digit
an editable space within a field
display
the backlit LCD screen on all SpectraSYSTEM™ instruments
E
elution time
the length of time needed to pass a particular sample through a packed LC
column
equilibration
the process used to bring a system (solvent, column, and so on) to a point of
equilibrium, where all thermal and chemical reactions occur at equal rates; a
stable baseline is a good sign of a well-equilibrated system
error message
a printed or displayed message that notifies the user of an error condition
error recovery
user selectable responses to error conditions detected by the instrument, such
as a power interruption or over-pressure
external event
an action performed by an external device that is under the control of the
current instrument (see also timed event)
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Thermo Scientific
F
field
an area in a display, screen, or menu where an entry is required or a choice
must be made
file protect
a setting which allows files to be edited when "off" and protects files from
being changed by editing when "on"
flow parameters
flow rate, run time and solvent composition
flow rate
the rate at which solvent flows through a system
G
gradient
changes the percentage composition of two or more solvents over time;
changes may be continuous or in steps
gradient curve
pre-programmed gradient time line accessed in the Develop file of the P4000
pump. Also refers to the shape of the solvent composition curve between
two time lines.
ground terminal
a terminal used to connect the ground or earth lead of a signal or contact
closure cable; generally green and/or black
H
helium manifold
a pneumatic assembly containing valves and switches for regulating helium
sparging
helium sparging
see sparging
I
inert
see biocompatible
injection
the manual or automatic introduction of a sample into a chromatography
system
integrator
the instrument used to analyze data and produce a chromatogram
isocratic
constant flow and solvent composition
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169
K
Kel-F seal
the translucent seal, made of Kel-F material, inside the pump head which
faces the piston seal
keypad
all of the keys by which you can communicate with an instrument or
computer
L
LC
Liquid Chromatography
linear
a gradient curve that follows a straight line
liquid end
the inlet or outlet assemblies of the pump consisting of the head, piston,
seals, and sometimes a check valve
M
maintenance log
a place to record dates, service, and cumulative solvent volume pumped
menu
a list of choices
method
the set of parameters that define how one or more analyses will be
accomplished
method development
the process of specifying the parameters under which an instrument will
perform a particular function
N
NOVRAM
Non-volatile RAM (random access memory). Computer memory into which
the user can enter information and instructions and from which the user can
recall information. Data in NOVRAM are saved even when the instrument
is switched off
P
parameter
a value or set of values used to define the characteristics or behavior of an
instrument or system
PEEK
polyetheretherketone; a material frequently used in fabricating
inert/biocompatible components
piston
the short cylinder piece which moves inside the sealed cylindrical opening
and used to pressurize fluid
170
Thermo Scientific
piston holder housing
the shaft into which the piston and its holder are housed
piston flush seal
the low-pressure spring seal inside the liquid ends, facing the piston
assembly
piston seal
also called pump seal, a high-pressure spring seal located inside the pump
head
plot
the presentation of analytical data in a graphical manner; typical plots
include chromatogram traces and calibration curves
prime
to flush the solvents contained in a new pump in order to prepare the pump
for solvents chosen by the user
proportioning
the process of opening and closing solenoid valves in sequence to create a
desired solvent composition; usually makes a clicking sound as the solvent
composition is generated
pump
the instrument used to push a liquid solvent through a chromatography
system
purge
to flush the system with fresh, degassed solvent
Q
quaternary
capable of mixing or switching between as many as four solvents
queue
a set of files in a prearranged order
R
RAM
Random Access Memory (computer)
real-time
the current, actual time
reproducibility
the precision with which a piece of data can be repeated; a good measure of
a system's overall performance
retaining screw
also retaining cap screw; the screw which holds the piston assembly into the
piston holder housing
run
a complete analytical operation cycle of the chromatographic system
run file
the file that has been loaded and that the pump is currently operating by
run time
the duration of a sample run, from injection to separation
Thermo Scientific
171
S
sample
a known or unknown substance in a small quantity
seal holder
a metal part used for pumps fitted with standard parts which contains two
seals, (piston seal and piston flush seal) and allows the pump head and
piston holder housing to be joined
shutdown file
a special file used by the pump after the pump has been in a READY state
for a period of time set by the user
solvent
a substance that can completely dissolve another; the mobile phase of an LC
system
solvent filter
a small cylindrical attachment for inlet tubing used to filter a solvent prior to
the solvent entering a pump
solvent program
a set of time lines indicating pump flow and solvent composition at specific
times during a run
solvent-strength gradient
a gradient in which the secondary solvents' composition ratios remain
constant during the run
sparging
a degassing technique in which solvent gases are replaced with an inert gas
such as helium or nitrogen
status
the current condition
status lock
a feature used to prevent a run file from being changed from the Status
Menu
step gradient
a gradient created in step-wise fashion using two solvents, pumped through a
binary, isocratic, solvent delivery system
stroke
one complete revolution of the pump's cam which displaces both pistons
system
a set of chromatography instruments that operate together in a concerted
manner to produce an analytical result
T
timed event
an instrument action triggered to occur at a specific, preset time during a run
or analysis
trace
a chromatogram
transducer check valve
the valve which attached to the inlet of the pressure transducer
172
Thermo Scientific
V
vacuum degassing
the technique of removing dissolved gasses from solvents by passing the
solvent through tubing made of gas-permeable membrane, and creating a
vacuum around the tubing, thus allowing gasses to be evacuated out of the
solvent and into the surrounding chamber
viscosity
the degree to which a fluid resists flow
Thermo Scientific
173
C
Introduction
Kits and Parts Lists
This chapter contains unpacking lists and information for several kits
and accessories available from Thermo Fisher Scientific for use with
your SpectraSYSTEM pump. Described in this chapter are:
• Accessory Kit
p/n A4070-010
• Standard Maintenance Kit
p/n A4050-010
• Standard LC Fittings Kit
p/n A4051-010
• Piston Flush Seal Kit (10 mL)
p/n A4114-010
• Piston Seal Kit (30 mL)
p/n A4084-010
• Solvent Inlet Tube Kit
p/n A4074-010
• Solvent Tube Extension Kit
p/n A4117-010
• Manual Injection Valve Bracket Kit
p/n A4054-010
(also included with A4052-010, the
Rheodyne 7125 Standard Bracket Kit,
and A4053-010)
• Narrow-bore Upgrade Kit
p/n A5190-060
Accessory Kit
Your kit consists of:
2
1
2
2
1
2
1
1
1
1
1
1
1
1
1
3
4 amp, 250V fuse
12-inch piece stainless steel tubing
(0.06 OD x 0.02 ID) (27.5 cm length)
nuts, 0.06 OD
ferrules
seal removal tool
piston seals
hex/ball wrench
4-connection cable
external function connector
20 mL (cc) priming syringe with Luer LOK® tip
Luer adapter
waste tube kit:
48-inch Teflon tubing, 0.031 ID (123 cm)
washer and finger-tight fitting
solvent bottle label
stainless steel tubing, 0.06 OD x 0.007 ID
Thermo Scientific
175
Standard Maintenance Kit
The Standard Maintenance Kit contains the following:
4
1
1
2
1
1
1
6
2
6
1
1
inlet filter cartridges
inlet check valve
transducer check valve
sapphire pistons (0.125 inch-diameter)
barbed fitting (nylon)
piece piston flush tube (Tygon®), 0.0655 ID (approx. 76 cm
long)
syringe (20 cc)
piston seals
piston flush seals
Kel-F seals
seal removal tool
seal insertion tool
Standard LC Fittings Kit
Your kit consists of:
1
1
2
4
4
3
1
storage/carrying case
120-inch piece Teflon tubing, 0.063-inch ID
adapter fittings (10-32M)
Rheodyne nuts/ferrules
Parker type nuts/ferrules
wrenches (sizes: 1/4" - 5/16", 3/8" - 7/16", 1/2" - 9/16")
1/8-inch tube adapter for gas regulator assorted tubing:
stainless steel 0.020-inch ID
stainless steel 0.010-inch ID
Fittings kits contain tubing and fittings commonly needed for LC
systems. These fittings can be used for several different instruments.
If you change your LC configuration or damage connections, the
fittings provided in this kit should allow you to make changes or
replacements quickly and easily. All fittings can be kept in the
compartmentalized storage box.
176
Thermo Scientific
Piston Flush Seal Kit (10 mL)
Your kit consists of:
1
1
2
1
1
1
piece tubing, 30-inch, Tygon (76 cm length)
barbed fitting (nylon)
piston flush seals
seal removal tool
seal insertion tool
syringe, 20 cc
Piston Seal Kit (30 mL)
Your kit consists of:
1
3
1
1
piston (with small O-ring, high pressure)
PEEK wash rings
large O-ring
Piston flush seal (low-pressure)
Solvent Inlet Tube Kit
Your kit consists of:
1
1
bottle cap
tubing, Teflon, 0.063 ID
Solvent Tube Extension Kit
Your kit consists of:
4
1
60-inch extension tubing, with washer and union
(152 cm length)
tubing Teflon, 0.063 ID
Thermo Scientific
177
Manual Injection Valve Bracket Kit
Your kit contains:
1
2
2
2
1
4
2
2
2
1
1
11-3/16 inch steel mounting rod
rod brackets
short column brackets
long column brackets
manual injector valve mount
short set screws (6-32 x 1/4-inch)
flat-head screws (8-32 x 3/8-inch)
screws (6-32 x 7/16-inch)
long set screws (6-32 x 5/8-inch)
Allen wrench (1/16-inch)
12 inches stainless steel tubing, 0.06 OD x 0.01 ID, 12 inches
If you purchased a Rheodyne valve, you also received the valve,
accompanied by Rheodyne's documentation.
Installing the Holder
onto the Valve
Refer to page 187 to install the valve onto the bracket:
178
1.
Move the injector valve handle to the "LOAD" position.
Using the Allen wrench supplied with your Rheodyne valve,
loosen the two set screws and remove the injection valve
handle.
2.
Hold the valve mounting bracket so that the two set screw
holes are on the left. Place the injector valve into the bracket
from the rear. If your injector valve has a remote start cable
attached to it, place the cable into the cut-out on the left side
of the bracket. When aligned correctly the "V" made by the
two flats of the valve shaft will point to the upper left-hand
mounting hole.
3.
Fasten the valve securely with the two flat-head (Phillips)
screws.
4.
Attach the handle to the valve by tightening the two set
screws, making sure that each screw is positioned over a flat
edge of the valve shaft.
5.
Install a long (5/8-inch) set screw into each of the valve
bracket's mounting holes.
6.
Install the rod through the valve bracket so that the notched
portion of the rod faces you, as you look at the front of the
injection valve.
Thermo Scientific
7.
Slide the valve bracket so that it is within the top quarter of
the rod. Tighten the two set screws, securing the valve
bracket to the rod.
8.
Brackets for long (22 cm - 25 cm) and short (3 cm - 10 cm)
columns are provided. Choose the bracket size that matches
the type of column you use. Both brackets can be mounted, if
desired.
9.
Install a short (1/4-inch) set screw into each column bracket
you wish to use. Slide the brackets onto the rod for
positioning. Temporarily tighten the set screws to hold the
brackets in place (Figure C.1).
10.
Remove the two top cover screws from the right side of your
SpectraSYSTEM pump or detector. Install a rod bracket into
the lower hole, using a 7/16-inch Phillips-head screw.
11.
Rest the rod in the lower rod bracket (with the injector valve
facing toward the front) and place the upper rod bracket on
top of the rod. Attach the upper rod bracket to the pump or
detector, using the other 7/16-inch screw.
12.
Re-position the column brackets or manual injector valve as
needed, and tighten the set screws.
Thermo Scientific
179
Slotted Rod
Valve Holder
RUN
STATUS
MENU
STOP
ENTER
SpectraSYSTEM
UV2000
Wrench
Column
Bracket
Rod Bracket
Figure C.1 Manual Injection Valve/Column Bracket mounted to a SpectraSYSTEM instrument
180
Thermo Scientific
DT-Z018/FM
ZERO
Narrow-bore Upgrade Kit
Your kit consists of:
• 0.01” ID bypass, crossover, and transducer stainless steel tubes
• 0.03” ID Teflon solvent inlet tube
• Low-volume outlet liquid-end assembly
• 3 μL static mixer
• 0.007” stainless steel tubing
• Nuts and ferrules for all tubes
• Instruction sheet
Thermo Scientific
181
INDEX
KEYS
[+] and [-] keys, 25, 28
[>] key, 28
[ENTER] key, 28
[PURGE] key, 28
[RUN] key, 27, 58
[STOP] key, 28, 59
[^] key, 28
A
AC Power Fail, 66
accessories, 175
Accessory Kit, basic, 3, 175
active tests, 109
alphanumeric entries, 29
increasing and decreasing, 25
arrow keys
defined, 28
illustrated, 27
rules, 25
asterisks, 30
B
BELOW MINIMUM, 164
binary switching valve, see switching valve:, 156
blank key, see PURGE key, 28
bypass valve
illustrated, 12, 13
C
Calibration Menu, 114
illustration, 114
cautions
defined, 133, 158
maintenance, 133
troubleshooting, 158
Check Valve test, 112
results, 113
check valves
inlet, illustration, 137
inlet, maintenance, 151
maintenance, 150
transducer, illustration, 151
transducer, maintenance, 151
CHECKSUM ERROR, 164
CODE ERROR, 164
column holder, see kits, manual injection valve
bracket, 178
commands
Continue, 60
Copy, 39, 53
Delete (files), 40
Delete (FILES), 53
Delete (QUEUE), 72
display, 60
Hold, 58, 60
Load (FILES), 39, 52
Load (QUEUE), 72
Menu, 29, 60
pause, 75
Pause, 73
Reset, 59, 60
Software Version, 110
Confirmation messages, 30
Continue, 60
conventions used in manual
caution, 133, 158
Current History, 118
cursor
movement, 26
square, 26
triangular, 25
underscore, 26
Cursor Speed, 68
Cycle Step Count test, 119
D
damage
shipping, 3
Degassing, 10
Delay Volume, 68
Delete (FILES), 40, 53
Delete (QUEUE), 72
Develop File
#Curves, 85
corresponding shapes, 85
#Develop Cycles, 84
#Runs/Curve, 87
#Runs/Interval, 84
common fields in Setup Menu, 79
Thermo Scientific
index-i
creating, 78
Edit Menu, 78
illustration, 78
equilibration time, 83
examples, 88
curves selected, 94
grad/2 solvents, 92
iso/2 solvents, 89
run time increment, 96
general description, 76
gradient programs, 84
isocratic programs, 79, 80
Options Menu
gradient, 87
gradient, illustrated, 87
isocratic, 83
isocratic, illustrated, 83
Preview Record, 106
Preview Screen, 88
program modes, 78
Run Time Increment, 87
Setup Menu, 78
illustration, 78
sub-loops, 82
Total # of Runs, 84
Diagnostics Menu
illustration, 110
display
contrast adjustment, 161
Display Solvents, 49
Display test, 120
down-arrow ( ), right side of display, 26
drip tray, 153
E
Edit Menu (Develop file), 78
Edit Menu (FILES), 40
Edit Menu (QUEUE), 71
editing parameters during a run, 30
Editing parameters during a run, 61
End %, 79
ENTER
KEY DEFINED, 28
EQUIL, 61
Equilibration Time, 44
importance of in Develop File, 83
Error messages, 30
Error Recovery Menu, 65
EXCEEDS FLOW RANGE, 164
exclamation points, 30
external events
connection for, 9
External Inputs Test, 120
index-ii
F
fields
#Develop Cycles (Develop File), 84
#Run/Total#Runs (Status Menu of a Develop
File), 99
#Runs, 71
#Runs/Curve (Develop File), 87
#Runs/Interval (Develop File), 84
% Interval (Develop File), 80
@ Maximum Pres, 67
A, B, C, or D, 43
AC Power Fail, 66
Cursor Speed, 68
DATE, 129
Delay Volume, 68
Display Solvents, 49
DUE, 129
Edit File, 40
End % (Develop File), 79
Equilibration Time, 44
Event, 50
File Name (FILES), 41
File Name (QUEUE), 71
File Protection, 69
Fixed Solvent % (Develop File), 83
Fixed Solvent (Develop File), 83
Flow, 41
Flow (Purge), 55
Flow Range, 130
Flow Rate (Develop File), 80
Gradient Curve, 44
ITEM, 129
Liquid End Type, 130
Maximum Pressure, 44
Measured Time, 117
Measured Vol, 116, 117
Minimum Pressure, 44
Order, 71
Pres, 55
Pressure Units, 67
Program (Develop File), 78
Purge, 55
Purge Mode, 68
Ready Output Active, 69
Relative % Interval (Develop File), 81
Relative End % (Develop File), 81
RelativeStart% (Develop File), 81
Run Time (Develop File), 80
Run Time Increment (Develop File), 87
Select Solvents, 69
Solvent 1 (Develop File), 79
Solvent 2 (Develop File), 79
Solvent Not Used (Develop File), 80
Solvents 2,3 (Develop File), 80
Thermo Scientific
Start % (Develop File), 79
Status Lock, 68
Time (Purge), 55
Time (Solvent Program), 41
Time (Timed Events), 50
Time from READY, 51
Total # of Runs (Develop File), 84
Transducer Range, 121
VOL, 129
file linking, see Queue:, 70
File Name (FILES), 41
file numbers, 40
File Protection, 69
files
initializing, 58
Files
Menu, 29
FILES
Copy, 39, 53
Delete, 40, 53
Edit, 40
Load, 39, 52
Menu, 39
Fixed Solvent, 83
Fixed Solvent %, 83
Flow
field of [PURGE] Menu, 55
Flow Calibration Test, 114
Flow Correction, 131
flow stability test
example of running, 34
Flow Stability test, 110
front panel
illustration, 27
fuse
power entry module replacement, 155
G
gradient
curves (P4000 only)
examples, 45
illustration, 45
operation, 45
delay volume, 68
programs (Develop File), 84
solvent strength, 85
step, 48
H
Hardware modifications
autosampler, 37
pump, 37
Hardware Series test, 121
results, 124
high voltage, defined, 133, 158
Hold, 60
HOLD, shown in Status, 61
hot surface
defined, 133, 158
I
Increment (+) and decrement (-)
keys
defined, 28
illustrated, 27
INIT, 61
initializing a file, 58
injection valve/column bracket, see kits, manual
injection valve bracket:, 178
inlet bracket, 156, 162
installation
bench space needed, 8
external events connection, 9
power-on response, 8
setting voltage, 5
unpacking, 3
instrument control, 27
isocratic
programs (Develop File), 79
K
Kel-F seal, 138
keypad
illustrated, 27
instrument control, 27
moving around, 25
keys
[+] and [-], 25, 28
[<], 28
[>], 28
[ENTER], 28
[MENU], 28
[PURGE], 28
[RUN], 27
[STATUS], 28
[STOP], 28
[^], 28
BLANK, 28
kits, 175
maintenance, 10 mL/min piston flush seal, 177
maintenance, standard, 176
manual injection valve bracket
illustration, 180
Manual Injection Valve Bracket, 178
Thermo Scientific
index-iii
Piston Flush Seal, 177
Solvent Inlet Tube, 177
Solvent Tube Extension, 177
Standard LC Fittings, 176
L
Lifetime History, 118
lifting, 5
linking, see Queue:, 70
liquid end
piston seal, 143
Liquid End Type, 130
liquid ends
assembly, 143
components, illustration, 142
disassembly, 138
inlet, illustration, 149
installation, 147
maintenance, 134
removal, 136
Load (FILES), 39, 52
Load (QUEUE), 72
M
Main Menu
illustration, 29, 30, 31, 32, 33
maintenance
check valves, see also check valves, 150
drip tray, 153
kits
contents of 10 mL/min piston flush seal, 177
contents of standard, 176
liquid end, see also Liquid Ends, 134
period, extending, 132
piston, 141
piston flush seal, 144
preparing the pump for, 134
tips, 153
Maintenance Log, 128
DATE, 129
DUE, 129
ITEM, 129
menu, 128
illustration, 129
message, 128, 130
setting intervals, 129
table, 129
using, 129
VOL, 129
Maintenance Position, 130
Manual Injection Valve Bracket, 178
MAX PRESSURE, 164
index-iv
Measured Parameters, 111
MENU
KEY DEFINED, 28
menus
Calibration, 114
COMMANDS, 60
Current History, 118
Diagnostics, 110
Edit (FILES), 40
Edit (QUEUE), 71
Error Recovery, 65
FILES, 39
Flow Correction, 131
Lifetime History, 118
Liquid End Type, 130
More (OPTIONS), 67
OPTIONS, 65
Options (Develop File), 83
Options (FILES), 43
QUEUE, 70
Service, 118
Setup (Develop File), 78
Solvent Program, 41
Status, 61
TESTS, 108
Timed Events, 50
Menus
Status, 30
Menus and Screens, general description of
Commands Menu, 29
Files Menu, 29
Main Menu, 29
Options Menu, 29
Queue Menu, 29
Status Screen, 29
menusDiagnostics, 110
messages, 30
confirmation, 30
error, 30
user, 30
More Menu (OPTIONS), 67
MOTOR STALLED, 164
Motor Step/Valve test, 121
N
numeric entries, 29
O
Options Menu, 29, 65
under Files, Edit, 43
Options Menu (Develop File), 83
Thermo Scientific
Order (QUEUE), 71
OVER MAXIMUM, 164
P
passivation of stainless steel parts, 154
passive tests, 109
pause (QUEUE), 73, 75
pausing a queue, 75
piston
flush seal, 144, 177
holder housing, 138
scratches, 127
illustration, 143
seal, 143
Piston flush seal kit, 177
power entry module
fuse replacement, 155
POWER FAILURE, 164
power selection
see voltage selection, 5
power-on response, 8
preset choices
scrolling through, 28
selecting from, 28
pressure
setting limits, 44
pressure units, 67
example of changing, 32
Pressure Units, 67
Preview Screen, 88
Priming the Pump, 12
program modes (Develop File), 78
Program Version, 110
pump
front panel, illustration of, 27
gradient basics, 35
LC basics, 35
lifting and carrying, 5
list of routine operations, 39
operating statistics, 118
P4000
description, 36
placement, 8
pressure, 44
rear panel, illustration of, 6
repair, 155
rules for operation, 25
state changes, 118
stopping, 58
theory of operation, 157
Pump
carrying, 5
purge
field, of Purge Menu, 55
mode
example of changing, 32
starting, 56
stopping, 56
PURGE
key, 54
key defined, 28
MENU, 54
Purge Mode, 67
purging
after priming the pump, 16
example, 32
solvents, 54
Q
Q RUN, 62
QEQUIL, 61
QREADY, 62
QSTOP, 62
queue
#Runs, 71
adding lines, 71
Delete command, 72
deleting entire, 72
deleting lines, 72
description, 70
editing, 71
illustration, 71
holding a file in the, 75
Load command, 72
Menu, 29
illustrated, 70
order, 71
Pause command, 75
running, 73
stopping, 75
R
RAM test, 119
rate of change, gradient curves, 45
READY, 58
Ready Output Active, 69
READY, shown in Status, 62
Rear panel
illustration, 6
records
Develop File, 100
Preview Screen, 106
Relative %
description, 82
fields, 81
Thermo Scientific
index-v
Relative % Interval, 81
Relative End %, 81
RelativeStart %, 81
repair
fuse replacement, 155
solvent switching valve replacement, 156
Reset, 60
reviewing parameters during a run, 30
ROM test, 119
RUN, 57, 58
key defined, 27
shown in Status, 62
run file, defined, 39
run time increment 87
example, 96
S
safety
icons, 133, 158
safety certification
see Safety Information:, 133
safety precautions
for troubleshooting, 158
saving an entry, 26
screens
Preview (Develop File), 88
Status, 61
Screens
Status
P4000 illustrated, 29
Status (P4000), 62
scrolling, 28
seals
Kel-F, 138
piston, 143
piston flush, 144, 177
Service Menu, 118
illustration, 118
Setup Menu (Develop File), 78
shipping
damage, 3
Shutdown File, 51
Software Version, 110
Solvent 1, 79
Solvent 2, 79
Solvent Not Used, 80
Solvent Program Menu, 41
Solvent Selection Menu, 69
solvents
changing the number displayed, 49
composition
example of, 43
connecting, 9
index-vi
drip tray, 153
entering percentages, 43
labeling
example, 31
labels, 69
naming, illustration, 69
purging, 54
selection, 69
solvent strength gradient, 85
Solvents 2,3, 80
stainless steel
passivation, 154
Start %, 79
state changes, chronological list of, 118
statistics, operating, 118
status
display, 62
Status, 61
Develop File, 98
P4000, 62
STATUS
KEY DEFINED, 28
Status lock
differences between file protection, 68
Status Lock, 68
Status Menu
described, 30
P4000 illustrated, 62
Status Messages, 61
Status Screen
described, 29
P4000 illustrated, 62
step gradients, 48
STOP
KEY DEFINED, 28
shown in Status, 62
sub-loops, 82
switching valve
replacement, 156
SYNC, 62
T
tests
active and passive, 109
Calibration, 114
Check Valve Test, 112
Cycle Step Count, 119
Display, 120
External Inputs, 120
Flow Stability, 110
Hardware Series, 121
initiating, 109
Measured Parameters, 111
Thermo Scientific
Motor/Step Valve, 121
RAM, 119
results, 109
ROM, 119
Service, 118
Software Version, 110
Transducer Range, 121
Tests Menu, 108
illustration, 108
theory of operation, 157
time
shown in Status, 61
Time
field of [PURGE] display, 55
time lines
adding, 41
chronological order of, 42
creating, 41
general rules, 41
Time lines
defined, 41
timed events
display, 50
Timed Events Menu, 50
Total # of Runs, 84
fields affecting, 98
modifying, 98
Transducer Range, 121
troubleshooting, 157
isolating the source of the problem, 158
safety precautions for, 158
U
unpacking, 4
V
Visual clues on display, 26
voltage select barrel, 5
voltage selection, 5
barrel, 7
illustration, 7
instructions for, 7
Z
ZERO FLOW RATE, 164
Zero Pressure, 114
Thermo Scientific
index-vii
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