SpectraSYSTEM UV/Vis Detectors Reference

SpectraSYSTEM UV/Vis Detectors Reference
Finnigan™
SpectraSYSTEM™
UV/Vis Detectors
Reference Manual
A0099-540 Revision G
Finnigan™ and SpectraSYSTEM™ are trademarks of Thermo Electron Corporation.
This manual and the instruments to which it applies have satisfied the requirements for CSA, FCC, the EMC,
and Low Voltage Directives. Use of non-approved components and repair methods may reduce or invalidate
the built-in protection that is required to meet the above certifications.
Technical information contained in this publication is for reference purposes only and is subject to change
without notice. Every effort has been made to supply complete and accurate information; however,
Thermo Electron Corporation assumes no responsibility and will not be liable for any errors, omissions,
damage, or loss that might result from any use of this manual or the information contained therein (even if this
information is properly followed and problems still arise).
This publication is not part of the Agreement of Sale between Thermo Electron Corporation and the purchaser
of an LC system. In the event of any conflict between the provisions of this document and those contained in
Thermo Electron Corporation’s Terms and Conditions, the provisions of the Terms and Conditions
shall govern.
System Configurations and Specifications supersede all previous information and are subject to change
without notice.
Printing History: Revision G printed in January 2004
The products of Thermo Electron San Jose are produced under ISO 9001 accredited quality management systems.
Australia: P.O. Box 239 Rydalmere • Unit 14, 38 – 46 South Street • Rydalmere, N.S.W. 2116 • [61] (02) 9898-9000
Austria: Wehlistrasse 27b • A-1200 Wien • [43] (01) 333 50 34-0
Belgium: Technologiestraat 47 • B-1082 Brussels • [32] (02) 482 30 30
Canada: 5716 Coopers Avenue, Unit 1 • Mississauga, Ontario • L4Z 2E8 • [1] (905) 712-2258
France: 16 Avenue du Québec • Silic 765 • Z.A. de Courtaboeuf • F-91963 Les Ulis Cédex • [33] (01) 60 92 48 00
Germany: Im Steingrund 4-6 • D-63303 Dreieich • [49] (06103) 408 0
Italy: Strada Rivoltana • I-20090 Rodano (Milano) • [39] (02) 95059 226
Japan: C-2F • 3-9, Moriya-cho, Kanagawa-ku • Yokohama, Kanagawa • 221-0022 • [81] (45) 453 9100
Japan: Esaka Grand Building • 2-3-1 Esaka-cho, Suita City • Osaka 564-0063 • [81] (06) 6387-6681
Netherlands: Takkebijsters 1 • NL-4817 BL Breda • [31] (076) 5878 722
P.R. China: Room 901, Ping-an Mansion • No. 23, Jin Rong Street • Xi Cheng District • Beijing 100032 • [86] (010) 6621 0839
Spain: Sepulveda 7 A • ES-28108 Alcobendas (Madrid) • [34] (091) 657 4930
Spain: Acer 30 – 32 • Edificio Sertram – Planta 2, Modulo 3 • ES-08038 Barcelona • [34] (093) 223 0918
Sweden: Pyramidbacken 3 • S-141 75 Kungens Kurva (Stockholm) • [46] (08) 556 468 00
United Kingdom: Stafford House • 1 Boundary Park • Boundary Way • Hemel Hempstead • Hertfordshire HP2 7GE • [44] (01442) 233 555
U.S.A.: 355 River Oaks Parkway • San Jose, CA 95134-1991 • [1] (408) 965-6000
Notes: The country code is enclosed in square brackets [ ]. The city code or area code is enclosed in parenthesis ( ). For countries other than the U.S.A.,
when you are dialing from within the specified country, dial the 0 of the city code. For countries other than Italy, when you are dialing from outside the country,
do not dial the 0 of the city code.
Published by Technical Publications, Thermo Electron Corporation, San Jose, California.
Copyright© 2004 Thermo Electron Corporation. All rights reserved. Printed in the United States of America.
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EDITOR, TECHNICAL PUBLICATIONS
THERMO ELECTRON SAN JOSE
355 RIVER OAKS PARKWAY
SAN JOSE, CA 95134-1991
UNITED STATES OF AMERICA
fold
Contents
Back Pocket:
Quick Reference Card
Menu Trees
Customer Support ................................................................................................................................ iii
Safety Information ............................................................................................................................ vii
Start-up Checklist............................................................................................................................. xix
List of Spare Parts, Consumables, and Kits.................................................................................... xxi
Chapter 1 Getting Started
Introduction ............................................................................................................................
Learning Your Way Around ...................................................................................................
Instrument Control .................................................................................................................
Manual Conventions ...............................................................................................................
What's Next? ..........................................................................................................................
1
1
2
6
8
Chapter 2 A Quick Example
Introduction ............................................................................................................................ 9
UV1000, An Example ............................................................................................................ 10
UV2000, An Example .......................................................................................................... 14
What's Next? ........................................................................................................................ 18
Chapter 3 Basic Operations
Introduction ..........................................................................................................................
Before You Begin .................................................................................................................
UV1000, Single-wavelength Operation .................................................................................
UV2000, Single- and Dual-wavelength Operation ...............................................................
More about Files ...................................................................................................................
Analog Outputs .....................................................................................................................
19
19
20
26
32
35
Chapter 4 Advanced Operations
Introduction ..........................................................................................................................
Wavelength Programming ....................................................................................................
Programmed Autozero .........................................................................................................
Automatic Lamp Operations ................................................................................................
Other Features ......................................................................................................................
UV2000, Scanning .................................................................................................................
UV2000, Automatic Scanning ...............................................................................................
The Develop File....................................................................................................................
Sample Queue.........................................................................................................................
K-Factor .................................................................................................................................
Absorbance Ratios..................................................................................................................
37
37
40
41
43
46
51
54
56
60
64
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Chapter 5 Required Maintenance
Introduction .......................................................................................................................... 67
Flowcells ............................................................................................................................... 67
Lamps ................................................................................................................................... 73
Appendix A Installation and Specifications
Introduction .......................................................................................................................... 79
Installation ............................................................................................................................ 79
Specifications ........................................................................................................................ 90
Appendix B Menu Reference
Introduction .......................................................................................................................... 93
Menu Trees ........................................................................................................................... 93
Menu Reference .................................................................................................................... 97
Appendix C Troubleshooting
Introduction ........................................................................................................................
Theory of Operation ...........................................................................................................
Common Problems .............................................................................................................
Troubleshooting Table ........................................................................................................
Error Messages ...................................................................................................................
Diagnostic Tests .................................................................................................................
105
105
107
107
110
112
Appendix D Glossary
Introduction ........................................................................................................................ 117
Appendix E Cuvette Holder Accessory
Introduction ........................................................................................................................
Installation ..........................................................................................................................
Using the Cuvette Holder ...................................................................................................
Maintenance ........................................................................................................................
121
122
127
128
Index
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Customer Support
Thermo Electron San Jose products are supported by
Thermo Electron Customer Service Engineers with customer support
available in North America, in Europe, and in Australasia and Asia.
IN NORTH AMERICA
In North America, Thermo Electron Customer Service Engineers are
available from each of the Thermo Electron field offices as follows:
Northeastern Region
Phone [1] (732) 627-0220
Fax
[1] (732) 627-0260
Southern Region
Phone [1] (770) 516-5589
Fax
[1] (770) 516-6916
Central Region
Phone [1] (847) 310-0140
Fax
[1] (847) 310-0145
Western Region
Phone [1] (408) 965-6000
Fax
[1] (408) 965-6123
Canada
Phone [1] (905) 712-2258
Fax
[1] (905) 712-4203
In the Americas, use the following telephone number or fax number
to order parts for all instruments:
Thermo Electron Customer Service Operations
1400 Northpoint Parkway, Suite 10
West Palm Beach, FL 33407
Phone: [1] (800) 532-4752
Fax: [1] (561) 688-8731
Thermo Electron Technical Support is available at the following
location:
Thermo Electron Technical Support Operations
1400 Northpoint Parkway, Suite 10
West Palm Beach, FL 33407
Phone: [1] (800) 685-9535
Fax: [1] (561) 688-8736
01/2004
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iii
IN EUROPE
In Europe, customer support, replaceable parts, and technical support
are available from each of the Thermo Electron offices as follows:
Wien (Vienna), Austria
Phone [43] (01) 333 50 34-0
Fax
[43] (01) 333 50 34-26
Brussels, Belgium
Phone [32] (02) 482 30 30
Fax
[32] (02) 482 30 31
Les Ulis Cédex, France
Phone [33] (01) 60 92 48 00
Fax
[33] (01) 60 92 49 00
Dreieich, Germany
Phone [49] (06103) 408 0
Fax
[49] (06103) 408 1222
Milano, Italy
Phone [39] (02) 95 059 226
Fax
[39] (02) 95 320 370
Breda, Netherlands
Phone [31] (076) 587 8722
Fax
[31] (076) 571 4171
Madrid, Spain
Phone [34] (091) 657 4930
Fax
[34] (091) 657 4937
Barcelona, Spain
Phone [34] (093) 223 0918
Fax
[34] (093) 223 0982
Stockholm, Sweden
Phone [46] (08) 556 468 00
Fax
[46] (08) 556 468 08
Hemel Hempstead, United Kingdom
Phone [44] (01442) 233 555
Fax
[44] (01442) 233 667
For all other countries, contact your local Thermo Electron dealer.
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IN AUSTRALASIA AND ASIA
In Australasia and Asia, customer support, replaceable parts, and
technical support are available from each of the Thermo Electron
offices as follows:
Rydalmere, N.S.W., Australia
Phone [61] (02) 9898-9000
Fax
[61] (02) 9898-9800
Yokohama, Japan
Phone [81] (45) 453-9100
Fax
[81] (06) 453-9110
Osaka, Japan
Phone [81] (06) 6387-6681
Fax
[81] (06) 6387-6641
Beijing, P.R. China
Phone [86] (010) 6621 0839
Fax
[86] (010) 6621 0851
For all other countries, contact your local Thermo Electron dealer.
01/2004
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v
Safety and EMC
Information
In accordance with Thermo Electron’s commitment to customer
service and safety, these instruments have satisfied the requirements
for the FCC and the European CE Mark including the Low
Voltage Directive.
Designed, manufactured and tested in an ISO9001 Registered facility,
this system has been shipped to you from our manufacturing facility
in a safe condition.
IDENTIFYING SAFETY
INFORMATION
This reference manual contains precautionary statements that can
prevent personal injury, instrument damage, and loss of data if
properly followed. All statements of this nature are called to your
attention through the use of bold type and the following icons:
CAUTION!
HOT
SURFACE!
HIGH
VOLTAGE!
Every instrument has specific hazards, so be sure to read and comply
with the following precautions. They will help ensure the safe, longterm use of your system.
1. Before plugging in any of the instrument modules or turning on
the power, always make sure that the voltage and fuses are set
appropriately for your local power supply.
2. Only use fuses of the type and current rating specified. Do not
use repaired fuses and do not short-circuit the fuse holder.
3. The supplied power cord must be inserted into a power outlet
with a protective earth contact (ground). When using an
extension cord, make sure that the cord also has an earth
contact.
4. Do not change the external or internal grounding connections.
Tampering with or disconnecting these connections could
endanger you and/or damage the system.
CAUTION! The instrument is properly grounded in accordance with
these regulations when shipped. You do not need to make any changes
to the electrical connections or to the instrument's chassis to ensure
safe operation.
CAUTION! Do not override the lamp cover safety interlock, which
turns the lamps off when the cover is removed, or personal injury
could result.
5. Never run the system without the top cover on. Permanent
damage can occur.
01/2004
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vii
6. Do not turn the instrument on if you suspect that it has incurred
any kind of electrical damage. Instead, disconnect the power
cord and contact a Thermo Electron Service Representative for
a product evaluation. Do not attempt to use the instrument
until it has been evaluated. (Electrical damage may have
occurred if the system shows visible signs of damage, or has
been transported under severe stress.)
7. Damage can also result if the instrument is stored for prolonged
periods under unfavorable conditions (e.g., subjected to heat,
water, etc.).
8. Always disconnect the power cord before attempting any type
of maintenance.
9. Capacitors inside the instrument may still be charged even if
the instrument is turned off.
10. Never try to repair or replace any component of the system that
is not described in this manual without the assistance of
Thermo Electron.
GOOD LABORATORY
PRACTICES
Keep Good Records
To help identify and isolate problems with either your equipment or
your methodology, we recommend that you keep good records of all
system conditions (e.g., %RSDs on migration times and peak areas,
peak shape and resolution). At a minimum, keep an
electropherogram of a typical sample and standard mixture, welldocumented with system conditions, for future reference. Careful
comparison of migration times, peak shapes, peak sensitivity, and
baseline noise can provide valuable clues to identifying and solving
future problems.
Chemical Toxicity
Although the large volume of toxic and flammable solvents used and
stored in laboratories can be quite dangerous, don't 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 electrolyte
buffer. Sample precipitation can plug the system by obstructing the
flow through the capillary. This obstruction may result in irreparable
damage to parts of the system. Particulate matter can be avoided by
filtering the samples through 0.45- or 0.2-micron (or less) filters.
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Solvent Requirements
Many chemical manufacturers provide a line of high-purity or
spectro-quality 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 will significantly prolong 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's compatible with the sample and
column you've selected for your separation. Remember that
some solvents are corrosive to stainless steel.
Inert/biocompatible instrument versions are also available from
Thermo Electron.
Degas the Eluents
Degas your eluents using either the vacuum degassing or the
helium sparging technique. Complete information for using
either of these techniques is found in separate documentation
provided with degas accessories.
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, but since liquids aren't
highly compressible, they do not store much energy. Thus, little
immediate danger arises from the high pressure in an LC system.
However, if a leak occurs, it should be corrected as soon as
possible. Finally, we recommend that you always wear eye and
skin protection when working on an LC system and that you
always shut down the system and return it to atmospheric
pressure before attempting any maintenance.
01/2004
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ix
Information sur
la sécurité et la
compatibilité
électromagnétique (CEM)
Selon notre engagement à assurer à nos clients service et sécurité, ces
instruments sont déclarés conformes aux normes de la FCC et à la
réglementation européenne (CE), y compris à la directive sur les
basses tensions.
Conçu, fabriqué et testé dans une installation homologuée ISO9001,
cet instrument a été livré à partir de notre usine de fabrication dans le
respect des règles de sécurité.
MISE EN GARDE ! Cet instrument doit être utilisé selon les
instructions figurant dans ce manuel. Le non respect des consignes
d’utilisation de cet instrument décrites dans le présent manuel risque
d’endommager l’instrument et/ou d’infliger des blessures à
l’opérateur.
IDENTIFICATION DES
INFORMATIONS SUR LA
SÉCURITÉ
Ce manuel de référence contient des précautions d’usage afin de
prévenir tout dommage corporel ou matériel ainsi que toute perte de
données lorsque l’opérateur se conforme aux instructions indiquées.
Ces instructions sont accompagnées des icônes suivantes et sont
affichées en caractères gras pour attirer l’attention de l’opérateur :
MISE EN
GARDE !
SURFACE
BRÛLANTE !
HAUTE
TENSION !
Chaque instrument présentant des dangers spécifiques, il incombe à
l’opérateur de lire les précautions suivantes et de s’y conformer, afin
de maintenir la durée de vie et la sécurité du système.
1. Avant de brancher un module d’instruments ou de le mettre
sous tension, toujours s’assurer que la tension et les fusibles
sont réglés de façon à correspondre à la tension locale du
secteur.
2. N’utiliser que des fusibles du type et du courant nominal
spécifiés. Ne pas utiliser de fusibles réparés et ne pas courtcircuiter le porte-fusible.
01/2004
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xi
3. Le cordon d’alimentation accompagnant l’instrument doit être
branché à une prise de courant avec mise à la terre. En cas
d’utilisation d’une rallonge électrique, s’assurer que celle-ci
comporte également une mise à la terre.
4. Ne pas modifier les connexions de mise à la terre internes ou
externes. La modification ou le débranchement de ces
connexions représente un danger pour l’opérateur et/ou risque
d’endommager le système.
MISE EN GARDE ! Cet instrument est mis à la terre conformément
aux règlements applicables lors de son expédition. Ne pas modifier
les branchements électriques ou le châssis de l’instrument afin
d’assurer un fonctionnement en toute sécurité.
MISE EN GARDE ! Ne pas abroger le contact de sécurité de
couverture de lampe (qui éteignez la lampe quand le couverture est
enlevé) ou les blessures pourraient résulter.
5. Ne jamais faire fonctionner le système sans son boîtier.
Des dommages permanents pourraient en résulter.
6. Ne pas mettre l’instrument sous tension si celui-ci a subi des
dommages électriques. Débrancher le cordon d’alimentation de
l’appareil et consulter un représentant du service technique
pour procéder à un examen du produit. Ne pas essayer
d’utiliser l’instrument avant qu’il n’ait été examiné. (Des
dommages électriques peuvent s’être produits si le système
montre des signes visibles d’endommagement ou si les
conditions de transport ont été extrêmement difficiles.)
7. L’instrument peut également être endommagé s’il est entreposé
pendant une période de temps prolongée, dans de mauvaises
conditions (par exemple, s’il est exposé à la chaleur, à
l’humidité, etc.).
8. Toujours débrancher le cordon d’alimentation avant d’effectuer
n’importe quel type d’entretien.
9. Les condensateurs présents à l’intérieur de l’instrument
peuvent toujours être chargés, même si l’instrument est hors
tension.
10. Ne jamais tenter de réparer ou de remplacer un composant du
système non décrit dans ce manuel sans obtenir de l’aide
auprès d’un représentant du service technique.
BONNES PRATIQUES DE
LABORATOIRE
Bonne tenue des dossiers
Pour permettre d’identifier et d’isoler les problèmes pouvant survenir
avec l’équipement ou la méthodologie utilisés, il est recommandé de
tenir correctement des dossiers de toutes les conditions du système (p.
ex., % CV sur les temps de rétention et les zones de pics, la forme et
la résolution des pics). Il est recommandé tout au moins de conserver
xii
Thermo Electron
01/2004
pour référence future un chromatogramme d’un échantillon type et
d’un mélange standard, bien documenté et accompagné des
conditions du système. Une comparaison précise des temps de
rétention, des formes et de la sensibilité des pics ainsi que des bruits
de référence peuvent fournir des indices précieux pour l’identification
et la résolution de problèmes futurs.
Toxicité chimique
Bien que l’utilisation et l’entreposage dans les laboratoires de grandes
quantités de solvants inflammables et toxiques puissent représenter un
danger, ne pas négliger les dangers potentiels posés par les
échantillons. Veiller particulièrement à lire et à suivre toutes les
précautions indiquées pour assurer la ventilation, le stockage, la
manutention et l’élimination des solvants et des échantillons. Se
familiariser avec les données sur la toxicité et les dangers potentiels
associés à tous les produits chimiques en consultant les fiches
techniques sur la sécurité des substances (FTSS) du fabricant.
Préparation des échantillons
Toujours considérer la solubilité de l’échantillon dans la phase
mobile. La précipitation des échantillons peut boucher la colonne, les
tubes et/ou la cellule de dilution, et en limiter le débit. Cette
obstruction peut endommager le système de façon irréparable.
L’accumulation de particules peut être évitée par la filtration des
échantillons à travers des filtres de 0,45 ou 0,2 µm (ou moins).
Caractéristiques des solvants
Un grand nombre de fabricants de produits chimiques fournissent des
réactifs de pureté élevée ou de qualité spectrographique dépourvue de
toute impureté chimique. La filtration systématique de tous les
solvants ou éluants à travers un filtre fluorocarboné de 0,45 ou 0,2 µm
(ou moins) avant de les placer dans le réservoir de solvants prolonge
de façon significative la durée de vie et l’efficacité des filtres
d’entrée, des clapets et des joints d’étanchéité, de l’injecteur et de la
colonne. De façon générale, les solvants pour chromatographie
liquide sous haute pression ne nécessitent pas de filtration.
Choisir une phase mobile qui est compatible avec l’échantillon et la
colonne sélectionnés pour la séparation. Noter que certains solvants
sont corrosifs pour l’acier inoxydable. Des versions inertes et
biocompatibles des instruments sont disponibles auprès de
Thermo Electron.
Dégazage des éluants
Effectuer le dégazage des éluants selon la méthode de dégazage par le
vide ou à l’hélium. Une description complète de ces méthodes est
disponible dans la documentation fournie séparément avec les
accessoires de dégazage.
01/2004
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xiii
Élimination des solvants
S’assurer qu’il existe un conteneur pour solvants à éliminer ou tout
autre système de vidange au niveau de la table de travail ou audessous de celle-ci. La plupart des solvants doivent être éliminés dans
des conditions particulières et ne doivent pas être évacués directement
par les canalisations. Respecter la réglementation en vigueur
concernant l’évacuation des produits chimiques.
Systèmes à haute pression et fuites
Les systèmes de chromatographie liquide (CL) fonctionnent à des
pressions élevées. Les liquides n’accumulent pas de grandes quantités
d’énergie car ils ne sont pas hautement compressibles. Par
conséquent, le risque d’un danger immédiat causé par les pressions
élevées dans un système CL est faible. En revanche, si une fuite
survient, il est nécessaire de la réparer le plus rapidement possible.
Enfin, il est recommandé à l’opérateur de se protéger en permanence
les yeux et la peau lorsqu’il travaille sur un système CL. De plus, il
doit toujours mettre le système hors tension et le ramener à la
pression atmosphérique avant de procéder à tout entretien.
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Informationen zu
Sicherheit und
Funkentstörung
Wir sind dem Dienst am Kunden und der Sicherheit des Kunden
verpflichtet. Diese Geräte entsprechen den Anforderungen
für die FCC-Zulassung und für das CE-Zeichen sowie den
Bestimmungen der Richtlinie für Niederspannungsgeräte.
Dieses Gerät wurde in einer nach ISO 9001 zertifizierten
Fertigungsstätte entwickelt, hergestellt und getestet und hat
unser Werk in sicherem Zustand verlassen.
VORSICHT! Dieses Gerät darf nur nach den Vorschriften
dieser Bedienungsanleitung benutzt werden. Wenn dieses
Gerät auf andere Weise als hier beschrieben benutzt wird,
kann dies zu Schäden am Gerät oder zur Verletzung des
Bedieners führen.
ERKENNEN VON
SICHERHEITSINFORMATIONEN
Dieses Handbuch enthält Warnhinweise, deren genaue Befolgung
Personenschäden, Schäden am Gerät oder Datenverluste verhindern
kann. Auf alle derartigen Warnhinweise wird durch Fettschrift und
durch
Verwendung der nachfolgenden Symbole gesondert
aufmerksam gemacht:
VORSICHT!
OBERFLÄCHE
HEISS!
HOCHSPANNUNG
Jedes Gerät kann unter bestimmten Umständen gefährlich
sein. Lesen Sie daher in jedem Fall die nachstehenden
Sicherheitshinweise, und ergreifen Sie die entsprechenden
Maßnahmen. Auf dieses Weise sorgen Sie für einen sicheren Betrieb
und eine lange Lebensdauer des Geräts.
1. Bevor Sie eines der Gerätemodule einstecken oder das
Gerät einschalten, überprüfen Sie in jedem Fall, ob die
Nennspannung und die Sicherungen der Netzspannung
der örtlichen Stromversorgung entsprechen.
2. Verwenden Sie nur Sicherungen des angegebenen Typs
und der angegebenen Amperezahl. Verwenden Sie keine
reparierten Sicherungen, und überbrücken Sie die
Sicherung nicht.
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3. Das mitgelieferte Netzkabel muß in eine Steckdose
mit Schutzleiter eingesteckt werden. Wird ein
Verlängerungskabel verwendet, muß auch hier
der Schutzleiter durchgeführt sein.
4. Verändern Sie nichts an den externen oder internen
Schutz- bzw. Erdungskontakten. Wenn Sie sich an
diesen zu schaffen machen oder sie unterbrechen,
können Sie sich selbst und andere gefährden, oder
das Gerät könnte beschädigt werden.
VORSICHT! Das Gerät ist bei der Auslieferung vorschriftsmäßig
geerdet. Es brauchen keine Veränderungen an der elektrischen
Verkabelung oder am Gerätechassis vorgenommen werden,
um einen sicheren Betrieb zu gewährleisten.
VORSICHT! Setzen Sie niemals den Sicherheitsschalter der
Lampenabdeckung ausser Kraft! Der Sicherheitsschalter schaltet die
Lampen aus, wenn die Abdeckung entfernt wird. Die Überbrückung
des Sicherheitsschalters kann zu Gesundheitsschäden oder
Verletzungen führen.
5. Nehmen Sie das Gerät nie mit geöffnetem Gehäuse in Betrieb,
da dies zu irreparablen Schäden führen kann.
6. Schalten Sie das Gerät nicht ein, wenn Sie den
Verdacht haben, daß an der Elektrik möglicherweise Schäden
eingetreten sind. Ziehen Sie in diesem Fall den Netzstecker
heraus, und lassen Sie das Gerät von einem
Kundendiensttechniker untersuchen. Versuchen Sie bis
zu dieser Untersuchung keinesfalls, das Gerät in Betrieb
zu nehmen. (Eine Beschädigung der Elektrik kann z.B.
eingetreten sein, wenn das Gerät äußere Schäden aufweist oder
unter problematischen Umständen transportiert wurde.)
7. Schäden können auch eintreten, wenn das Gerät längere Zeit
unter ungünstigen Umständen gelagert wurde (z.B. unter der
Einwirkung von Hitze oder Wasser).
8. Ziehen Sie vor allen Wartungsmaßnahmen immer zuerst
den Netzstecker aus der Steckdose.
9. Auch wenn das Gerät abgeschaltet ist, können die im Inneren
befindlichen Kondensatoren nach wie vor unter Spannung
stehen.
10. Versuchen Sie niemals, Gerätekomponenten zu reparieren oder
auszutauschen, die nicht in diesem Handbuch beschrieben sind,
ohne einen Kundendiensttechniker
zu Rate zu ziehen.
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GLP-VORSCHRIFTEN
(GOOD LABORATORY
PRACTICES)
Ordnungsgemäße Aufzeichnungen
Damit Probleme mit Geräten oder Methoden erkannt
und eingegrenzt werden können, empfehlen wir Ihnen,
ordnungsgemäße Aufzeichnungen sämtlicher Gerätezustände (z.B. %
RSDs zu Retentionszeiten, Kurvenflächen, Kurvenformen und
Auflösung). Archivieren Sie als Minimum ein Chromatogramm einer
typischen Probe und einer Standardmixtur mit umfassender
Dokumentation der Systembedingungen zum späteren Vergleich. Ein
sorgfältiger Vergleich von Retentionszeiten, Kurvenformen,
Empfindlichkeitswerten und Hintergrundrauschen liefert wertvolle
Hinweise für den Fall, daß zu einem späteren Zeitpunkt Probleme
auftreten und eingegrenzt und behoben werden müssen.
Chemische Toxizität
Die großen Mengen an toxischen oder brennbaren Lösungsmitteln,
die im Labor verwendet und aufbewahrt werden, können ein
erhebliches Gefahrenpotential darstellen, doch darf man hierüber
nicht die mögliche Gefährdung durch
die Proben selbst vergessen. Achten Sie insbesondere darauf,
sämtliche Warnhinweise hinsichtlich ausreichender Belüftung,
Lagerung, Handhabung und Entsorgung von Lösungsmitteln ebenso
wie von Proben sorgfältig zu lesen und zu befolgen. Machen Sie sich
mit den Toxizitätsdaten und den möglichen Gefahren sämtlicher
verwendeter Chemikalien anhand der betreffenden
Sicherheitsdatenblätter vertraut, die von den Produktherstellern zur
Verfügung gestellt werden.
Probenvorbereitung
Überprüfen Sie stets die Löslichkeit der Probe in der mobilen Phase.
Durch das Ausfällen von Feststoffen können die Säule, die Leitungen
oder die Durchflußzelle verstopfen und damit den Durchfluß
hemmen. Durch eine solche Verstopfung können irreparable Schäden
am System entstehen. Die Ablagerung von Partikeln läßt sich durch
Filtrieren der Proben durch ein Filter mit einer Porengröße von 0,45
oder 0,2 µm (oder weniger) vermeiden.
Anforderungen an das Lösungsmittel
Viele chemische Hersteller bieten eine Produktserie hochreiner
Reagenzien in spektroskopisch reiner Qualität an, die frei
von chemischen Unreinheiten sind. Die routinemäßige Filtrierung
aller Lösungs- und Extraktionsmittel durch ein
Fluorkohlenwasserstoff-Filter mit einer Porengröße von
0,45 oder 0,2 µm (oder weniger) vor dem Einfüllen in den
Lösungsmittelbehälter verlängert die Lebensdauer der Einlaßfilter,
der Ventile und Dichtungen, des Injektors und
der Säule beträchtlich. Spezielle HPLC-Lösungsmittel
brauchen normalerweise nicht filtriert zu werden.
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Wählen Sie eine mobile Phase, die zur Probe und zur für die
Separation verwendete Säule kompatibel ist. Dabei ist darauf zu
achten, daß Edelstahl durch bestimmte Lösungsmittel korrodiert wird.
Reaktionsträge, biokompatible Geräteausführungen werden ebenfalls
von Thermo Separation Instruments angeboten.
Entgasen des Lösungsmittels
Lösungs- und Extraktionsmittel sollten entgast werden, und zwar
entweder durch Vakuum oder Heliumdurchperlung. Eine umfassende
Beschreibung dieser Techniken finden Sie in dem separaten
Handbuch, das dem Entgasungszubehör beiliegt.
Entsorgung von Lösungsmitteln
Sorgen Sie dafür, daß ein Auffangbehälter für Lösungsmittel oder
eine andere Auffangvorrichtung in Höhe des Arbeitstisches oder
darunter zur Verfügung steht. Für die meisten Lösungsmittel gelten
besondere Entsorgungsvorschriften; eine Entsorgung über die
Abwasserleitung ist hier nicht zulässig.
Bei der Entsorgung von Chemikalien gleich welcher Art sind
die einschlägigen Vorschriften streng zu beachten.
Hochdrucksysteme und Undichtigkeiten
Flüssigchromatographen arbeiten unter hohem Druck. Da
Flüssigkeiten kaum komprimierbar sind, können sie nicht viel Energie
speichern. Dementsprechend stellt der hohe Druck in einem
Flüssigchromatographen auch kaum eine unmittelbare Gefahr dar.
Jedoch sollten auftretende Undichtigkeiten umgehend beseitigt
werden. Schließlich ist noch zu empfehlen, bei der Arbeit mit einem
Flüssigchromatographen stets Augen und Haut zu schützen und vor
allen Wartungsarbeiten darauf zu achten, daß das Gerät abgeschaltet
und druckfrei gemacht wurde.
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Startup Checklist
Use this checklist to ensure that you have completed all the steps
necessary for the proper installation of your Finnigan™
SpectraSYSTEM™ UV/Vis detector. Complete installation
information can be found in Appendix A.
UNPACKING
❒
❒
❒
Unpack and inspect your instrument. Check for damage.
Check your accessory kit and manual. Are they:
❍
present?
❍
complete?
Read the Safety Information Card.
POSITIONING DETECTOR
❒
Place on benchtop as close as possible to the column and at
least 5 inches (13 cm) from the wall.
POWER CHECKOUT
❒
❒
Set voltage for local requirements.
Check that the correct fuses are installed.
REAR PANEL CONNECTIONS
❒
❒
❒
❒
01/2004
Insert the 8- and 12-pin green connectors into their
corresponding sockets.
Connect the Analog Output and the corresponding ground
terminals to your data system/recorder.
Connect the desired remote communications terminals to
external devices:
❍
STOP (Input)
❍
RUN (Input)
❍
ZERO (Input)
❍
READY (Output)
Connect the power cord.
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xix
FLOWCELL CONNECTIONS
❒
❒
❒
Remove the detector's front panel.
Remove the flowcell assembly from the detector.
Connect the flowcell inlet directly to your LC column
outlet.
❒
❒
Connect the flowcell outlet to waste tubing and a waste
container.
Replace the detector's front panel.
INSTRUMENT POWER-UP
❒
❒
Install the power cord and turn on the instrument.
Check that self-tests are running and that no error messages
appear.
❒
Check that the Status Screen appears on display.
❒
Complete and return the registration card.
REGISTRATION CARD
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List of Spare Parts,
Consumables, and Kits
Shown below is a list of spare parts and consumables
available from Thermo Electron for use with your
Finnigan SpectraSYSTEM UV/Vis detector. Contact your local
Thermo Electron representative for current prices.
Flowcells
9550-0100
9550-0234
9550-0197
9550-0053
9550-0265
9550-0101
9550-0263
Analytical LC (6 mm)
Analytical LC (10 mm)
Biocompatible LC (6 mm)
Microbore (3 mm)
Microbore (6 mm)
Semi-preparative, Open Column (3 mm)
Cuvette Cell Holder
Options And Accessories
2103-9119
A4095-010
9551-0022
9551-0023
9051-0143
External Events Connector
Remote Interface Cable
Tungsten Lamp, prealigned
Deuterium Lamp, prealigned
Regulated Backpressure Accessory
Manuals
A0099-540
UV/Vis Detectors Reference Manual (English)
Maintenance Parts
A4051-010
A4061-010
Standard Fittings Kit
(Kit includes stainless steel fittings and tubing used in a Finnigan
SpectraSYSTEM LC system.)
Inert/Biocompatible Fittings Kit
(Kit includes PEEK fittings and tubing used in an
inert/biocompatible Finnigan SpectraSYSTEM LC system.)
Upgrade Kits
Upgrade kits are available for the Finnigan SpectraSYSTEM UV1000 detector. Contact your local
Thermo Electron Representative for details.
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1
Getting Started
Introduction
This Chapter provides you with the three basic rules you'll need for
using your Thermo Electron, Finnigan SpectraSYSTEM UV/Vis
detector. It also introduces you to the instrument's command center
and describes the conventions we'll use in this manual.
Before you start this chapter, be sure to read the Safety Information
section beginning on page v of this manual and to install your
detector as described in Appendix A.
Throughout our explanations, we encourage you to explore the
general architecture of the instrument's menus and screens. Use the
Menu Tree in Appendix B as your guide if you wish. Learning Your
Way Around
AS EASY AS 1-2-3!
It's easy to learn your way around a SpectraSYSTEM detector. Just
remember these three rules:
1. The arrow keys ([∧], [∨], [<], [>]) 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 change values. Depending on the field, you will
scroll up or down through preset choices, or change
alphanumeric entries one letter or digit at a time.
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.
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VISUAL CLUES
The following conventions are used on the detector's display:
1. Top-level menu choices are displayed in all-capital 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 display line
(without a solid down-arrow).
Instrument Control
Take a look at the keypad and two-line display located on the front
panel (Fig. 1.1). 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
SpectraSYSTEM UV2000
ZERO
DET\Z008\FM
ENTER
Figure 1.1 The detector's command center
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THE 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 detector, a blank key called [ZERO].
The remaining keys either access commands ([MENU] and
[ENTER]), or are used to set parameters and move around the display
([∧], [∨], [<], [>], [+], [-]). The function of each is explained below.
[RUN]
Pressing [RUN] starts a run. The detector must be in the READY
state (or QREADY if a queue is loaded), indicating that the detector is
stabilized and waiting to begin a run.
[STOP]
Pressing [STOP] halts a run, stops the internal clock, and returns the
detector to a READY state. If a wavelength program is operating,
pressing [STOP] halts the program and returns the detector to its
initial conditions.
[STATUS]
Pressing [STATUS] displays the Status Screen (Fig. 1.1). From the
Status Screen you can monitor the run in progress. You can also
access the Status Menu. See page 5 for more information.
[ZERO]
The unlabeled key is the only variable key in the whole
SpectraSYSTEM family. On the detector, the blank key is the
[ZERO] key. The key's name appears on the nameplate below the
key.
Pressing [ZERO] resets the detector output to zero volts, plus or
minus any offset.
[MENU]
Pressing [MENU] displays the Main Menu (Figs. 1.2 and 1.1). See
page 4 for more information.
[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.
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3
[∧], [∨], [<], and [>]
Pressing any arrow key (up, down, left, or right) moves the cursor in
the direction indicated on the key. The up- and down-arrow keys also
move the cursor between menus and displays.
[+] 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.
In fields that require numerical 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 character entries, such as
the File Name field, the [+] and [-] keys 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, SCREENS,
AND MESSAGES
Your detector's display can show you three kinds of information:
menus, screens, and messages. Menus require you to make selections
or enter specific values. Screens display information that cannot be
edited. Messages confirm actions and point out errors. The Menu
Tree in Appendix B outlines the structure and content of the detectors'
menus and screens.
Main Menu
The Main Menu is the top level of the menu structure. In the
UV1000, (Fig 1.2) the Main Menu gives you access to four other
menus: FILE, COMMANDS, OPTIONS, and TESTS. In the
UV2000, there is and additional menu choice, QUEUE (Fig. 1.3). To
see the Main Menu, press the [MENU] key at any time.
>
❑ COMMANDS
FILES
❑ OPTIONS
❑ TESTS
Figure 1.2 The UV1000's Main Menu
>
❑ QUEUE
FILES
❑ COMMANDS
❑ TESTS
❑ OPTIONS
Figure 1.3 The UV2000's Main Menu
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From the UV1000’s and the UV2000's File(s) Menu you can edit,
load, delete or copy files. The UV2000 also lets you copy files. The
Commands Menu lets you insert an event mark onto your
chromatogram, short outputs, or shut down the detector. The Tests
Menu lets you run built-in instrument tests and diagnostics. 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 of files in the sample queue. Refer to Chapters 3, 4, 5, and
Appendix B for more information on any of the instrument's menus.
Status Screen
The Status Screen (Fig. 1.4) displays the detector status, wavelength
setting(s), and the absorbance reading. It automatically appears
whenever the instrument is powered on or the [STATUS] key is
pressed. No entries are made on the Status Screen.
Status
λ
AU
READY
250
0.00001
T
Figure 1.4 The Status Screen
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 your
detector's display: user messages, confirmation messages, and error
messages.
User Messages
User messages, indicated on the display by double asterisks, tell you
about an existing instrument condition or ask for further actions.
Some of these will only appear on the display for three seconds. An
example of a message requiring further action is shown in Figure 1.5.
** Protected File **
No Editing Allowed
Figure 1.5 An example of a user message
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Confirmation Messages
Confirmation messages (Fig. 1.6), also indicated on the display by
asterisks, appear for one second after an operation has been carried
out successfully.
** File Loaded **
Figure 1.6 An example of a confirmation message
Error Messages
Error messages (Fig. 1.7), indicated on the display with capital letters
and exclamation points, are shown 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.
!!
RAM ERROR
!!
Figure 1.7 An example of an error message
Manual Conventions
This manual uses several conventions. Among them are menu
displays, text conventions (brackets, slashes, etc.), standard words,
and several different icons.
DISPLAYS
Figure 1.8 shows how we depict the two-line display. Note that, in
menu illustrations, the triangular cursor location is indicated by a
caret (>).
>
❑ COMMANDS
FILE
❑ OPTIONS
❑ TESTS
Figure 1.8 A two-line menu display
Frequently the two lines shown on the display are only part of a
longer menu. In this manual, menus having more than two lines are
represented as in Figure 1.9.
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01/2004
Zero on λ Change
Yes
Cursor Speed
Medium
--------------------------------------------------------Status Lock
Off
READY Output
Active Hi
Figure 1.9 A menu longer than two lines
TEXT
Three typographic conventions are used to differentiate between keys,
menus, and fields.
Brackets
Brackets, [ ], indicate instrument keys. For example: Press [MENU].
Slashes
Slashes, / /, are used around menu choices. For example: From the
Main Menu, select /FILES/.
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: Select /FILES/, /Copy/, Copy File #.
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
preset value.
enter
The word "enter" is used when you need to specify individual
alphanumeric digits. To "enter" a particular value, move the cursor to
the desired field and use the [+] and [-] keys to increment or
decrement each digit in the field until the desired value or letter
appears.
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ICONS
This manual uses the following icons to alert you to various
situations. Each is called out by an icon in the left margin.
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.
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.
What's Next?
Now you're ready to try the practice example in Chapter 2: A Quick
Example.
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2
A Quick Example
Introduction
In Chapter 1, you read about the three easy rules for using your
detector's command center and some of its menus and screens. In this
chapter, you will find an example procedure that shows you how the
rules and keys actually work as you move through the various menus.
Instructions begin on page 10 for the UV1000 and on page 14 for
the UV2000.
This quick example uses only a fraction of the features available on
your detector and is included only as a first step in becoming familiar
with your new instrument.
After experimenting with this example, you'll want to turn to
Chapters 3 and 4, which cover the detector's basic and more advanced
operations. It is in those chapters that you'll learn about the full
capabilities of your detector. First though, to give you a general
understanding of the detectors' capabilities and design, we will briefly
describe the features and benefits of the UV1000 and UV2000 here.
THE UV1000
The UV1000 detector is a time-programmable, variable-wavelength
UV/Vis (ultraviolet/visible) absorbance detector. It operates in
single-wavelength mode in either the UV range (using a deuterium
lamp), or in the visible range (with an optional tungsten lamp). The
UV1000’s optical system has a novel, high light-throughput design
that provides high sensitivity detection along with maximal
application versatility. The UV1000 detector can be upgraded to a
UV2000.
THE UV2000
The UV2000 detector is a full-featured, time-programmable, dualwavelength UV/Vis absorbance detector. It operates in both singleand dual-wavelength modes in the UV and visible ranges. The
UV2000 offers the same optical system design as the UV1000. In
addition to the features of the UV1000, the UV2000 also offers
spectral scanning, a Develop File (for method development),
multiple file storage, a Queue feature (that allows you to link files),
and more.
BEFORE
YOU BEGIN
Once the detector is installed in your chromatographic system
according to the procedures described in Appendix A and you have
completed the Startup Checklist, you are ready to begin your quick
example.
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UV1000
An Example
In this example, we will show you how to prepare an edit file and
how to load the edit file into the detector's run file. After a practice
run, we will add a stop-time.
HINT: You may wish to keep the Menu Tree in Appendix B on hand as you
work through this example. If you lose your place at any time, you can:
1.
2.
STARTUP
Press the [∧] key to move back to a previous screen.
Or, press [STATUS] to return to the Status Screen and retrace
your steps.
Set the power switch located on the detector's rear panel to On. After
a series of power-up tests, the Status Screen (Fig. 2.1) appears on the
display. (We will discuss the Status Screen after you have set up your
operating parameters.)
Status
λ
AU
READY
250
0.00001
T
Figure 2.1 The UV1000's Status Screen
SETTING
PARAMETERS
To set your parameters, you need to prepare an edit file. The
following steps will show you how to access the Edit Menu and
prepare the file:
1. Press the [MENU] key. The detector's Main Menu appears on
the screen (Fig. 2.2).
>
❑ COMMANDS
FILE
❑ OPTIONS
❑ TESTS
Figure 2.2 The UV1000's Main Menu
2. Now select /FILES/ to display the Files Menu (Fig. 2.3).
>
❑ Load
Edit
❑ Delete
Figure 2.3 The UV1000's Files Menu
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3. Select /Edit/ to display the Edit Menu (Fig. 2.4).
>
Wavelength Program
❑
Options
Figure 2.4 The UV1000's Edit Menu
Wavelength
You use the Wavelength program to set the monitoring wavelength.
Wavelength is an example of a field that requires a numeric entry.
To set the wavelength:
1. From the Edit Menu (Fig. 2.4), select /Wavelength Program/ to
display the Wavelength Program (Fig. 2.5).
Time
0.00
Wavelength
254
Figure 2.5 The UV1000's wavelength program
2. Using the [+] and [-] keys, edit the wavelength field to the
desired setting for your analysis. Remember that each digit
must be edited individually.
3. Press [ENTER] to accept the new wavelength setting.
Range
Range is an example of a field that has a preset list of choices. To set
the range:
1. Select /Options/ from the Edit Menu (Fig. 2.4) to display the
Options Menu (Fig. 2.6).
Rise Time
1.0
Autozero Time
0.00
--------------------------------------------------------Range
1.0
Figure 2.6 The UV1000's Options Menu
2. Scroll down in the Options Menu and move the cursor to
Range 1 using the [∨] key.
3. Using the [+] or [-] key, select the desired setting from the list
of choices.
4. Press [ENTER] to accept the new range setting.
We will use the rise time and autozero time default settings for this
example. You will learn more about setting these parameters in
Chapter 3.
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Loading the File
You are now ready to load the settings from the edit file into the
detector's operating parameters (its run file). To load the file:
1. Return to the File Menu (Fig. 2.3) using the [∧] key.
2. Select /Load/. The screen in Figure 2.7 appears.
>Load File
Figure 2.7 The Load File command
3. Press [ENTER] to execute. The confirmation message shown
in Figure 2.8 appears for one second.
** File Loaded **
Figure 2.8 The file-loaded message
You are automatically returned to the Status Screen and are ready to
run your detector.
A PRACTICE RUN
Now you're ready for a practice run! Note that the Status Screen
(Fig. 2.1) now displays your wavelength setting, the detector's status,
and the absorbance reading. If the Status reads READY, the required
lamp is lit; if it reads NRDY (Not Ready), there is an error or the
lamp isn't lit; and if it reads UVW, the ultraviolet (D2) lamp is still
warming up.
When the baseline is stabilized:
1. Press the [ZERO] key to zero the detector's analog output
signal.
2. Inject your sample.
During setup, you may have noticed that there was no stop-time
entered in the detector's parameters. In this case, the detector stays in
the READY state and continually monitors the column eluant. You
do not need to manually start or stop a run with this set-up.
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ADDING A
STOP-TIME
To add a stop-time, you need to modify the detector's operating
parameters as follows. We will then show you how to start and stop a
run using the new setting.
1. From the Status Screen, press the [∨] key to move down to the
Status Menu (Fig. 2.9), which is the programming area below
the Status Screen. The cursor appears on the “tens” digit of the
wavelength value.
Time
Wavelength
0.00
250T
----------------------------------------------------------Rise Time
1.0
Autozero Time
0.00
Range
1.0
Figure 2.9 The UV1000's Status Menu
2. Using the [∨] key, move the cursor to the blank line below the
0.00 time line and press [+]. This adds a second line, with a
time of 1.00 and the same wavelength setting as the first.
Change 1.00 to the desired stop-time for the run, and leave the
wavelength unchanged.
3. To save your edits, scroll down to the words "Save File" which
now appear below Range, and press [ENTER]. The
confirmation message shown in Figure 2.10 appears and you
are automatically returned to the Status Screen.
** File Saved **
Figure 2.10 The file-saved message
RUNNING WITH
A STOP-TIME
Now that you have entered a stop-time, you will need to start the run
with each injection.
1. Zero the detector's analog output signal by pressing the
[ZERO] key.
2. When the detector is stabilized, inject your sample and press
[RUN].
Notice that Status now shows the run time. If you wish to stop your
run before the set stop-time, simply press [STOP].
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UV2000
An Example
In this example, specifically designed for the UV2000, we will show
you how to prepare a file and how to load it into the detector's
operating parameters. After a practice run, we will add a stop-time.
To keep the instructions simple, we will use the single-wavelength
mode.
HINT: You may wish to keep the Menu Tree in Appendix B on hand as you
work through this example. If you lose your place at any time, you can:
1.
2.
STARTUP
Press the [∧] key to move back to a previous screen.
Or, press [STATUS] to return to the Status Screen and retrace
your steps.
Set the power switch located on the detector's rear panel to On. After
a series of power-up tests, the Status Screen (Fig. 2.11) appears on the
display. (We will discuss the Status Screen after you have set up your
operating parameters.)
Status
λ
AU
READY
250
0.00001
T
Figure 2.11 The UV2000's Status Screen
SETTING
PARAMETERS
To set your parameters, you need to prepare an edit file. The
following steps will show you how to access the Edit Menu and
prepare the file:
1. Press the [MENU] key. The detector's Main Menu appears on
the screen (Fig. 2.12).
>
❑ QUEUE
FILES
❑ COMMANDS
❑ TESTS
❑ OPTIONS
Figure 2.12 The UV2000's Main Menu
2. Now select /FILES/ to display the Files Menu (Fig. 2.13).
>
Edit
❑ Load
❑ Copy
❑ Delete
Figure 2.13 The UV2000's Files Menu
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3. Select /Edit/ to display the Edit Menu (Fig. 2.14).
Edit File
1
File Name
--------------------------------------------------------> Wavelength Program
❑
Options
Figure 2.14 The UV2000's Edit Menu
For this example, we will use a file designation of 1 and leave the File
Name field blank.
Wavelength
Wavelength is an example of a field that requires a numeric entry. To
set the wavelength:
1. From the Edit Menu (Fig. 2.14), select /Wavelength Program/
to display the Wavelength Program (Fig. 2.15).
Program
Single λ
--------------------------------------------------------Time
Wavelength
0.00
254
Figure 2.15 The UV2000's wavelength program
2. Scroll down to the wavelength field.
3. Using the [+] and [-] keys, edit the wavelength field to the
desired setting for your analysis. Remember that each digit
must be edited individually.
4. Press [ENTER] to accept the new wavelength setting.
Range
Range is an example of a field that gives you a preset list of choices.
Note that Range 1 and 2 correspond to Analog Outputs 1 and 2 on the
rear panel of your detector. To set the range:
1. Select /Options/ from the Edit Menu (Fig. 2.14) to display the
Options Menu (Fig. 2.16).
Rise Time
1.0
Autozero Time
0.00
--------------------------------------------------------Range 1
1.0
Range 2
1.0
Figure 2.16 The UV2000's Options Menu
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2. Scroll down in the Options Menu and move the cursor to
Range 1 using the [∨] key.
3. Using the [+] or [-] key, select the desired setting from the list
of choices.
4. Press [ENTER] to accept the new Range 1 setting.
We will use the rise time, autozero time, and range 2 default settings
for this example. You will learn more about setting these parameters
in Chapter 3.
Loading the File
You are now ready to load the settings from File 1 into the detector's
operating parameters. To load the file:
1. Return to the Files Menu (Fig. 2.13) by pressing either
[ENTER] or the [∨] key.
2. Select /Load/. The screen in Figure 2.17 appears.
>
Load File
1:(filename)
Figure 2.17 The Load File command
3. You will be able to select from among several files in the Load
File field. Depending on whether or not your detector has ever
been used before, these files will either contain previously
stored settings or default settings. Use the [+] and [-] keys to
scroll through available choices. When the file you wish to
load appears (we're using the default settings for this example),
press [ENTER] to execute the load command.
4. The confirmation message shown in Figure 2.18 appears for
one second, after which you are automatically returned to the
Status Screen.
** File Loaded **
Figure 2.18 The file-loaded message
A PRACTICE RUN
16
Now you're ready for a practice run! Note that the Status Screen
(Fig. 2.11) now displays your wavelength setting, the detector's
status, and the absorbance reading. If the Status reads READY, the
required lamp is lit; if it reads NRDY (Not Ready), there is an error
(for example, you may have chosen a wavelength outside the selected
lamp's range) or the lamp isn't lit; and if it reads UVW, the ultraviolet
(D2) lamp is still warming up.
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When the detector is stabilized:
1. Press the [ZERO] key to zero the detector's analog output
signal.
2. Inject your sample.
During setup, you may have noticed that there was no stop-time
entered in the detector's parameters. In this case, the detector stays in
the READY state and continually monitors the column eluant. You
do not need to manually start or stop a run with this set-up.
ADDING A
STOP-TIME
To add a stop-time, you need to modify the detector's operating
parameters as follows. We will then show you how to start and stop a
run using the new setting.
1. From the Status Screen, press the [∨] key to move down to the
Status Menu (Fig. 2.19), which is the programming area below
the Status Screen.
File 1:
--------------------------------------------------------Time
Wavelength
0.00
250
T
Rise Time
Autozero Time
Range 1
Range 2
1.0
0.00
1.0
1.0
Figure 2.19 The UV2000's Status Menu
2. Using the [∨] key, move the cursor to the blank line below the
0.00 time line and press [+]. This adds a second line, with a
time of 1.00 and the same wavelength setting as the first.
Change 1.00 to the desired stop-time for the run, and leave the
wavelength unchanged.
3. To save your edits, scroll down to the words "Save File" which
now appear below Range 2, and press [ENTER]. The
confirmation message shown in Figure 2.20 appears and you
are automatically returned to the Status Screen.
** File Saved **
Figure 2.20 The file-saved message
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RUNNING WITH
A STOP-TIME
Now that you have entered a stop-time, you will need to start the run
with each injection.
1. Zero the detector's analog output signal by pressing the
[ZERO] key.
2. When the detector is stabilized, inject your sample and press
[RUN].
Notice that Status now shows the run time. If you wish to stop your
run before the set stop-time, simply press [STOP].
What's Next?
Once you have completed this example and are comfortable with the
keypad and display, proceed to Chapter 3, Basic Operations, to learn
more about your detector.
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3
Basic Operations
Introduction
This Chapter provides step-by-step instructions for the most
frequently used detector operations, including setup and run
procedures for single- and dual-wavelength modes, detector file
management and protection, and analog output operations.
To keep the instructions easy to follow, we have divided the Chapter
into two sections. Instructions for the UV1000 begin on page 20.
Instructions for the UV2000 begin on page 26. You may wish to
keep the Menu Tree and the Menu Reference from Appendix B on
hand as you work through this chapter.
NOTE: You should be aware that your display's values may differ from
those presented in this manual, especially if the detector has been previously
programmed.
Before You Begin
Before you begin this chapter, your detector should be installed in a
chromatographic system (see Appendix A), and you should have
completed the Startup Checklist located at the front of this manual.
We also recommend that you review Chapter 1, Getting Started,
which includes general instructions for using the detector keypad and
which lists the conventions used throughout this manual.
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UV1000
Single-wavelength Operation
The UV1000 uses a standard deuterium lamp to operate in a singlewavelength mode in the ultraviolet (UV) range. Adding an optional
tungsten lamp increases the detector’s capabilities to the visible (Vis)
range.
To perform a single-wavelength operation, you first enter the desired
detector parameters into an edit file. You then load the edit file into
the run file, which contains the detector's current operating
parameters. These instructions will show you how to start and stop a
run, and how to modify the detector's operating parameters.
SETTING
PARAMETERS
You set up the UV1000’s parameters by using the File Menu to
prepare an edit file. You then load the edit file into the run file.
To access the File Menu, first press [MENU]. The Main Menu
appears on the screen. From the Main Menu, select /FILE/. The
menu shown in Figure 3.1 will appear.
>
❑ Load
Edit
❑ Delete
Figure 3.1 The UV1000's Files Menu
From the File Menu, select /Edit/ to display the Edit Menu. The Edit
Menu (Fig. 3.2) selections are /Wavelength Program/, which contains
time and wavelength fields, and /Options/, which contains the Rise
Time, Autozero Time, and Range fields.
❑
Wavelength Program
❑
Options
Figure 3.2 The UV1000's Edit Menu
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Wavelength
Program
Select /Wavelength Program/ from the Edit Menu. The Wavelength
Program is a Table containing the Time and Wavelength fields
(Fig. 3.3).
Time
0.00
Wavelength
254
Figure 3.3 The UV1000's Wavelength Program
In the single-wavelength mode, you can operate with either a one-line
or a two-line wavelength program. Using a one-line program, the
detector is always in the READY state and you can continually
monitor the baseline. Using a two-line program, you can use a stopline and you can start and stop the detector during a chromatographic
run. (Stop-lines are useful, for example, in an automated series of
runs where you want to autozero the detector's baseline after each
injection.)
For a one-line program, enter the wavelength(s) for your analysis in
the Wavelength field that corresponds to the time of 0.00.
For a two-line program, add an additional line (the stop-line) by
scrolling down to the blank line below the time 0.00 line and pressing
[+]. The second line will automatically have a time setting of 1.00
and the same wavelength setting(s) as the first. Change 1.00 to the
desired stop-time for the run, and leave the wavelength value
unchanged.
An example of a two-line wavelength program for a nine-minute run
at 283 nm is shown in Figure 3.4.
Time
Wavelength
0.00
283
--------------------------------------------------------9.00
283
Figure 3.4 An example of a two-line wavelength program with
a programmed stop-time
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Options Menu
Select /Options/ from the Edit Menu to display the Options Menu
(Fig. 3.5). Use this menu to set the detector's rise time, autozero time,
and range.
Rise Time
1.0
Autozero Time
0.00
------------------------------------------------------Range
1.0
Figure 3.5 The UV1000's Options Menu
Rise Time
This field controls the detector's response time. Rise time is inversely
proportional to the amount of baseline noise. For example, the longer the
rise time, the less noise detected. The one-second default value is
appropriate for most applications.
HINT: To minimize baseline noise while retaining maximum resolution, select a
rise time that is at least one-tenth of the peak width at the base of the narrowest
peak of interest.
Autozero Time
This parameter tells the detector when to perform an automatic zero
of the baseline. If you do not wish to set an autozero and you are
using a stop-line in your wavelength program, simply set the autozero
time to a value greater than your stop-time.
HINT: It is good practice to zero the detector automatically at the start of
each run. This will keep the detector output in range throughout an
automated series of runs.
Range
This parameter ranges the signal from the Ranged Output. (labeled as
“Ranged Output” on the detector's rear panel). Set the range to an
appropriate full-scale absorbance for your sample. For more
information on the use of ranges and analog outputs, see pages 25
and 84.
HINT: We recommend a range of 1.0 when you are using an integrator or
data system.
Loading the Edit File
When you are ready to load the settings from the edit file into the
detector’s run file, select /Load/ from the File Menu. The screen will
display the words "Load File." Press [ENTER] to accept the settings.
The confirmation message shown in Figure 3.6 will appear for one
second. You are then returned to the Status Screen.
** File Loaded **
Figure 3.6 The file-loaded message
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RUNNING YOUR
DETECTOR
Once you've set your detector parameters in the edit file and have
loaded the parameters into the run file, you’re ready to run your
analysis. First, check the detector's status by viewing the Status
Screen. If you’re using a stop-line in your wavelength program, you
will start and stop the run with each injection.
Status Screen
You can check the detector's status, wavelength setting, and
absorbance reading from the Status Screen (Fig. 3.7). To access the
Status Screen, press [STATUS].
Status
λ
READY
254
AU
+0.00001 T
Figure 3.7 The UV1000's Status Screen
If the Status reads READY, the detector is stabilized and ready to run.
If NRDY appears, the detector's lamps may need additional time to
warm up, or a wavelength outside the selected lamp's range may have
been chosen.
Inject your Sample
When the detector is stabilized and you are ready to inject your
sample, first manually zero the detector by pressing the [ZERO] key.
If you are not using a stop-line in the wavelength program, the
detector remains in the READY state throughout your
chromatographic runs. If you are using a stop-line, you must start and
stop the run with each injection, following the procedures below.
Starting a Run
If you are using a stop-line in your wavelength program, you need to
start the run with each injection. There are two ways to start a run
using the UV1000:
1. Manually, by pressing [RUN] each time you make an injection.
2. Automatically, by interfacing the detector with a remote runsignal from the injector (see Appendix A for details). In this
scenario, a signal that is equivalent to pressing [RUN] is
automatically sent from the injector to the detector with each
injection.
During the run, you can monitor the run time from the Status Screen.
Stopping a Run
There are two ways to stop a run:
1. Manually, by pressing [STOP] before the programmed stoptime.
2. Automatically, by allowing the run to finish at the programmed
stop-time.
In either case, the detector returns to its READY state.
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CHANGING RUN
PARAMETERS
There are two ways to change the detector's run parameters:
1. You can use the Files Menu and follow the procedures outlined
under "Setting Parameters" on page 20.
2. Or you can use the Status Menu, which is the programming
area below the Status Screen.
Each method has a distinct advantage. Programming in the Status
Menu allows you to change the detector's current operating
parameters, even while the detector is running. Programming in the
Files Menu allows you to prepare an edit file containing the changes
without altering the current detector settings. The file may then be
loaded later.
Status Menu
From the Status Screen, scroll down to the Status Menu (Fig. 3.8).
The Status Menu contains the Wavelength Program, Rise Time,
Autozero Time, and Range.
Time
0.00
Wavelength
254
T
--------------------------------------------------------Rise Time
1.0
Autozero Time
0.00
Range
1.0
Figure 3.8 The UV1000's Status Menu
The parameters are set using the same instructions given under
“Wavelength Program” and “Options Menu,” starting on page 21.
When you use the Status Menu to change the UV1000 settings, each
change is effective immediately upon leaving the field.
Notice the words “Save File” below the Range field. Press [ENTER]
when the cursor is in the Save File field to save the new settings to the
run file. The confirmation message shown in Figure 3.9 will appear
briefly.
** File Saved **
Figure 3.9 The File Saved message
NOTE: When you change the detector settings from the Status Menu, the
contents of the edit file do not change. Only the run file values are modified.
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To return to your previous setting without saving the new ones, do not
press [ENTER]. Instead, you may reenter the unaltered file, as follows::
1. Press [MENU].
2. Select /FILE/.
3. Select /Load/.
4. The words "Load File" will appear on the screen. Press
[ENTER].
A confirmation message (Fig 3.6) will appear for one second. You
are then returned to the Status Screen, and all settings will contain
their original values.
DELETING THE FILE
To delete the edit file, select /Delete/ in the File Menu. The words
“Delete File” will appear on the screen. When you press [ENTER],
the confirmation message shown in Figure 3.10 appears briefly, and
the display returns to the File Menu. The edit file parameters return
to their default settings.
** File Deleted **
Figure 3.10 The File- Deleted message
Analog Outputs
There are two analog outputs for the UV1000: Analog Output 1and
Analog Output 2. On the detector’s rear panel, they appear as
“Unranged output” and “Ranged Output.” Analog Output 1 is set at 1
V/AU and is intended for an integrator interface. Analog Output 2 is
range-selectable and is used for recorders and other devices. Rearpanel connections for both outputs are discussed on page 84.
ANALOG OFFSETS
Analog offsets may be used when there is a high background
absorbance reading, or when there is considerable baseline drift from
your chromatographic system and you are unable to keep your
integrator’s (recorder’s) signal on-scale.
Because integrators have very limited capacity for handling negative
signals, you may wish to set a small positive offset (1%) when using
an integrator.
Use negative offsets with recorders, where you may wish to set the
pen at either side of the strip chart.
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Offset options are selectable from the Analog Outputs Menu.
To access these options:
1. Press [MENU].
2. Select /OPTIONS/
3. Select /Analog Outputs/.
The Analog Outputs Menu is shown in Figure 3.11.
Analog 1 Offset (mV)
Analog 1 Offset (%)
0
0
Figure 3.11 The UV1000's Analog Outputs Menu
HINT: Although the default for the Analog 1 offset is set at zero, we
recommend a 1 mV setting for use with your data system or integrator.
UV2000
Single- and Dual-wavelength Operation
You can operate the UV2000 in either a single- or a dual-wavelength
mode. In the dual-wavelength mode, the detector simultaneously
monitors two wavelengths in either the UV range or the visible range
in a single run.
To perform a single- or dual-wavelength operation, you need to be
able to identify and enter a file, load that file into the detector's
current operating parameters, and start and stop a run. This section
will also show you how to modify the detector's current operating
parameters.
SETTING
PARAMETERS
Before you set any detector parameters, you need to access the Files
Menu to identify the file you wish to edit.
To access the Files Menu, first press [MENU]. The Main Menu
appears on the screen. From the Main Menu, select /FILES/. The
menu shown in Figure 3.12 will appear.
>Edit
❑ Load
❑ Copy
❑ Delete
Figure 3.12 The UV2000's Files Menu
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Select /Edit/ from the Files Menu to display the Edit Menu
(Fig. 3.13).
Edit File
1
File Name
--------------------------------------------------------❑
Wavelength Program
❑
Options
Figure 3.13 The UV2000's Edit Menu
File Identification
Enter the number of the file you wish to edit in the Edit File field.
The UV2000 can store up to four files in memory, so file numbers
from 1 to 4 are allowed. You may also enter a name of up to eight
characters in the File Name field.
While in the Edit File, you will see file choices of "S" and "D" that
represent the Scan and Develop files, respectively. These files are
some of the UV2000's advanced features that you will learn about in
Chapter 4.
Wavelength
Program
From the Edit Menu, select /Wavelength Program/. The Wavelength
Program designates dual- or single-wavelength operation, and also
contains a Table of time and wavelength. A wavelength program for
dual-wavelength operation appears in Figure 3.14.
Program
Dual λ(190-450)
--------------------------------------------------------Time
λ1
λ2
0.00
254
280
Figure 3.14 The UV2000's Wavelength Program
in dual-wavelength mode
Select Single λ, Dual λ(190-450), or Dual λ(366-700) in the Program
field. The Table for time and wavelength(s) will appear. (For singlewavelength operation, there is only one wavelength field.)
You can operate with either a one-line or a two-line wavelength
program. Using a one-line program, the detector is always in the
READY state and you can continually monitor the chromatographic
eluant. Using a two-line program, you can use a stop-line and you
can start and stop the detector during a chromatographic run. (Stoplines are useful, for example, in an automated series of runs where
you want to autozero the detector's baseline after each injection.)
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For a one-line program, enter the wavelength(s) for your analysis in
the λ1 and λ2 (or Wavelength) fields that correspond to the time
of 0.00.
For a two-line program, add an additional line (the stop-line) by
scrolling down to the blank line below the time 0.00 line and pressing
[+]. The second line will automatically have a time setting of 1.00
and the same wavelength setting(s) as the first. Change 1.00 to the
desired stop-time for the run, and leave the wavelength value(s)
unchanged.
An example of a dual-wavelength, nine-minute run at 254 and
283 nm is shown in Figure 3.15.
λ1
Time
λ2
0.00
254
283
--------------------------------------------------------9.00
254
283
Figure 3.15 A wavelength program with
a programmed stop-time
Options
Select /Options/ from the Edit Menu to display the Options Menu
(Fig. 3.16). Use this menu to set the detector's rise time, autozero
time, and ranges.
Rise Time
1.0
Autozero Time
0.00
-------------------------------------------------------Range 1
1.0
Range 2
1.0
Figure 3.16 The UV2000's Options Menu
Rise Time
This field controls the detector's response time. Rise time is inversely
proportional to the amount of baseline noise. For example, the longer
the rise time, the less noise detected. The one-second default value is
appropriate for most applications.
HINT: To minimize baseline noise while retaining maximum resolution,
select a rise time that is at least one-tenth of the peak width at the base of the
narrowest peak of interest.
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Autozero Time
This parameter tells the detector when to perform an automatic zero
of the baseline. If you do not wish to set an autozero and you are
using a stop-line in your wavelength program, simply set the autozero
time to a value greater than your stop-time.
HINT: It is good practice to zero the detector automatically at the start of
each run. This will keep the detector output in range throughout an
automated series of runs.
Range 1 and 2
These parameters range the signal from Analog Output 1 and Analog
Output 2 (shown as ANLG 1 Output and ANLG 2 Output on the
detector's rear panel). Set each range to an appropriate full-scale
absorbance for your sample. For more information on the use of
ranges and analog outputs, see pages 35 and 84.
HINT: We recommend a range of 1.0 when you are using an integrator or
data system.
Loading a File
When you are ready to load a file into the detector settings, select
/Load/ from the Files Menu. The screen will display the words "Load
File 1:(filename)." Use the [+]/[-] keys to view the number and name
of available files. When the desired file number appears, press
[ENTER].
The confirmation message shown in Figure 3.17 will appear for one
second. You are then returned to the Status Screen.
** File Loaded **
Figure 3.17 The message that's displayed
when a file is loaded
NOTE: When a dual-wavelength program is loaded, you'll hear the motor
start to operate in dual-wavelength mode even though you didn't press
[RUN].
RUNNING YOUR
DETECTOR
01/2004
Once you've set your detector parameters in the designated file and
have loaded the file into the detector's operating parameters, you are
ready to run your analysis. First, check the detector's status by
viewing the Status Screen. If you are using a stop-line in your
wavelength program, you will start and stop the run with each
injection.
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Status Screen
You can check the detector's status, wavelength setting(s), and
absorbance reading(s) from the Status Screen. To access the Status
Screen, press [STATUS]. The Status Screen for the UV2000 in dualwavelength mode appears below (Fig. 3.18). Note that, in the singlewavelength mode, the third line does not appear.
Status
λ
AU
READY
254
+0.00001 T
--------------------------------------------------------280
-0.00001
Figure 3.18 The UV2000's Status Screen
for dual-wavelength operation
If the Status reads READY, the detector is stabilized and ready to run.
If NRDY appears, the detector's lamps may need additional time to
warm up, or a wavelength outside the selected lamp's range may have
been chosen.
Inject your Sample
When the detector is stabilized and you are ready to inject your
sample, first manually zero the detector by pressing the [ZERO] key.
If you are not using a stop-line in the wavelength program, the
detector remains in the READY state throughout your
chromatographic runs. If you are using a stop-line, you must start and
stop the run with each injection, following the procedures below.
Starting a Run
If you are using a stop-line in your wavelength program, you need to
start the run with each injection. There are two ways to start a run
using the UV2000:
1. Manually, by pressing [RUN] each time you make an injection.
2. Automatically, by interfacing the detector with a remote runsignal from the injector (see Appendix A for details). In this
scenario, a signal that is equivalent to pressing [RUN] is
automatically sent from the injector to the detector with each
injection.
During the run, you can monitor the run time from the Status Screen.
Stopping a Run
There are two ways to stop a run:
1. Manually, by pressing [STOP] before the programmed stoptime.
2. Automatically, by allowing the run to finish at the programmed
stop-time.
If you're conducting a dual-wavelength run, you can also stop the run
by loading a single-wavelength file.
Regardless of how you stop a run, the detector returns to READY.
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CHANGING RUN
PARAMETERS
If you wish to change the detector's parameters:
1. You can use the Files Menu and follow the procedures outlined
under "Setting Parameters" on page 26.
2. Or you can use the Status Menu, which is the programming
area below the Status Screen.
Each method has a distinct advantage. Programming in the Status
Menu allows you to change the detector's current operating
parameters, even while the detector is running. Programming in the
Files Menu allows you to prepare an edit file containing the changes
without altering the current detector settings. The file may then be
loaded later.
Status Menu
From the Status Screen, scroll down to the Status Menu (Fig. 3.19).
The Status Menu contains the loaded file identification (its number
and name), Wavelength Program, Rise Time, Autozero Time, and
Ranges.
File 1:
--------------------------------------------------------Time
λ1
λ2
0.00
254
280
T
Rise Time
Autozero Time
Range 1
Range 2
1.0
0.00
1.0
1.0
Figure 3.19 The UV2000's Status Menu
for dual-wavelength operation
The Status Menu shown in Figure 3.19 is typical for dual-wavelength
operation. In the single-wavelength mode, only one wavelength field
appears in the wavelength program.
The detector's parameters are set following the same instructions
given under "Wavelength Program" and "Options Menu," starting on
page 27. However, you cannot modify either the file identification or
the wavelength mode (dual or single) from the Status Menu.
NOTE: When you modify a file's parameters from the Status Menu, you do
not change the contents of the same file number stored in the detector's
memory. Only the copy of the active file is modified.
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Saving the File
When you change the UV2000's settings from the Status Menu, each
change is effective as soon as you leave the field. You'll also see that
the File identification on the first line of the Status Menu (Fig. 3.19)
now reads "File N:xxxx-changed" (where N:xxxx is the file number
and name) and that the words "Save File" now appear below Range 2.
To save the changed file, press [ENTER]. The confirmation message
shown in Figure 3.20 will appear briefly.
** File Saved **
Figure 3.20 The File Saved message
To keep the original file without saving the changes, don't press
[ENTER]. Instead, reload the unaltered file using the Files Menu as
follows:
1. Press [MENU].
2. Select /FILES/.
3. Select /Load/.
4. The words "Load File" will appear on the screen. Enter the
desired file number and press [ENTER].
A confirmation message (Fig 3.17) will appear for one second. You
are then returned to the Status Screen, and all settings will contain
their original values.
More about Files
On page 26, you learned how to edit and load files from the Files
Menu. The UV2000 also allows you to copy and delete files (and to
protect files from being edited, copied to, or deleted) in a few, easy
steps.
COPYING FILES
To copy a file:
1. Press [MENU].
2. Select /FILES/ to display the Files Menu (Fig. 3.21).
> Edit
❑ Copy
❑ Load
❑ Delete
Figure 3.21 The UV2000's Files Menu
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3. Select /Copy/. The Copy Menu will appear on the screen
(Fig. 3.22).
>
Copy File
1: (filename1)
❑
To File
2: (filename2)
Figure 3.22 The UV2000's Copy Menu
4. Enter the identification number for the file you wish to copy in
the Copy File field.
5. Enter the number of the file to which you wish to copy in the
To File field.
6. Press [ENTER]. The confirmation message shown in
Figure 3.23 appears briefly, and you are returned to the Files
Menu.
** File Copied **
Figure 3.23 The message that's displayed
when a file is copied
If you attempt to copy to a protected file (see the section below, titled
"Protecting Files"), you will get the message shown in Figure 3.24. If
a file is not protected, make sure it's empty or unwanted before you
copy to it, as it will be overwritten.
** Protected File **
Cannot Be Copied To
Figure 3.24 The message that is displayed
when you attempt to copy to a protected file
You cannot use Copy for the Scan or Develop files. (You will learn
more about these files in Chapter 4.)
DELETING FILES
To delete a file:
1. Press [MENU].
2. Select /FILES/ to display the Files Menu (Fig. 3.21).
3. Select /Delete/. The Delete File field will appear on the screen.
4. Enter the identification number of the file you wish to delete.
When you press [ENTER], the confirmation message shown in
Figure 3.25 appears briefly and the display returns to the Files
Menu. (The parameters in the file you have just deleted return
to their default values.)
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** File Deleted **
Figure 3.25 The message that's displayed
when a file is deleted
If you attempt to delete a protected file (see the next section,
"Protecting Files"), you will get the message shown in Figure 3.26.
** Protected File **
Cannot Be Deleted
Figure 3.26 The message that's displayed
when you try to delete a protected file
PROTECTING FILES
The UV2000 allows you to protect files from being edited, copied to,
or deleted. To access the file protection operation, follow these steps:
1. Press [MENU].
2. Select /OPTIONS/. The Options Menu appears in Figure 3.27.
>
Lamps
❑
Analog Outputs
--------------------------------------------------------❑
More
Figure 3.27 The UV2000's Options Menu
3. Select /More/. The More Menu appears in Figure 3.28.
Zero on λ Change
Yes
Cursor Speed
Medium
--------------------------------------------------------Status Lock
Off
READY Output
Active Hi
File Name
Protect
1:
Off
2:
Off
3:
Off
4:
Off
Figure 3.28 The UV2000's More Menu
4. Scroll down to the Table containing the fields File Name and
Protect. To protect a file, select On in the Protect field
corresponding to the appropriate file number. To remove the
file protection, select Off.
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Analog Outputs
There are two analog outputs on the UV2000, Analog Output 1 and
Analog Output 2. On the detector's rear panel, they appear as
ANLG 1 Output and ANLG 2 Output. Rear-panel connections for
both outputs are discussed on page 84.
ANALOG OUTPUT 1
By default, Analog Output 1 is either the absorbance reading for
single-wavelength operation, or the absorbance reading of wavelength
one (λ1) for dual-wavelength operation.
ANALOG OUTPUT 2
Analog Output 2 is selectable (AU, AU1-K*AU2, and AU1/AU2),
and so can be used to monitor several different outputs. To access
these options:
1. Press [MENU].
2. Select /OPTIONS/.
3. Select /Analog Outputs/. The Analog Outputs Menu shown in
Figure 3.29 appears.
Analog 1 Offset (%)
0
Analog 2 Offset (%)
0
--------------------------------------------------------Analog 2
AU
K Factor
1.000
Figure 3.29 The UV2000's Analog Outputs Menu
4. Scroll down to Analog 2. The selections are:
01/2004
•
AU, which is either the same absorbance reading you got
from Analog Output 1 in single-wavelength operation, or
the absorbance reading of wavelength two (λ2) for dualwavelength operation.
•
AU1-K*AU2, which is the readout of the suppressed signal
using the K-Factor technique. See page 60 for more
details.
•
AU1/AU2, which is the ratio of the dual-wavelength
absorbance values. This ratio is sometimes used to check
peak purity. See page 64 for more details.
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ANALOG OFFSETS
Both analog outputs 1 and 2 can be offset on the UV2000. Analog
offsets may be used in cases where there is a high background
absorbance reading, or when there is considerable baseline drift from
your chromatographic system and you are unable to keep your
integrator's (recorder's) signal on-scale.
Because integrators have very limited capacity for handling negative
signals, you may wish to set a small positive offset (1%) when using
an integrator.
Negative offsets are available for use with recorders, where you may
wish to set the pen at either side of the strip chart.
The offset options are selectable from the Analog Outputs Menu
shown in Figure 3.29.
HINT: Although the offset for each output is set at 0% of full-scale readout
by default, we recommend a 1% setting for use with your data system or
integrator.
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4
Advanced Operations
Introduction
In this chapter, you will learn to use the more advanced capabilities of
your detector. The chapter’s first section covers the capabilities
offered by both the UV1000 and the UV2000 detectors; the second
section, beginning on page 46, contains those functions specific to the
UV2000. You should be familiar with the instructions presented in
Chapter 3, Basic Operations, before you begin.
UV1000 and UV2000
Wavelength Programming
Your detector can change wavelength as a function of time, a feature
we call Wavelength Programming. This feature gives you maximum
detection sensitivity for each component of a mixture without making
multiple injections of the sample.
NOTE: A wavelength program can be built in either the Status Menu or the
File(s) Menu.
BUILDING THE
PROGRAM
In wavelength programming, you enter time lines into a "Wavelength
Program." Each time line specifies the time at which you want a
wavelength change to occur.
The following instructions are for single-wavelength operation, but if
you have a UV2000, you can build a dual-wavelength program using
the same procedure.
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Initial Conditions
Access the Wavelength Program (Fig. 4.1) through either the Status
Menu or the Files Menu.
Time
0.00
Wavelength
250
Figure 4.1 The wavelength program for single-wavelength operation
The initial time entry is 0.00. Move the cursor to the corresponding
Wavelength field, and enter the initial wavelength for your analysis.
Adding Lines
To add a second time line, scroll down to the first blank line and press
[+]. The second line will automatically have a time setting of 1.00
and the same wavelength setting as the first. Change the Time and
corresponding Wavelength fields to the desired values. Subsequent
lines are added in the same fashion.
A wavelength program may contain as many as ten lines for a single
run. On the UV1000, all of the lines’ wavelengths must be in the
same range (either UV or visible). On the UV2000, however, you
can cross between the UV and visible ranges (in single-wavelength
mode only).
If you enter time lines out of sequence, the detector will automatically
sort the lines and place them all in chronological order.
The Stop-line
The last line of the program (the stop-line) lists the time at which the
detector will automatically end the run and return to initial conditions.
Since wavelength is not important in the stop-line, it can be set to any
value.
NOTE: Remember, the last line of the program is always the detector's
signal to end a run; it is not a programmed wavelength change!
Deleting a Line
38
To delete an entire time line, place the cursor in the Time field and
press [-] repeatedly until the value goes blank. When you leave the
line, it will be deleted.
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An Example
Figure 4.2 shows a completed wavelength program for singlewavelength operation.
Time
Wavelength
0.00
254
--------------------------------------------------------5.00
280
7.00
265
10.00
265
Figure 4.2 An example of a completed
wavelength program
In our example, the initial detection wavelength is 254 nm. At 5.00
minutes into the run, the wavelength changes to 280 nm. At 7.00
minutes, it changes to 265 nm. The run ends at 10.00 minutes, and
the detector returns to its initial wavelength of 254 nm and to its
READY state.
RUNNING THE
PROGRAM
After you set the rest of your parameters, the detector is ready to run.
It is good practice to zero the detector at the beginning of every run
and at each wavelength change. See the next section, titled
"Programmed Autozero," for details.
Once you start the run, you may edit any timed event (wavelength
change, autozero, or stop-time) that has not yet taken place. These
edits can only be made from the Status Menu however! Each edit is
entered immediately into the detector's operating wavelength
program.
For example, for the program displayed in Figure 4.2, the stop-time is
10.0 minutes. If, at 7.00 minutes into the run, you determine that the
run should be 9.00 minutes long, you can edit the last line of the
program such that the current run will stop at 9.00 minutes.
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Programmed Autozero
The detector can be programmed to perform an automatic zero with
each wavelength change during a run using the Zero on λ Change
field. To access this feature:
1. Press [MENU] and select /OPTIONS/ to access the Options
Menu (Fig. 4.3).
>
Lamps
❑
Analog Outputs
--------------------------------------------------------❑
More
Figure 4.3 The Options Menu
2. Select /More/ to display the More Menu.
3. Place the cursor on the Zero on λ Change field. This field
appears on the first line of the More Menu for both the
UV1000 and the UV2000.
4. Select Yes, to automatically zero the detector response with
each wavelength change during a run, or No, to turn this
feature off.
You can also use this automatic zero feature to add autozeros into
your wavelength program without changing the detector's wavelength
settings. To do this, simply add additional time lines. Adding
autozeros in this way is convenient in cases such as solvent
programming, where the detector's baseline may drift due to changes
in solvent background.
An example program is shown in Figure 4.4.
Time
Wavelength
0.00
254
--------------------------------------------------------2.00
254
5.00
280
7.00
280
10.00
280
Figure 4.4 An example of a wavelength program
with automatic autozeros
With Zero on λ Change set to Yes, the detector will autozero at 2.00,
5.00, and 7.00 minutes into the run, even though the wavelength will
only change once (at 5.00 minutes into the run).
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Automatic Lamp Operations
THE LAMPS MENU
The Lamps Menu (Fig. 4.5) allows you to select lamps, track lamp
life, and turn the lamps on and off automatically. Field descriptions
for this menu follow.
To access the Lamps Menu:
1. Press [MENU] and select /OPTIONS/.
2. Select /Lamps/.
Lamp
D2 (190-365)
D2 Lamp Hours
0
--------------------------------------------------------W Lamp Hours
0
Current Time
0:00
Startup
Manual
Startup Time
0:00
Shutdown
Manual
Shutdown Time
0:00
Time from READY
1:00
Figure 4.5 The UV2000's Lamps Menu
Lamp
The Lamp field allows you to select from the following:
•
D2 (190-365), for deuterium
[the UV1000 reads D2 (190-380)],
•
W (366-800), for tungsten,
•
D2+W (190-800), for dual-lamp operation
•
or Off, to shut the lamp(s) off.
Actually though, the wavelength setting in the loaded file
automatically selects the appropriate lamp for you. In fact, the
wavelength setting you choose in your file has priority over any
selection you make here in the Lamp field!
For example, if the loaded file designates a wavelength in the UV
range, but you manually selected W (366-800) in the Lamp field, the
detector's display will read NRDY (not ready) for the deuterium lamp.
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Lamp Hours (W and D2 fields)
These fields automatically track the number of hours each lamp has
been in operation. For the value to be accurate, set the appropriate
Lamp Hours field to zero each time you install a new lamp.
HINT: If you switch lamps before they are burned out (with the intention of
using them again at a later date), keep a record of how many hours they
have been in operation.
Startup and Shutdown
When you set the Startup and Shutdown fields to "Manual," the lamp
designated in the Lamp field turns on and off when the detector
power is switched on and off, respectively.
Startup and Shutdown Times
When you set the Startup and Shutdown fields to "Time" (see above),
the designated lamp will automatically turn on and off at the local
time set in the Startup Time and Shutdown Time fields, respectively.
NOTE: For the detector to perform automatic lamp startup and shutdown
correctly, the detector's 24-hour clock must be set to your local time. Set the
clock in the Current Time field. Since the clock resets to zero each time the
detector is turned off, it will have to be reset prior to performing automatic
lamp startup and shutdown unless the detector has been left on continuously.
Time from READY
If you prefer, you can program the detector to shut the lamp off after a
series of automated runs by using the Time from READY feature.
Time from READY is a preset time interval that automatically begins
each time the detector returns to its READY state. If the Time from
READY interval elapses without a run signal being received from
either the keypad or the detector's Run(Input) terminal, the detector's
lamp turns itself off.
To use the Time from READY feature:
1. Select Time from READY in the Shutdown field.
2. In the Time from READY field, enter the length of time during
which a run signal must be received by the detector before the
lamp turns off.
For example, let's say your chromatographic system is set up for an
automated run and the autosampler signals the detector to run after
each injection. With the detector settings shown in Figure 4.6, the
lamp will turn off ten hours after the last run is completed.
Shutdown
Time from READY
Shutdown Time
00:00
--------------------------------------------------------Time from READY
10:00
Figure 4.6 An example of the Time from Ready feature
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You can also program the UV2000's lamps to turn off at the end of a
queue by selecting End of Queue in the Shutdown field. For more
information on the Queue feature, see page 57.
Other Features
Additional features offered by the UV1000 and UV2000 include the
abilities to lock the Status Screen, to short the detector outputs, to
place an event mark on the chromatogram, and to send a ready signal
to external devices. You can also control the display's contrast and
cursor speed, and do a quick shutdown of the detector's lamps and
motors.
STATUS LOCK
You can lock the detector's display using the Status Lock field. This
feature lets you prevent accidental changes to a file that is currently
being run. With the lock on in the UV1000, only the Status Screen
appears. In the UV2000, you can scroll down from the Status Screen
as far as the Status Menu's File Name field. You will still be able to
access the Main Menu and the rest of the menu structure using the
[MENU] key however.
To access Status Lock:
1. Press [MENU].
2. Select /OPTIONS/.
3. Select /More/.
4. Scroll down to Status Lock. Select On or Off to turn the lock
on or off, respectively.
5. Press [STATUS].
SHORT OUTPUTS
When zeroing a readout device such as an integrator or recorder, it's
convenient to be able to short the detector outputs. You can do this
using the Short Outputs feature.
To access Short Outputs:
1. Press [MENU].
2. Select /COMMANDS/. The Commands Menu (Fig. 4.7)
appears.
>
Event Mark
❑
Short Outputs
--------------------------------------------------------❑
Shutdown Detector
Figure 4.7 The Commands Menu
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When you select Short Outputs, the detector's analog outputs are
shorted together (zero volts) and the field name changes to "Unshort
Outputs." To remove the short and return the outputs to their normal
operating state, select Unshort Outputs, and the field changes back,
now reading "Short Outputs." (When you leave this screen, the field
returns automatically to Short Outputs.)
EVENT MARK
Using the event mark feature, you can place an event mark on your
chromatogram to note various occurrences, such as the turning of a
sampling valve. The event mark is a spike (15% of full-scale for one
second) in both detector output signals.
To access Event Mark:
1. Press [MENU].
2. Select /COMMANDS/. The Commands Menu (Fig. 4.7)
appears.
3. Place the cursor on Event Mark. Press [ENTER] each time you
wish to place an event mark on your chromatogram.
NOTE: You may not want to use event marks if your data will be analyzed by
an integrator. Integrators can misinterpret event marks as peaks!
READY OUTPUT
Using the READY(Output) terminal on the detector's back panel, the
detector can send a signal to other devices each time it goes to its
READY state. This feature is frequently used with autosamplers to
signal that the detector is ready for the next injection.
To access the READY Output field:
1. Press [MENU].
2. Select /OPTIONS/.
3. Select /More/.
4. Scroll down to the READY Output field. Select Active Hi or
Active Lo, depending on which signal you wish to send.
HINT: All SpectraSYSTEM instruments are set to receive high
signals, so select Active Hi if you are hooking up to this type of
chromatograph. For any other type of instrument, refer to the
appropriate reference manual.
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DISPLAY
CONTRAST
You can vary the display's contrast to make it easier to read.
To change the display's contrast, first press [STATUS] to access the
Status Screen. Then simultaneously press the [>] key and the [+] key
to increase the contrast, or the [>] key and the [-] key to reduce the
contrast.
CURSOR SPEED
You can control the display's cursor speed to make it easier to use.
To access Cursor Speed:
1. Press [MENU].
2. Select /OPTIONS/.
3. Select /More/.
4. Scroll down to Cursor Speed and select Fast, Medium, or
Slow.
SHUTDOWN
DETECTOR
This feature offers a quick shutdown, and subsequent startup, of the
detector's lamps and motors. The electronics stay on to maintain the
detector's memory.
To shut down the detector:
1. Press [MENU].
2. Select /COMMANDS/.
3. Scroll down to the Shutdown Detector field.
4. Press [ENTER]. The confirmation message shown in
Figure 4.8 appears on the display.
** Detector Shutdown **
Figure 4.8 Shutdown confirmation message
To start the detector up again, press any key on the keypad. The
detector will come up under the same conditions present when the
Shutdown Detector command was activated.
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UV2000 only
Scanning
The UV2000 can perform a spectral scan on eluting peaks without
stopping the eluant flow. This unique feature greatly simplifies the
determination of wavelength maxima for individual compounds in your
sample during method development work.
HOW IT WORKS
When a scan is initiated, the monochromator moves from the runwavelength to the scan's start-wavelength. The detector scans by
stepping through a defined spectral range at specified wavelength
increments. Individual absorbance values are read at each increment
until the monochromator has reached the last wavelength.
The UV2000 can collect and store as many as ten spectra from a
single chromatographic run in its memory. The actual number of
spectra is determined by the number of data points in each scan.
Since the number of data points varies with the wavelength interval
and the scanning range, first calculate the number of data points using
Equation 1, then use either Equation 2 or Equation 3 to determine the
number of spectra you will be able to collect.
Equation 1. Use this equation to calculate the number of data points
for any scan between λ1 (the lower wavelength), and λ2 (the higher
wavelength):
λ2 - λ1
# of data points
=
⎯⎯⎯⎯⎯ + 1
λ interval
Equation 2. Use this equation to calculate the number of spectra you
can collect when using wavelength intervals of 2 nm or greater.
Round the resulting number down to the nearest integer.
5000 - (# of data points * 12)
# of spectra
=
⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯
(# of data points * 4) + 14
Equation 3. Use this equation to calculate the number of spectra you
can collect when using wavelength intervals of 1 nm. Round the
resulting number down to the nearest integer.
5000 - (# of data points * 4)
# of spectra
=
⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯⎯
(# of data points * 4) + 14
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For example, if you want to scan from 190 to 564 nm in 2-nm steps,
there would be 188 data points and the UV2000 would be able to
store up to 3 spectra:
5000 - (188 * 12)
# of spectra
=
⎯⎯⎯⎯⎯⎯⎯⎯ =
(188 * 4) + 14
2744
⎯⎯⎯ = 3.58 = 3
766
Each scan is corrected for baseline absorbance before being played
back either as individual data points, or as a smoothed, continuous
spectrum.
SELECTING
THE SCAN FILE
To select spectral scanning, follow these step-by-step instructions.
1. Press [MENU]. Select /FILES/.
2. Select /Edit/.
3. Use the [+] key to increment the Edit File field until an "S" is
displayed (Fig. 4.9). The File Name field is automatically
named SCAN. (You cannot edit the Scan File's name.)
Edit File
S
File Name
SCAN
--------------------------------------------------------❑
Setup
❑
Replay
Figure 4.9 The Scan File's Edit Menu
4. Select /Setup/ to set up your spectral scanning parameters.
SETTING UP
THE SCAN FILE
The Scan File's Setup Menu is shown in Figure 4.10.
Start λ
220
End λ
365
--------------------------------------------------------λ Interval
Run λ
5
250
Rise Time
Scan Zero Time
Range 1
Range 2
1.0
0.00
1.0
1.0
Figure 4.10 The Scan File's Setup Menu
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Use these steps to set the parameters for scanning:
1. In the Start λ field, enter the wavelength at which each scan
should start.
2. In the End λ field, enter the wavelength at which each scan
should end.
3. In λ Interval, enter the wavelength interval to be used. To
perform a scan, the UV2000 takes individual absorbance
readings at wavelengths incremented by the interval you
specify.
HINT: Five nanometers is an excellent wavelength interval for most
applications. At this interval, you get very rapid scans and you can
still display the λMax to 1 nm accuracy.
4. In Run λ, enter the wavelength at which the chromatographic
run will be monitored.
5. In /Scan Zero Time/, enter the runtime at which you wish the
detector to perform an automatic baseline scan. If you use an
automatic baseline scan, make sure no peaks are eluting during
the designated scan time.
6. Fill in entries for Rise Time, Range 1, and Range 2 as you
would for any chromatographic run.
When you are finished setting up the Scan File, you are ready to load
it and run.
RUNNING
THE SCAN FILE
When the Scan File is loaded, you will notice the fields Zero and
Scan in the Status Screen (Fig. 4.11).
Status
λ
AU
❑ Scan
READY
250
0.0001
> Zero
T
Figure 4.11 The Status Screen with the Scan File loaded
Zero
/Zero/ is used to perform baseline scans of the solvent's background
absorbance. With the detector's baseline stabilized and the cursor on
the Zero field, press [ENTER]. The UV2000 performs and stores a
baseline scan using the parameters you set in the Scan File. While the
detector is performing a baseline scan, the Status field displays
SCAN 0.
Baseline scans may be taken at any time during the run, as long as no
peak is eluting at that time. Subsequent sample scans are corrected
using the last baseline scan taken. This is especially advantageous for
gradient elution, where the background absorbance of the eluant may
be constantly changing.
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For example, let's say you perform a baseline scan before you initiate
a run, and then again at 5.00 minutes into the run. You also perform
sample scans of your eluting peaks at 2.4 and 5.6 minutes into the
run. The sample scan taken at 2.4 minutes will be corrected using the
baseline scan taken before the run began. The sample scan taken at
5.6 minutes will be corrected using the baseline scan taken at 5.0
minutes.
Scan
Once you begin the run, the cursor will move from Zero to Scan in
the Status Screen. Each time you wish to perform a sample scan,
press [ENTER].
NOTE: There is a one-second delay from the time the detector takes its
absorbance readings to the time you see the same reading on the analog
readout. Keep this in mind when choosing your scan times.
Each time you perform a sample scan, the detector's monochromator
moves from the run wavelength to the start wavelength. The detector
performs each scan (from the start wavelength to the end wavelength)
by taking individual absorbance readings at wavelengths incremented
by the interval you set in the Scan File. When the scan is finished, the
monochromator returns to the run wavelength.
For example, using the default Scan File Setup Menu shown in
Figure 4.10, the detector would monitor the run at 250 nm. Each scan
would include absorbance readings for wavelength settings of 220,
225, 230, 235, and so on, up to 350 nm.
NOTE: If you chose starting and ending wavelengths that were not an exact
multiple of your wavelength interval, the ending spike (event mark) on your
chromatogram would be placed at the last multiple of the wavelength
interval that falls within the scanning range. For example, with a starting
wavelength of 200 nm, an ending wavelength of 365 nm, and a wavelength
interval of ten, the end spike on your chromatogram would be at 360 nm, the
last full wavelength multiple within the range.
While the detector is scanning, the Status field displays SCAN.
NOTE: During scanning, the output signal will hold at the last absorbance
value taken before the scan was initiated until the scan is finished. For this
reason, quantitative analysis should never be performed when scanning.
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Scan Summary
Data Screen
When the Scan File is loaded, the normal Status Menu no longer
appears below the Status Screen. Instead, several new lines that we
call the "Scan Summary Data Screen" appear. The Scan Summary
Data Screen is useful in setting up the parameters to replay your
stored spectra.
An example of the Scan Summary Data screen as it appears after two
sample scans is shown in Figure 4.12.
File S: SCAN
--------------------------------------------------------Time
10.50
11.66
λMax
λMaxAU
λMin
280
255
1.6668
0.7768
230
220
Figure 4.12 An example of the Scan Summary Data Screen
The Scan Summary Data Screen has four fields:
•
Time, which is the run time at which the scan was initiated
•
λMax, which is the scan wavelength where the maximum
absorbance occurred
•
λMaxAU, which is the maximum absorbance
•
λMin, which is the scan wavelength where the minimum
absorbance occurred
If no maximum was found, the λMax and λMaxAU fields read 0
(zero). The summary information is updated as each sample scan is
completed.
NOTE: The UV2000 uses a second derivative to find the "local" λMax.
In our example (Fig. 4.12), scans were taken at 10.50 and 11.66
minutes into the run. The scan taken at 10.50 minutes has a
maximum absorbance of 1.6668 AU at 280 nm. The minimum
absorbance occurred at 230 nm. To replay your 10.50-minute scan,
you would use a range of 2.0 AUFS to keep the absorbance values
on-scale.
STOPPING THE
SCAN FILE
50
There is no programmed stop in the Scan mode. The run will
continue until it reaches 99.99 minutes, or until you press [STOP].
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Automatic Scanning (Model UV2000 only)
DT\Z155E\DT
If you've set the Auto Scan field in the Setup Menu to On, your
detector will perform an automatic scan whenever there are at least
three consecutive data points with positive slopes followed by three
consecutive data points with negative slopes. The absorbance values
for all these data points must exceed 5 percent of the value set in the
/Range 2/ field. In our example chromatogram (Fig. 4.13), a scan
would occur automatically for Peak A, since it has at least three data
points with positive slopes followed by at least three data points with
negative slopes, all of which exceed 5% of the value set in /Range 2/.
Conversely, no scan would occur for peak B, since none of its
absorbance values exceeds the 5% threshold, even though it may
satisfy the consecutive-slope criteria.
Peak A
5% of Range 2
Peak B
Time
Figure 4.13 An example of how automatic scanning works
An automatic baseline scan will occur at the time specified in the
Spectra Menu's /Scan Zero Time/ field.
NOTE: Make sure that no peaks are eluting at the specified scan-zero time
or your baseline scan will be zeroed erroneously for the eluting peak's value
at the moment when the scan zero occurs. This will produce a baseline scan
that is heightened artificially.
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REPLAYING YOUR
SPECTRA
When you have completed your run, you can retrieve your stored
sample spectra using the Replay Menu (Fig. 4.14).
To access the Replay Menu:
1. Press [MENU]. Select /FILES/.
2. Select /Edit/ to display the Scan File's Edit Menu (Fig. 4.9).
3. Select /Replay/.
Range 1
1.0
Range 2
1.0
--------------------------------------------------------Replay Rate (nm/sec)
5
Spectra Time
10.50
❑
Replay Spectra
❑
Display AU,λ
Figure 4.14 The Replay Menu
Setting Replay
Parameters
To set the parameters for replay:
1. Set Range 1 and Range 2 for Analog Output 1 and Analog
Output 2. If you are using only one output, disregard the
appropriate range.
2. Enter the Replay Rate (nm/sec). This is the rate at which the
detector will read out the spectral data to your chart. You will
use this value and an appropriate chart speed to calculate
wavelength increments on your printed sample spectrum.
For example, if your sample scan were taken between 190 and
340 nm (a span of 150 nm), a replay rate of 5 nm/sec would
print the spectrum in 30 seconds. A chart speed of 30 cm/min
would give you a scan of 15 centimeters in increments of
10 nm/cm.
3. Select the spectrum you want to replay by selecting its start
time in the Spectra Time field. Each spectrum taken during the
run is individually identified by the run time at which it was
initiated.
When you finish setting your replay parameters, you are ready to send
the spectral data to your chart using the Replay Spectra command.
Running Replay
52
To initiate the Replay Spectra command in the Replay Menu, press
[ENTER]. While the replay is occurring, the screen shown in
Figure 4.15 appears on the display.
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Replay
λ
AU
10.50
220
0.00001
Figure 4.15 The display as it appears
while spectra are being replayed
The screen's Replay field displays the start time of the spectrum being
replayed. The λ and AU fields display the individual data points
being plotted.
The UV2000 uses advanced curve-fitting algorithms to present a
smooth, continuous plotted spectrum. The spectrum is replayed in
1-nm steps regardless of the wavelength interval selected. To change
the appearance of replayed spectra from 1-nm stepped curves to
smooth curves (or vice versa), vary the recording device's replay rate
and response time.
If no spectra are stored in memory when you activate the Replay
Spectra command, the message shown in Figure 4.16 will appear on
the display. When the replay is finished, the display returns to the
Replay Menu.
** No Scans Stored **
Figure 4.16 The message that appears
when no spectra are stored in memory
Stopping Replay
You may stop a replay at any time by pressing [STOP].
SPECTRAL
DATA STORAGE
Spectral data are stored in the UV2000's memory until a new file or
queue is loaded or the detector is turned off.
Viewing Data
You can display the individual data points of your stored spectra by
selecting the Display AU, λ field in the Replay Menu (Fig. 4.14). A
screen similar to that shown in Figure 4.17 will appear on the display.
Display
λ
AU
10.50
220
0.00001
Figure 4.17 The Display AU, λ screen
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NOTE: Only actual data points (separated by the proper wavelength
interval) can be displayed.
The Display AU, λ screen shows the time at which the scan was
initiated, along with each wavelength and absorbance reading
collected. You can scroll through the data using the [+] and [-] keys.
To return to the Replay Menu, press [∧].
The Develop File
The Develop File is unique to the UV2000. It allows you to make
sequential sample injections at different wavelengths automatically.
This automation makes method development much easier because
you can use an automated run to determine the optimum wavelength
of detection for each component in your sample. You can also use
the Develop File to troubleshoot chromatographic problems, or to
confirm method transfer from laboratory to laboratory.
SELECTING THE
DEVELOP FILE
Use the following instructions to select the Develop File.
1. Press [MENU]. Select /FILES/.
2. Select /Edit/.
3. Use the [+] key to increment the Edit File field until a "D" is
displayed. The File Name field will read DEVELOP. (You
cannot edit the Develop File's name.)
EDITING THE
DEVELOP FILE
Follow these instructions to edit Develop File parameters:
1. Once you've selected the Develop File as described above,
press either the [ENTER] or the [∨] key to access the Develop
File's Edit Menu (Fig. 4.18).
Edit File
D
File Name
DEVELOP
--------------------------------------------------------Start λ
220
End λ
350
λ Interval
Run Time
Runs per λ
Rise Time
Autozero Time
Range 1
Range 2
5
10.00
2
1.0
0.00
1.0
1.0
Figure 4.18 The Develop File's Edit Menu
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2. In the Start λ field, enter the wavelength at which the first
chromatogram is to be monitored.
3. In the End λ field, enter the wavelength at which the last
chromatogram is to be monitored.
4. In λ Interval, enter the wavelength increment that the detector's
monochromator should use for each wavelength change.
5. In Run Time, enter how long each run should last.
6. In Runs per λ, enter the number of injections to be made at
each wavelength setting.
7. Enter Rise Time, Autozero Time, Range 1, and Range 2, as you
would for a typical run. Note that Range 1 and Range 2 are the
corresponding ranges for Analog Outputs 1 and 2, respectively.
As an example, we will use the Develop File shown in Figure 4.18.
The UV2000 would make its first two ten-minute runs at 220 nm.
The monochromator would then change to 225 nm, and the detector
would make two runs at this wavelength. This pattern would
continue in five-nanometer increments until the detector has made
two runs at the last wavelength, 350 nm.
After setting up your Develop File, you are ready to load it and run.
RUNNING THE
DEVELOP FILE
When the Develop File is loaded, you will notice an additional field
in the Status Screen, #Runs (Fig. 4.19).
Status
λ
AU
#Runs
READY
220
+0.0001
1/3
T
Figure 4.19 The Status Screen with the Develop File loaded
#Runs
The #Runs field in the Status Screen shows the current run number,
followed by a forward slash and the total number of injections for the
wavelength specified in the λ field. For example, if the file is set up
to make three injections per wavelength, and the detector is in the
second run for the 250-nm setting, the #Runs field would appear as
2/3. The field is updated with each injection.
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Status Menu
The Status Menu looks the same for a Develop File as it does for a
typical chromatographic file (Fig. 4.20).
File D:
DEVELOP
--------------------------------------------------------Time
Wavelength
0.00
250
10.00
250
T
Rise Time
Autozero Time
Range 1
Range 2
1.0
0.00
1.0
1.0
Figure 4.20 The Status Menu with the Develop File
NOTE: You can change any of the parameters in the Status Menu while the
detector is running, but the changes will be effective only until the next
wavelength is loaded.
REPEATING THE
DEVELOP FILE
After the last wavelength is run, the detector is reset automatically to
the starting wavelength in the Develop File. The file can be run as
many additional times as you wish, as long as the detector continues
to receive run signals.
Sample Queue
Sometimes it's convenient to group samples together under different
detector conditions in an automated run. For these occasions, the
UV2000 offers a queuing feature. Using a queue, you can program
the detector to load and run a specified file for your first group of
samples, and then automatically load a second file to run your next
group of samples. The queue feature allows you to run as many as
ten groups in a single queue.
QUEUE MENU
To access the Queue Menu, follow these steps:
1. Press [MENU].
2. Select /QUEUE/.
When no queue is loaded, the Queue Menu appears as shown in
Figure 4.21. On page 58, we'll see how the menu appears when a
queue is loaded.
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>
❑ Load
Edit
❑ Delete
Figure 4.21 The Queue Menu with no queue loaded
SETTING UP
A QUEUE
To set up a queue, select /Edit/ from the Queue Menu. For an empty
queue, the display appears as shown in Figure 4.22.
Order
1
File:Name
#Runs
Figure 4.22 An empty queue
Entering a Line
A "1" is automatically placed in the Order field of the first file to be
run. You can't change that, so the cursor appears under the first
editable field, File:Name. Scroll through the available files and press
[ENTER] when your choice appears.
NOTE: You may only select numbered files. The Scan and Develop files are
not available in the Queue mode.
Enter the number of injections to be made in the #RUNS field and
press [ENTER]. You can have as many as 999 injections per file.
Adding More Lines
After completing the first line, a second line appears automatically.
The Order field reads 2, and the rest of the line is blank. Select the
proper file and the number of injections to be made for that file. You
can have as many as ten groups in the queue.
Deleting a Line
To delete a line, use the [-] key in the File:Name field until it goes
blank. When you leave the line, it is deleted and the queue is resorted
automatically.
An Example
An example of a queue appears in Figure 4.23.
Order
File:Name
1
2
3
2:THEOPHYL
3:ABCD
1:BARBITUA
#Runs
5
25
10
Figure 4.23 An example of a queue
In our example, we have programmed the detector to run File 2 for
the first five injections, File 3 for the next 25 injections, and File 1 for
the last ten injections.
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LOADING A QUEUE
To load a queue, select /Load/ in the Queue Menu. When the words
"Load Queue" appear, press [ENTER]. The confirmation message in
Figure 4.24 appears for one second.
** Queue Loaded **
Figure 4.24 The confirmation message
when a queue is loaded
When a queue is loaded, the letter "Q" appears at the extreme left of
the Status Screen (Fig. 4.25).
Status
Q READY
λ
250
AU
+0.00001 T
Figure 4.25 The Status Screen when a queue is loaded
If you attempt to load a queue when no queue exists, the message
shown in Figure 4.26 will appear on the display.
** No Queue Available **
Figure 4.26 The message that is displayed
when no queue is available
RUNNING A QUEUE
When the detector receives its first start signal, it loads and runs the
file designated in Order 1. It will continue to run this file each time it
receives a start signal until the file has run the number of times
specified in the #Runs field. The detector will then load and run the
file designated in Order 2 and run it the number of times specified in
that line, and so on, until the entire queue has run.
Viewing its
Progress
To view a queue's progress while it is running:
1. Press [MENU].
2. Select /QUEUE/. Note that when a queue is loaded, the Queue
Menu (Fig. 4.27) looks different. The Load field has been
replaced by "Pause," which we will discuss on page 59.
>
❑ Pause
Edit
❑ Delete
Figure 4.27 The Queue Menu with a queue loaded
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3. Select /Edit/ to display the queue. (Refer to Figure 4.23 for an
example queue.)
While the queue is running, the #Runs field automatically decreases
by one with each injection. When a particular file's last injection is
made, the queue is automatically resorted. In other words, the
information for Order 2 is now moved up to Order 1, the information
for Order 3 is moved up to Order 2, and so forth. This process
continues until the queue becomes empty, is paused, or is deleted.
Loading other Files
When a queue is loaded or running, you may not load any other file
from the Files Menu without first pausing or deleting the queue. If
you forget to pause or delete the queue and attempt to load a different
file, you will get the message shown in Figure 4.28. You are then
returned to the Files Menu.
** Queue Loaded **
Cannot Load File
Figure 4.28 The message that appears when you attempt to load a
file when a queue is already loaded or running
EDITING A QUEUE
To edit an existing queue, follow the procedures outlined in "Setting
Up a Queue" on page 57. You are allowed to edit the Queue while it
is running, but if you want to edit anything in Order 1, you'll have to
pause the queue first.
PAUSING A QUEUE
To pause a queue:
1. Select /Pause/ from the Queue Menu.
2. When the words "Pause Queue" appear, press [ENTER]. If a
file is running, the run continues until it is completed, at which
point the detector returns to its READY state. The letter Q no
longer appears in the Status Menu.
When you wish to continue, you must reload the queue. When the
detector receives a start signal, the queue will resume operation at the
point where it left off.
DELETING/
STOPPING A
QUEUE
Use the following steps to delete an existing queue or to stop a
running queue:
1. Display the Queue Menu.
2. Select /Delete/.
3. When the words "Delete Queue" appear, press [ENTER]. If a
file is running, the run continues until it is completed. The
confirmation message shown in Figure 4.29 appears for one
second and you are returned to the Queue Menu.
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** Queue Deleted **
Figure 4.29 The queue-deleted message
You may delete or stop a queue at any time, but remember that the
queue will be subsequently erased from the detector's memory. It is
good practice to delete an existing queue prior to designing a new
one.
K-Factor
The K-factor calculates a factored response that can be used to
eliminate, add, or subtract absorbances. This technique is useful for
suppressing peaks when there are two co-eluting, or poorly resolved,
peaks in your chromatogram. It is also useful in applications where
you want to add or subtract absorbances at two different wavelengths
in real-time.
For example, if you want to quantitate a peak without interference
from another peak, you would use the K-factor to calculate a response
of zero.
More specifically, let's say you want to analyze for Compound A in
the presence of Compound B. If both absorb at the monitoring
wavelength, λ1, but only Compound B absorbs at a second
wavelength, λ2, you can calculate a K-factor for Compound B using
its absorbances at λ1 and λ2. You can then use the K-factor to
calculate the absorbance due only to Compound A at the monitoring
wavelength (λ1), by subtracting Compound B's contribution from the
total absorbance. The UV2000 uses the algorithm:
Absorbance due to A at λ1 = TAbs(λ1) - K x TAbs(λ2)
where TAbs(λ1) is the sum of the absorbances of A and B at the
monitoring wavelength, K is the K-factor, and TAbs(λ2) is the total
absorbance obtained at λ2.
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Figure 4.30 shows a chromatogram of a mixture of toluene and butyl
paraben where the two compound peaks overlap. Toluene (Peak A) is
the compound of interest. Butyl paraben (Peak B) is the peak we
want to suppress. We will use this example throughout the following
steps for determining and using the K-factor.
DT-Z023E\FM
AN EXAMPLE
A
B
1.35 1.51
Time (in Minutes)
Figure 4.30 A chromatogram of two unresolved peaks:
toluene (A) and butyl paraben (B)
Choosing a Pair of Wavelengths
The first step in determining the K-factor is to choose a pair of
wavelengths for your analysis.
1. Take an absorbance spectrum of each compound. You can do
this by injecting samples of compound A and compound B
alone, separately, under the same chromatographic conditions
as your analysis, and using the UV2000's scanning feature.
(See "Scanning" on page 46.)
For the compounds in our example, we get the spectra shown
in Figure 4.31.
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DT-Z024E/FM
Toluene
200
380
200
380
Butyl Paraben
Wavelength (nm)
Figure 4.31 Spectra of individual compounds
2. Label the wavelength maximum for your peak of interest as λ1.
3. From the spectra, pick a wavelength for which compound B
absorbs and compound A does not. This wavelength is labeled
λ2. For our example, we have chosen 254 nm as λ1 and 280
nm as λ2.
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Calculating the K-factor
Use the UV2000's Display AU, λ screen (page 53) to obtain the
individual absorbance value data from your scan of compound B.
Calculate the K-factor using the following equation:
K = AU1/AU2
where AU1 and AU2 are the absorbance values for compound B at λ1
and λ2, respectively.
For our example, the absorbance values are 0.0144 and 0.0032 (for
254 and 280 nm respectively), so our K-factor is 4.5, calculated as
follows:
K = 0.0144 / 0.0032 = 4.5
Using the K-Factor
To use the K-factor, set the parameters in the Analog Outputs Menu,
inject your sample, and monitor the results as follows:
1. Press [MENU].
2. Select /OPTIONS/.
3. Select /Analog Outputs/.
The menu shown in Figure 4.32 will appear.
Analog 1 Offset (%)
0
Analog 2 Offset (%)
0
--------------------------------------------------------Analog 2
AU
K Factor
1.000
Figure 4.32 UV2000's Analog Outputs Menu
4. Scroll down to Analog 2 and select AU1-K*AU2.
5. Scroll down to K-factor and enter your calculated value
(4.5, for our example).
6. Inject your sample.
HINT: Make sure your file was set to dual-wavelength mode as
described in Chapter 3. Also remember that in this example, AU1
(λ1) is 254 nm and AU2 (λ2) is 280 nm.
7. Use Analog Output 2 on the detector's rear panel to monitor the
chromatograms for your peak of interest.
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DT-Z025E/FM
1.51
Time (in Minutes)
Figure 4.33 Chromatogram of toluene with butyl paraben suppressed
Our example chromatogram would now appear as shown in
Figure 4.33, with a slightly lowered response for toluene and no
absorbance contribution from butyl paraben. Using the K-factor in
this way, we can quantitate toluene in the presence of butyl paraben
without altering the chromatography.
Absorbance Ratios
Ratioing the detector's outputs from two different wavelengths can be
a useful way of confirming peak purity. When a peak is pure, the
ratio of the absorbances should remain constant. Thus, the ratio for a
pure compound produces a relatively square wave, while the ratio for
an impure compound produces a distorted wave (see the plots at 1.57
and 0.97 minutes, respectively, in Figure 4.34).
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DET\Z17V\FM
Mixture of
Two Compounds
Chromatogram
Pure
Compound
0.97
1.57
Pure
Compound
Mixture of
Two Compounds
AU1/AU2 Plot
0.97
1.57
Time (in Minutes)
Figure 4.34 Using absorbance ratios to determine the purity
of two peaks in a chromatogram
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To use absorbance ratioing, you need to select AU1/AU2 for the
Analog 2 Output field in the Analog Outputs Menu. You also need to
select the two wavelengths you want to ratio.
To select the most appropriate wavelengths, use the UV2000's Scan
File to collect a spectrum across a range of wavelengths. Then select
/Display AU, λ/ from the Replay Menu and examine the collected
data.
The data shown in Figure 4.35 are typical.
Display
λ
AU
1.50
220
0.00001
--------------------------------------------------------1.50
250
1.66681
1.50
280
0.28831
Figure 4.35 The Display AU, λ screen
Ratioing only occurs when the absorbance value for each wavelength
exceeds 12.5% of the corresponding range value. So, in our example,
if Ranges 1 and 2 were set to 1.0 in the /FILES/, /Edit/, Options
Menu, the 250 and 280 nm wavelengths could be ratioed. [Twelve
and a half percent of 1.0 (the range) is 0.125. Absorbance values less
than 0.125 are too low for ratioing.] No ratio output is produced
when the absorbance values fall below 7.5% of the range values.
Generally, good wavelengths to choose are:
1. the lambda max of the main peak (AU1)
2. a wavelength with an absorbance value less than the lambda
max but greater than 12.5% of the corresponding range (AU2)
HINT: A good rule of thumb is to select a second wavelength that is
either half the height of the lambda max or more than ten nanometers
removed from the lambda max.
Whichever wavelengths you choose, don’t select a wavelength that
has a low absorbance value. Low absorbance values decrease the
signal-to noise ratio, thus making the absorbance ratios less
meaningful. Similarly, a small fluctuation in AU2 results in a big
difference in the absorbance ratio if AU2 is very small. Fortunately,
by relying on the preset range values, the UV 2000 has a built-in
safeguard that prevents the ratioing of low absorbance values.
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5
Required Maintenance
Introduction
Finnigan SpectraSYSTEM detectors are finely-tuned scientific
instruments that we at Thermo Electron are proud to stand behind.
Even so, routine maintenance is necessary to ensure peak
performance, so we can only guarantee our detectors' performance if
you follow proper care and maintenance procedures.
This Chapter shows you how to replace and clean your detector's
flowcell and lamps.
Also included in this Chapter is a procedure for testing the detector's
absorbance linearity. This characteristic is particularly useful if your
laboratory's standard operating procedures require periodic detector
validation. You will need the optional cuvette holder to perform the
test.
If you have any questions on proper maintenance or would like to
arrange for a preventive maintenance program, please contact your
local Thermo Electron representative.
Flowcells
This section describes the changing and general cleaning of your
detector's flowcell. For other flowcell problems, such as a cracked
window or leaks that occur in locations other than at the inlet/outlet
fittings, contact your Thermo Electron representative.
NOTE: Flowcells are factory-assembled units that should not be
disassembled under any circumstance.
CHANGING THE
FLOWCELL
The flowcell needs to be removed whenever you replace a broken
cell, change specialized applications, or clean the cell with nitric acid.
For a list of available flowcells, see "Specifications - Flowcells" in
Appendix A. All flowcells are shipped premounted in a holder for
easier installation and alignment.
To access the flowcell, remove the front panel of the detector. The
flowcell assembly is located behind the lower housing (Fig. 5.1).
Once the housing is removed, the flowcell is easily identified by the
tubing that extends from the fittings on either side of the cell body
(Fig. 5.2).
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Display
RUN
STATUS
MENU
STOP
Keypad
ENTER
ZERO
SpectraSYSTEM
UV2000
Lamp
Housing
Flowcell
Housing
Figure 5.1 Detector flowcell and lamp housings
Flowcell Removal
Use the following steps to remove the flowcell:
1. Disconnect the power cord from the rear panel of the detector
and make sure that the instrument is turned off.
2. If you have not already done so, remove the detector's front
panel by grasping the bottom of the panel firmly with one hand
and pulling back.
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3. Loosen the knurled thumbscrew that holds the flowcell housing
in place, and remove and set aside both the thumbscrew and the
housing.
4. Disconnect the flowcell inlet tube from the chromatograph and
free the flowcell outlet tubing from the waste reservoir.
5. Loosen the two thumbscrews on the photodiode mount and
carefully pull the mount straight back (Fig. 5.2). The cable that
connects the photodiode mount to the detector is sufficiently
long to allow the mount to rest on the bench top.
NOTE: Avoid putting fingerprints or scratches on the flowcell
windows, photodiode surface, or monochromator lens, all of which
are exposed during these procedures. If dirty, the surfaces should be
cleaned with spectroscopic-grade methanol (or isopropanol) and lintfree lens paper only.
Flowcell
Outlet
Flowcell
Inlet
Tubing
Clamp
Photodiode
Mount
Flowcell Assembly
Thumbscrews
DT-Z019E\FM
Flowcell
Assembly
Figure 5.2 Removing the cell cover to expose the flowcell
and the photodiode mount
6. Loosen the thumbscrew that holds the tubing clamp in place.
Gently pull the clamp toward you just far enough to disengage
the tubing.
7. Loosen the two thumbscrews that hold the flowcell assembly.
Carefully pull the assembly toward you to remove it from the
detector.
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Flowcell Installation
To install a flowcell, follow these steps:
1. With the inlet tube on the bottom, slide the flowcell assembly
onto the alignment pins (Fig. 5.3) and securely fasten it in place
with the two thumbscrews.
Tubing
Clamp
Flowcell
Alignment
Pins
Flowcell
Assembly
DT-Z020E\FM
Photodiode
Mount
Figure 5.3 Installing the flowcell assembly
2. Slip the flowcell's inlet and outlet tubes into the slots of the
tubing clamp and tighten the thumbscrew that holds the clamp
in place.
3. Replace the photodiode mount and fasten it securely with the
two thumbscrews.
4. Connect the inlet tubing to the chromatographic column and
the outlet tubing to the waste reservoir.
5. Taking care not to pinch the cable or tubing, replace the
flowcell housing and secure it with the knurled thumbscrew.
Replace the detector's front cover.
6. Connect the power cord to the rear detector panel.
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CLEANING THE
FLOWCELL
The exterior and/or interior surfaces of the flowcell can become
contaminated. When flowcell contamination occurs, it is usually
caused by precipitation or solubility problems, such as when the
quality of your mobile phase solvent components and the cleanliness
of your samples are variable. Signs of a contaminated flowcell are
increased baseline noise, signal spiking, erratic or drifting baselines,
and increased backpressure.
Cleaning with
Organic Solvents
If you suspect that your flowcell needs to be cleaned, start with the
following procedure using organic solvents.
NOTE: Flowcells are factory-assembled units that should not be
disassembled under any circumstance. If you encounter contamination
problems that are not remedied by this cleaning procedure, contact your
local Thermo Electron representative to arrange for repair or replacement.
1. Make certain that the cleaning solvent(s) you plan to use is/are
miscible with the solvent already present in the flowcell and
pump. Isopropanol is a good choice for most applications.
NOTE: If the last solvent in the pump was an aqueous buffer
solution, be sure to pump 25 - 40 mL of HPLC-grade water (or
equivalent) through the system to remove any salts before flushing
with the cleaning solvent(s).
2. Flush the flowcell with 40 - 50 milliliters of solvent (HPLCgrade water, methanol, or isopropanol). You can either pump
the solvent through the flowcell with the chromatographic
pump, or you can draw the solvent through the flowcell using a
large-volume syringe.
If you use an LC pump to flush the flowcell, first remove the
column from your chromatographic system to avoid column
degradation. Replace the column with an appropriate length of
tubing, ensuring that all connections are snug and leak-free.
If you use a syringe, always draw the solution through the
flowcell.
CAUTION! Never use a syringe to force solvent through a
flowcell. Pressurizing the syringe could cause a leak or rupture
that would result in an extremely dangerous, uncontrolled
spraying of solvent.
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Cleaning with
Nitric Acid
Methanol or isopropanol is generally sufficient for cleaning a
flowcell. However, if the flowcell is still contaminated after flushing
with organic solvents, follow this nitric acid procedure.
CAUTION! Nitric acid is extremely corrosive and can react explosively
with alcohols (especially methanol). Be sure to adhere to your
company's safety procedures for handling and disposal of corrosive
acids. Flush the flowcell with water to remove all traces of alcohol prior
to flushing with nitric acid!
1. Remove the flowcell assembly from the detector housing
(following the procedure on page 68) before cleaning with a
nitric acid solution. This will prevent possible leaks from
harming the mechanical or electronic components of the
detector.
2. Flush the flowcell with water before proceeding. This step is
very important!
3. Prepare a 20% (v/v) solution of nitric acid in HPLC-grade
water.
4. Pump the nitric acid solution through the flowcell with the
chromatographic pump or draw it through with a large-volume
syringe.
If you use an LC pump, replace your column with tubing and
make sure water was the last solvent in the pump and solvent
reservoir. If you use a syringe, always draw the solution
through the flowcell.
CAUTION! Never use a syringe to force nitric acid through a
flowcell. Pressurizing the syringe could cause a leak or rupture
that would result in an extremely dangerous, uncontrolled
spraying of nitric acid.
5. After you have finished the cleaning procedure and before
returning to the buffer solution, pump another 25 - 40 mL of
water through the flowcell to remove all traces of nitric acid
before returning to your chromatographic solvents. Reinstall
the flowcell assembly.
NOTE: Flush the pump with water immediately after the nitric acid
flush. Leaving nitric acid solution in the pump for prolonged periods
can damage pump seals.
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Lamps
As lamps age, there is a reduction in light output that results in
increased baseline noise. If the noise level on your detector's output
signal is increasing and cleaning the flowcell doesn't help, you should
change the appropriate lamp, using the procedures in this section.
Remove the front panel of the detector. The deuterium and tungsten
lamps are located in the upper housing (Fig. 5.4). Both lamps are
supplied prealigned in their individual assemblies to make them easy
to install and align.
NOTE: Never loosen the screws that hold the lamp to its assembly or
attempt to rotate or move the lamp up or down in the assembly. Either of
these actions can cause a loss of alignment and degrade the system's
performance.
Display
RUN
STATUS
MENU
STOP
Keypad
ENTER
ZERO
SpectraSYSTEM
UV2000
Lamp
Housing
Flowcell
Housing
Figure 5.4 Location of lamp housing
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73
THE DEUTERIUM
LAMP
The deuterium (D2) lamp typically requires a warm-up time of twenty
to thirty minutes. However, for applications that demand great
sensitivity, you may want to allow a warm-up period of up to an hour.
The deuterium lamp's lifetime is usually at least 1000 hours. Each D2
lamp assembly is equipped with a chronometer (Fig. 5.5) that tracks
the total hours of lamp operation. To read the chronometer, note the
position of the "gap" in the mercury tube against the graduated
background. You can also track lamp life automatically. (See
"Automatic Lamp Operations" on page 41 for details.)
NOTE: The lamp surface must be kept free of fingerprints and smudges.
If the surface needs cleaning, use a lint-free lens paper moistened with
methanol or isopropanol.
Plug-In
Connector
Gap
0
Hours
2000
DT-Z038E/DG
Lamp Leads
Figure 5.5 Deuterium lamp chronometer
D2 Lamp Removal
These four steps explain how to remove the deuterium lamp.
1. Disconnect the power cord from the detector's rear panel and
make sure that the instrument is turned off.
CAUTION! Intense UV light can damage your eyes. Always
disconnect the power cord before exposing the lamp and always
allow sufficient time for the lamp to cool before removing it, as it
gets quite hot when lit.
2. If you have not already done so, remove the detector's front
panel by grasping the bottom of the panel firmly with one hand
and pulling back.
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RUN
STATUS
MENU
STOP
ENTER
ZERO
SpectraSYSTEM
UV2000
Tungsten
Lamp Assembly
Deuterium
Lamp Assembly
Flowcell
Inlet
Flowcell
Outlet
DT-Z004E\FM
Flowcell
Assembly
Figure 5.6 Deuterium and tungsten lamp assemblies
3. Remove the lamp housing by loosening the thumbscrew and
pulling the cover straight back to expose the lamp assemblies
(Fig. 5.6).
4. Unplug the deuterium lamp lead from the detector, taking care
not to twist the connector as you gently pull it out.
5. Loosen the two thumbscrews that hold the lamp assembly in
place and pull the assembly straight out.
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D2 Lamp Installation
Follow these steps to install a new D2 lamp.
1. Hold the deuterium lamp assembly so that the leads are at the
top. Slide the assembly onto the alignment pin shown in
Figure 5.7. (The alignment pin is located directly below the
detector's monochromator aperture.)
2. Securely fasten the assembly in place with the two
thumbscrews and aluminum standoffs.
3. Connect the lamp lead to the right-hand terminal in the lamp
compartment.
4. Replace the lamp housing and secure it with the knurled
thumbscrew. Replace the detector's front cover.
5. Connect the power cord to the rear detector panel.
Tungsten
Lamp
Alignment
Pins
DT-Z021E/M
Deuterium
Lamp
Alignment
Pin
Figure 5.7 Deuterium and tungsten lamp alignment pins
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THE TUNGSTEN
LAMP
The tungsten (W) lamp typically requires only fifteen minutes of
warm-up time. Its lifetime is approximately 2500 hours. You can
track lamp life automatically. (See "Automatic Lamp Operations" on
page 41 for details.)
W Lamp Removal
Follow the steps below to remove the tungsten lamp.
1. Disconnect the power cord from the detector's rear panel and
make sure that the instrument is turned off.
CAUTION ⎯ Hot Surface! Avoid burns. Always allow
sufficient time for the lamp to cool before removing it.
2. If you have not already done so, remove the detector's front
panel by grasping the bottom of the panel firmly with one hand
and pulling back.
3. Remove the lamp housing by loosening the thumbscrew and
pulling the cover straight back to expose the lamp assembly
(Fig. 5.7).
4. Unplug the tungsten lamp lead from the detector, taking care
not to twist the connector as you gently pull it out.
5. Loosen the thumbscrew and the aluminum standoff that hold
the lamp assembly in place and pull the assembly straight out.
W Lamp Installation
These five steps explain how to replace the tungsten lamp.
1. Hold the lamp assembly so that the leads are at the top. Slide
the assembly onto the two alignment pins shown in Figure 5.7.
(The alignment pins are located on either side of the detector's
monochromator aperture.)
2. Securely fasten the assembly in place with the thumbscrew and
aluminum standoff.
3. Connect the lamp lead to the left-hand terminal in the lamp
compartment.
4. Replace the lamp housing and fasten securely with the
thumbscrew.
5. Connect the power cord to the rear detector panel.
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A
Installation and
Specifications
Introduction
This Appendix covers the initial installation of your UV/Vis detector,
including hookup to other chromatographic instrumentation. As you
go through unpacking and installation, you may want to use the
Start-up Checklist located at the beginning of this manual. The
checklist is an abbreviated version of this Appendix and is supplied as
a quick reference of how to conduct a successful installation. After
installation, verify that the detector is working properly by running
the two tests described on page 116.
Also included in this Chapter is a list of your detector's specifications.
Installation
UNPACKING
Carefully remove the detector from the shipping container and inspect
both the detector and packing for any signs of damage. If you find
any damage, immediately contact the shipping company.
The shipping container should contain the detector, an accessory kit,
any options you ordered for your detector, and this manual. The
accessory kit should contain the following items:
01/2004
•
8-pin connector
•
12-pin connector
•
Nut and ferrule tubing set
•
Teflon tubing
•
1/16 nut fitting (1/4-28 thread)
•
1/16 ferrule fitting
•
Union
•
External run/autozero cable
•
Analog cable
•
Extra cap screws (2)
•
LC test mix vial
•
3-foot, 4-conductor cable
Thermo Electron
79
Carefully check to make sure you received all the items listed on the
packing list. If any items are missing, contact your Thermo Electron
representative immediately.
You will need the following tools for installation:
•
a narrow-tip screwdriver (2 mm wide)
•
a #2 Phillips screwdriver
Place the detector on the benchtop as close as possible to the
chromatographic column outlet (thus minimizing the length of tubing
necessary for connection to the flowcell inlet). Allow at least five
inches (13 cm) of clear space between the detector's rear panel and
any wall or obstruction. This provides both access to the rear-panel
connectors and a free flow of cooling air.
CHECKING THE POWER
The detector is shipped with the voltage and fuses preset for your
location. To verify the correct setting, look through the cut-out
window on the voltage selector cover (Figure A.1). (The cover is
located on the detector’s rear panel but, if your instrument is new, it
may be hidden behind a precautionary sticker.) If the voltage setting
satisfies you local site requirements, skip to “Fuses” on page 82. If
not, proceed to the next section, “Voltage Selection.”
NOTE: Do not plug in the instrument without first verifying that the voltage
is properly set for your location! And never run the detector at more than
10 % below the nominal line voltage!
Voltage Selection
If the preset voltage does not satisfy your local requirements, select
the correct voltage by following these steps:
PRY OPEN HERE
DT-Z033E/FM/CT
1. Insert a small flat-blade screwdriver into the slot at the top of
the voltage selector cover (Fig. A.1).
220Vac
Figure A.1 Opening the voltage selector cover
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Current
Voltage
Selection
VoltageSelector
Drum
DT-Z026E/DG
Fuse
Holder
Figure A.2 Voltage selector barrel and fuse holders
2. Gently pry open the cover. Once unlatched, the cover will
swing downward to reveal the voltage selector barrel and the
fuses.
3. Remove the voltage selector barrel from the detector. The
selector resembles a wheel with four settings: 100, 120, 220,
and 240 V (Fig. A.2)
4. Rotate the barrel such that the desired voltage setting will be
visible through the cut-out in the cover when it is replaced.
5. Replace the barrel in the detector. Before closing the cover,
check the fuses according to the procedure on page 82.
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81
Fuses
To verify that your detector is fitted with the correct fuses, follow
these steps. (If you haven't already done so, first open the voltage
selector cover according to step 1 in the "Voltage Selection"
procedure listed above.)
1. Pull each fuse holder straight towards you. The fuse holders
are the black squares with arrows located directly beneath the
voltage selector (Fig. A.2).
2. Remove each fuse from its holder. Check the fuse amperage,
voltage, and type according to the following description. You
should have either:
•
two T2A/250V fuses, for 100 - 120 VAC operation, or
•
two T1A/250V fuses, for 220 - 240 VAC operation
Door of
Power-Entry Module
Fuse
DT-Z027E\DG
Fuse Holders
Figure A.3 Fuses
3. Assuming that you have the proper fuses on hand, reinsert the
fuses and fuse holders, making sure that the arrows on the
holders are oriented in the same direction as the arrow inside
the cover panel (Fig A.3).
4. Close the cover panel by swinging it upward and pressing it in
until it snaps closed. The correct voltage should appear in the
cut-out panel.
NOTE: To avoid damaging the instrument, verify that the new voltage
setting (displayed in the cut-out window) is correct before you turn it on!
Power Cord
82
Attach the power cord at the lower left of the detector’s rear panel.
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01/2004
MAKING
REAR PANEL
CONNECTIONS
Locate the two connectors (8-pin and 12-pin) in your accessory kit
and insert them in the appropriate sockets on the detector's rear panel
(Fig. A.4). Note that the connectors are both labeled and keyed to the
sockets, making it impossible to insert them incorrectly.
UV1000
I
ON
O OFF
TERMINAL CONFIGURATION
12. STOP (Input)
11. GROUND
10. RUN (Input)
9. GROUND
8. ZERO (Input)
7. GROUND
6. CHASSIS GND.
5. GROUND
4. READY (Output)
3.
2.
1.
H.
G.
F. 1AU/V
E. GROUND
D.
C.
B. 1.O. VF/S
A. GROUND
TERMINAL CONFIGURATION
THIS PRODUCT IS COVERED BY ONE OR MORE OF THE FOLLOWING PATENTS:
498, 812
220Vac
TERMINAL CONFIGURATION
12. STOP (Input)
11. GROUND
10. RUN (Input)
9. GROUND
8. ZERO (Input)
7. GROUND
6. CHASSIS GND.
5. GROUND
4. READY (Output)
3.
2.
1.
ANALOG 1
OUTPUT
H.
G.
F. 1.O. VF/S
E. GROUND
ANALOG 2
OUTPUT
D.
C.
B. 1.O. VF/S
A. GROUND
THIS PRODUCT IS COVERED BY ONE OR MORE OF THE FOLLOWING PATENTS:
498, 812
DT-Z170E/DT
UV2000
Figure A.4 Analog output and remote communications connectors
The upper connector holds the detector's analog output terminals.
The lower connector allows the detector to communicate with other
devices in your liquid chromatographic system. There is also a
communications port, labeled COMM.
Use the cables supplied with your detector to make the connections
described in this section. For each connection, insert the cable's bare
wire into the appropriate detector terminal. Hold the wire in place
while you tighten the small setscrew located next to each opening.
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83
UV2000 Analog
Output Connections
The terminals on the UV2000's analog output connector are labeled
ANLG 1 Output and ANLG 2 Output (Fig. A.4). Each output has
four terminals, labeled H through E for Output 1, and D through A for
Output 2. These terminals correspond to:
•
0.01 V full-scale (terminals H and D)
•
0.10 V full-scale (terminals G and C)
•
1.0 V full-scale (terminals F and B)
•
Ground (terminals E and A)
NOTE: Analog outputs are driven to twice their range. In other words,
their maximum output is twice the selected range. To avoid clipping the
voltage, be sure to connect integrators and data systems to the 1.0 V
terminal and to use caution when connecting recorders to the 0.01 or 0.10 V
terminals.
UV1000 Analog
Output Connections
The terminals on the UV1000's analog output connector are labeled
UNRANGED Output and RANGED Output (Fig. A.4). The
UNRANGED Output is a 1 AU/VOLT unrangeable output primarily
used for integrators and workstations. The UNRANGED Output can
be connected to terminal F with a ground at terminal E or A. The
RANGED Output has four terminals:
•
0.01 V full-scale (terminals H and D)
•
0.10 V full-scale (terminals G and C)
•
1.0 V full-scale (terminals F and B)
•
Ground (terminals E and A)
Integrators/Workstations
For the UV2000, connect your integrator/workstation to the 1.0VF/S
(F or B) and corresponding ground (E or A) terminals. For the
UV1000, connector your integrator/workstation to the 1AU/V output
terminal (F) and the ground (E) terminal.
Recorders
Connect the positive input from your recorder to the full-scale voltage
(0.01, 0.10, or 1.0 V) appropriate for your recorder. Connect the
recorder's floating ground input to the corresponding GROUND
terminal.
NOTE: Do not connect the detector's GROUND to any earth ground on
your recorder. This would lead to an increased noise level and a subsequent
decrease in sensitivity.
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Remote
Communications
Connections
Your detector can accept inputs from, and send inputs to, remote
devices through the remote communications connector (Fig. A.4).
For example, if your chromatographic system has programmable
timed events (contact closures or TTL), you can use one to
automatically zero the detector signal during a run.
The terminals available on the detector's remote communications
connector are labeled STOP (Input), RUN (Input), ZERO (Input), and
READY (Output), each with an associated ground terminal. The
terminals are labeled 12 through 1.
STOP (Input)
You can use a timed event from your chromatographic system to take
the detector out of run by connecting the system's event to the
detector's STOP (Input) and GROUND terminals (terminals 12
and 11).
RUN (Input)
You can use the remote-start event on your injector or autosampler to
automatically put the detector into run whenever an injection occurs
by connecting the event to the detector's RUN (Input) and GROUND
terminals (terminals 10 and 9).
ZERO (Input)
You can zero the detector signal automatically by connecting a timed
event on your chromatograph to the detector's ZERO (Input) and
GROUND terminals (terminals 8 and 7).
READY (Output)
The detector is capable of driving one TTL load each time it goes to
its READY state through the READY(Output) terminal. This ability
to signal other instruments is particularly useful with autosamplers,
where the detector can signal that it is ready for the next injection in
an automated series of runs. To hook up the READY(Output)
terminal, connect the input from the other instrument to the detector's
READY(Output) and GROUND terminals (terminals 4 and 5). For
more information on accessing this feature through the detector's
keypad, see page 44.
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85
CONNECTING TO
THE FLOWCELL
Use the following steps to connect the flowcell:
1. Remove the front panel of the detector. Although the flowcell
assembly is located behind the lower housing (Fig. A.5), the
housing does not need to be removed to connect your inlet and
outlet lines.
Display
RUN
STATUS
MENU
STOP
Keypad
ENTER
ZERO
SpectraSYSTEM
UV2000
Lamp
Housing
Flowcell
Housing
Figure A.5 The flowcell assembly is located behind the flowcell housing
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2. Use the finger-tight fitting and ferrule sets included with the
installation kit to connect the column outlet directly to the
detector's flowcell (fluid) inlet. Figure A.6 shows how the inlet
line enters the detector from the left side, and winds around the
flowcell before entering the flowcell from the bottom.
NOTE: If additional tubing is required to reach the inlet, use a zero
dead-volume union.
3. Connect the detector's fluid outlet to the low-pressure union
and waste tubing supplied in the installation kit.
HINT: If you have several detectors (fluorescence, refractive index,
electrochemical, etc.) hooked up in series, place your UV1000 or
UV2000 detector closest to the column outlet, as its flowcell can
withstand the greatest backpressure.
4. Replace the front panel of the detector; making sure that the
tubing passes through the slots without being pinched.
Flowcell
Outlet
Flowcell
Inlet
Tubing
Clamp
Photodiode
Mount
Flowcell
Assembly
DT-Z019E\FM
Flowcell Assembly
Thumbscrews
Figure A.6 The flowcell assembly showing thumbscrews,
photodiode mount, and flowcell inlet
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OPTIONAL FLOWCELLS
Thermo Electron offers several different flowcells for use in different
applications. Each flowcell possesses distinct design characteristics
and performance specifications. These characteristics are compiled in
Table A.1. Contact your Thermo Electron representative for details.
Table A.1 Design and performance specifications for Finnigan SpectraSYSTEM flowcells*
Path
Length
(mm)
Path
Flowcell
Analytical LC
Analytical LC
Microbore
Microbore
Semi-Prep,
Open Column
Volume (µL)
Tubing
Diam. (in.)
Max. Flow
Material** (mL/min)
Max.
Press.
(psi)
6
10
3
6
9
15
1.2
7.0
.01
.01
.005
.007
SS1
SS1
SS1
SS1
50
50
10
20
1000
1000
1000
1000
3
4.5
.02
SS1
100
1000
* All cells use sapphire for windows. All but the preparative flowcells have a heat exchanger.
** SS1 = Stainless Steel with TFE Gaskets.
Flowcell Orientation
The flowcells listed above are configured for use with any
SpectraSYSTEM detector. These detectors are vertically-oriented
detectors that have the tubing clamp located above the flowcell as
shown in Figure A.7.
If you plan to use any of these flowcells on non- SpectraSYSTEM
detectors (primarily horizontally-oriented detectors that have their
tubing clamps located to the left side of the flowcell as shown in
Figure A.8), you must realign the cell holder as described in the
following instructions.
Figure A.7 shows the flowcell as it is shipped.
NOTE: Figures A.7 and A.8 show the tubing clamp as an aid to the
proper positioning of the inlet and outlet tubes. The tubing clamp,
however, is actually mounted onto the detector and is not part of the
flowcell itself.
HINT: To ensure proper alignment, always hold the cell holder and
flowcell in the orientation shown in the illustrations.
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DET\Z014\FM
Inlet Tube
Outlet Tube
Tubing Clamp
(on
verticallyoriented
detectors)
Photodiode
Standoffs
Mounting
Screws
A.
Figure A.7 The alignment of UV1000 and UV2000 flowcells
as they are shipped
1. Remove the mounting screws and set them aside.
2. Rotate the cell holder 90° counterclockwise. Do not rotate the
cell body! Part B of Figure A.8 shows the cell holder in its
new position. Note the new position of the photodiode
standoffs.
Photodiode
Standoffs
Tubing Clamp
(on horizontallyoriented detectors)
Outlet Tube
Inlet Tube
Cell Holder
B.
Mounting Screw
Holes
C.
Figure A.8 Changing the alignment of a flowcell so that it can be used on other detectors.
(Turn the cell holder as shown in Part B. Align the inlet and outlet tubes
with the tubing clamp as shown in Part C.)
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89
3. Reattach the cell body by replacing and securing the mounting
screws.
4. Gently bend the inlet and outlet tubes as shown in Part C of
Figure A.8. The inlet tube (wound around the cell body)
should always enter at the bottom of the flowcell; the outlet
tube should always exit at the top of the flowcell.
Specifications
Wavelength:
D2 lamp: 190 to 380 nm (UV1000)
190 to 365 nm (UV2000)
W lamp: 366 to 800 nm
Lamp(s):
UV1000: D2 standard, W optional
UV2000: D2 and W standard
Bandwidth:
6 nm
Wavelength Accuracy:
± 1.0 nm
Wavelength Precision:
± 0.1 nm
Range Selections:
3.0, 2.0, 1.0, 0.5, 0.2, 0.1,
0.05, 0.02, 0.01, 0.005,
0.002, 0.001, 0.0005 AUFS
Absorbance Range:
0.0005 to 3.0 AUFS
Absorbance Linearity
@ 254 nm:
Better than 1% to 2.0 AU
Analog Outputs:
UV2000:
Outputs 1 and 2:
Range-selectable over entire
absorbance range
Communications:
Remote Inputs:
Remote Outputs:
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Run, Stop, and Zero
Ready
01/2004
Specifications (cont.)
Noise:
Single-wavelength Mode:
@ 254 nm,
1.0-sec rise time:
< ± 1.0 × 10 -5 AU
Dual-wavelength Mode:
@ 254 280 nm,
1.0-sec rise time
(UV2000 only):
< ± 2.5 × 10 -5 AU
Drift:
after warm-up @ 254 nm < 2 × 10 -4 AU/hour
Display:
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2 × 24 character, high-contrast LCD
Dimensions:
14.5” (37 cm) × 6” (15 cm) ×
18.5” (47 cm)
Weight:
40 lb. (18 kg)
Power Requirements:
100/120/220/240 VAC
50/60 Hz
1.5/1.6/8.0/0.8A, 225VA
Environmental:
10-40°C
5-95% RH noncondensing
Thermo Electron
91
B
Menu Reference
Introduction
This Appendix provides you with two Menu Trees, one each for the
UV1000 (page 94) and the UV2000 (page 95). It also provides you
with an alphabetical description of all the instrument's display fields.
Fortunately, it is not necessary to read this Appendix in order to learn
how to use your detector. It is included in the manual simply as a
quick reference and aid to using your instrument.
The Menu Trees are a representation of the detector’s overall menu
structure. They show the location and interrelation of all the menus
for your detector and, as such, they are a good reference to keep on
hand while you work through the operating instructions in Chapters 3
and 4. The menu trees will also help if you become "lost" while
moving through the detector's menus. Separate, removable copies can
be found in the pocket at the front of this manual.
The Menu Reference is an alphabetical listing of each menu field and
command. Included in each listing is the field's definition and, where
appropriate, all allowable and default values.
Menu Trees
The UV1000 and UV2000 Menu Trees are useful tools for learning
your way around your detector. You may wish to keep one handy
while you learn where each display is located in the overall menu
structure.
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93
Thermo Electron
[MENU]
FILE
COMMANDS
OPT IONS
T EST S
Load
Delete
Software Version
Light Levels
Diode Offsets
Self-Tests
T EST S
Lamps
Analog Outputs
More
OPT IONS
Event Mark
Short Output
Shutdown Detector
COMMANDS
Edit
FILE
UV1000 Menus
Zero on λ Change
Cursor Speed
Status Lock
READY Output
More
10
10
Yes
Medium
Off
Active Hi
Analog 1 Offset (mV)
Analog 2 Offset (%)
Analog Outputs
D2 (190-380)
Lamp
0
D2 Lamp Hours
0
W Lamp Hours
0:00
Current Time
Manual
Startup
0:00
Startup Time
Manual
Shutdown
0:00
Shutdown Time
1:00
Time From Ready
Lamps
Delete File
Delete
Load File
Load
Wavelength Program
Options
Edit
Status
λ
READY
250
Time
Wavelength
0.00
250
Rise Time
Autozero Time
Range
[STATUS]
1.0
0.00
1.0
1.0
0.00
1.0
AU
0.00001
Wavelength
250
Options
Rise Time
Autozero Time
Range
Time
0.00
Wavelength Program
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FILES
QUEUE
TESTS
COMMANDS
OPTIONS
[MENU]
pg 1 of 2
L oad
Delete
Thermo Electron
(see page 2)
QUEUE
Software Version
L ight L evels
Diode Offsets
Self-Tests
TESTS
Lamps
Analog Outputs
More
OPTIONS
Event Mark
Short Outputs
Shutdown Detector
COMMANDS
Edit
Copy
FILES
UV2000 Menus
More
Analog Outputs
Lamps
Delete File 1:
Delete
Copy File 1:
To File 1:
Copy
Load File 1:
Load
Edit File 1:
File Name
Edit
1
1
AU
+01.000
Yes
Medium
Off
Active Hi
Protect
Off
Off
Off
Off
Zero On λ Change
Curs or Speed
Status Lock
READY Output
File Name
1:
2:
3:
4:
D2 (190-365)
0
0
0:00
Manual
0:00
Manual
0:00
1:00
Analog 1 Offset (%)
Analog 2 Offset (%)
Analog 2
K Factor
Lamp
D2 Lamp Hours
W Lamp Hours
Current Time
Startup
Startup Time
Shutdown
Shutdown Time
Time From Ready
(see page 2)
(see page 2)
Wavelength Program
Options
File D: Develop
File S: Scan
Files 1-4
Program Single λ
(190-450)
λ2
280
Rise Time
Autozero Time
Range 1
Range 2
Options
Program Dual λ
Time
λ1
0.00
366
(366-700)
λ2
700
Program Dual λ (366-700)
Program Dual λ
Time
λ1
0.00
250
Program Dual λ (190-450)
Program Single λ
Time
Wavelength
0.00
250
DT-Z042E\DT
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Setup
Replay
[MENU]
QUEUE
Replay
Setup
Pause
Delete
Edit
Queue is Loaded
Edit
Load
Delete
No Queue Loaded
220
Start λ
350
End λ
λ Interval
5
10.00
Run Time
1
File D: Develop Runs Per λ
1.0
Rise Time
Autozero Time 0.00
1.0
Range 1
1.0
Range 2
File S: Scan
FILES QUEUE TESTS
COMMANDS OPTIONS
pg 2 of 2
UV2000 Menus
(filename)
1
Pause Queue
Pause
Delete Queue
Delete
File Name
Order
Load Queue
Edit
1
# Runs
1.0
1.0
5
0.00
Range 1
Range 2
Replay Rate (nm/sec)
Spectra Time
Replay Spectra
Display AU, λ
Load
220
350
5
250
1.0
0.00
1.0
1.0
Start λ
End λ
λ Interval
Run λ
Rise Time
Autozero Time
Range 1
Range 2
λ
[STATUS]
λ
Single λ
Dual λ
Scan File Loaded
Develop File Loaded
Display
Display AU, λ
Replay
AU
AU
Replay Spectra
Single λ
Scan
Zero
λMaxAU lMin
AU
0.0001
Status
λ
AU #Runs
READY
250 0.0001 1/3
File D:Develop
Time
Wavelength
0.00
250
Rise Time
1.0
Autozero Time
0.00
Range 1
1.0
Range 2
1.0
Develop File Loaded
Status
λ
READY 250
File S:Scan
Time
λMax
Scan File Loaded
Status
λ
AU
READY 250 0.0001
READY 280 0.0001
File 1:
Time
λ1
λ2
0.00
250
280
Rise Time
1.0
Autozero Time
0.00
Range 1
1.0
Range 2
1.0
Dual λ
Status
λ
AU
READY 250
0.0001
File 1:
Time Wavelength
0.00
250
Rise Time
1.0
Autozero Time
0.00
Range 1
1.0
Range 2
1.0
DT-Z043E/DT
Menu Reference
For quick reference, we have included this alphabetical list of each
field, including a short definition, and allowable and default values.
For a more detailed explanation of the functions of your detector, you
should refer to Chapter 3: Basic Operations, and Chapter 4: Advanced
Operations.
Some fields are common to both the UV1000 and the UV2000, so we
have indicated the detector name for fields that appear in only one
detector.
Analog 1 Offset (mV)
This field offsets the Analog 1 output signal by 0, 1, 2, 5, 10, 20, or 50 mV.
The default setting of 10 mV is appropriate in most cases. UV1000 only.
Analog 1 Offset (%)
This field offsets the Analog 1 output signal by a positive or negative 50, 20,
10, 5, 2, 1, or 0 percent of the full-scale range. Default is 0%. UV2000 only.
Analog 2
This field allows you to select the output signal from the Analog Output 2
terminal. The selections are AU (the absorbance signal for wavelength one
in single-wavelength operation or from wavelength two in dual-wavelength
operation), AU1-KxAU2 (a calculated peak response using the K Factor
technique), and AU1/AU2 (the absorbance ratio of wavelength 1 to
wavelength 2). Default is AU. UV 2000 only.
Analog 2 Offset (%)
This field offsets the Analog 2 output signal by a positive or negative 50, 20,
10, 5, 2, 1, or 0 percent of the full-scale range. Default is 0%.
Analog Outputs
This menu allows you to offset the signals from the analog output terminals
located on the back panel of the instrument. For the UV2000, you can also
select the output signal for Analog Output 2 and input a K factor.
AU
This field, located in the Status Screen, shows the detector's current
absorbance reading. It is a six-digit number, ranging from -3.00000 to
+3.00000 AUFS.
Autozero Time
This field tells the detector when to perform an automatic zero. Allowable
values are 0.00 to 999.99 minutes. Default is 0.00 minutes.
COMMANDS
The Commands Menu lets you put an event mark into your chromatogram,
short detector outputs, and shut down the detector.
Copy
This field accesses the Copy File field. UV2000 only.
Copy File
This field, along with the To File field, allows you to copy from the specified
file to another file designation. UV2000 only.
Current Time
This field displays local time ranging from 0:00 to 23:59.
Cursor Speed
This field may be set to Slow, Medium, or Fast according to your need.
Default is Medium.
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Delete
Under the top-level menu FILE(S), this field accesses the Delete File
command.
Under the top-level menu QUEUE, this field accesses the Delete Queue
command. UV2000 only.
Delete File
This field deletes the designated file, setting all fields to their default values.
After pressing [ENTER], the confirmation message **File Deleted**
appears for one second.
Delete Queue
This field deletes the queue. After pressing [ENTER], the confirmation
message **Queue Deleted** appears for one second. UV2000 only.
D2 Lamp Hours
This field tracks the total number of hours the detector's deuterium lamp has
been in operation (up to 9999). When a new lamp is installed, you must set
this parameter to zero.
Diode Offsets
This field displays the analog-to-digital (A/D) conversion frequencies of the
sample and reference diodes when both lamps are turned off. These values
can be used to measure the detector's digital noise level.
Display AU, λ
This command calls up the Display AU, λ screen, a screen that shows the
incremental wavelength versus absorbance data for the selected spectral
scan.
Edit
Under the top-level FILE(S) menu, the Edit Menu allows you to set up or
edit files. The edits do not change the current settings of the detector until
the file is loaded.
Under the top-level QUEUE menu, the Edit Menu allows you to set up or
edit a Queue. Edits may not be made to Order 1 while a queue is loaded or
running unless you pause the queue first. UV2000 only.
Edit File
This field allows you to identify the file for set up or edit. Allowable
designations are 1 to 4, S for the Scan file, and D for the Develop file.
Default is 1. UV2000 only.
End λ
In the Scan File Setup, this field defines the wavelength at which the detector
should finish the scan. Allowable values are 191 to 800 nm. Default is 350
nm. UV2000 only.
In the Develop File Setup, this field defines the wavelength at which the
detector should run its last set of injections. Allowable values are 191 to 800
nm. Default is 350 nm. UV2000 only.
Event Mark
The Event Mark field applies a 15% of full-scale spike on the detector's
output signals.
FILE(S)
The UV1000’s FILES Menu and the UV2000’s FILES Menu allow you to
edit, load, and delete files. On the UV2000, the FILES Menu also lets you
copy files.
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File Name
This field allows you to enter a file name for a designated file (numbered
1 to 4). The name can contain up to eight characters from the following list:
A to Z, 0 to 9, /, -, and blank. Default is blank. UV2000 only.
For Files S and D, the file names are automatically designated as SCAN and
DEVELOP, respectively. No editing of these file names is allowed.
UV2000 only.
K Factor
This field is used in the technique. Allowable values are -99.999 to 99.999.
Default is 1.000. UV2000 only.
λ (λ1, λ2)
The wavelength field (λ in the UV1000, λ1, λ2 in the UV2000) located in
the Status Screen shows the current detector wavelength setting(s).
For the UV2000 only, these fields also appear in the Wavelength Program
for dual-wavelength operation.
λ Calibration
The wavelength calibration screen located in the Tests Menu shows the
current detector wavelength setting(s).
λ Interval
In the Scan File Setup, this field defines the wavelength interval at which the
detector should perform the scan. Allowable values are 1, 2, 3, 4, 5, and 10
nm. Default is 5 nm. UV2000 only.
In the Develop File Setup, this field defines the wavelength increment the
detector monochromator should use for wavelength changes between each
set of injections. Allowable values are 1, 2, 3, 4, 5, 10, and 20 nm. Default
is 5 nm. UV2000 only.
λMax
This field is the wavelength maximum in a spectral scan. UV2000 only.
λMaxAU
This field is the absorbance value for the corresponding wavelength
maximum in a spectral scan. UV2000 only.
λMin
This field is the wavelength minimum in a spectral scan. UV2000 only.
λ Offset
The lambda offset screen lets you choose a number of steps, each
representing 0.25 nm, by which you want to offset the wavelength. This
field is used to check the detector's wavelength accuracy. Allowable entries
are: 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, -1, -2, -3, -4, -5, -6, -7, -8, -9, and -10.
The default is 0.
Lamp
The Lamp field allows you to choose from among several selections: D2
(190-380) for the UV1000's deuterium lamp; D2 (190-365) for the UV2000's
deuterium lamp; W (366-800) for the tungsten lamp; D2+W (190-800) for
dual-lamp operation (UV2000 only); or Off to shut the lamp(s) off. Default
is D2 (190-380) or D2 (190-365), for the UV1000 and UV2000,
respectively.
Lamps
The Lamps Menu allows you to control the detector's lamp operations.
Light Levels
This field displays the analog-to-digital (A/D) conversion frequencies of the
light detected by the sample and reference diodes when the D2 lamp is on.
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Load
Under the top-level menu FILE(S), the Load selection accesses the Load File
command.
Under the top-level menu QUEUE, the Load selection accesses the Load
Queue field. UV2000 only.
Load File
The Load File field loads the designated file settings into the active runfile.
After pressing [ENTER], the confirmation message **File Loaded**
appears for one second.
Load Queue
The Load Queue field loads the queue. After pressing [ENTER], the
confirmation message **Queue Loaded** appears for one second. UV2000
only.
More
This menu allows you to access the Zero on λ Change, Cursor Speed, Status
Lock, and READY Output fields. In the UV2000, you can also protect files
from this menu.
OPTIONS
Found in the Main Menu, the Options Menu allows you to perform lamp and
analog output operations.
Options
The Options selection in the Edit Menu of FILE(S) allows you to edit Rise
Time, Autozero Time, and Range.
Order
This field designates the order in which the selected files in a queue will be
run. UV2000 only.
Pause
This field accesses the Pause Queue command. UV2000 only.
Pause Queue
This field pauses an active queue. If a file is running, it continues until
completed, and the detector returns to a READY state. UV2000 only.
Program
This field allows you to select single- or dual-wavelength operation. The
selection toggles between Single λ, Dual λ(190-450), and Dual λ(366-700).
Default is Single λ.
Protect
This field, along with the File Name field, protects a specified file from
being edited, copied to, or deleted. The field toggles between On, allowing
no changes to the file, and Off, where changes may be made. Default is Off.
UV2000 only.
QUEUE
The Queue Menu allows you to edit, load, delete, or pause a queue. A queue
is a series of files that are run in a specific order, and is typically used for
automated runs. UV2000 only.
Range
The Range field controls the full-scale output range for the UV1000’s
Analog Output 2 terminal. Allowable full-scale ranges are 3.0, 2.0, 1.0, 0.5,
0.2, 0.1, 0.05, 0.02, 0.01, 0.005, 0.002, 0.001, and 0.0005 AUFS. Default is
1.0 AUFS. UV1000 only.
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Range 1, Range 2
The Range 1 and Range 2 fields control the full-scale output ranges for the
UV2000's Analog Output 1 and Analog Output 2 terminals. Allowable fullscale ranges are 3.0, 2.0, 1.0, 0.5, 0.2, 0.1, 0.05, 0.02, 0.01, 0.005, 0.002,
0.001, and 0.0005 AUFS. Default is 1.0 AUFS. UV2000 only.
READY Output
This field is used to communicate with other devices through the detector's
READY(Output) terminal. This TTL terminal switches the transistor
between high and low states whenever the detector starts a run. Select
"Active Hi" or "Active Lo," for the high or low state, respectively. Default
is Active Hi.
Replay
The Replay command sends you to the Replay Menu, from which you can
set up the parameters for replaying stored spectra. UV2000 only.
Replay Spectra
This command is used to initiate replay of the designated spectrum. UV2000
only.
Replay Rate
This field designates the rate at which the detector replays a stored spectrum.
Allowable values are 1, 2, 5, 10, and 20 nm/sec. Default is 5 nm/sec.
UV2000 only.
Rise Time
This field controls the detector's response time. Rise time is inversely
proportional to the amount of baseline noise. Allowable values are 0.0, 0.1,
0.2, 0.5, 1.0, 2.0, 3.0, 4.0, and 5.0 seconds. Default is 1.0 second.
#Runs
When this field appears in the Status Screen, the current run and the total
number of injections to be made at the displayed wavelength appear directly
below it. The field is updated at the beginning of each injection. UV2000
only.
When this field appears in a Queue setup, it displays the number of times
each file runs in a queue. UV2000 only.
Run λ
This field designates the monitoring wavelength to be used when running the
Scan file. Allowable values are 190 to 800 nm. Default is 254 nm. UV2000
only.
Run Time
Located in the Develop file, this field is the amount of time designated for
each chromatographic run. Allowable values are 0.01 to 999.99 minutes.
Default is 10.00 minutes. UV2000 only.
Runs per λ
Located in the Develop file, this field designates the number of injections to
be performed at each wavelength increment. Allowable values are 1 to 9.
Default is 1. UV2000 only.
Scan
This field appears in the Status Screen when the Scan file is loaded. To
initiate a scan, move the cursor to this field and press [ENTER]. UV2000
only.
Scan Zero Time
This field allows you to set a runtime at which the detector will perform a
baseline scan automatically. Allowable values are 0.00 to 99.99 minutes.
Default is 0.00. UV2000 only.
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Self-Tests
This command tells the detector to run through its internal diagnostic tests.
Setup
The Setup Menu allows you to set up the parameters in the Scan file.
UV2000 only.
Short Outputs
This command allows you to short the detector's outputs together. When
you select Short Outputs, the detector's analog outputs are shorted together
(zero volts) and the field changes to Unshort Outputs. To remove the short
and return the outputs to their normal operating state, select Unshort
Outputs, and the field changes back to Short Outputs. When you leave this
screen, the field returns automatically to Short Outputs.
Shutdown
This field toggles between Manual (you turn off the lamp manually), Time
(the lamp turns off automatically at a preset time), Time from READY (as
explained under the Time from READY field), and End of Queue (the lamp
turns off when the queue is finished, UV2000 only). Default is Manual.
Shutdown Detector
This command shuts down the detector's lamps and motors, leaving the
electronics on to preserve memory. Press any key to return the detector to
the same settings as when this field was activated.
Shutdown Time
This field displays the local time, ranging from 0:00 to 23:59, at which you
have programmed the lamp to turn off automatically. Default is 0:00.
Software Version
This field displays the EPROM version of your detector's software.
Spectra Time
This field displays a list of the scans that are currently stored in memory.
Each scan is identified by the runtime at which it was initiated. UV2000
only.
Start λ
In the Scan File Setup, this field defines the wavelength at which the detector
should begin the scan. Allowable values are 190 to 799 nm. Default is 220
nm. UV2000 only.
In the Develop File Setup, this field defines the wavelength at which the
detector should run its first set of injections. Allowable values are 190 to
799 nm. Default is 220 nm. UV2000 only.
Startup
The Startup field toggles between Manual, where you manually turn on the
lamp, and Time, where the lamp automatically powers up at a preset time.
Default is Manual.
Startup Time
This field displays the local time, ranging from 0:00 to 23:59, at which you
have programmed the lamp to start up automatically. Default is 0:00.
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Status
This field in the Status Screen shows the current condition of the detector.
The possible conditions are: READY (the selected lamp is lit and ready for
initiation of a run), NRDY (the detector is set to the wrong lamp for the runwavelength requested, is performing internal tests, or has a possible internal
problem), or UVW (the deuterium lamp is warming up). The run time is
displayed when the running file has a programmed stop-time.
In the UV2000, the letter Q appears at the beginning of this field when a
queue is loaded.
Status Lock
The Status Lock field limits accessibility to the Status Menu (the
programming section below the Status Screen). When set to On, only the
Status Screen appears on the display and the down-arrow icon is not seen.
Default is Off.
TESTS
The allows you to perform the detector's internal diagnostic, light level, and
diode offset tests.
Time, Wavelength
The Wavelength Program contains the Time and Wavelength fields. It
allows you to program changes in the detector's wavelength as a function of
time.
Time refers to the run time, in minutes, when a timed event (wavelength
change, autozero, or run stop) is to occur. Allowable values range from 0.00
to 999.99 minutes. Default is 0.00 minutes.
Wavelength refers to the wavelength that will be set at a specified time.
Allowable values for the UV1000 are: 190 to 380 nm with the deuterium
lamp, and 366 to 800 nm with the tungsten lamp. Allowable values for the
UV2000 are: 190 to 365 nm with the deuterium lamp, and either 366 to
700 nm or 366 to 800 nm with the tungsten lamp (depending on whether the
detector is operating in the dual-wavelength or the single-wavelength mode,
respectively). Default is 250 nm.
Time from READY
A preset time interval from the Ready state of the detector, after which the
detector lamp will turn off if a start signal has not been received from the
keypad or external Run(Input) terminal. Allowable values range from 0:30
to 9:59 hours. Default is 1:00.
To File
This field, along with the Copy File field, allows you to copy a file to the
specified file identification. UV2000 only.
W Lamp Hours
This field tracks the total number of hours the detector's tungsten lamp has
been in operation (up to 9999). When a new lamp is installed, you must set
this parameter to zero.
Wavelength Program
This command allows you to access the Wavelength Program. See the
"Time, Wavelength" description above for details.
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Zero
This field appears in the Status Screen when the Scan file is loaded. To
initiate a background scan, move the cursor to this selection and press
[ENTER]. UV2000 only.
Zero on λ Change
This field toggles between Yes, where the detector baseline automatically
zeroes at each timed event during a programmed run, and No. Default
is Yes.
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C
Troubleshooting
Introduction
This Appendix provides you with helpful information for
troubleshooting possible detector and chromatographic system
problems. We have divided it into four sections:
•
a brief theory of operation
•
a troubleshooting guide that lists symptoms, possible problems,
and remedies
•
error messages you might see on the detector's display
•
a description of internal and external diagnostic tests
Theory of Operation
This brief Theory of operation is included to aid you in
troubleshooting problems and performing maintenance for your
detector. For more detailed information, you should contact your
Thermo Electron representative.
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OVERVIEW
Figure C.1 shows the optical system used in both the UV1000 and
UV2000 detectors. The detector operates in a double-beam mode
using a fiber-optic beam-splitter that creates sample and reference
beams. The reference beam is directed to a reference photodiode.
The sample beam is lens-focused prior to passing through the flowcell
to a sample photodiode.
An analog PCB processes the signals from the photodiodes and
provides analog output signals through an 8-pin external connector.
The digital PCB contains the EPROM (the built-in software),
provides digital processing circuitry, and interfaces with the
keyboard/display and the remote communications devices.
(Additional software is held on an EPROM PCB.) The Motherboard
provides all the necessary interconnections and power supplies.
Lamps
Deuterium
"D2"
Grating
Shutter
Baffles
Tungsten
"W"
Reference
Photodiode
Beam
Splitter
Lens
DT-Z001E/FM
Sample Cell
Sample Photodiode
Figure C.1 The optical system for the UV1000 and UV2000 detectors
(Only the UV2000 has a shutter.)
The deuterium and tungsten lamps are continuum light sources that
provide high light intensity over the UV and visible wavelength
ranges. Two sets of baffles minimize stray light. A concave
holographic grating actuated by a microprocessor-controlled stepper
motor provides wavelength selection.
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Common Problems
This next section contains a table of symptoms, possible causes, and
remedies for some common problems you may observe in detector
response. Many of the problems attributed to the detector may
actually be due to other components in the chromatographic system,
so we have included references to these types of problems and
solutions as well.
Troubleshooting Table
Symptom
Cause
Remedy
1. Spikes on baseline.
a. Gas bubbles in the flowcell.
a. Degas mobile phase. Supply
backpressure device to flowcell (check
back-pressure rating). Check for leaks at
high-pressure fittings.
b. Immiscible solvent bubbles
following mobile phase
changeover.
b. Flush flowcell with 2-propanol, then
with mobile phase.
c. Electrical interference.
c. Check electrical lines for good
connections and/or interference from
broadcast radiation. Check for ground
loops.
d. Extremely large fluctuations
in voltage on AC line.
d. Remove systems (e.g., ovens) that cause
voltage fluctuations, isolate the detector
to "quiet" circuit, or use UPS (UPS,
uninterruptible power supply).
a. Contaminated flowcell.
a. Flush flowcell with cleaning solvents as
described in Chapter 5. Check for leaks.
b. Leak in sample inlet line.
b. Check all fittings from column outlet to
flowcell inlet for leaks.
c. Bubble trapped in flowcell.
c. Increase flow rate until bubble is
removed. Supply backpressure device to
flowcell (check back-pressure rating to
avoid rupturing flowcell).
d. Leaking flowcell.
d. Replace flowcell.
2. Random noisy baseline.
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Troubleshooting Table (cont.)
Symptom
Cause
Remedy
2. Random noisy baseline,
cont'd.
e. Insufficient lamp warm-up.
e. Allow a 30 minute warm-up for normal
operation; one hour for maximum
sensitivity.
f. Lamp aging or defective.
f. Replace lamp.
g. Ground loop problem
between integrator and
detector.
g. Check for proper cable connections for
detector output; do not ground at both
ends of cable.
h. Flowcell, lamp, lenses, or
photodiode dirty.
h. Clean dirty component as described in
Chapter 5.
i. Integrator input voltage
does not match detector
output voltage.
i. Connect integrator to appropriate output
connectors on detector (see
Appendix A). Check attenuation setting
on integrator.
a. Flowcell contaminated.
a. Flush flowcell with cleaning solvents as
described in Chapter 5. Check for leaks.
b. Mobile phase
contamination.
b. Replace with fresh mobile phase made
with high-purity solvents.
c. Material bleeding from
column.
c. Clean or replace column.
d. Leaks in system, or
flowcell.
d. Check all fittings for leaks. Replace
flowcell.
e. Tiny bubble trapped in
flowcell.
e. Increase flow rate until bubble is
removed. Connect backpressure device
to flowcell outlet (check back-pressure
rating to avoid rupturing flowcell).
f. Large temperature
fluctuations.
f. Remove system from drafts. Thermostatically control column temperature.
3. Excessive baseline drift.
See Baseline problems.
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Troubleshooting Table (cont.)
Symptom
Cause
Remedy
4. No peaks, or peaks much
smaller than expected.
a. Incorrect wavelength setting.
a. Check wavelength setting. Make sure
the correct file is selected.
b. Lamp not on or defective.
b. Make sure lamp is lit. Run detector's
diagnostic tests to check lamp. Replace
lamp if necessary.
c. Integrator input voltage
does not match detector
output voltage.
c. Connect integrator to appropriate output
connectors on detector (see
Appendix A). Check attenuation setting
on integrator.
d. Insufficient sample reaching
the detector.
d. Check entire chromatographic system for
leaks. Verify sample injection volume.
a. Rise time is too large (too
slow).
a. Lower the rise time selection.
b. Flowcell volume too large.
b. Change to a flowcell with smaller
volume.
6. Clicking sound when
UV2000 is in dualwavelength mode.
a. Noise comes from grating
motor, and is normal.
a. No action necessary.
7. Detector won't power up.
a. Tripped circuit breaker at AC
wall outlet.
a. Resolve problem, reset circuit breaker.
b. Blown detector fuse.
b. Resolve problem, replace fuse.
c. Incorrect voltage selected.
c. Reset detector for correct incoming linevoltage (see Appendix A).
d. Power cord not connected.
d. Connect power cord.
5. Broad, tailing peaks.
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Error Messages
There are three types of error messages that you may see on your
detector's display:
•
System errors
•
Real-time errors
•
User-input errors
Each type of error is explained below in further detail.
SYSTEM ERRORS
REAL-TIME
ERRORS
System errors are indicated on the display by a double set of
exclamation points (!! !!). They occur whenever an undesirable
condition exists that prevents the detector from operating. If one of
these messages appears, try turning the detector's power switch off
and on. If the message recurs, contact your Thermo Electron
representative.
•
SYSTEM RESET
•
RAM ERROR
•
ADDRESS ERROR
•
BUS ERROR
•
DIVIDE BY ZERO
•
LOW L0 ERROR
•
LOW L1 ERROR
•
DISTANT QUEUE ERROR
Real-time error messages indicate that you need to correct a certain
hardware condition. Possible messages are:
Lamp Cover Open
Check that the detector's lamp housing is in place and properly
installed.
Low Light Detected From Deuterium Lamp
This message indicates that the deuterium lamp may not be on, may
be improperly installed, or needs to be replaced due to low light
energy. It can also appear if the lamp cover is replaced while the
lamp is on.
Using the Lamps Menu (see "Automatic Lamp Operations" on
page 41), turn the lamp state to off, wait five seconds, and then switch
the lamp on. If the error message recurs, check for proper lamp
installation according to the procedure outlined in Appendix A.
If the lamp is installed correctly, its surface is clean, and the message
still appears, replace the lamp.
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Low Light Detected From Tungsten Lamp
This message indicates that the tungsten lamp may not be on, may be
improperly installed, or needs to be replaced due to low light energy.
Using the Lamps Menu (see "Automatic Lamp Operations" on
page 41), turn the lamp state to off, wait five seconds, and then switch
the lamp on. If the error message recurs, check for proper lamp
installation according to the procedure outlined in Appendix A.
If the lamp is installed correctly, its surface is clean, and the message
still appears, replace the lamp.
INPUT ERRORS
The following error messages indicate improper use of the detector's
menu system.
A File Is Already Running
You cannot start a different file while a file is already running.
Invalid Parameters, Spectrum Not Allowed
Invalid scanning setup parameters have been entered, so the detector
cannot perform a spectral scan.
No More Available Memory
All available system memory is full.
No Queue Available
You cannot load a queue if none has been set up first. (Because
queues are not available on the UV1000, this message only appears
on the UV2000.
No Spectra Available
You cannot run Replay Spectra when no spectra are available in
memory. Because there is no scanning feature on the UV1000, this
message only appears on the UV2000.
Protected File, Cannot Be Copied To
You cannot copy to a protected file. (File protection is not offered on
the UV1000, so this message can only appear on the UV2000.
Protected File, Cannot Be Deleted
You cannot delete a protected file. (File protection is not offered on
the UV1000, so this message can only appear on the UV2000.
Protected File, Cannot Be Edited
You cannot modify a protected file. (File protection is not offered on
the UV1000, so this message can only appear on the UV2000.
Queue Loaded, Cannot Load File
When a queue is loaded, you cannot load any other file. Because
queues are not available on the UV1000, this message only appears
on the UV2000.
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Run In Progress, Testing Not Allowed
You cannot run the detector's built-in diagnostics while a run is in
progress.
Run Not In Progress, No Scanning Allowed
A spectral scan can only be performed when a run is in progress.
Because there is no scanning feature on the UV1000, this message
only appears on the UV2000.
Detector Shutdown
This message occurs when you use the Shutdown Detector field to
turn off the detector. (See "Shutdown Detector" in Chapter 4.) Press
any key on the keypad to turn on the detector.
Scan Memory Full
This message occurs when the Scan File is loaded and the scan data
memory storage is full. Because there is no scanning feature on the
UV1000, this message only appears on the UV2000.
Run In Progress, No Replay Allowed
The UV2000 does not allow you to replay stored spectral scans when
the Scan file is loaded and a run is active. Because there is no
scanning feature on the UV1000, this message only appears on the
UV2000.
Diagnostic Tests
This section describes the internal diagnostic tests supplied with your
detector. It also references two external tests that you can run. Use
these tests if you suspect that your detector is not working properly.
INTERNAL
DIAGNOSTIC
TESTS
You can access the detector's internal diagnostic tests by following
these steps:
1. Press [MENU].
2. Select /TESTS/.
3. The Tests menu appears in Figure C.2.
❑
Software Version
❑
Light Levels
--------------------------------------------------------❑
Diode Offsets
❑
λ Calibration
❑
Self-Tests
Figure C.2 Detector's Tests Menu
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Software Version
Select this field to display the EPROM version of your detector's
software (Fig. C.3).
Version 1.01
Figure C.3 The software version
Light Levels
The Light Levels test displays numbers related to the level of light
intensity seen by the sample and reference photodiodes. When you
select /Light Levels/, the screen in Figure C.4 appears.
S1:
S2:
nnnnn.n
nnnnn.n
R1:
R2:
nnnnn.n
nnnnn.n
Figure C.4 The Light Levels screen
The sample (S1, S2) and reference (R1, R2) numbers may differ
considerably between instruments. A five- or six-digit number is
typical. If you get an unusual reading, check the photodiodes and the
analog PCB. These components are the ones that are the most likely
to affect light intensity. If any of the numbers is zero, call
Thermo Electron.
Diode Offsets
The Diode Offsets test presents numbers related to the level of
background signal received from the sample and reference
photodiodes when the lamps are off (dark current). When you select
/Diode Offsets/, the screen in Figure C.5 appears.
>C S1:
nnnn.n
R1: nnnn.n
S2: nnnn.n
R2: nnnn.n
Figure C.5 The diode offsets screen
The sample (S1, S2) and reference (R1, R2) numbers may vary
considerably between instruments. A three- or four-digit number is
typical. As with the Light Levels test, check the photodiodes and the
analog PCB, the components most likely to affect light intensity, if
you get an unusual reading. If any of the numbers are zero, call
Thermo Electron.
To recalculate the diode offsets, select C. The offsets may need to be
recalculated if the light levels are less than the diode offsets. This
situation normally occurs after slight diode offset drift or while
working with extremely low light.
λ Calibration
01/2004
Selecting /λ Calibration/ brings up the screen shown in Figure C.6.
You can use this screen (in combination with the optional Cuvette
Thermo Electron
113
Holder Accessory) to offset the factory-calibrated wavelength to more
closely match an FDA, industry, or in-house calibration standard.
NOTE: If you wish to conduct your calibration using the Cuvette Holder,
the following procedure has also been detailed in Appendix E for your
convenience.
λ Offset (steps)
0
Figure C.6 The lambda offset screen
NOTE: The UV1000 and UV2000 detector is calibrated using a mercury
lamp fixture. This provides a very narrow emission line at 254 nm. Broadband calibration standards, such as holmium oxide and didymium filters,
make calibration more difficult and less accurate.
To offset the factory-calibrated wavelength, select the number of
"steps" by which you want the wavelength to be offset. Each step
represents approximately 0.25 nm, so if you choose "2" for the
number of steps, you will have offset the wavelength by + 0.5 nm.
You can offset the wavelength by as much as ± 2.5 nm.
NOTE: The offset value is not cleared upon resetting the RAM memory. It
can only be changed from the lambda offset screen.
Self-Tests
The detector automatically runs eight internal diagnostic tests when it
is powered up. To run the tests at any other time, simply select
/Self-Tests/.
If any test (other than the two lamp tests) fails, you'll see a message to
that effect on the display. Clear the message and run the remainder of
the self-tests by pressing [ENTER]. Repeat this process as many
times as necessary until all self-tests are completed and the Status
Screen appears. If any test has failed, the Status Screen will read
"NRDY" (Not Ready).
Although you can frequently get back to the Ready state on your own
(e.g., you can manually turn on the lamps from the Options Menu, or
load a file), the detector may not function properly and your results
may be affected. For this reason, and to help you troubleshoot the
detector on your own, we have listed the MLF (most likely failure)
for each test. Problems that are not readily resolved should be
referred to your Thermo Electron representative.
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The eight self-tests are:
1. RAM. This test checks both non-volatile and volatile RAM
with a read/write test. The "Testing RAM" message only
appears during self-initiated testing. On power-up, the test
occurs without any special message. A failure during either
type of testing is indicated by the messages "Bad DRAM" or
"Bad NOVRAM." MLF: Digital PCB.
2. Voltages. This test checks the circuitry-supply voltages.
MLF: Motherboard.
3. Analog Outputs. This test checks the scale and linearity of the
output signal (recorder/integrator). Failure is indicated by a
"Fail" or a "Bad Analog Linearity" message. MLF: Analog
PCB.
4. Diode Offsets. This test checks the diodes (photodiodes) with
the lamp(s) off (dark current). Either a “Bad Sample Diode” or
an “Intense Light Detected” message indicates failure. You
should verify that the sample photodiode is fastened securely to
the flowcell and that light is actually passing through the
flowcell. If "Fail" or a "Bad Ref. Diode Detected" appear, call
your Thermo Electron representative. MLF: Photodiode or
Analog PCB.
5. Motor. The Motor Test checks the monochromator motor and
its voltages. MLF: Motor.
6. Deuterium Lamp. This test checks the D2 lamp and its
voltages when the lamp is on and when it is off. If the message
"D2 Not Detected" appears, the lamp voltages are good, but the
lamp is either not present or not functioning properly. Try
replacing the deuterium lamp and retrying the test. If the word
"Fail" appears, call your Thermo Electron representative.
MLF: Lamp or Motherboard.
7. Tungsten Lamp. This test checks the W lamp and its voltages
when the lamp is on and when it is off. If the message "W Not
Detected" appears, the lamp voltages are good, but the lamp is
not present or is not functioning properly. Try replacing the
tungsten lamp and retrying the test. If the word "Fail" appears,
call your Thermo Electron representative. MLF: Lamp or
Motherboard.
8. Lamp and Shutter. (UV2000 only)This test actually has
several parts, each of which checks a different part of the
lamps' and shutter's operation. If either of the lamps fails, an
appropriate message will be displayed. Try replacing the lamp
and retrying the test. If a "Bad Shutter" message appears, call
your Thermo Electron representative. MLF: part listed on
display.
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EXTERNAL
DIAGNOSTIC
TESTS
This section describes two external diagnostic tests that can be used to
verify that your detector is working properly.
LC Test Mix
An ampule of prepared LC Test Mix is included as part of your
detector's accessory kit. An instruction sheet describing the
parameters for running the test mix and showing the resulting
chromatogram is also enclosed. This is a good test to run when you
first set up your LC system.
HINT: Keep the chromatogram that you generate with the LC Test Mix.
It can be a useful baseline for troubleshooting problems later on.
Absorbance
Linearity
Use the optional cuvette holder (see Appendix E) and certified
standards to test the absorbance linearity of your detector in the UV
range (approximately 235 nm to 350 nm). For your convenience, the
following procedure is also detailed in Appendix E.
HINT: This procedure is particularly useful for laboratories that require
periodic detector validation.
To perform the test, you will need procedure E 925 from the
American Society for Testing and Materials (ASTM) and standard
potassium dichromate (SRM 930) from the National Institute of
Standards and Technology (NIST; formerly the National Bureau of
Standards, NBS). The test involves the preparation of acidic
solutions of potassium dichromate at four concentrations and the
absorbance measurement of each solution at four wavelengths
between 235 and 350 nm. After correcting for an absorbance blank,
the linearity deviation of a plot of absorbance versus concentration
should be less than 1%.
If you want more information on this test, or find that your instrument
does not conform to these specifications and requires service, contact
your local Thermo Electron representative.
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D
Glossary
Introduction
We have included a glossary to define certain technical terms used
throughout the manual's text. These terms are consistent with
standard definitions used throughout the analytical industry, and are
added here as a quick reference only.
A-C
A/D
Analog-to-digital. Converts a detector's analog signal to a digital signal.
AUFS
Absorbance units full-scale; a measure of sensitivity.
absorbance
A process where the intensity of light shining through a sample is decreased;
the transmitted light is measured in absorbance units, which are directly
proportional to the concentration of the absorbing sample.
analog offset
A voltage applied to the output signal in order to keep the signal "on-scale"
throughout a run.
background scan
The reference spectrum of the mobile phase. It is subtracted from the sample
spectral scans to correct for baseline absorbances. Also called baseline scan.
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.
chronometer
A gauge for measuring the total amount of time something has been in
operation.
D-F
defaults
The values or choices built-in to a system. If no specific choice is made, the
detector will run using the default settings.
develop file
A feature that allows you to make multiple injections of a sample at different
wavelengths, automatically.
degassing
The practice of removing air from the mobile phase, usually by sparging or
applying a vacuum.
diagnostics
Methods used to detect and isolate problems.
display
The two-line screen on all SpectraSYSTEM instruments.
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edit file
A copy of the file used for editing. Once loaded, the parameters set in the
edit file are transferred to the runfile.
error message
A displayed message that notifies you of a problem.
field
The area in a display, screen, or menu where an entry is required or a choice
must be made.
file
A list of detector parameters that contains the desired settings for an
analysis.
G-K
gradient elution
A liquid chromatographic technique where the mobile phase composition
changes over time; changes may be continuous or in steps.
ground terminal
A terminal used to connect the ground or earth lead of a signal or contact
closure cable.
K-factor
A factor used to calculate a response of zero for one of two coeluting or
poorly resolved peaks; also known as peak suppression.
keypad
All of the keys that you use to communicate with your instrument.
M-Q
menu
A list of choices.
miscible
Two solvents are miscible if they combine with each other to form a single
phase.
parameter
A value or set of values used to define the characteristics of behavior of an
instrument or system.
peak broadening
The dilution of a peak as it moves through the chromatographic system.
peak suppression
A technique that uses a factor (the K-factor) to calculate a response of zero
for one of two coeluting or poorly resolved peaks.
photodiode
The detector component that measures light intensity.
queue
A set of items (i.e., samples, files) in a prearranged order.
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R-S
RAM
Random Access Memory.
range
A detector parameter that controls the full-scale range for the output signal.
replay
Retrieves a stored spectrum that can then be played back as either individual
data points or a smoothed spectrum.
rise time
A detector parameter that controls its response time; rise time is inversely
proportional to the amount of baseline noise.
runfile
The runfile is the file that contains the current detector parameter settings.
run time
The duration of a sample run, from injection to detection.
signal to noise
A measurement of the sensitivity of a detector; the ability to measure a very
small sample response over the baseline noise.
solvent programming
See gradient elution.
spectral scan
A sample spectrum.
status
The current condition.
T-Z
timed events
An instrument action triggered to occur at a specific, preset time during a run
(e.g., autozero, wavelength change, stop-time).
troubleshooting
Locating the cause of problems with equipment or procedures, and solving
these problems.
wavelength programming
Programming the detector to change the monitoring wavelength as a function
of time during a run.
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E
Cuvette Holder Accessory
Introduction
This Appendix provides information on the installation, use, and
maintenance of the Cuvette Holder Accessory (Fig. E.1). This
accessory is available to simplify calibration/standardization of your
UV2000 UV/Vis detector using FDA, industry, or in-house
calibration standards. The cuvette holder is a modular accessory that
installs in place of the detector flowcell. It allows analysis of
calibration standards (for example, potassium dichromate) to ensure
your detector's compliance with FDA, industry, and/or in-house
regulations.
D T-Z045/DG
To use the cuvette holder, prepare your calibration standard according
to the instructions provided with the sample. Then place the sample
in a standard 10.0 mm I.D. (12.5 mm O.D.) quartz cuvette. Analyze
the sample and compare its measured maxima to its certified maxima.
If there's a discrepancy in the measured wavelength, the detector can
be recalibrated using the procedure described on page 114.
Cuvette
Photodiode
Standoffs
Mounting
Screws
Figure E.1 Cuvette Holder accessory
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Installation
The cuvette holder attaches to your detector using the standard
flowcell mounting hardware.
Use the following steps to remove the flowcell and install the cuvette
holder:
1. Remove the front panel of the detector (Fig. E.2) to gain access
to the flowcell mounting area. Note that the front panel is a
friction-grip mount and will snap free if you pull outward on its
lower edge.
2. Remove the thumbscrew that secures the flowcell cover to the
front of the detector (Fig. E.3). Remove the flowcell cover and
set it aside.
3. Remove the two thumbscrews that secure the photodiode
assembly to the front of the flowcell (Fig. E.4) and then
reposition the photodiode assembly out of the way to provide
access to the flowcell.
4. Remove the two thumbscrews (top left, lower right) that secure
the flowcell mount (Fig. E.5) to the front of the detector and
then remove/reposition the flowcell.
NOTE: You needn't disconnect the flowcell's tubing connections to
your LC system if the cuvette holder is only going to be used long
enough to conduct a calibration. Simply reposition the flowcell and
omit Step 7 of this procedure.
5. Position the cuvette holder and secure the two thumbscrews (at
top left and lower right) that secure the holder to the threaded
holes on the detector's front panel.
6. Replace the photodiode assembly and secure it to the cuvette
holder's standoffs with its two thumbscrews (at top right and
lower left).
7. Snap the detector's front panel back in place.
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RUN
STATUS
MENU
STOP
ENTER
ZERO
SpectraSYSTEM
UV2000
Removable
Front Panel
DT-Z046\FM
Grip and Pull
Here
Figure E.2 Detector front panel
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RUN
STATUS
MENU
STOP
ENTER
ZERO
SpectraSYSTEM
UV2000
DT-Z047\FM
Flowcell
Housing
Figure E.3 Detector with front panel removed to expose flowcell housing
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RUN
STATUS
MENU
STOP
ENTER
ZERO
SpectraSYSTEM
UV2000
Flowcell
Inlet
Flowcell
Outlet
Tubing
Clamp
Rear of
Photodiode
Assembly
DT-Z048\FM
Mounting
Thumbscrews
Figure E.4 Detector with flowcell assembly exposed to show
photodiode assembly-mounting thumbscrews
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RUN
STATUS
MENU
STOP
ENTER
ZERO
SpectraSYSTEM
Flowcell
Inlet
UV2000
Flowcell
Outlet
Tubing
Clamp
Photodiode
Assembly
DT-Z049\FM
Mounting
Thumbscrews
Figure E.5 Detector with photodiode assembly repositioned to expose flowcell
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Using the Cuvette Holder
The two procedures that follow allow you to use the Cuvette Holder
to test the linearity of your detector's absorbance and to recalibrate the
detector, if necessary.
NOTE: Be sure to insert the cuvette inside the holder so that its transparent
sides (rather than the frosted ones) are in line with the beam of light from
the detector's lamp. Failure to insert the cuvette properly may result in
insufficient light levels for accurate analyses.
ABSORBANCE
LINEARITY
You can use the optional Cuvette Holder and certified standards to
test the absorbance linearity of your detector in the UV range
(approximately 235 to 350 nm).
HINT: This procedure is particularly useful for laboratories that require
periodic detector validation.
To perform the test, you'll need procedure E 925 from the American
Society for Testing and Materials (ASTM) as well as standard
potassium dichromate (SRM 930) from the National Institute of
Standards and Technology (NIST; formerly the National Bureau of
Standards, NBS). The test involves the preparation of acidic
solutions of potassium dichromate at four concentrations and the
absorbance measurement of each solution at four wavelengths
between 235 and 350 nm. After correcting for an absorbance blank,
the linearity deviation of a plot of absorbance versus concentration
should be less than 1%.
If you want more information on this test, or find that your instrument
does not conform to these specifications and requires service, contact
your local Thermo Electron representative.
λ CALIBRATION
Selecting /λ Calibration/ brings up the screen shown in Figure E.6.
You can use this screen to offset the factory-calibrated wavelength to
more closely match FDA, industry, or in-house standards.
λ Offset (steps)
0
Figure E.6 The lambda (wavelength) offset screen
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NOTE: The UV2000 detector is calibrated using a mercury-lamp fixture.
This provides a very narrow emission line at 254 nm. Broad-band
calibration standards, such as holmium oxide and didymium filters, make
calibration more difficult and less accurate.
To offset the factory-calibrated wavelength, select the number of
"steps" by which you want the wavelength to be offset. Each step
represents approximately 0.25 nm, so if you choose "2" for the
number of steps, you will have offset the wavelength by + 0.5 nm.
You can offset the wavelength by as much as ± 2.5 nm.
NOTE: The offset value isn't cleared upon resetting the RAM memory. It
can only be changed from the lambda offset screen.
Maintenance
The cuvette holder contains no user serviceable components;
however, cleanliness of the cuvettes is critical to obtaining accurate
analyses. Therefore, these instructions are provided for inspecting
and cleaning the cuvettes.
Inspecting a Cuvette
Cuvettes, whether previously used or new, should always be visually
inspected before use. Use the following steps to inspect a cuvette:
1. Grasp the cuvette by its two frosted sides and hold it up in front
of a bright light source such as a fluorescent fixture,
incandescent bulb, or sunny window.
2. Carefully observe the cuvette's two transparent glass sides.
Look for physical damage such as chips, cracks, scratches, etc.
Also look for dirt, smudges, fingerprints, and so forth.
3. Based on the results of your inspection, you can do one of the
following three things:
a. If no optical-surface damage or contamination is noted,
you can fill the cuvette with sample and use it for your
analysis.
b. If you see physical damage or severe contamination on the
optical surfaces, you may wish to replace the cuvette with a
new one.
c. If no physical damage is noted and only light to moderate
contamination, clean the cuvette using the procedure that
follows in the next section of this appendix.
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Cleaning a Cuvette
If the visual inspection reveals contamination of or damage to the
cuvette's optical surfaces (the inner and/or outer surfaces of the
cuvette's two transparent faces), then the cuvette should be cleaned
before use. Use the following steps to clean a cuvette:
1. Immerse the cuvette in a small beaker filled with an
appropriate cleaning solution. Use detergent and water to clean
cuvettes that are contaminated with residue from water-based
solutions. Use an appropriate organic solvent (e.g., methanol,
ethanol, isopropanol, etc.) for cuvettes contaminated with
residue from organic-solvent-based samples.
2. Place the beaker containing the cuvette(s) and cleaning solution
in an ultrasonic bath and set the timer. Use a time setting that's
appropriate for the amount of contamination that has to be
removed.
3. Remove the cuvette from the beaker, handling it by its frosted
sides only. Rinse it thoroughly with clean deionized water
until all traces of detergent and dirt have been flushed away.
4. Dry the cuvette thoroughly with a lint-free wiper, exercising
care to handle the cuvette only by its two frosted (non-optical)
sides.
5. Carefully inspect the cuvette for residual contamination using
the steps detailed in the preceding section of this appendix. If
any is noted, repeat Steps 1 through 4 until the cuvette is
completely clean and dry.
NOTE: In cases of serious contamination that resists removal, it may
be easier to simply replace a dirty cuvette than to spend a lot of time
cleaning it.
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INDEX
The asterisks (*) in this index refer to topics that are common to both the UV1000 and the UV2000
detectors. For each asterisked item, there are two page numbers. The first refers to the appropriate
page number for the UV1000 detector; the second refers to the page number for the UV2000.
KEYS
[+] and [-] keys 1, 4
[<] key 4
[>] key 4
[ENTER] key 3
[MENU] key 3
[RUN] key 3
[STATUS] key 3
[STOP] key 3
[ZERO] key 3
[∧] key 4
[∨] key 4
A
A File Is Already Running message 111
A/D defined 117
Absorbance
defined 117
linearity 116, 127
ratioing (UV2000 only) 35, 64
Accessory kit 79
Alphabetical entries, changing 1
Alphanumeric entries, changing 4
Amperage requirements, see Fuses
Analog offset, see Analog outputs
Analog Output Connections
UV1000 84
UV2000 84
Analog outputs
adjusting for different devices* 25, 36
connections to 84
recorders 84
description of* 25, 35
event mark 44
fields defined 97
illustration of connectors 83
K-factor 63
offsets* 25, 36
term defined 117
range* 22, 29
scanning 52
shorting the 43
testing the 115
use of negative and positive offsets* 25, 36
Analog Outputs Menu
accessing the* 26, 35
general description of 97
illustration of* 26, 35
ANLG, see Analog outputs
Arrow keys 1, 4
Asterisks, use of 5, 6
ASTM Procedure E 925 127
AU
analog output choice 35
entries (for analog 2) 35
field described 97
AU entries (for analog 2) 97
AUFS, acronym defined 117
Auto Scan
illustration of 51
Autozero
example of 40
programming 39, 40
time field described 97
time* 22, 29
B
Back-pressure
causes of 71
detectors in series 87
increasing 71
specifications for flowcells 88
Background scan
defined 117
zeroing 48
Baseline problems
baseline defined 117
correcting for background absorbance 48
Thermo Electron
index-i
drifting
causes of 71, 108
correcting with autozero 40
correcting with offsets* 25, 36
erratic, causes of 71
noise, causes of 71, 107
spikes, causes of 71, 107
Blank key 3
Brackets, reason for 7
C
Calibration 114, 116, 127, 128
Capitalization, standard use of 7
CAUTIONS!, what they signify 8
Changing the fuses 82
Chemical hazard, icon defined 8
Chronometer
defined 117
illustration of 74
tracking deuterium lamp life 74
Cleaning procedures
deuterium lamp surface 74
flowcell 71
optical surfaces 69
Clicking sound (UV2000 only) 109
Clock, internal 42
Column bleed 108
COMM port 83
Commands Menu
accessing the 43
general description of 5, 97
illustration of 43
Communications to external devices 83, 85
Confirmation messages 6
Connectors, see External contacts
Conventions used in manual 6
brackets 7
capitalization 7
icons 8
slashes 7
standard words 7
two-line display 6
Copy Menu
accessing the 32
illustration of 33
Copying files 32, 103
copy field described 97
Copy File command 97, 103
Develop File 33
index-ii
protected files 33
Scan File 33
CSA, see Safety Information
Current time field 42, 97
Cursor
blinking square 1
movement 1
speed, setting the 45, 97
triangular 1
Curves, smoothing spectral replays 53
Cuvette holder
cleaning a cuvette 129
cuvette size defined 121
function of 121
inspecting a cuvette 128
installation of 122
maintenance of 128
testing absorbance linearity with 127
wavelength calibration using 127
D
D, see Develop File
D2 Lamp Hours 98
Dark current, test for 115
Defaults
see Appendix B
term defined 117
Degassing 117
Delete commands
Delete File 98
Delete Queue 98
fields defined 98
Deleting
a queue 59
files* 25, 33
protected files 34
time lines 38
Detector
calibration 114, 128
command center 2
front panel, illustration of 2
modes described 3
rules for operation 1
Shutdown message 45
Deuterium lamp
alignment of 76
changing the 74
chronometer 74
cleaning the surface 74
Thermo Electron
01/2004
D2 lamp hours field 97
illustration of 75
lamp life 74
Low Light Detected message 110
testing the 115
tracking D2 hours 74
warm-up time 74
Develop File (UV2000 only)
#Runs field 55, 101
accessing the 54
advantages of 54
defined 54, 117
Edit Menu 54
editing parameters 54
editing while running 56
example of 55
no copying 33
repeating the file 56
setting parameters 54
Status Menu 56
Status Screen 55
DEVELOP, see Develop File
Diagnostic tests
analog outputs 115
deuterium lamp 115
diagnostics defined 118
external
absorbance linearity 116, 127
LC test mix 116
internal
diode offsets
self-test 115
user-selected 113
light levels 113
software version 113
wavelength calibration test 114, 127
lamp and shutter (UV2000 only) 115
light levels 98
motor 115
RAM 115
self-tests 114
software version 102
Tests Menu
illustration of 112
tungsten lamp 115
voltages 115
Diode Offsets
field described 98
self-test 115
user-selected test 113
01/2004
Display
changing the contrast 45
changing the cursor speed 45
illustration of the 2
term defined 118
the Display AU, l screen 53, 98
Down-arrow (t) key, right side of display 2
Drift, see Baseline problems
Dual-wavelength operation 26
analog output 2 35
selecting 100
Status Screen 30
stopping a run 30
wavelength program 27
E
Edit File 20, 98, 118
Edit Menu
accessing the* 20, 27
general description of 98
illustration of* 20, 27
while in the Develop File 54
while in the Scan File 47
Editing
a file
from the File(s) Menu* 20, 26
from the Status Menu* 24, 31
a queue 59
parameters during a run 5
the Develop File 54
time lines 39
Electrical interference, causes of 107
End λ 98
End of queue 43
Enter, as a standard word 7
ENTER key 3
Equipment damage, avoiding 8
(see also Safety Information)
Error messages
conventions of 6
definition of 118
see also Messages
Event mark 44, 98
accessing 44
placement of 49
Exclamation points, use of 6
Thermo Electron
index-iii
External contacts 83
analog outputs
connections to 84
scanning 49
shorting 43
ANLG 84
connectors 83
illustration of 83
making connections 83
READY(Output) 85
accessing command 44
RUN(Input) 42, 85
STOP(Input) 85
ZERO(Input) 85
F
FCC, see Safety Information
Fields
alphabetical listing of 97
changing entries in 1, 4
description of all 97
term defined 118
File Menu (UV1000 only)
accessing the 20
general description of 5, 98
illustration of 20
loading the edit file 22
setting parameters 20
Files
copying (UV2000 only) 32, 97, 103
deleting* 25, 33
edit file 20
editing from File(s) Menu* 20, 26
editing from Status Menu* 24, 31
escaping without saving changes* 25, 32
identification of (UV2000 only) 27, 99
loading
an example* 12, 16
fields described 100
technique described* 22, 29
while a queue is running 59
managing (UV2000 only) 32
messages
Already Running 111
Copied 33
Deleted* 25, 34
Loaded* 22, 29
Protected, Cannot Be Copied To 33
Protected, Cannot Be Deleted 34
index-iv
Saved* 24, 32
name field (UV2000 only) 99
protecting (UV2000 only) 34, 100
run file 20
saving* 24, 32
term defined 118
Files Menu (UV2000 only)
accessing the 26
general description of 5, 98
illustration of 14, 26
loading the file 29
setting parameters 26
Floating ground 84
Flowcell
bubbles in the 107, 108
changing the 67
cleaning the
precautions during 71
safety precautions 72
using a pump 71, 72
using a syringe 71, 72
windows 69
with nitric acid 72
with organic solvents 71
connecting to inlet/outlet lines 86
contamination of 71
different types of 88
disassembly of 67
installing the 70
leaks 107
location of 67
maintenance of 67
orientation for non-Finnigan
SpectraSYSTEM detectors 88
removing flowcell and housing 68
specifications 88
Fluid connections 86
Front panel, illustration of 2
Fuses
changing the 82
illustration of fuses 82
illustration of holders 81
requirements 82
G
Glossary 117
Good laboratory practices, see Safety
Information
Gradient elution 48, 118
Thermo Electron
01/2004
Ground loop problems, causes of 108
Ground terminal 118
hooking to recorders 84
Grounding 1
illustration of 2
moving around 1
Keys, description of 3
H-I
L
High voltage warning 8
Hints, defined 8
Icons, how they're used 8
Identification of files 27, 99
Injecting a sample* 23, 30
Input errors 111
Installation
accessory kit 79
analog outputs 84
connecting to recorders 84
connecting to inlet/outlet lines 86
flowcell connections 86
fluid connections 86
fuses 82
illustration of 82
illustration of holders 81
requirements 82
positioning the detector 80
power cord 82
power requirements 80
rear panel connections 83
tools for 80
unpacking 79
voltage selection 80
Instrument control 2
Integrators/Workstations
rear panel connections to 84
Invalid Parameters, Spectrum Not Allowed
message 111
Laboratory safety, see Safety Information
Lambda (λ)
fields defined 50
max (lMax) 50
maxAU (lMaxAU) 50
min (lMin) 50
Lambda (λ)
fields defined 99
Lamps
aging 73
alignment illustrated 76
and shutter tests 115
changing the 73, 77
Cover Open message 110
deuterium, see Deuterium lamp 74
illustration of assemblies 75
installation of 76, 77
lamp hours fields 42, 98
lamp life 74, 77
location of 73
removal of 74, 77
selecting the 41, 99
priority over 41
shutting down the
at end of queue 43
automatically 42
manually 42
shutdown fields described 102
Time from READY 42, 103
tungsten, see Tungsten lamp 77
turning on the 102
automatically 42
manually 42
Lamps Menu
accessing the 41
general description of 99
illustration of 41
LC test mix 116
LCD, see Display
Light Levels test 99, 113
Linearity of response 116, 127
Load fields
described 100
used/illustrated
J-K
K-factor 60
algorithm for 60
allowable values of 98
calculation of 63
choosing wavelengths for 61
defined 118
operating in the K-factor mode 63
setting analog output for 35
uses for 60
using Display AU, λ screen 63
using the Scan File 61
Keypad 3, 118
01/2004
Thermo Electron
index-v
Load command* 12, 16
Load File command* 12, 16
Load Queue command (UV2000 only)
58
Loading
a file
an example* 12, 16
technique described* 22, 29
while a queue is running 59
a queue (UV2000 only) 58
Locking the status screen 43
Low Light Detected messages
from deuterium lamp 110
from tungsten lamp 111
M
Main Menu
accessing the* 20, 26
general description of 4
illustration of* 10, 14
Maintenance, see Chapter 5
Manual, conventions used in 6
MENU key
function of 3
used to access Main Menu* 20, 26
Menu Reference 97
Menu Tree 93
Menus
display conventions of 4
general description of upper-level 5
illustration of longer 6
moving between/within 1, 4
term defined 118
Messages
A File Is Already Running 111
Detector Shutdown 45, 112
File Copied 33
File Deleted* 25, 34
File Loaded* 22, 29
File Saved* 24, 32
Invalid Parameters, Spectrum Not Allowed
114
Lamp Cover Open 110
Low Light Detected 110, 111
No More Available Memory 111
No Queue Available 58, 111
No Scans Stored 53
No Spectra Available 111
NRDY (for one of the lamps) 41
index-vi
Protected File
Cannot Be Copied To 33, 111
Cannot Be Deleted 34, 111
Cannot Be Edited 111
Queue Deleted 60, 98
Queue Loaded 58, 100
Queue Loaded, Cannot Load File 59, 111
real-time errors 110
Run In Progress, No Replay Allowed 112
Run In Progress, Testing Not Allowed 112
Run Not In Progress, Spectrum Not
Allowed 112
Scan Memory Full 112
system errors 109
types of messages 5, 110
user-input errors 111
Method development, see Develop File
Method transfer, see Develop File
Miscible 118
MLF (most likely failure) 114
Monochromator
cleaning the lens 69
illustration of apertures 70
motor, testing the 115
movement during a scan 49
More Menu
accessing the* 34, 40
general description of 100
illustration of 34
Motor test 115
Moving within/between menus 1, 4
N
Negative signals 25, 36
Nitric acid
safety precautions 72
used to clean the flowcell 72
No More Available Memory message 111
No Queue Available message 58, 111
No Scans Stored message 53
No Spectra Available message 111
Notes, what they signify 8
NRDY (not ready message) 23, 30, 41, 103
Numerical entries, changing 1, 4
O
Offsets, see Analog outputs
Optical system 106
Thermo Electron
01/2004
Options Menu
(chosen from the Edit Menu) 100
accessing the* 22, 28
description of* 22, 28
illustration of* 22, 28
(chosen from the Main Menu) 100
accessing the* 34, 40
illustration of 40
illustration of (UV2000 only) 34
general description of 5
Order 1
defined 100
editing 59, 97
in a queue 58
Order field 57
Organic solvents, used to clean flowcells 71
Outputs, see Analog outputs
P
Parameters
allowable and default values 97
defined 118
setting* 20, 27
Pausing a queue 59
pause command 100
pause queue command 100
resuming 59
Peak
problems
broadening 109, 118
size 109
tailing 109
purity 35, 64
suppression
analog output for 35
defined 118
see also K-factor
Photodiode
cleaning the surface 69
defined 118
mount
illustration of 69, 87
removing the 69
Positioning the detector 80
Power
cord 82
requirements 80
switch, location of* 10, 14
01/2004
Preset choices, selecting from 4
Preventative maintenance 67
Program field 27, 100
Programmed autozero 39, 40
example of 40
illustration of 40
Protected files 34, 100
copying to 33
deletion of 34
illustration of 34
messages
Cannot Be Copied To 33, 111
Cannot Be Deleted 34, 111
Cannot Be Edited 111
Pumps, used to clean flowcells 71, 72
Purity determinations 35, 64
Q
Q, see Queue
Queue (UV2000 only) 56
a "Q" in the display 58, 103
defined 118
deleting a line 57
deleting a queue 59, 98
description of 56
editing a 59
example of 57
field described 100
file name 57
illustration of an empty queue 57
lamp shutdown at end of 43
Loaded message 59
loading a 58, 99
messages
No Queue Available 58, 111
Queue Deleted 60
Queue Loaded 58, 100
Queue Loaded, Cannot Load File 111
monitoring while running 58
number of files per queue 57
number of injections per file 57
order 1 58
order field 57, 100
pausing a 59, 100
resuming after a pause 59
running a 58
setting up 57
Status Screen 58
stopping a 59
Thermo Electron
index-vii
Queue Menu
accessing the 56
general description of 5
illustrations
with no queue loaded 57
with queue loaded 58
pause field 58, 100
R
RAM
defined 119
testing 115
Range
example of use* 11, 15
fields defined 100
setting the* 22, 29
term defined 119
Ratioing absorbance values (UV2000 only)
35, 64
READY
output
accessing the command 44
connecting to terminal 85
field described 101
use of 44
state* 23, 30
time from ready field 42
Real-time error messages 110
Recorders
connecting to analog outputs 84
Remote communications 85
Replay
fields described 101
rate 52
term defined 119
Replay Menu (UV2000 only)
accessing the 52
replay fields described 101
replay spectra
command 52
illustration of display 53
stopping replay 53
setting parameters 52
Reviewing parameters during a run 5
Rise time*
defined 119
field described 101
incorrect setting, results of 109
suggestions for use* 22, 28
index-viii
Run
fields
fields described 101
used in a queue 57
used in the Develop File 55
input terminal 85
shutting down lamps 42
parameters* 20, 27
programming during a* 24, 31
progress messages
No Replay Allowed 112
No Scanning Allowed 112
Testing Not Allowed 112
ready signal for external devices 85
starting a
example of* 13, 18
from an external device 85
technique described* 23, 30
stopping a
example of* 13, 18
from an external device 85
technique described* 23, 30
the run file 20
the run file, defined 119
time, defined 119
RUN key
example of use* 13, 18
functions of 3
Runfile, defined 119
S
S, see Scan File 47
Safety
certification, see Safety Information
definition of icons 8
general precautions, see Safety Information
nitric acid precautions 72
Sample Queue, see Queue
Save File command* 24, 32
Saving an entry 1
Scan File (UV2000 only) 46
accessing
summary data 50
the Scan File 47
analog output 49
background, defined 117
baseline scan
automatic 48
defined 117
Thermo Electron
01/2004
manual 48
subtracting 48
choosing scan times 49
data storage 53
Display AU, l screen 53
illustration of display 53
Display AU, λ screen
K-factor calculation 63
Edit Menu for 47
fields described 101
file name 47
how the detector scans 46
an example 49
identification of stored spectra 52, 102
initiating a scan 49
K-factor 61
Memory Full message 112
min/max wavelength fields 99
no copying 33
No Scans Stored message 53
quantitative analysis 49
Replay Menu
accessing the 52
illustration of 52
replay fields described 101
replay rate 52
replay spectra command 52
setting parameters 52
replay spectra command
illustration of display 53
stopping replay 53
running replay 52
running the 48
setting the wavelength
end of scan 48, 98
interval for scan 48, 99
monitoring the run 48, 102
start of scan 48, 102
setting up the 47
Setup Menu for 47, 102
spectral scan, defined 119
Status Screen for 48
stopping a scanning run 50
subtracting baseline scans 48
summary data screen, illustration of 50
wavelength min/max fields 99
zero command 48
zero time field 48
SCAN, see Scan File
Scan Summary Data Screen
accessing the 50
01/2004
illustration of 50
Scan zero time 48, 101
Scanning
automatically 51
how it works 51
number of spectra stored 46
replaying spectra
corrected for baseline absorbance 47
see also Scan File 47
Scrolling through choices 4
Select, as a standard word 7
Self-Tests 116
analog outputs 115
command described 102
deuterium lamp 115
diode offsets 115
lamp and shutter (UV2000 only) 115
motor 115
RAM 115
tungsten lamp 115
voltages 115
Setting parameters
current time 42
Develop File 54
example of 55
File(s) Menu* 20, 26
replaying spectra 52
sample queue 57
scanning 47
Status Menu* 24, 31
Setup Menu
general description of 102
illustration of 47
Short outputs
accessing 43
command described 44, 102
Shortcuts, icon for 8
Shutdown detector
accessing the command 45
command described 102
message described 112
restarting the detector 45
Shutdown lamps
how 102
when 41, 102
Signal-to-noise defined 119
Single-wavelength operation
analog output 2 (UV2000 only) 35
selecting 100
using* 20, 26
Slashes, reason for 7
Thermo Electron
index-ix
Software version 102, 113
Solvents
disposal of wastes, see Safety Information
programming 40, 119
used to clean the flowcell 71
Specifications
flowcells 88
instrument 90
Spectra
changing the appearance of 53
corrected with baseline scan 48
getting ready to replay 50
identification of 52
replaying 52
Spectra Time field defined 102
storage of 53
Spectral scanning, see Scan File
Spiking 71
Standard words 7
Start λ 102
Starting a run* 23, 30
from an external device 85
Startup fields defined 102
Status
field defined 103
lock feature 43, 103
term defined 119
STATUS key
brings up Status Screen 5
functions of 3
Status Menu
accessing the* 24, 31
general description of 5
illustration of* 24, 31
setting parameters* 24, 31
when Develop File is loaded 56
when Scan File is loaded 50
Status Screen
accessing the* 23, 30
general description of 5
illustration of* 23, 30
locking the 43, 103
when a queue is loaded 58
when Develop File is loaded 55
when Scan File is loaded 48
STOP key
example of use* 13, 18
functions of 3
to stop replaying spectra 53
used to stop a run* 23, 30
STOP(Input) terminal 85
index-x
Stop-line
adding a* 13, 17
affects how you start a run* 23, 30
in a wavelength program 21, 27, 38
used to stop a run* 23, 30
Stop-time, see Stop-line
Stopping
a queue 59
a run
from an external device 85
techniques for* 23, 30
with a stop-line* 21, 27
a scan 50
dual-wavelength files 30
System error messages 110
T
Temperature fluctuations, results of 108
Terminals, see External contacts
Tests Menu
accessing the 112
general description of 5, 103
illustration of 112
Tests, see Diagnostic tests
Text, conventions of 7
Theory of operation 105
Time
current time field 42
field in Wavelength Program 103
setting the 42
stop-times 38
Time from READY 103
accessing 42
description of 42
example of 42
illustration of 42
Time lines, see Wavelength programming
Timed events 85
autozero 40
defined 119
lamp startup and shutdown 42
on wavelength change 40
scanning 51
stop-time 38
Tools needed for installation 80
Troubleshooting
clicking sound 109
flowcell contamination 71
MLF (most likely failure) 114
Thermo Electron
01/2004
Table of common problems 107
term defined 119
Tungsten lamp
alignment of 76
changing the 77
illustration of 75
lamp life 103
Low Light Detected message 111
testing the 115
W lamp hours field 103
warm-up time 77
TÜV/GS certification, see Safety Information
U
Unpacking your detector 79
Unshort outputs 44
UPS, uninterrupTable power supply 107
User messages 5
UV1000
allowable and default values 97
analog outputs 25, 84
building a wavelength program 21
checking the status 23
deleting a file 25, 98
diagnostic tests 112
features 9, 43
installation 79
lamp operations 41
locking the Status Screen 43
maintenance 67
periodic validation 67
programmed autozero 40
quick example 10
running the detector 23
setting detector parameters 20, 24
single-wavelength operation 20
software version 113
starting a run 23
stopping a run 23
troubleshooting 107
wavelength programming 37
UV2000
allowable and default values 97
analog outputs 35, 84
building a wavelength program 27
checking the status 30
copying files 32, 97
deleting a file 98
Develop File 54
diagnostic tests 112
01/2004
dual-wavelength operation 26
features 9, 43
file identification 27, 99
installation 79
K-factor 60
lamp operations 41
loading a file 29
locking the Status Screen 43
maintenance 67
periodic validation 67
programmed autozero 40
protecting files 34
quick example 14
running the detector 29
sample queues 56
scanning ability 46
setting detector parameters 26, 31
single-wavelength operation 26
software version 113
starting a run 30
stopping a run 30
troubleshooting 107
wavelength programming 37
UVW message 103
V
Validation
of linearity 116, 127
of wavelength 114, 127
VDE, see Safety Information
Voltage
AC line problems 107
clipping 84
preset 80
selector barrel
illustration of 81
location of 80
setting the 80
setting 80
testing the 115
W
W Lamp Hours 103
WARNINGS!, what they signify 8
Wavelength
(λ) fields defined 99
calibration test 99, 114, 127
ending (in Develop file) 55
Thermo Electron
index-xi
example of setting the* 11, 15
field in the Status Screen 99
field in Wavelength Program 103
interval 55
offset field described 99
recalibration using Cuvette Holder 127
starting 55
testing accuracy 114, 127
Wavelength Program 103
accessing from the Edit Menu* 21, 27
accessing from the Status Menu* 24, 31
command described 103
description of 21, 27, 37
example of a stop-line* 21, 28
illustration of* 21, 27
single-line* 21, 27
two-line* 21, 27
using a stop-line* 21, 27, 38
Wavelength programming
adding time lines 38
automatic ordering 38
autozeroing with or without a wavelength
change 40
building a program 37
deleting a line 38
described 37
editing while running 39
example of 39
illustration of 39
last line 38
sorting time lines 38
term defined 119
using a stop-line 38
zeroing the baseline 39
index-xii
X-Z
Zero
(Input) terminal 85
example of use* 12, 17
fields described 104
from an external device 85
key, functions of 3
programming an autozero 40
used for baseline scans 48
.End Index
Thermo Electron
01/2004
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