SpectraSYSTEM Autosamplers Reference

SpectraSYSTEM Autosamplers Reference
Finnigan™
SpectraSYSTEM™
Autosamplers
Reference Manual
A0099-587 Revision E
Finnigan™, SpectraSYSTEM™, SpectraNET™, and PushLoop™ are trademarks of Thermo Electron Corporation. Teflon® is a registered trademark of
E.I. du Pont de Nemours & Company.
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 E Printed 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.
Contents
Back Pocket:
Quick Reference Cards
Menu Trees
Technical and Customer Support ........................................................................................................................v
In North America.......................................................................................................................................v
In Europe ..................................................................................................................................................vi
In Australasia and Asia............................................................................................................................vii
Safety and EMC Information ..............................................................................................................................ix
Manual Conventions...........................................................................................................................................xxi
Installation and Specifications..............................................................................................................................1
Introduction ...............................................................................................................................................1
Startup Checklist .......................................................................................................................................2
Site Requirements......................................................................................................................................4
Unpacking .................................................................................................................................................5
Installation .................................................................................................................................................6
Priming ....................................................................................................................................................19
Communication .......................................................................................................................................21
Ready Participation .................................................................................................................................31
Instrument Configurations and Other Features........................................................................................32
Performance Verification ........................................................................................................................33
Specifications ..........................................................................................................................................40
A Quick Example .................................................................................................................................................41
Introduction .............................................................................................................................................41
Learning Your Way.................................................................................................................................41
Instrument Control...................................................................................................................................42
Practice Examples....................................................................................................................................47
Basic Operations ..................................................................................................................................................55
Introduction .............................................................................................................................................55
Theory of Operation ................................................................................................................................56
SpectraSYSTEM Autosamplers ..............................................................................................................62
Run Preparation .......................................................................................................................................64
Starting Your Run ...................................................................................................................................83
Changing the Run in Progress .................................................................................................................87
File Management .....................................................................................................................................91
Needle Height..........................................................................................................................................93
Optimization ............................................................................................................................................94
Special Commands: Commands Menu...................................................................................................96
Advanced Operations ........................................................................................................................................101
Introduction ...........................................................................................................................................101
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Sample Preparation ............................................................................................................................... 101
Gradient Delay ...................................................................................................................................... 127
Diagnostics (Tests Menu)...................................................................................................................... 129
Required Maintenance ...................................................................................................................................... 135
Introduction.......................................................................................................................................... 135
Maintenance Schedule........................................................................................................................... 136
Empty flush-solvent tray ................................................................................................................ 136
Inspect the sample needle module.................................................................................................. 136
Check the solvent tubing and connections for leaks. ..................................................................... 136
Run a standard sample. .................................................................................................................. 136
Clean the drop catch....................................................................................................................... 136
Replace the flush-solvent inlet filter. ............................................................................................. 136
Replace needle assembly................................................................................................................ 136
Replace the injector valve's rotor seal............................................................................................ 136
Clean and lubricate the syringe drive mechanism.......................................................................... 136
Clean and lubricate the XYZ arm. ................................................................................................. 136
Verify the XYZ arm alignment. ..................................................................................................... 136
Procedures ............................................................................................................................................. 137
Maintenance Options............................................................................................................................. 149
Troubleshooting................................................................................................................................................. 153
Introduction.......................................................................................................................................... 153
General Chromatography Troubleshooting........................................................................................... 153
Display Messages.................................................................................................................................. 165
Testing for Tubing Restrictions............................................................................................................. 169
Glossary.............................................................................................................................................................. 171
Introduction.......................................................................................................................................... 171
Kits and Parts Lists ........................................................................................................................................... 177
Index
List of Tables
Table 1.1
Table 1.2
Table 1.3
Table 1.4
Table 1.5
Pin Connections for the Recommended Configuration.......................................................... 26
Input Signal Levels ................................................................................................................ 28
Output Signal Levels.............................................................................................................. 29
Reproducibility Test Sample Queue Parameters .................................................................... 34
%RSD Results for the Reproducibility Test........................................................................... 35
Table 3.1 Recommended Injection Size For Selected Sample Loop Sizes* .......................................... 95
Table 3.2 Maximum Injection Size for Syringe Size ............................................................................. 95
Table 4.1 When Changes Affect the Run in Progress.......................................................................... 111
Table 5.1 Maintenance Schedule ......................................................................................................... 136
Table A.1 General LC Troubleshooting Table..................................................................................... 154
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Table A.2 Autosampler-specific Chromatographic Problems ..............................................................156
Table A.3 Autosampler Hardware Problems....................................................................................... 159
Thermo Electron
iii
Technical and Customer
Support
This manual contains procedures for installing your equipment and
verifying that it is operating within specifications. It will also help
you understand how to use and care for your equipment. For
additional support, contact one of the customer service offices listed
below.
In North America
In North America, Thermo Electron San Jose Customer Service
Engineers are available from the following offices:
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
REPLACEABLE
PARTS
Contact Customer Service Operations to order replaceable parts. The
location and telephone and fax numbers for North America are as
follows:
North America Customer Service Operations
1400 Northpoint Parkway, Suite 10
West Palm Beach, FL 33407
Phone: [1] (800) 532-4752
Fax: [1] (561) 688-8731
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v
TECHNICAL
SUPPORT
You can contact Technical Support at the following location,
telephone and fax numbers, and e-mail address:
North America Technical Support Operations
1400 Northpoint Parkway, Suite 10
West Palm Beach, FL 33407
Phone: [1] (800) 685-9535
Fax: [1] (561) 688-8736
E-mail: [email protected]
In Europe
In Europe, customer support, replaceable parts, and technical support
are available from each of the following offices.
Technical support is also available from North America Technical
Support Operations at the following phone number and e-mail
address:
Phone [1] (561) 688-8700
E-mail [email protected]
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, 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) 95059 226
Fax
[39] (02) 95320 370
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Thermo Electron
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 San Jose
products dealer.
In Australasia and Asia
In Australasia and Asia, customer support, replaceable parts, and
technical support are available from each of the following offices.
Technical support is also available from North America Technical
Support Operations at the following phone number and E-mail
address:
Phone [1] (561) 688-8700
E-mail [email protected]
Rydalmere, N.S.W., Australia
Phone [61] (02) 9898-9000
Fax
[61] (02) 9898-9800
Yokohama, Japan
Phone [81] (45) 453-9100
Fax
[81] (45) 453-9110
Osaka, Japan
Phone [81] (06) 6387-6681
Fax
[81] (06) 6387-6641
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vii
Beijing, P.R. China
Phone [86] (010) 6621 0839
Fax
[86] (010) 6621 0851
For all other countries, contact your local Thermo Electron San Jose
products dealer.
viii
Thermo Electron
Safety and EMC
Information
In accordance with Thermo Finnigan’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.
5. Never run the system without the top cover on. Permanent
damage can occur.
Thermo Electron
ix
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 Finnigan 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 Finnigan.
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, well-documented 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).
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Thermo Electron
Sample Preparation
Always consider the solubility of your sample in the mobile phase.
Sample precipitation can plug the system by obstructing the flow
through the injector and/or the column. 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 (see above).
Solvent Requirements
Many chemical manufacturers provide a line of high-purity or HPLCgrade reagents. Routine filtration of all solvents or eluants through a
0.45- or 0.2-micron (or less) fluorocarbon filter before placing them
in the solvent reservoir will prolong the life and effectiveness of the
inlet filters, check valves, seals, injectors, and columns.
Choose a mobile phase that is compatible with the sample and column
you have selected for your separation. Remember that some solvents
are corrosive to stainless steel. Inert/biocompatible instrument
versions are also available from Thermo Finnigan.
Degas the Eluants
Degas your LC system eluants by vacuum degassing or the helium
sparging. Complete information for using Thermo Finnigan
equipment to perform 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 are not highly
compressible, they do not store much energy. Thus, little immediate
danger arises from the high pressure in an LC system unless a
dangerous mobile phase is used, such as THF, CH3CN, DMSO,
H2SO4, etc. 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.
NOTE: If, for any reason, you need to return your autosampler to
Thermo Finnigan, flush the lines with methanol.
Thermo Electron
xi
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.
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xiii
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.
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
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
xiv
Thermo Electron
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
Finnigan.
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.
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
Thermo Electron
xv
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.
xvi
Thermo Electron
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.
Thermo Electron
xvii
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.
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.
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
xviii
Thermo Electron
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.
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
Thermo Electron
xix
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.
xx
Thermo Electron
Manual Conventions
This manual uses several conventions. Among them are menu
displays, text conventions (brackets, slashes, etc.), standard words,
and several different icons.
DISPLAYS
The following figure shows how we will depict the two-line display.
Note that, in menu illustrations, the triangular cursor location is
indicated by a caret (>).
>FILES
QUEUE
COMMANDS
TESTS
OPTIONS
A two-line menu display
Frequently, the two lines shown on the display are only part of a
longer menu. In this manual, menus having more than two lines are
represented as shown here.
Injection Volume
1.0 (AS3000)
Injections/Sample
1
-------------------------------------------------------Cycle Time
Col Oven:
TrayTemp:
2.5
OFF_
OFF_
Temp
18
Temp
20
A menu longer than two lines
COMMENTS
Fields that appear only on some SpectraSYSTEM™ and
SpectraSERIES autosampler models or only when certain options are
installed are documented with a comment to the right of the field. For
example, in the menu shown above, the Injection Volume field
appears only on the variable-injection volume (VIV)
AS3000/AS3500 or AS300 autosamplers.
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/.
Thermo Electron
xxi
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/, /Edit/, /Injection/.
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.
xxii
Thermo Electron
ICONS
The icons located in the left margin of this manual will alert you to
the following situations.
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.
Thermo Electron
xxiii
1
Installation and
Specifications
Introduction
This chapter describes the site requirements, installation procedures,
and performance testing for the Thermo Electron, Finnigan
SpectraSYSTEM and SpectraSERIES autosamplers. It covers the
AS1000, AS3000, and AS3500 SpectraSYSTEM autosamplers, and
the AS100 and AS300 SpectraSERIES autosamplers. The following
topics are included in this chapter:
•
Installation
•
Instrument Startup and Priming Procedures
•
System Communication
•
Communication
•
Ready Participation
•
Instrument Configurations and Other Features
•
Performance Verification
BEFORE YOU
START
Before you start, review the site requirements and assemble the tools
you'll need to install your instrument:
•
One Phillips screwdriver
•
Two 1/4" open-end wrenches
•
One small, standard screwdriver
Thermo Electron
1
Startup Checklist
This list is a brief summary of tasks that should be completed to
install your autosampler. Complete installation information is
contained in this chapter.
NOTE: Items marked with an asterisk (*) pertain to instruments with
the sample preparation option installed. Items marked with two
asterisks (**) are optional advanced installation procedures that
should be performed only by very experienced users with specialized
applications.
UNPACKING
Unpack and inspect your instrument
Remove cardboard/foam packing insert from sample tub
Check for parts shortages
Read the Safety Information Section on page ix
REAR PANEL
Place the autosampler with rear panel accessible
Install flush bottle/bracket
Install filter/solvent line and flush bottle
Connect the external solvent tubing to tubing extensions (sample
preparation only)*
Connect heater/mixer to an air supply*
Connect the power cord
Hardwire external events, make connection to other
SpectraSYSTEM instruments**
2
Thermo Electron
FRONT PANEL
Install solvent tray/waste container
Connect autosampler to pump
On instruments without the column oven or
inert/biocompatible versions, connect pump outlet line to port
6 on injector valve
Instruments with column oven installed, connect pump outlet
line to solvent preheat tube
Mount column
On instruments without the column oven, connect inlet end to
injector valve port 5
On instruments with the column oven installed, install the
column between the column mounts, resecure column mounts
to oven, and connect inlet end to injector valve port 5.
Connect autosampler to detector
(Connect 12-inch 0.010 ID tubing to inlet side of flowcell)
STARTUP/PRIMING
Turn on, check initial response to power-on
Using the 2.5 mL syringe, flush lines with 5000 µL from the
Btl reservoir.*
Using the 250 µL syringe, flush lines with 500 µL
PERFORMANCE
VERIFICATION
Run a standard sample (a test mix is shipped with the
instrument)
Date and retain run results with instrument documentation
Customize READY participation**
Run an unretained sample to determine gradient delay time**
Optimize needle height**
Installed by:____________________________ Date:_______________________________
Thermo Electron
3
Site Requirements
SPACE
The standard autosampler weighs 16 kg. with all options installed, the
autosampler weighs 24 kg. The instrument requires a bench space at
least 37 cm by 41 cm by 51 cm. Be sure to allow adequate ventilation
away from heating or air conditioning ducts. Avoid exposure to
direct sunlight. If used with the SpectraSYSTEM pump and detector,
place the autosampler in the middle (Figure 1.1).
HINT: You may wish to set your instrument on the bench such that you
have easy access to the rear panel.
SpectraSYSTEM
STATUS
RUN
MENU
STATUS
MENU
PURGE
Membrane
Degasser
RUN
STOP
Pump
P4000
STATUS
MENU
STOP
ENTER
ENTER
SpectraSYSTEM
SAMPLES
SpectraSYSTEM
AS3000
Autosampler
ENTER
ZERO
SpectraSYSTEM
UV2000
Detector
SYS-Z001E\DT
RUN
STOP
Figure 1.1 The SpectraSYSTEM or SpectraSERIES configuration
CAUTION! Be sure to use the proper venting apparatus if you use
hazardous solvents.
ELECTRICAL
Your autosampler automatically configures itself to operate from the
local line voltage (100 - 260 VAC at 50 or 60 Hz). You do not need
to set the line voltage. The instrument is shipped with the necessary
fuses installed.
4
Thermo Electron
You should provide a stable, spike-free power source. This will avoid
intermittent failures and unexplained occurrences (lockup, loss of
memory, incorrect operation) that are directly related to poor power.
We also recommend that you use an uninterruptable power supply
(UPS) for maximum protection from power-related failures.
Unpacking
Your autosampler was shipped in a specially designed shipping
container to protect it from damage during transit. Upon arrival,
inspect all containers and immediately report any damage in transit to
the transportation company. Have the carrier note the shipping crate's
condition on both the delivery receipt and the freight bill. The carrier
is responsible for all damage incurred in shipment.
All SpectraSYSTEM autosamplers are shipped with the sample trays
and syringe(s) installed. A cardboard shipping insert on top of the
sample trays restrains the XYZ arm and holds the sample trays in
place during shipment. Remove the shipping insert. Retain both the
shipping insert and shipping crate should future shipment become
necessary.
After unpacking, inspect your autosampler and its accessories for
missing parts and/or physical damage. If damage is found, notify
both the carrier and your Thermo Finnigan representative. Please do
not return goods without prior authorization from Thermo Finnigan.
The shipping kit contains the following:
•
Autosampler
•
Accessory Kit (see Appendix C)
•
Vial Kit (see Appendix C)
•
Declaration of Conformity (p/n A0099-813)
•
SpectraSYSTEM and SpectraSERIES Autosamplers
Reference Manual (p/n A0099-587)
Thermo Electron
5
Installation
HINT: If you are setting up your autosampler for the first time, you may
wish to follow the procedures in the basic installation section and skip the
advanced section until you are more familiar with your instrument.
The easiest way to install your SpectraSYSTEM or SpectraSERIES
autosampler is to unpack it and set it on the bench, and then complete
the plumbing, electrical, priming, and communication procedures.
Additional or modified procedures required for instruments with one
or more of the integrated options are incorporated into this section.
NOTE: For a list of all available options, upgrades, and accessories for
your autosampler, see Appendix C or contact your Thermo Finnigan sales
representative. Note that all upgrades require specialized installation.
Contact your Thermo Finnigan field service representative for further
information.
PLUMBING
Use the procedures below to install your bottles, solvent tray and
solvent lines. Then connect your autosampler to the pump and
detector and install your column and sample trays. Specific plumbing
procedures include:
•
Flush solvent and bottle bracket
•
External solvents (with Sample Preparation Option only)
•
Heater/mixer air supply (with Sample Preparation Option
only)
•
Solvent tray/waste container installation
•
Pump-autosampler connection
•
Column installation
•
Autosampler-detector connection
The 250 µL syringe, 40-vial sample trays, sample needle, and a
100 µL sample loop are already installed.
NOTE: The AS1000 and AS100 come with the 20 µL sample loop
installed.
If you purchased the Sample Preparation Option, the large (2.5 mL)
syringe) is also factory installed. If you purchased the Tray
Temperature Control Option, your trays have only 35 vial positions
(to accommodate the tub insulation). If you purchased a
inert/biocompatible instrument, the inert/biocompatible needle is
already installed.
6
Thermo Electron
Bottles
Flush Solvent Bottle Bracket
Your accessory kit includes a 250 µL flush-solvent bottle and cap and
a bottle bracket. The bracket attaches to the upper-right corner of the
rear panel (as you are facing the rear of the instrument) and holds the
flush bottle in the correct position and elevation for optimal flow
through the lines.
NOTE: If you wish to use your own solvent bottle or other container, be
sure that it is clean and chemically inert. We recommend using glass
bottles.
HINT: If you purchased the SpectraSYSTEM Solvent Conditioning Module
(SCM400), you may want to store your flush solvent in one of the bottles in
your SCM instead of installing a separate flush bottle and bracket.
The bottle bracket fastens to the rear panel with two Phillips-head
screws included in your accessory kit. To install the flush-bottle
bracket:
1. Align the bracket's mounting holes with the corresponding
screw holes on the autosampler's rear panel (Figure 1.2).
2. Insert the screws into the holes and tighten them in place. Be
sure that the bracket is fastened securely onto the back of the
instrument.
Thermo Electron
7
6-Port Solvent-Selection
Valve
COMM Port
12-Pin Terminal
1
6
3
4
2
Bottle-Bracket
Screws
5
TERMINAL CONFIGURATION
Fuse Compartment
AC Power
Air-Supply Port
Figure 1.2 The autosampler's rear panel connections (with the Sample Preparation
Option installed)
8
Thermo Electron
AS-Z004E\DT
WARNING
Flush Solvent Bottle
The flush solvent bottle rests in the bracket and connects to the
factory-installed flush line by means of a Cheminert® union. The cap
assembly comes with the 1/8" OD x 1/16" ID Teflon inlet line and
high-molecular-weight polyethylene filter attached.
Use the following steps to install the flush bottle:
1. Rinse the bottle with LC-grade solvent to remove any dust.
Then fill with appropriate LC-grade solvent.
2. Insert the filter into the solvent bottle and screw on the cap.
Make sure that the inlet filter rests on the bottom of the solvent
bottle.
3. Run the vent line to an appropriate exhaust apparatus,
particularly when solvents with harmful vapors are used.
CAUTION! Chemical Hazard! Many LC solvents are hazardous
and must be handled properly. Always dispose of waste solvents in
an environmentally correct manner.
4. Place the flush bottle in the bracket.
NOTE: If you did not purchase the Sample Preparation Option, proceed
to “Solvent Tray/Waste Collection.”
External Solvents (S1-S4) (Sample Preparation Option)
If you purchased the Sample Preparation Option, the internal solventline plumbing is already installed. You only need to set up your
external solvent bottles and cap assemblies (not supplied), and
connect the inlets and filters to the solvent-line extensions on the rear
panel (Figure 1.2). Your accessory kit contains four solvent lines and
four filter assemblies. Each solvent line has a flared (flanged)
Cheminert nut on one end; the other end is flareless. To connect a
solvent line, you simply connect the Cheminert nut to the Cheminert
union on the appropriate solvent-line extension and attach the filter
assembly to the other (flareless) end.
Use the following procedure to connect each external solvent line to
its extension.
1. Designate four bottles as your solvent bottles (S1-S4), and plan
the required space near your autosampler.
2. Remove the solvent lines and numbered labels from your
accessory kit.
3. Clip the tie-wrap that coils the solvent-line tubing during
shipment.
4. Remove the red shipping cap from the end of the appropriate
solvent-line extension.
NOTE: You don't have to use all four solvent bottles. Leave the red
shipping cap on the end of any solvent line you're not planning to use.
Thermo Electron
9
5. Connect the Cheminert union to the Cheminert nut on the end
of the appropriate solvent-line extension.
NOTE: If you are going to use bottle caps, thread the other (flareless) end
through the bottle cap.
6. Remove a filter assembly from the accessory kit.
7. Slide the filter over the tubing with the threaded portion toward
the end of the tubing.
8. Repeat steps 3 - 7 for the remaining solvent bottles and lines.
Heater/Mixer Air
Supply
The Sample Preparation Option also requires that you connect the
heater/mixer to an air supply. The heater/mixer requires a clean,
filtered, regulated air supply (40 - 100 psi) to prevent particulates
from obstructing the narrow tubing in the heater.
NOTE: You can also use dry, filtered house air.
Connect the autosampler's air input line to your air supply. The air
inlet line is located in the lower right-hand corner of the rear panel
(Figure 1.2). It is flared and has a Cheminert nut on the end.
Use the following steps as guidelines and adapt as necessary when
connecting your autosampler to your air supply.
1. Remove the red cap from the end of the air input line.
2. Attach the flared end of the air inlet line to your air supply.
NOTE: Depending upon your regulator, you may need to cut off the
Cheminert nut and flared end of the air supply line and use a standard 1/4"
brass fitting in order to make the connection.
3. Connect the brass connector to your regulator and use a wrench
to tighten it in place.
Waste Container
Solvent Tray/Waste-Collection Container
The solvent tray (the flat, white plastic container included in your
accessory kit) can hold 100 mL of waste. To install it, simply slide it
into the compartment under the front-left side of the instrument.
NOTE: The back of the compartment prevents the tray from sliding too far
under the instrument.
Orient the tray so that the black-plastic, barbed elbow fitting is
directed through the hole in the liquid collection trough (across the
front of the instrument) (Figure 1.3).
10
Thermo Electron
2x24 Alpha-numeric Display
RUN
STATUS
MENU
STOP
ENTER
SAMPLES
AS3000
Oven
(Optional)
To
Flush
Valve
To
Detector
WARNING
From
Pump
Liquid
Collection
Trough
Elbow
Fitting
Solvent
Pre-heat
Tube
Waste
Container
Aluminum
Mounting
Plates
(Oven Models
Only)
Power
Switch
AS\Z006E\DT
Single-piece
Injection Needle
Figure 1.3 The autosampler's front panel connections (the optional column oven
installed)
Thermo Electron
11
NOTE: Sometimes the barbed fitting may become dislodged during
shipping. Be sure that the end of the barbed fitting is directed through the
hole (not into the trough).
Other Waste-Collection Container
For larger-injection applications, or for those requiring frequent
flushing, you may wish to install a piece of Tygon® tubing onto the
waste port to allow the waste to flow to a carboy or other large
container for waste-collection.
To hook up your waste container, locate the Tygon tubing included in
your accessory kit and connect it to the end of the black, barbed
fitting. To prevent backflushing into the lines, place the container
lower than the instrument. Direct the Tygon tube into your container.
Pump-Autosampler
Connection
The way you connect the pump depends upon whether or not a
column oven is installed. For instruments without the column oven or
for inert/biocompatible instruments, the pump is connected directly to
the injector valve. If the column oven is installed, the pump is
connected instead to the factory-installed solvent-preheating tube, a
tube that routes behind the heating mantle (plate), up through the
service loop, and then to the injector valve.
NOTE: Your standard accessory kit contains two pieces of 0.010 ID
stainless-steel tubing (one 4-inch piece and one 12-inch piece). The 12inch piece is included to make the column-detector connection, but may be
used instead to make the pump-autosampler connection.
Without the Column Oven Installed
The instrument is shipped with the needle and flush lines installed.
To connect your pump to the autosampler's injection valve, use the
following steps:
1. Remove the column compartment cover (if you have not
already done so) and set it aside.
NOTE: The cover fits snugly. To remove, place your fingers or thumb in
the indentation on the top edge of the column door and pull back toward
you firmly.
2. Route the pump outlet line through one of the two notches on
the left side of the autosampler.
3. Connect the line to port 6 (Figure 1.4) on the injector valve.
Finger-tighten, then use a 3/16" wrench to tighten the fitting an
extra 1/8 turn.
NOTE: Do not under- or over-tighten the screws, or damage to the
connections could occur. Undertightening causes leaks; overtightening
may cause leaks or damage the connections.
12
Thermo Electron
2
3
1
4
6
5
RUN
STATUS
MENU
STOP
ENTER
SAMPLES
SpectraSYSTEM
AS3000
AS-Z007/FM
WARNING
Figure 1.4 The autosampler's injector valve
With the Column Oven Installed
The instrument is shipped with the column oven assembly and the
flush and needle lines already installed. The stainless-steel, lowdead-volume union that connects the pump outlet tubing to the
column oven tubing is also factory-installed. You will have to
connect the pump to the zero dead-volume union (ZDV), however.
The preheat line runs behind the column oven, through the oven's
mantle, up through the service loop, and into the injector valve. For
standard analytical configurations, this provides approximately
180 µL of preheated solvent. Instruments with 0.007” narrow-bore
preheat tubing provide approximately 22.7 µL preheated solvent.
Connect the pump to the preheat tube (Figure 1.5) as follows:
1. Remove the column oven cover (if you have not already done
so) and set it aside.
NOTE: The cover fits snugly. To remove, place your fingers or thumb in
the indentation on the top edge of the column door and pull back toward
you firmly.
2. Route your pump outlet line through the lower notch on the left
side of the autosampler.
Thermo Electron
13
3. Connect the line to the low-dead-volume union on the preheat
tube as follows:
a. Locate the ferrule included in your accessory kit.
b. Thread the tubing through the fitting, and place the ferrule
on the end of the preheat tube (Figure 1.5).
c. Then use two 1/4" wrenches to tighten the connection. Use
one wrench to hold the union body and one to tighten the
nut 1/8-turn past the stop.
Refer to Chapter 3 for instructions on operating the oven.
14
Thermo Electron
RUN
STATUS
MENU
STOP
ENTER
SAMPLES
AS3000
WARNING
Low Dead-Volume Union
Ferrule
Nut
AS-Z013E/KF
Pump Outlet Line
Figure 1.5 The pump-to-preheat tube connection
Sample Loop
Installation
The AS3000, AS3500, and AS300 autosamplers come with a 100 µL
sample loop installed; the AS1000 and AS100 autosamplers come
Thermo Electron
15
with a 20 µL loop. If you wish to change sample loops, refer to
Chapter 5, page 147.
Column Installation
Without the column oven installed, the column requires no special
installation. If the column oven is installed, the column is placed
between two aluminum column mounts that attach to the column
oven's plate. The aluminum mounts increase the oven's temperature
stability during ambient temperature changes, thereby ensuring a
stable column temperature. They also provide excellent heat transfer
from oven to column. Both mounting procedures are described here.
Without the Column Oven Installed
Use this procedure for mounting a column on instruments without the
Column Oven Option and for inert/biocompatible versions.
NOTE: Flush/purge the pump and autosampler lines before connecting the
column. This will ensure that no air gets into the column.
Mount a column on your autosampler as follows:
1. Locate the 4-inch length of stainless steel tubing (0.010) and
fittings included in your accessory kit.
2. Connect the 4-inch tubing to the inlet end of the column.
3. Connect the other end of the 4-inch tubing to port 5 on the
injector valve (Figure 1.5).
NOTE: Thermo Finnigan columns are connected such that liquids flow
from left to right as you read the label. If you're not using a Thermo
Finnigan column, refer to your column documentation to orient your
column in the correct direction of flow.
4. Use a 1/4 - 3/16" wrench to tighten the fittings. If you're
installing the column onto the autosampler for the first time,
tighten the fitting 1/8-turn past finger-tight. Otherwise, tighten
the fittings 1/16-turn past finger-tight.
NOTE: Do not overtighten the fittings. You could damage, or cause
leaks to form around, the column connections.
With the Column Oven Installed
If you purchased the column oven, the column secures directly onto
the front of the oven plate. The instrument is shipped with the column
mounts already installed. To remove the front portion of the mounts:
NOTE: Flush/purge the pump and autosampler lines before connecting the
column. This will ensure that no air gets into the column.
1. Use a flat-blade screwdriver to loosen the mounting screws and
to remove and separate the column mounts.
NOTE: Thermo Finnigan columns are usually connected such that liquids
flow from left to right as you read the label. If you are not using a Thermo
Finnigan column, refer to your column documentation to orient it in the
correct direction of flow.
16
Thermo Electron
2. Place the column between the column mounts, and replace the
mounting screws to secure the column in place.
AutosamplerDetector
Connection
The 12-inch piece of 0.010 ID tubing is provided in your accessory
kit to connect your autosampler to your detector. Simply, connect the
outlet end of the column to your detector (Figure 1.5).
NOTE: If you are using a Thermo Finnigan detector, you won't need the
12-inch tubing because the detector's inlet line is long enough to reach to
the outlet end of your column.
You have completed the front-panel connections. Replace the
column oven cover. It snaps firmly into place.
HINT: To replace the door, place the bottom edge into place and then
snap the top closed.
Install Sample Trays
Your autosampler comes with three sample trays and a supply of
vials, silicone and Teflon® septa, and screw-top vial caps.
Additional vials may be ordered from Thermo Finnigan. Be sure to
use standard size vials to ensure smooth autosampler operation.
NOTE: Be sure to use a 40-60 mil (1.0-1.5 mm) rubber or silicone
septum with a 5 mil (0.1 mm) Teflon liner on one side.
Each tray contains 40 (35, with Tray Temperature Control) vials
arranged in eight (seven) rows of five. The first vial position is A01;
the last is C40. The position labels are on the left of each position.
After your vials are in place, install sample trays A, B, and C as
follows:
1. Hold the tray handle, tilting the back end down (Figure 1.6).
2. Insert the tray's "runners" into the slots in position A at the rear
of the tub.
3. Lower the front of the tray into place and press down firmly to
seat the tray.
HINT: Pull the handle back toward you until the tray snaps into
place. Try moving each tray from side to side to be sure that it's
seated completely.
Thermo Electron
17
4. Insert the other two sample trays into positions B and C.
NOTE: Be sure that each tray is properly seated. Each tray's upper
front edge should fit under the upper lip of the tub. If a tray is not
completely seated, the autosampler will not be able to correctly pick
up or return vials to that sample tray.
Sample Tray Handle
(front)
A
B
Tub
Tray Positions
Runner
Slots
AS-Z003E\FM
C
Figure 1.6 Installing a sample vial tray
POWERUP
Locate the power cord in the shipping container (not in your
accessory kit). Insert one end into the AC power receptacle on the
rear panel and the other end to a grounded power outlet. You don't
have to worry about the fuses. Your autosampler is shipped with the
correct fuses installed. Turn on your instrument by pressing the
on/off button located in the lower right-hand corner just below the
front (Figure 1.1).
During startup, the instrument's self-diagnostic sequence
• Completes a series of ROM checks
• Completes a series of RAM checks
• Activates the injection valve
• Verifies the proper connections on all installed options
• Verifies the proper operation of all sensors
• "Homes" (returns to starting position) all syringes and motors
18
Thermo Electron
Priming
After you have completed the plumbing and powerup procedures and
started your instrument, you need to prime your autosampler to wet
all of the lines. To do so, you'll need to flush all syringes and lines
with an appropriate solvent.
NOTE: The autosampler is shipped with methanol in the lines. Be
sure to use a miscible solvent.
HINT: If you purchased the Sample Preparation Option, proceed to the
"2.5 mL" section to beginning priming the lines using the prep syringe. It
will require less time. Then return to the "250 µL Syringe" section.
250 µL Syringe
To prime the autosampler:
1. Fill your flush solvent bottle, if you have not already done so.
2. Press [MENU] and select /COMMANDS/, (Figure 1.7).
Flush Sample Syringe
Initialize Hardware
Injector
Arm
Figure 1.7 The Commands Menu
Flush Sample Syringe
Flush Prep Syringe
Initialize Hardware
Injector
Arm
Figure 1.8 The AS3000 Commands Menu with the Sample
Preparation Option installed
3. Select /Flush Sample Syringe/ to access the menu shown in
Figure 1.9.
4. Use the [+] and [-] keys to select 5000 µL. Then press
[ENTER] to initiate the flush operation.
Flush 5000 µL
Figure 1.9 The Flush Sample Syringe Menu
The autosampler automatically flushes the sample syringe and flush
lines with the specified volume of flush solvent and returns the
syringe to its starting position. The recommended 5000 µL flush
takes approximately ten minutes. During the flush sequence, a
confirmation message is displayed.
Thermo Electron
19
2.5 mL Syringe
(AS3000 and
AS3500 only)
If you purchased the Sample Preparation Option for your AS3000 or
AS3500 autosampler, you must also flush the large (2.5 mL) sample
preparation syringe and external solvent lines as follows:
1. Fill each solvent bottle with an appropriate solvent.
2. Press [MENU] and select /COMMANDS/.
3. Select /Flush Prep Syringe/ (Figure 1.10).
4. Use the [+] and [-] keys to select 5000 µL. Then move the
cursor to the Solv field and select /Btl/. Press [ENTER] to
initiate the flush sequence.
Flush 5000 µL Solv Btl
Figure 1.10 The Flush Prep Menu
NOTE: If there is residual air in the solvent-holding loop, flushing
may expel solvent out the open port. To eliminate residual air that
may be present during shipping, be sure to flush the lines with Btl
selected before flushing with S-1 through S-4 selected. Be sure that
the S1-S4 solvent lines are connected.
5. Repeat this step for S-1 through S-4 with 1000 µL from the
flush bottle (Btl).
The autosampler automatically flushes the selected solvent line with
the specified volume of flush solvent and then returns the syringe to
its starting position.
NOTE: If you are replacing syringe(s), be sure to install the
2.5 mL syringe on the left. Injecting small sample volumes with the
large syringe can adversely affect reproducibility. Also, be sure to
verify the correct syringe size in the Options/Configuration Menu.
20
Thermo Electron
Communication
The SpectraSYSTEM or SpectraSERIES autosamplers can be
configured to send information to other Thermo Electron
SpectraSYSTEM modules or to older SpectraSYSTEM and nonSpectraSYSTEM LC instruments, as follows:
•
SpectraNET Mode: For AS1000 and AS3000, and AS3500
instruments only, SpectraNET mode as a part of a
SpectraSYSTEM chromatograph connected to other
SpectraSYSTEM modules through the SN4000, and controlled
by your chromatography data system
•
Stand Alone Mode: For any autosampler, stand-alone mode
hardwired to other non-SpectraSYSTEM HPLC modules
through the remote input/output connectors or connected to
older SpectraSYSTEM and non-SpectraSYSTEM instruments
by means of specialized cables and/or communication
protocols. These options are described below.
NOTE: To connect to SpectraSYSTEM and non-SpectraSYSTEM
integrators using your chromatography data system, refer to your
chromatography data system Reference Manual.
SpectraNET MODE
(AS1000, AS3000,
AND AS3500 ONLY)
Configured as a SpectraSYSTEM module using SpectraNET
communication and the SN4000, your autosampler is automatically
controlled by your chromatography data system. The chromatgraphy
data system assumes almost all keyboard control and coordinates the
majority of the autosampler's operations. To configure your
autosampler to use SpectraNET communication:
1. Turn off your autosampler.
2. Connect your autosampler to the SpectraNET Interface Module
as described in your chromatography data system Reference
Manual.
3. Turn on your autosampler, wait for the powerup sequence to
complete, and then press /OPTIONS/Configurations/ to access
the Configurations Menu (Figure 1.11).
Thermo Electron
21
Sample Syringe
250
Prep Installed
Yes
(With sample prep)
--------------------------------------------------------Oven Installed
Yes
TrayTemp Installed
Yes
Key Repeat Rate
Door Interlock
File Name
Medium
On
Protect
1: VITAMINS
On
2: BARBITUA
Off
3: EXPER 3
Off
4: STAT
Mode
Solv Viscosity
Off
SpectraNet
Normal
Figure 1.11 The Configurations Menu in SpectraNET mode
NOTE: If you are running the RAM Hardware Test with "No System
Control" selected in the Mode Menu (38), the Controller (ISM100) will
cause the autosampler to reinitialize the hardware.
4. Move the cursor to the Mode field, and use the [+] and [-] keys
to select SpectraNET. Then press [ENTER] to accept the field
value and exit the Configurations Menu.
Your autosampler is now configured as a SpectraSYSTEM module.
HARDWIRED
STAND-ALONE
COMMUNICATION
Stand alone mode is used to operate SpectraSYSTEM or
SpectraSERIES modules without data system control. Stand alone
mode is also used to connect older and non-SpectraSYSTEM
laboratory equipment.
The 12-pin, in-line connector on the instrument's rear panel allows
communications between your autosampler and other components in
an LC system (Figure 1.12). This simplistic communications system,
known as "hardwiring," provides injection synchronization between
your autosampler and the other components of your LC system.
A mating connector (provided in the accessory kit) allows you to
attach control wires from other LC devices to the 12-pin terminal by
loosening the appropriate screw, inserting the wire, and then
tightening the screw. Once all connections are made, you can quickly
make and break connections without disturbing individual wires.
Hardwire communications allow the autosampler to start and stop a
pump or detector, to begin a gradient program (gradient pumps only),
or to control other features of the externally connected instruments.
In addition, the terminal has four timed-function outputs that can be
22
Thermo Electron
used to control valve-switching devices or other devices on a timed
basis. (Refer to Chapter 3, "Timed Events Program," page 72 to set
up a timed-events program.)
This section describes the pin assignments, the recommended wiring
configuration, and some optional wiring configurations.
12
11
10
9
8
7
6
5
4
3
2
1
TERMINAL CONFIGURATION
WARNING
C
O
M
M
.
TERMINAL CONFIGURATION
DATE OF MANUFACTURE:
DESCRIPTION
SIGNAL
12. TIMED EVENT 1
11. TIMED EVENT 2
10. TIMED EVENT 3
9. TIMED EVENT 4
8. A/S READY
7. INJ HOLD
6. INJ OUT
5. PUMP READY
4. GRAD START
3. PUMP STOP
2. +5V 150MA
1. GROUND
OUT
OUT
OUT
OUT
OUT
IN
OUT
IN
OUT
OUT
OUT
AS\Z009\FM
THIS PRODUCT IS COVERED BY ONE OR MORE OF THE FOLLOWING PATENTS:
4,957,009
4,953,075
4,478,095
Figure 1.12 The 12-pin terminal
Thermo Electron
23
Pin Assignments
Outputs
All output signals, including Time Functions 1 through 4 (TF1-4), are
open-collector outputs capable of sinking (handling) currents up to
600 mA at voltages up to 24 VDC. The active state is user
programmable in the Timed Events Menu (see Chapter 3).
Inputs
All inputs accept TTL-level inputs, with a transition activating each.
The two inputs are tied to +5 VDC through pull-up resistors. A "Lo"
level is from 0 to 0.8V; a "Hi" level is from 2 to 5.5V.
NOTE: All inputs and outputs are referenced to ground.
Pin 1 - GROUND: The ground reference used for all inputs and
outputs.
Pin 2 - +5VDC: This regulated +5 VDC supply can be used to power
pull-up resistors or external relays. The current is limited to a
maximum of 300 mA to protect against external short circuits
affecting the autosampler's operation.
Pin 3 - PUMP STOP: (output)This pulsed output is used to stop the
pump after the last sample has been processed. It is activated when
the last sample in the queue has completed its last injection, or when
the shutdown file has been activated (using the special shutdown
command). See Chapter 5 for more information on the Shutdown
command.
Pin 4 - GRADIENT START OUTPUT: This signal activates the
pump's gradient program. It occurs at a specified amount of time
before the Inject Out line is activated. The time is specified as a
Gradient Delay value in the More Menu (under Files). This
momentarily active, pre-injection signal permits the gradient to start
prior to sample injection to compensate for system delay volume.
The result is an injection that occurs just as the gradient front reaches
the injection valve. See the last section in this appendix for more
information on the gradient delay function.
Pin 5 - PUMP READY (input): Through this input, the pump
signals the autosampler that the mobile-phase flow rate is stable and
that the pump is ready for an analysis to begin. (Other devices may
also be connected here.) This input must be continuously active for
the autosampler to proceed with injections. From the Input Polarity
Menu you can select whether a high or low level on this line means
"ready." The default is Hi.
24
Thermo Electron
Pin 6 - INJECT OUT (output): This line sends a momentary signal
each time the autosampler makes an injection. This signal allows the
synchronization of external devices (pumps, detectors, integrators,
etc.) to the autosampler's injection. See "Outputs" below for the
specifications of this output.
Pin 7 - INJECT HOLD (input): This signal puts the autosampler in
the hold state with sample in the loop. Releasing the hold will cause
an immediate injection. The signal must remain continuously active
to hold injections. That is, when active, this signal will delay an
imminent injection until this line goes to the inactive state. This
function can be used with external equipment to precisely control the
timing of the injection valve activation. The active level (Hi or Lo) of
this input can be selected in the Input Polarity Menu (see Advanced
Installation) at the end of this appendix.
Pin 8 - A/S READY: This line is used to signal to other devices that
the autosampler is in a READY condition. See "Outputs" below for
the specifications of this output.
Pins 9 thru 12 - TF1-TF4 (timed outputs): These user-programmed
outputs are useful for triggering column switching valves or other
external hardware. These outputs are controlled by the time functions
defined in the currently active file (see Chapter 3 "Timed Events"
page 72).
Recommended
Hardwire
Configuration
The recommended configuration requires a pump, autosampler,
UV/VIS or fluorescence detector, and an integrator. We recommend
that you connect the following signals:
•
Pump Ready
•
Inject Hold (detector ready)
•
Detector Run/Start
•
Integrator Run/Start
Refer to Table 1.1 to wire the appropriate pin connections. Pin
connections for the SpectraSYSTEM modules are indicated in
parentheses. Optional connections are indicated by asterisks (*).
Thermo Electron
25
Table 1.1 Pin Connections for the Recommended Configuration
Autosampler (Pin
Connection)
Pump
Detector
Integrator
Ground (1)
Ground (3)
Ground (9)(11)
Ground (B1)
Pump Stop (3)
(pulsed output)
Pump Stop (5)
N/A
N/A
Pump Ready (5)
(input)
Pump Ready (1)
READY (4)
N/A
Inject Out (6)
N/A
RUN/START (10)
RUN/START "A" (B5)
RUN/START "B" (B14)
*Inject hold (7)
Sync (8)
N/A
N/A
Gradient Start (4)
Run Grad (7)
N/A
N/A
Figure 1.13 illustrates the hardwire connections for the
SpectraSYSTEM modules.
J4
CH1
F
1.0 V F/S
E
Ground
B
1.0 V F/S
A
Ground
CH2
TB1
1
3
P2
Pump Ready
Pump Ready
Ground
Ground
5
Pump Stop
7
Run Grad
8
Sync
Pump Stop
Gradient
Start
Inject Hold
5
1
J2
Ready
Pump Ready
Ground
Ground
3
4
+
CH. A
9
11
Ground
4
+
CH. B
Run Start Cable
Integrator
7
"A" Run Start
6
Run/Start
Inject Out
Autosampler
10
"B" Run Start
Detector
Figure 1.13 Hardwire connections for SpectraSYSTEM Modules
26
Thermo Electron
AS-Z046E/DT
Pump
Communications
Menu Parameters
For normal stand-alone operation, you do not need to need to change
any of the values in the Communications Menu. Leave the
communications parameters set to the default values (Figure 1.14).
Details provided below for changing the polarity are provided for
reference only.
Baud Rate
9600
Parity
NONE
Flow Control
NONE
Data Bits
8
Stop Bits
1
Echo
On
Figure 1.14 The Communications Menu default values used for
normal stand-alone operation
The allowable entries and default values are listed below for each of
the communications parameters.
Baud Rate: Allowable entries: 1200, 2400, 4800, or 9600 Bd;
default: 9600Bd.
Parity: Allowable entries: None, Even, or Odd; default:
NONE.
Flow Control: Allowable entries: NONE or XON/OFF;
default: NONE.
Data Bits: Allowable entries: 8 or 7; default: 8.
Stop Bits: Allowable entries: 1 or 2; default: 1.
Echo: Allowable entries: On or Off; default: On.
NOTE: If the Communications Menu doesn't appear as one of the
OPTIONS Menu choices, return to the OPTIONS Configurations Menu,
and verify that Mode is set to Stand Alone. The Communications field will
not appear in the OPTIONS Menu if Mode is set to SpectraNet.
Setting the Polarity
From the Options Menu you can change the sense of your
autosampler's input and output signals as required by your other LC
system modules. For example, when shipped, the Autosampler Ready
Output is set to send a positive voltage (Hi) when the instrument is
ready. If you change the Autosampler Ready Output Polarity to Lo,
the instrument will send a 0-volt signal (Lo) when it's ready. The
procedure for changing the polarity of the input and output signals is
described below.
Thermo Electron
27
Input Polarity
To change the polarity of an input signal, select /Input Polarity/ from
the Options Menu. Use the [+] and [-] keys to select either Hi or Lo
for the desired input signal shown in Figure 1.15.
Pump Ready Active
Hi
Inj Hold Active
Lo
Figure 1.15 The Input Polarity Menu
This menu shows the level for the active state for each input signal.
NOTE: If you want to determine the current state of a particular signal,
return to the Main Menu, and select /TESTS/Display Input States/.
Output Polarity
To change the polarity of an output signal, select /Output Polarity/
from the Options Menu. Use the [+] and [-] keys to select either Hi or
Lo for the desired output signal shown in Figure 1.16.
Autosampler Ready
Lo
Inject Out Active
Lo
Grad Start Active
Lo
Pump Stop Active
Lo
Figure 1.16 The Output Polarity Menu
This menu shows the level for the active state for each output signal.
NOTE: If you want to determine the current state of a particular signal,
return to the Main Menu, and select /TESTS/Display Output States/.
Table 1.2 and Table 1.3 show the output signal levels relative to the
polarity settings and instrument status for the autosampler's input and
output signals, respectively..
Table 1.2 Input Signal Levels
Pump Ready
Ready
Not Ready
Inj Hold
Active
Inactive
Note: The asterisked (*) values indicate default settings.
28
Thermo Electron
---------Polarity------Hi
Lo
Hi*
Lo
Lo
Hi
Hi
Lo
Lo*
Hi
Table 1.3 Output Signal Levels
---------Polarity------Hi
Lo
Hi
Lo*
Lo
Hi
Autosampler Ready
Ready
Not Ready
Inject Out
Active
Inactive
Hi
Lo
Lo
Hi*
Gradient Start
Active
Inactive
Hi
Lo
Lo
Hi*
Hi
Lo
Lo
Hi*
Pump Stop
Active
Inactive
Note: The asterisked (*) values indicate default settings.
STAND ALONE
COMMUNICATION
TO INTEGRATORS
Two specialized communication setups are available:
•
Via an autosampler-to-integrator cable to ChromJet or DataJet
integrators
•
Via BCD interface to older Thermo Finnigan integrators or to
non-Thermo Finnigan Integrators
Before using either of these communications setups, it is necessary to
configure your autosampler as a stand-alone module as follows:
1. Turn on your autosampler, wait for the powerup sequence to
complete, and then press /OPTIONS/Configurations/ to access
the Configurations Menu (Figure 1.17).
Sample Syringe
250
Prep Installed
Yes
--------------------------------------------------------Oven Installed
Yes
TrayTemp Installed
Yes
Key Repeat Rate
Door Interlock
File Name
Medium
On
Protect
1: VITAMINS
On
2: BARBITUA
Off
3: EXPER 3
Off
4: STAT
Mode
Solv Viscosity
Off
Stand Alone
Normal
Figure 1.17 Configurations Menu in stand-alone mode
Thermo Electron
29
2. Move the cursor to the Mode field, and use the [+] and [-] keys
to select Stand Alone. Then press [ENTER] to accept the field
value and exit the Configurations Menu.
Your autosampler is now configured as a stand-alone LC module.
ChromJet or
DataJet
Communication
Using an autosampler-to-integrator cable available from
Thermo Electron, you can set up your autosampler to send vial
information to a Thermo Finnigan ChromJet® (SP4400) or a DataJet
(SP4600) integrator. For this connection you will need an RJ-45-toRJ-45 cable and an integrator adapter (p/n A3981-010).
To make this connection:
1. Turn off your integrator.
2. Insert the RJ-45 connector (telephone connector) into the
COMM. Port (telephone jack) on the autosampler's rear panel.
3. Connect the adapter to the other end of the cable. It orients in
only one direction.
4. Plug the adapter-cable end into your DataJet or ChromJet
integrator.
5. Turn on your integrator and type in the following:
BAUD 9600
POKE#C12D,#FF
6. Hold the [CTL] key and press [CALIB].
NOTE: You must reenter this sequence after any integrator soft
reset!
This sequence provides you with the four-line report from your
autosampler shown in Figure 1.18.
Sample Vial: A02 Inj Volume :
Inject Vial: A01 Inj/Vial
:
Vial Type : SAMPLE
Cycle Time
File 1: FILENAME
Set Number :
5.0
1/1
0.8
Oven Temp :
26
Tray Temp:
0
Heater Temp: 24
1
Figure 1.18 The vial location and injection report
30
Thermo Electron
BCD
Communication
The BCD Communication Option provides BCD communication
from the AS1000/AS3000/AS3500 SpectraSYSTEM autosamplers to
older Thermo Electron and non-Thermo Electron integrators and
computers. This option provides tray- and/or vial-location
information to devices requiring either binary or binary-coded
decimal data. The BCD assembly must be installed by a Thermo
Finnigan service representative if ordered after initial instrument
delivery. Installation instructions are provided in the BCD Option
Kit. Contact your representative for further information.
Ready Participation
When the instrument is shipped, the following conditions must be
satisfied in order for the Status Screen to indicate that the autosampler
is "ready" for operation:
•
The door must be closed.
•
The oven must be at the set temperature (if installed).
•
The heater must be at the set temperature (with the Sample
Preparation Option only).
•
The tray must be at the set temperature (with the Tray
Temperature Control Option installed).
The Options Ready Participation Menu allows you to customize the
"autosampler ready" requirements to fit your own needs. For
example, you may wish to prepare your first sample set (without
injection) while the column oven is still warming up. In this case,
you would want to disable the column oven condition in order to
complete the sample preparation run. Or, you may wish to run your
autosampler with the door removed. In this case you would want to
disable the door interlock.
To access the Ready Participation Menu (Figure 1.19), select
/OPTIONS/Ready Participation/.
Door Interlock
ENABLE
Col Oven Ready
ENABLE
--------------------------------------------------------Heater Ready
ENABLE
Figure 1.19 The Ready Participation Menu
To change any of the above requirements, move the cursor to the
appropriate line, and use the [+] and [-] keys to change ENABLE to
DISABLE.
Thermo Electron
31
Instrument Configurations and Other Features
Your AS300, AS3000, or AS3500 autosampler is shipped with a
100 µL sample loop (the AS1000 or AS100 instruments come with a
20 µL loop), a 250 µL sample syringe, and a 2.5 mL sample
preparation syringe (with the Sample Preparation Option installed).
Should you wish to change your syringe sizes, you can do so from the
Configurations Menu (Figure 1.20). This menu also reflects your
other installed options (oven and tray temperature control). If you
would like to adjust the rate at which fields cycle through the
available choices, you can change the Key Repeat Rate entry.
Chapter 3 discusses the File Protect function also available from this
menu.
Sample Syringe Size
250
Prep Syringe Size
2500
--------------------------------------------------------Prep Installed
Yes
Oven Installed
Yes
TrayTemp Installed
Yes
Key Repeat Rate
Medium
File
Protect
1:
Off
2:
Off
3:
Off
4:
Off
Solv Viscosity
Normal
Figure 1.20 The Configurations Menu
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 [>] and [+] to increase the
contrast, or [>] and [-] to reduce the contrast.
32
Thermo Electron
Performance Verification
After you have installed your SpectraSYSTEM autosampler, we
suggest that you run a standard sample, such as a 0.5% mixture of
toluene and methanol, to verify the instrument's proper operation. In
addition, you can use the following procedures to test your
instrument's reproducibility and sample carryover, and to troubleshoot
and maintain the instrument in the future.
NOTE: Before beginning the performance verification runs, you may wish
to familiarize yourself with the autosampler’s menu structure and keypad.
This information is found in Chapter 2 or on the quick reference and menu
cards in the front pocket of the manual.
REPRODUCIBILITY
Use this procedure to measure your autosampler's reproducibility. In
this procedure, you will inject several volumes of a standard "sample"
of 0.5% toluene:MeOH, determine the percent relative standard
deviations (%RSD) for the injection volumes, and compare them with
the desired results included in this section. The test will require
approximately three hours. Retain these results for comparison
should troubleshooting be necessary.
Use the following HPLC column, solvents, and test "sample":
Column:
4.6 x 100 mm Spheri-5 RP18, 5 micron
Flow rate:
1.5 mL/minute
Mobile phase:
100% HPLC-grade methanol
Flush solvent:
100% HPLC-grade methanol
Test Sample:
0.5% toluene in methanol
Detection:
254 nm
Test Setup
Set up the test run as follows:
1. Fill five vials with the test sample and place them in tray
positions A01-A05.
2. Create a file using the Injection Menu parameters shown in
Figure 1.21.
Thermo Electron
33
Injection Volume
10
Injections/Sample
10
-------------------------------------------------------Cycle Time
1.5
Col Oven
OFF
Temp
18
TrayTemp
OFF
Temp
0
Figure 1.21 The Injection Menu performance verification parameters
3. Return to the Edit Menu.
4. Access the More Menu and verify that the Needle Height is set
to 2.0. (Do not change any other parameters in this menu.)
5. Return to the Main Menu and Load your file.
6. Press the [SAMPLES] key and set up a sample queue that
contains five sample sets of one vial each. For each sample set,
enter the following Sample Menu values:
Inj/Sample 10
Cycle Time 1.5
No. Vials
1
Vary the injection volume (Inj Vol) for each set using the
values listed in Table 1.4.
Table 1.4 Reproducibility Test Sample Queue Parameters
Sample
Inj
Sample
Set
Vol. (µL)
Vial
1
10
A01
2
1
A02
3
3
A03
4
7
A04
5
10
A05
7. Return to the Main Menu.
8. Flush the syringe with 500 µL of flush solvent. (Select
/COMMANDS/, /Flush Sample Syringe/).
9. Inspect all sample lines for bubbles. (Be sure to check the
Teflon line that connects to port 3 of the injection valve.) If
bubbles are present, repeat step 8 until all bubbles are removed.
10. Check for leaks.
11. Initialize your pump.
12. Prepare your integrator for a run.
13. Press [RUN].
The instrument should perform 50 injections. The first group of ten
injections is used to equilibrate the system, so don't use the data from
these injections.
34
Thermo Electron
%RSDs
Determine the percent relative standard deviations (%RSD) for the
injection volumes and compare them with the desired results in
Table 1.5.
Table 1.5 %RSD Results for the Reproducibility Test
Volume (in µL)
%RSD
1
<1*
3
<0.7*
7
<0.5*
10
* Typical value
** Instrument specification
<0.5**
If your results are other than expected, refer to Appendix A,
Troubleshooting.
SAMPLE
CARRYOVER
This procedure will allow you to verify that the sample carryover is
below the 0.01% specification for your autosampler. It requires the
use of silicon septa, a deflected point needle (p/n 4719-010, -020), the
appropriate volume of the required flush solvent, and clean (not
previously used) blanks. The procedure minimizes the effects of
sample concentration or chemistry, chromatographic conditions,
detector linearity, or other hardware characteristics that might affect
sample carryover.
This procedure is independent of sample- and system-specific
characteristics. It reduces sample effects including extinction
coefficients, chemical interaction with other sample or system
components (the column), and sample degradation. It also minimizes
system effects including detector linearity, pump and mixing
anomalies, and column and guard-column contamination.
In this procedure you will inject your sample, a subsequent blank
(BLANK1), additional blanks to return the system to baseline, and
then a standard (STANDARD) that is 0.01% of your sample. You
will determine the % carryover by multiplying the ratio of the
carryover by the 0.01% specification:
Carryover = [Peak AreaBLANK1 / Peak AreaSTANDARD] x 0.01%
If desired, run this procedure three times to replicate the results.
Thermo Electron
35
Materials
To perform this procedure you'll need:
•
Your sample
•
7 vials, septa, and caps for each run
•
Autosampler with deflected point needle
•
Any HPLC pump, detector, and data system
Sample Preparation
Prepare your SAMPLE and STANDARD as follows:
1. Place appropriate volumes of sample and starting mobile phase
into a vial labeled SAMPLE.
2. Dilute your sample 10,000 times in starting mobile phase and
pipette into a vial labeled STANDARD.
Example:
1. The sample is 100 mg protein in 1 mL phosphate buffer
(SAMPLE)
2. Transfer 1 µL into 9.99 mL of phosphate buffer
(STANDARD)
Blank Preparation
Prepare your blanks as follows:
1. Pipette fresh starting mobile phase into five vials.
2. Label vials as BLANK, BLANK1, BLANK2, BLANK3, and
BLANK4 for each test set.
Test Procedure
System Setup
Set up your system as follows:
1. Prepare the appropriate mobile phase(s).
2. Set up the appropriate chromatographic conditions for your
sample.
3. Set up your integrator/data system so that it displays retention
times and peak area information.
Autosampler Setup
Set up your autosampler as follows:
1. Set the flush volume to 2000 µL:
a. Press [MENU], /Files/Flush Volume/
b. Select 2000 µL
NOTE: The flush solvent should be the starting mobile phase or other
solvent that completely dissolves the sample.
36
Thermo Electron
2. In the Injection Menu, set the injection volume (loop size) to 5,
10, or 20 µL as appropriate.
3. Set the number of injections to 1.
4. In the /Files/More/ Menu, set the injection type to Push.
5. Set the vial positions as follows:
A01: BLANK
A02: SAMPLE
A03: BLANK1
A04: BLANK2
A05: BLANK3
(Add additional Blanks as necessary to allow the system to
return to baseline before running the STANDARD.)
B01: STANDARD
B02: BLANK4
(Add additional Blanks as necessary to allow the system to
return to baseline before replicating the test.)
6. Set up two additional groups of seven vials to run the carryover
test in triplicate (optional, but recommended).
7. Run your HPLC system using the appropriate chromatographic
conditions for your sample.
Carryover
Calculation
To calculate the % sample carryover:
1. Divide the peak area of BLANK1 by the peak area of the
STANDARD. This gives you the ratio of the carryover to the
0.01% specification.
2. Multiply this value by 0.01% to obtain the percent carryover.
Example:
If the peak area for BLANK1 is 110,958 counts,
and the Peak area for the STANDARD is 656,283 counts,
the ratio of the carryover to the specification is
110,958/656,283 or 0.1691
% carryover = 0.1691 x 0.01%
% carryover = 0.001691
Specification
Since 0.001691% < 0.01%, the autosampler is within specification.
Thermo Electron
37
DILUTION
REPRODUCIBILITY
(AS3000 AND AS3500
ONLY)
This procedure is used to determine your autosampler's dilution
reproducibility. In this procedure you will prepare and inject five
1/100 dilution from a standard sample of 50% Toluene:Methanol.
The percent relative standard deviation of peak areas will then be
determined across the five samples and compared to the specification
presented in this section. The test will require approximately
1.5 hours. These results should be retained for comparison should
troubleshooting be necessary.
Use the following HPLC column, solvents, and "test" sample:
Column:
Spheri-5 RP18, 5 micron, 100 x 4.6 mm
Flow Rate:
1.5 mL/min
Mobile Phase:
100% HPLC grade Methanol (MeOH)
Flush Solvent (Btl and S-1):100% HPLC grade Methanol (MeOH)
Test Sample:
50% Toluene in Methanol
Detection:
254 nm
Test Setup
Setup the test run as follows:
1. Fill five vials with the test sample and place them in tray
positions A01, A03, A05, A07, and A09.
2. Place empty vials in tray positions A02, A04, A06, A08, and
A010.
3. Advance to the Sample Prep menu in Method 1. Enter 1 for
the dilution template and setup the parameters as follows:
- Template 1
- Dilution
- # of dilution cycles 1
- 1 - Load 1000 µL Solv S-1
- 2 - Pickup 10 µLSample
- 3 - Pickup 0 µL Sample
- 4 - PickLG 0 µL Sample
- 5 - Add 1000 µL to Sam + 1
- Mix for 0.5 minute
- Overlap enable - Yes
38
Thermo Electron
4. Flush sample syringe with 500 µL. Next flush the prep syringe
with 4800 µL from Btl and S-1. Check lines for air bubbles,
repeat if necessary.
5. Press the [SAMPLES] key and setup a sample queue that
contains five sample sets of one vial each. For each sample set,
enter the following Sample Menu values:
Inj/Sample
3
Cycle Time
1.5
No. Vials
1
Vary the sample vial position for each sample set:
Sample Set
Inj. Vol.
Sample Vial
1
10
A01
2
10
A02
3
10
A03
4
10
A04
5
10
A05
6. Initialize your pump and check for leaks.
7. Prepare the integrator for a run.
8. Press [RUN].
The autosampler will prepare 5 dilution of the test sample that make a
0.5% Toluene in methanol sample. It will then inject 10 µL of the
diluted sample three times.
% RSD
When the test run is complete, gather the area counts for each of the
sample set's third injection. Calculate the average of these five area
counts and their standard deviation. Divide the standard deviation by
the average area count and multiply by 100 to obtain the % RSD.
The specification is < 1%.
If your results are not within the specification range, refer to
Troubleshooting, Appendix A.
Thermo Electron
39
Specifications
Vial capacity
120 vials in three removable trays
105 vials with tray temperature control
Variable-Volume
Injection Precision
Injection Volume
<0.5% RSD @ 10 µL or greater
0.1 - 100 µL injection standard (250 µL syringe)
up to 1000 µL injections with larger loops and syringes
Fixed-Loop
Injection Precision
Injection Volume
<0.5% RSD @ 10 µL or greater
20 µL standard (250 µL syringe)
1000 µL injections with larger loops and syringes
Sample Carryover
Typically <0.01% at 400 µL flush volume
Typically 5 mL flush volumes can be set
Minimum Sample Volume
1 µL can be injected from 10 µL with a standard needle
Needle Height
Column Oven (optional)
Programmable in 0.1 mm increments
Settable in 1° increments from 20° - 80°C, starting at 5°C
above ambient
Temperature stability typically ± 0.2°C
Accommodates up to 30 cm columns
Tray Temperature Control (optional)
Sample Preparation (optional)
Reagent Vials
Dilution Solvents
Heating/Mixing
Dispensing Precision
Method Files
Communications
Outputs
Three vials can be specified per file
Four external solvents can be selected
Single vial, vortex heater/mixer
Settable in 1°C increments from 30° - 100°C
Typically <1.0% RSD for a 1/100 dilution (combined
dilution and injection)
Typically <0.2% RSD for 1 mL
Protected in non-volatile memory
Pump Ready, Inject Hold
Pump Stop
Autosampler Ready
Inject
Gradient Start
Four timed-event outputs
BCD (optional)
SpectraNET/RS-232 port
Dimensions
Weight
14.5” (37 cm) x 16” (41 cm) x 20”(51 cm) (H x W x D)
35-53 lb. (16 -24 kg) depending on options installed
Power requirements
Environmental
Safety/EMC Compliance:
100-120/200-240VAC 50/60 Hz, 450VA
10-40°C; 5-95%RH noncondensing
CSA, TÜV, FCC, CE Mark
Dilution Precision
40
Settable in 1°C increments from 0° - 60°C
Temperature stability typically ± 1°C
Typical vial temperature from 4° - 50°C
Thermo Electron
2
A Quick Example
Introduction
This chapter provides you with the three basic rules you'll need for
using your AS3500, or AS3000, AS300, or AS100 SpectraSYSTEM
or SpectraSERIES autosampler. 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 that you read the Safety
Information section beginning on page vii and install your
autosampler as described in Chapter 1.
NOTE: Throughout our explanations, we encourage you to explore the
general architecture of your instrument's menus and screens. Use the
Menu Tree on the menu card in the front pocket of this manual as your
guide if you wish.
Learning Your Way
AS EASY AS 1-2-3!
It's easy to learn your way around a SpectraSYSTEM or
SpectraSERIES autosampler. 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 scroll up or down through preset choices, or to
increase or decrease alphanumeric entries.
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]
Thermo Electron
41
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.
VISUAL CLUES
The following conventions are used on the autosampler'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 (Figure 2.1). This is the command center from which you'll
access menus and control the instrument's operations. A brief
explanation of the keys and the main menus and screens follows.
RUN
STATUS
MENU
STOP
SAMPLES
SpectraSYSTEM AS3000
Figure 2.1 The autosampler's front panel
(With the exception of the model number, the front panel is identical
for all SpectraSYSTEM and SpectraSERIES autosamplers.)
42
Thermo Electron
AS\Z008\FM
ENTER
The keypad of each SpectraSYSTEM instrument consists of twelve
keys. Four keys directly control the instrument's operation: [RUN],
[STOP], [STATUS], and, on the autosampler, a blank key called
[SAMPLES]. 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 the run.
The specific [RUN] operation depends on the contents of the sample
queue:
1. If there are sample sets (groups of sample and associated
calibration vials) in the sample queue, pressing [RUN] begins
processing the first vial in the sample set.
2. If the sample queue is empty, pressing [RUN] prompts you to
press [SAMPLES] to display the Samples Menu so that you
can add sample sets to the sample queue.
[STOP]
Pressing [STOP] pauses the sample set currently running, aborts
subsequent sample sets, and returns the autosampler to the initial run
conditions. Refer to Chapter 3 for more information on the [STOP]
key's function.
[STATUS]
Pressing [STATUS] displays the Status Screen. From the Status
Screen you can monitor the run in progress
NOTE: Unlike the other SpectraSYSTEM instruments, on the
autosamplers, you can make changes to the run in progress from the
[SAMPLES] key rather than from the [STATUS] key (see below).
[SAMPLES]
The unlabeled key is the only variable key on the SpectraSYSTEM
and SpectraSERIES instruments. (It has a different function on each
of the SpectraSYSTEM modules.) On the autosampler, the blank key
is the [SAMPLES] key. The key's name appears on the nameplate
below the key.
The [SAMPLES] key displays the Samples Menu that lets you define
the resources (samples, reagents, and standards) and assign a file to
each sample set. From the Samples Menu you also add sample sets to
the sample queue.
Thermo Electron
43
Because you manage resources and files from the Samples Menu, the
[SAMPLES] key gives you the real-time editing capability provided
by the [STATUS] key on the other SpectraSYSTEM instruments.
That is, from the Samples Menu you can make changes to the file that
is currently running. Pressing the [SAMPLES] key during a run lets
you obtain information about, or edit the locations and injection
volumes of, the sample and calibration vials. Chapter 3 contains
more information on the [SAMPLES] key.
[MENU]
Pressing [MENU] displays the Main Menu (Figure 2.2). See below
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.
[∧], [∨], [<], 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 alphabetical or 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.
44
Thermo Electron
MENUS AND
SCREENS
Your autosampler has two kinds of displays: menus and screens.
Menus require you to make selections or enter specific values.
Screens display information that cannot be edited. The Menu Tree in
the front pocket of your manual illustrates the structure and content of
the autosampler's menus and screens.
Main Menu
The Main Menu (Figure 2.2) is the top level of the menu structure. It
gives you access to five other menus: FILES, COMMANDS,
QUEUE, OPTIONS, and TESTS. To see the Main Menu, press the
[MENU] key at any time.
FILES
QUEUE
COMMANDS
TESTS
OPTIONS
Figure 2.2 The Main Menu
From the Files Menu you can edit, load, or delete files. From the
Queue Menu you can edit or change the order of sample sets in the
sample queue. The Tests Menu gives you access to the instrument's
built-in diagnostics. The Commands Menu lets you run built-in
instrument routines. In the Options Menu, you can set up or change
your instrument's configuration. Refer to Chapters 3 and 4 for more
information on any of the instrument's menus.
Status Screen
The Status Screen automatically appears whenever you turn on the
instrument or press the [STATUS] key. The Status Screen
(Figure 2.3) displays the status of the sample queue, the current
sample set, the current sample vial, and the total number of samples in
the set. Pressing the down-arrow once displays the Inject Status
Screen (Figure 2.4). Pressing the down-arrow again displays the
Oven/Tray Temperature Status Screen (Figure 2.5), if these options
are present. Chapter 3 contains more information on the Status
Screen.
QStatus
Set
IDLE
1
#Samples
1 / 1
Figure 2.3 The Queue Status Screen
Status
Vial
Vol
#Inj
READY
A01-S
1000
1 / 1
Figure 2.4 The Injector Status Screen
Thermo Electron
45
Oven
TrayTemp
OFF
23
OFF
25
Figure 2.5 The Oven/Tray Temperature Status Screen
MESSAGES
There are three different kinds of messages that can appear on your
autosampler's display: user messages, confirmation messages, and
error messages.
User messages
User messages 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 2.6.
* File Protected *
No Editing Allowed
Figure 2.6 An example of a user message
Confirmation messages
Confirmation messages (Figure 2.7), indicated on the display by
asterisks, appear for one second after an operation has been carried
out successfully.
* *
File Loaded
* *
Figure 2.7 An example of a confirmation message
Error messages
Error messages (Figure 2.8), indicated on the display by 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.
!!
Unexpected Vial in Hook
!!
Figure 2.8 An example of an error message
46
Thermo Electron
Practice Examples
In the first part of this chapter, you read about the three easy rules for
using your autosampler's command center and some of its menus and
screens. In this section you'll work through two examples. In the
first example, you'll enter your run parameters from the autosampler's
Samples Menu without first creating a file. In the second example,
you'll create a file using the Edit Menu, load, and run that file. Each
of these examples injects a 10 µL sample (distilled water) from the
vial in position A01. In these examples you won't be changing any
menu values. Instead, you'll use the default values already entered
into the autosampler's menus as your run parameters. Use this
practice run to make yourself comfortable with your autosampler's
menus and file-entry procedures, and to become familiar with the
seven steps for setting up a run.
The steps for setting up a run are as follows:
•
Prepare your sample, solvents, mobile phase, vial(s), and
sample tray(s)
•
Enter your parameters into the Samples Menu or create an
autosampler file
•
Load your autosampler file
•
Build your sample queue
•
Enter your pump, detector, and data-output device parameters
•
Install your sample tray(s)
•
Start your run
The following practice examples use only a fraction of the features
available on your autosampler, so we've included a more complete
description of your autosampler's features and benefits first.
Before you start the practice runs, be sure that you review the Safety
Information section and complete the Startup Checklist. This portion
of the chapter assumes that you have installed your instrument as
described in Chapter 1 and that your sample trays are installed.
NOTE: Your instrument is shipped with the sample trays installed. If, for
some reason your trays are not installed, refer to Chapter 3 or to your
Quick Reference Card for more information on installing your trays.
Thermo Electron
47
Use these examples to get acquainted with your autosampler's menus.
To keep these examples simple, yet still introduce you to as many
menus as possible, we'll use the instrument's default parameters.
HINT: If at any time you lose your place, you can do one of the following:
•
Press [∧] to return to a previous screen,
OR
•
Press [MENU] to return to the Main Menu (the top of the menu
structure) and retrace your steps.
PREPARE YOUR
CHEMICALS
Prepare your sample, solvents, mobile phase, vial, and sample tray as
follows:
1. Fill a vial from your accessory kit with distilled water. Cap it
securely, and place it in position A01, in the front left corner of
tray A (Figure 2.9).
NOTE: Be sure to use either: 1) a 40-60 mil (1.0-1.5 mm) rubber
or silicone septum with a 5 mil (0.1 mm) Teflon® liner on one side
OR 2) a single 10 mil (0.25 mm) Teflon liner by itself.
2. Fill the flush-solvent bottle with a 70:30 MeOH:H20 solution.
Refer to Chapter 1 for the flush-bottle installation procedure.
ENTER YOUR
PARAMETERS
There are two ways to enter your run parameters into your
autosampler: from the Samples Menu and from the Edit Menu. If
you have an AS1000 or AS100, you will use the Sample Menu
method most often. Most likely you will want to enter into the Edit
Menu those parameters that are least likely to change and those that
change with each run into the Samples Menu. The Samples Menu
allows you to vary individual parameters with successive runs.
Should you decide to enter your parameters into the Edit Menu, there
is no need to re-enter them into the Samples Menu. They are
automatically copied into the Samples Menu. Refer to Chapter 3 for
more information on entering your run parameters. For now, you
may wish to familiarize yourself with both approaches for entering
your parameters.
48
Thermo Electron
Hook
Vial Position
A01
Sample Tray Handle
(front)
On/Off Switch
AS\Z001E\DT
Samples
Key
Figure 2.9 Front of the autosampler
Thermo Electron
49
From the Samples
Menu
To enter your parameters from the Samples Menu:
1. Turn on your autosampler by pressing the on/off switch located
in the lower right-hand corner of the front of the autosampler
(Figure 2.9).
When the startup sequence is complete, the Status Screen appears
(Figure 2.10).
QStatus Set
#Samples
IDLE
0
0
/
0
T
Figure 2.10 The Status Screen
2. Press the [SAMPLES] key to display the Samples Menu
(Figure 2.11 and Figure 2.12). When the STATUS Menu first
appears, the file name field is blank. Figure 2.11 and
Figure 2.12 show how this field will appear when a file name is
entered.
Sample Set
1
File 1:FILENAME
--------------------------------------------------------Loop Size
Injections/Sample
Cycle Time
First Sample Vial
Number of Samples
Add to Queue?
20
1
10.0
A01
1
Yes
Figure 2.11 The Samples Menu (AS1000 and AS100)
Sample Set
1
File 1:
--------------------------------------------------------Injection Volume
Injections/Sample
Cycle Time
First Sample Vial
Number of Samples
Add to Queue?
10
1
10.0
A01
1
Yes
Figure 2.12 The Samples Menu (AS3000 and AS300)
3. Use the [∨], [<], [>], [+] and [-] keys to change the Cycle Time
to 1.0.
4. Press the [∨] key to move down the Samples Menu and press
[ENTER] to select "Yes" to add your sample set (1) to the
queue.
50
Thermo Electron
NOTE: We'll tell you more about building your queue later on in this
chapter. For now, proceed to step 5.
5. Press [RUN] to start your run. During the run, the autosampler
hook picks up the vial, carries it to the sample tower, injects the
sample, returns the vial to the sample tray, and flushes the
lines. When the run is complete, the instrument's status returns
to "IDLE."
From the Edit Menu
In this example you'll use the Edit Menu to "create" and run the
practice file. You'll use this method when you want to create more
complex files and/or to create more than one file before starting a run.
You will first specify a file name and then press [ENTER] in each
menu field to accept its default value.
Use the following steps to create your autosampler file.
NOTE: If you worked through the last example, the autosampler returned
you to the Status Screen at the completion of the run. Go on to step 1.
NOTE: If you skipped the last example, turn on your autosampler and wait
for the Status Screen to appear. Then proceed to step 1.
1. Press [MENU] and select /FILES/, /Edit/ to display the Edit
Menu (Figure 2.13). Notice that File 1 is already entered into
the Edit File field. Press [ENTER] to edit File 1.
>
Edit File
1_
File Name: PRACTICE
--------------------------------------------------------Injection
Calibration
Timed Events
Sample Prep
(Appears only with the Sample Preparation Option installed)
More
Figure 2.13 The Edit Menu
2.
Use the [+], [-], and arrow keys to specify each character in the
file name of your choice. We've chosen "PRACTICE" as our
file name.
3. Move the cursor to each field in the Injection, Calibration,
Timed Events, and More Menus and press [ENTER] to accept
their default values. (If you were running a file other than this
practice file, you would, of course, select other entries while
moving through these menus.)
NOTE: For the practice example, press the [∨] key to skip the
Sample Prep Menu.
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51
Notice that after the last field in each menu, the autosampler returns
you to the next most probable choice in the menu structure. For
example, after you accepted the Calibration Menu's default values, the
autosampler automatically returned you to the Edit Menu with the
cursor in the Timed Events field. Similarly, after you accepted the
More Menu's default values, the autosampler places the cursor in the
Load field of the Edit Menu. This is a pattern that occurs throughout
the user interface. The instrument "anticipates" the menu choice you
are mostly likely to need next. You can, of course, override that
choice at any time by moving the cursor wherever you like.
LOAD YOUR FILE
Now that you've created your practice file, use the following steps to
load your practice file manually.
1. Select /Load/ from the Edit Menu.
2. Verify that File 1 is entered, and press [ENTER] to load it into
the autosampler's active memory. The message "**File
Loaded**" appears briefly. Then the Status Screen is
displayed (Figure 2.14).
Loading the file manually activates any installed options (column
oven or tray temperature control) that you set in the Injection Menu.
QStatus
Set
IDLE
0
#Samples
0 / 0
Figure 2.14 The Queue Status Screen with the queue idle
BUILD
YOUR QUEUE
Build your sample queue by adding Sample Set 1 (that contains your
practice sample) to the sample queue as follows:
1. Press [SAMPLES] to display the Samples Menu (Figure 2.11
and Figure 2.12).
2. Press [ENTER] in each field to accept the Samples Menu's
default values.
If you were injecting an actual sample instead of water, you would
enter your pump, detector, and integrator/recorder or data-system
parameters now. For this example, however, simply load your trays,
and close the autosampler's cover.
52
Thermo Electron
START YOUR RUN
To start your run, press [RUN]. If everything's running properly, the
Status Screen's QStatus field will read "RUN" (Figure 2.15). The
injection status field displays the time elapsed since the run was
started (Figure 2.16).
QStatus
Set
RUN
1
#Samples
1 / 1
Figure 2.15 The Queue Status Screen with the queue running
Status
0.05
Vial
Vol
#Inj
A01-S
10
1 / 1
Figure 2.16 The Injector Status Screen with a run in progress
As you know from the first example, the autosampler injects vial
A01. After the run is complete and the sample queue is empty, the
vial is returned to the tray, and the instrument's queue status (QStatus)
returns to "IDLE."
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53
3
Basic Operations
Introduction
This chapter describes the routine operation of the SpectraSYSTEM
and SpectraSERIES autosamplers. In this chapter you will learn the
steps required for setting up and performing a run on your
autosampler as follows:
•
Prepare your chemicals
•
Create a file that contains your autosampler run parameters
•
Load your file
•
Build your sample queue using the [SAMPLES] key and
Samples Menu
•
Start and stop a run
•
Monitor the run status
•
Change the parameters of a run in progress
•
Insert a priority sample set into the queue
There are four options available for SpectraSYSTEM autosamplers:
•
Column oven (standard or narrow-bore configuration)
•
Tray temperature control (AS3000 and AS3500 only)
•
Sample preparation (AS3000 and AS3500 only)
•
BCD communication
The menus and fields used to control these options appear only if they
are installed. This chapter includes the column oven and tray
temperature control procedures. Chapter 4, Advanced Operations,
describes the sample preparation menus (templates) and procedures.
If you purchased the Sample Preparation Option, you'll first want to
learn the procedures described in this chapter and then proceed to
Chapter 4 to learn how to use the sample preparation templates and
the heater/mixer. Refer to Chapter 1 and contact your representative
for information about the BCD option since this option requires
specific installation procedures.
BEFORE YOU
START
Before beginning this chapter, be sure that you've completed the
Startup Checklist found in the beginning of this manual and installed
your instrument according to the procedures described in Chapter 1.
Thermo Electron
55
Theory of Operation
BASIC
AUTOSAMPLER
The autosampler consists of a number of subsystems. It is important
for you to be familiar with how these subsystems contribute to the
instrument's operation in order for you to be able to trace a problem
or symptom to its probable cause.
The basic autosampler consists of the following components: 1) flush
valve, 2) small (250 µL) syringe, 3) injector valve, 4) sample loop,
5) needle and hook, 6) flush funnel, 7) pump output line, 8) column
input line, and 9) sample vial. Figure 3.1 illustrates the autosampler's
major components without the Sample Preparation Option.
Figure 3.2 illustrates the major components with sample preparation
installed.
Vials filled with samples to be analyzed are loaded into three trays
that hold up to 40 vials each (35 with the Tray Temperature Control
Option installed). During operation, a mechanical arm (the XYZ arm)
locates a sample vial. The hook on the end of the arm moves under
the selected vial and lifts it from the sample tray. The XYZ arm
transports the vial to the sample tower and the sample needle for
sample injection. The injection valve switches between the Inject and
Fill positions (Figure 3.3) as necessary to draw sample or solvent into
the autosampler lines and to inject sample onto the column. All of the
SpectraSYSTEM autosamplers perform full-loop injection. The
AS300, AS3000, and AS3500 offer both pull-loop injection and
Thermo Electron’s PushLoop™ injection modes. The injection
routines are described here.
Full-loop injection
In full-loop injection, an air bubble is drawn into the needle to isolate
the incoming sample slug from the flush solvent already present in the
lines (Figure 3.4). With the injector valve in the FILL position, the
syringe retracts to pull the sample from the vial into the sample loop.
The injector valve rotates to the INJECT position, and the pump
pushes the sample from the sample loop onto the column.
Pull-loop injection
The pull-loop injection routine is useful when you have a limited
amount of sample. Only the desired sample amount is pulled from
the vial into the sample loop. Specifically, an air bubble is drawn into
the needle to isolate the incoming sample slug from the flush solvent
already present in the lines (Figure 3.5). With the injector valve in
the FILL position, sample is drawn into the needle. The sample vial
56
Thermo Electron
is lowered from the end of the needle and sufficient air drawn into the
needle to pull the sample slug into the sample loop. The injector
valve then rotates to the INJECT position and sample is expelled from
the sample loop onto the column.
Sample Syringe Valve
Flush
Solvent
Needle
Sample
Vial
Injection Tower
Sample
Syringe
Sample Loop
2
1
Pump Output Line
3
6
4
5
Injector Valve
AS-Z022E\DT
Column Input Line
Sample Syringe
Injector
Valve
Figure 3.1 The autosampler's major components without the Sample Preparation Option
Thermo Electron
57
Solvent
Selector
Valve
6
1
2
Selector-Flush
Tube
5
3
Solvent
Holding
Loop
4
Flush
Solvent
Prep-Flush
Tube
1
6
2
5
Injection
Tower
4
Sample
Loop
2
1
3
Injector
Valve
6
4
Prep
Syringe
Valve
5
To
Pump/Oven
Sample
Syringe
To
Column
3
Flush-Inject
Tube
Sample Syringe
Valve
(6-pos.)
Prep
Syringe
KEY
AS-Z010E/DT
Buffer
Pickup
Pickup
PickLg
Flush Solvent
Figure 3.2 The autosampler's major components with the Sample Preparation
Option installed
58
Thermo Electron
2
Inject Position
2
1
3
From
Pump
1
3
From
Pump
6
4
AS-Z032E/FM
Fill Position
6
4
5
5
To
Column
To
Column
Flush
Solvent
Flush Solvent
Sample Syringe
Valve
(3-Port)
Air
Air
Injection
Tower
2
(Fill Position)
KEY
6
4
Sample
Syringe
(Contains Flush
Solvent)
1
3
Injector
Valve
5
AS-Z027E/DT
Figure 3.3 The injector valve's inject and fill positions
From
Pump/Oven
Sample
Mobile Phase
Flush Solvent
To Column
Figure 3.4 Full-loop injection just prior to sample injection
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59
Sample Syringe
Valve
(3-Port)
Air
Injection
Tower
2
(Fill Position)
KEY
6
4
Sample
Syringe
1
3
Injector
Valve
AS-Z028E/DT
Flush
Solvent
5
From
Pump/Oven
Sample
Mobile Phase
Flush Solvent
To Column
Figure 3.5 Pull-loop injection sequence
PushLoop Injection
The concept behind PushLoop injection is that it is more accurate to
push a column of fluid than it is to pull it. Pulling liquids through a
small tube can cause "cavitation" within the tubing. Cavitation is the
outgassing of dissolved gases caused by reducing the pressure on a
column of liquid. This forms bubbles within the column of liquid,
resulting in inaccuracies and nonreproducible volumes.
With PushLoop injection, sample volume is separated from the
previous flush solvent by an air bubble. With the valve in the Inject
position, the sample slug is pulled up to and past the injection valve.
After compensating for the syringe backlash, the valve is switched to
the Fill position and the desired volume of sample is pushed into the
sample loop. In PushLoop injection, sample volumes are "pushed"
into the sample loop from solution directly adjacent to the loop. The
sample solution is essentially at the "bottom" of the column of liquid.
Any residue from the previous solvent has been minimized. The
starting boundary of the delivered sample slug is well defined by the
valve. There is no uncertainty about the air bubble or solvent droplets
that may affect accuracy or reproducibility. Finally, the injector
valve is then switched back to the Inject position, and sample is
expelled from the sample loop onto the column. Figure 3.6,
Figure 3.7, and Figure 3.8 illustrate this routine.
60
Thermo Electron
AS-Z029E/DT
Flush
Solvent
Sample Syringe
Valve
(3-Port)
Injection
Tower
2
KEY
1
3
6
Injector
Valve
4
5
(Inject Position)
Sample
Syringe
Sample
Mobile Phase
Flush Solvent
From
Pump/Oven
To Column
Figure 3.6 PushLoop injection
Flush
Solvent
AS-Z030E/DT
(step 1: The sample slug is pulled up to and past the injection valve.)
Sample Syringe
Valve
(3-Port)
Injection
Tower
2
KEY
1
3
Injector
Valve
(Fill Position)
6
4
Sample
Syringe
5
From
Pump/Oven
Sample
Mobile Phase
Flush Solvent
To Column
Figure 3.7 PushLoop injection
(step 2: The injection valve is switched to the Fill position, and the desired volume of
sample is pushed into the sample loop.)
Thermo Electron
61
AS-Z031E/DT
Flush
Solvent
Sample Syringe
Valve
(3-Port)
Injection
Tower
2
KEY
1
3
Injector
Valve
(Inject Position)
6
4
Sample
Syringe
5
From
Pump/Oven
Sample
Mobile Phase
Flush Solvent
To Column
Figure 3.8 PushLoop injection
(step 3: The injection valve is switched back to the Inject position, and sample
is expelled from the sample loop onto the column.)
SpectraSYSTEM Autosamplers
The SpectraSYSTEM autosamplers have been designed for ease of
use and unsurpassed performance. Two different SpectraSYSTEM
autosamplers are available each with a customized set of options and
capabilities to fit a wide variety of HPLC analytical applications.
SpectraSYSTEM autosamplers may be purchased with or without the
built-in column oven and tray temperature control options. These
options are available at the time of purchase or as upgrades.
Instruments purchased without the tray temperature control option
hold 120 vials; those purchased with the tray temperature control
option hold 105 vials.
AS1000
The AS1000 fixed-loop autosampler can function as a stand-alone
HPLC module or through the COMM. port as part of
Thermo Electron’s SpectraSYSTEM. In this configuration, all HPLC
modules are controlled by a system controller via SpectraNET™
communication. This instrument offers additional optimization
capability: it holds four user-programmable built-in files and the
sample queue can process up to 39 sample sets plus one priority
sample set.
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Thermo Electron
AS3000
The AS3000 variable-injection volume (VIV) autosampler offers
three injection methods: 1) full-loop, 2) traditional pull-loop where
sample conservation is of primary concern, and 3) Thermo Electron’s
patented PushLoop® injection. Similar to the AS1000, it holds 39
user-programmable built-in files and the sample queue can process up
to 39 sample sets plus one priority sample set. In addition to the
built-in tray temperature control and column oven options, an
automated sample-preparation option with accompanying
heater/mixer is also available.
AS3500
The AS3500 autosampler has the same capabilities and available
options as the AS3000. However, all "wetted" parts are made of
corrosion-resistant, biocompatible materials.
AS100
The AS100 is a fixed-loop autosampler that comes with a 20 µL
sample loop and a built-in file that allows you to quickly and easily
enter and modify your run parameters. The sample queue holds up to
39 sample sets plus one priority set that can be run automatically.
AS300
The AS300 is a variable-volume injection autosampler. This
instrument offers three injection methods: 1) full-loop, 2) traditional
pull-loop where sample conservation is of primary concern, and
3) Thermo Electron’s patented PushLoop injection. The AS300 is
shipped with a 100 µL sample loop and built-in customizable file.
The AS300 sample queue can hold and process up to 39 sample sets
plus an additional priority set.
OPTIONS
Column Oven
The column oven contains a resistive thermal device that heats up to
stabilize the column temperature. A temperature sensor detects the
temperature and sends the temperature reading back to the
microprocessor, which turns the resistive device on or off to regulate
the temperature. This is a closed assembly and requires no routine
maintenance.
Thermo Electron
63
Tray Temperature
Control
The tray temperature control unit consists of four Peltier devices, two
high-speed fans, and a heat sink. Heat is pumped from the sample
compartment into the heat sink and then blown out the back of the
instrument by the two fans. A temperature sensor sends the
temperature reading back to the microprocessor, which controls the
operation of the four Peltier devices to regulate the sample
compartment temperature. Similar to the column oven, the tray
temperature control unit is a closed assembly and requires no routine
maintenance.
Sample Preparation
The sample preparation hardware consists of the solvent selection
system and the heater/vortex mixer. (Refer to Chapter 4 for
information on the sample preparation software.)
The solvent selection system is located at the back of the instrument
and includes the sample preparation syringe(s) and the solvent
selection valve.
The heater/mixer or vortex mixer, located behind the sample tower,
uses compressed air to spin the sample vial creating the vortex inside
the vial that mixes the vial's contents. The microprocessor clock
tracks the run time and turns the mixer on and off at the desired times.
A resistive thermal device (heater) and a temperature sensor, similar
to the column oven, maintain the desired temperature during the
vortex-mixing process.
Run Preparation
The SpectraSYSTEM and SpectraSERIES autosamplers allow you to
process up to 39 non-priority sample sets (groups of sample vials and
associated calibration vials) within a given run. To prepare for a run,
use the following steps:
•
Prepare your samples, solvents, and mobile phase
•
Create your file by entering the parameters for each sample set
into one of the four built-in files
•
Load your file
•
Build your sample queue
•
Start your run
Once your run has started, you can:
•
64
Monitor the progress of the run
Thermo Electron
•
Modify the currently running file (the run file) to affect
subsequent vials in the active sample set
•
Change the run order of, add, or delete sample sets from the
sample queue
•
Add a priority sample set to the queue without disrupting the
calibration routine of the active sample set
These procedures, along with the instrument's file-management
features, are described in this section.
PREPARE YOUR
CHEMICALS
Prepare your samples, solvents, and vials. Be sure that your
sample(s) is (are) completely soluble in the mobile phase and that you
have filtered your samples and solvents through a 0.45-micron filter.
These techniques minimize sample precipitation in the lines and
remove any particulate matter that could obstruct the flow through the
autosampler's injector or column. Fill the solvent reservoir with flush
solvent.
NOTE: If you are using the automated Sample Preparation Option, you can
use up to four solvents. Refer to Chapter 4 for instructions on filling your
external solvent reservoirs and for setting up solvent selection in your
autosampler file.
CREATE YOUR FILE
The group of autosampler parameters applied to a specific sample set
is contained in a file. When your AS1000, AS3000, or AS3500 is
shipped, each of its four built-in files contains the default values. The
AS100 and AS300 have only a single method file. To create your
own file, simply edit one of the existing files by entering your own
parameters into the autosampler's Edit Menu. From the Edit Menu
you can name your file and access additional menus to enter your
injection, calibration, timed events, sample preparation, and other run
parameters.
NOTE: When you have entered all of your parameters in the Edit Menu,
you'll have finished creating your edit file. You can't run your edit file by
pressing the [RUN] key. Instead, you first have to load your edit file into the
run file. For more information, see "Load your file" on page 77.
To create your edit file:
1. Press the [MENU] key to display the Main Menu. From there,
select /FILES/, /Edit/ to display the Edit Menu (Figure 3.9).
Thermo Electron
65
Edit File
1
File Name
VITAMINS
--------------------------------------------------------Injection
Calibration
Timed Events
Sample Prep (If installed)
More
Figure 3.9 The Edit Menu
Edit File
An autosampler file is identified by its number (1-4) and name, which
you supply. (See "File Name," below.) The Edit File field indicates
the number of the selected file. File 1 automatically appears in this
field when the Edit Menu is first displayed. If you wish to edit a
different file, press the [+] and [-] keys to select a different file
number.
File Name
The File Name field allows you to name your file. When you first
display the Edit Menu, the File Name field contains eight blank
characters. Use the [+]/[-] and [<]/[>] keys to create a file name made
up of the following characters: A-Z, 0-9, \, blank, dash, or hyphen.
Injection Menu
Selecting /Injection/ from the Edit Menu accesses the Injection Menu
(Figure 3.10 and Figure 3.11) from which you can enter your
injection parameters. The next section describes the Injection Menu's
fields.
Loop Size
10.0
Injections/Sample
1
--------------------------------------------------------Cycle Time
10.0
(If installed)
Col Oven: OFF_ Temp 18
TrayTemp:
OFF
Temp
0
(If installed)
Figure 3.10 The Injection Menu (AS1000 and AS100)
Injection Volume
10.0
Injections/Sample
1
--------------------------------------------------------Cycle Time
66
10.0
Oven:
OFF_
Temp 18
(If installed)
TrayTemp:
OFF
Temp
(If installed)
Thermo Electron
0
Figure 3.11 The Injection Menu (AS3000 and AS300)
Injection Volume (Loop Size)
Use the first field of the Injection Menu to specify the sample Loop
Size (if you are using an AS1000 or AS100) or the sample injection
volume (if you are using an AS3000, AS3500, or AS300). The
injection volume value depends upon the injection method and range
that you entered in the More Menu's Injection Type and Injection
Range fields, respectively, (see page 74). To optimize injection
volume, see page 94.
Full-loop injection: If you're using either full-loop injection on the
autosampler, you can use the [+] and [-] keys to select one of the
following values: 5, 10, 20, 50, 100, 200, 500, and 1000 µL.
NOTE: Be sure that the Loop Size field value reflects the size of the sample
loop installed.
Variable-Loop Injection: You can choose either Pull or PushLoop
injection mode from the More Menu. These methods are described
on page 75. If you use pull-loop injection, you can select a sampleinjection volume from 1-1500 µL. If you use PushLoop injection,
you can specify a sample-injection volume from either 0.1-10.0 µL or
1-200, depending upon which Push type (Hi or Lo) you selected in
the More Menu's Injection Type field. You should limit the partial
injection volume to approximately one-half the sample loop size.
Injections/Sample
In this field, specify the number of injections to be made from each
vial (1-99). If you have selected a sample preparation template, and
want sample preparation without injections, enter 0 in this field.
NOTE: You can't enter a 0 unless you have first selected a sample
preparation template.
Cycle Time
In the Cycle Time field enter the desired time (0-655.3 minutes)
between the autosampler's injections (from the injection to the end of
the run). In calculating this value, be sure to allow enough time for
the column to equilibrate (if you are running a gradient), all peaks to
elute from the column, and for the integrator to print any post-run
reports and perform any post-run calculations.
The autosampler is capable of making an injection approximately
every 90 seconds, depending upon the injection and flush volumes
selected. If your cycle time is controlled by a Pump Ready contact
closure, you may set the Cycle Time to 0.0.
The default flush volume is 400 µL, so that the total injection cycle time
(including the flush cycle) is approximately two minutes.
Thermo Electron
67
Column Oven
This field appears only if you purchased the Column Oven Option
and set it to ON in your Configurations Menu. To use the column
oven, move the cursor to the Col Oven field and press the [+] and [-]
keys to toggle the oven's state between on and off. In the Temp field
in the same line, you can specify the oven temperature in one-degree
increments from 15 - 80°C.
NOTE: You can specify temperatures starting at 5°C above ambient.
NOTE: Even if you set the oven's state to On, the oven isn't activated
(turned on) until you load your file.
HINT: The autosampler automatically delays the start of the run until the
set temperatures have been reached. Once activated, the oven requires
about 15 minutes to reach a temperature of 70°C. To save time, you may
wish to load your file so that the oven can be warming up while you're
completing your run preparation and entering the file parameters for your
run. See page 77 for more information on loading your file.
NOTE: The vial temperature will typically reach 4°C with a set temperature
at 0°C and 58°C with a set temperature at 60°C.
Tray Temperature Control
This option allows you to heat or cool the sample tray within a range
of 0°C to 60°C (in 1° increments). This field appears only if you
purchased the tray temperature control option and specified Yes in the
Configurations Menu Tray Temp field. If you plan to use tray
temperature control, use the [+] and [-] keys to turn on the tray
heater/cooler in the Tray field. Then specify the temperature in its
Temp field. If set to "On" in the Injection Menu, the heater/cooler
will not be activated until you load the file manually (by pressing
Load from the Files Menu), or automatically when you press [RUN]
to start your queue.
HINT: The autosampler automatically delays the start of the run until the
set temperatures have been reached. Once activated, if empty, the tray
requires about 30 minutes to reach a temperature of 0 °C and about
10 minutes to reach a temperature of 60 °C. To save time, you may wish to
load your file so that the heater/cooler can be coming to temperature while
you are completing your run preparation. See page 77 for more information
on loading your file. You can also save time by preheating or precooling
your samples.
The autosampler's Tray Temperature Control Option is primarily
designed to maintain the temperature of the sample compartment
within ± 2 °C. The actual temperature inside the autosampler vial is
dependent upon the specified tray temperature, type of vial, sample
buffer, etc. The following graph (Figure 3.12) indicates the
approximate vial temperature given a specified tray set temperature.
68
Thermo Electron
55
50
45
40
Actual Temperature
-10
35
30
25
20
15
10
5
0
Set Temperature
-
AS-Z127E/DT
-
60
-
0
-
-
10
-
-
-
20
-
-
Vial Temperature
-
30
Tray Temperature
-
-
-
40
-
-
-
50
Vial vs.Tray Temperature
-
-
-
60
Figure 3.12 Plot showing the actual vial and tray temperature at a
specified set temperature
The Tray Temperature Control Option can also be used to heat and
cool samples. The time lag in heating and cooling is due to variations
in ambient temperatures, indirect contact with temperature control
units, and vial type. The accompanying graph (see Figure 3.13)
indicates the time required to heat (to 60°C) or cool (to 0°C) the tray
and vials from an ambient temperature of 23°C.
-
50
-
40
-
30
-
20
-
10
-
0
-
-
-
-
-
-
-
-
-
-
-
-
-
-
0
5
10
15
20
25
30
35
40
45
50
55
60
Vial Cooling
Vial Heating
Tray Cooling
-10
Tray Heating
AS-Z128E/DT
Temperature
Heating and Cooling Times
60
Time (min.)
-
Figure 3.13 Plot of heating and cooling times of a standard glass
vial and the autosampler tray. The starting ambient temperature was
23°C
HINT: You can save time by preheating or cooling your samples before
placing them into the autosampler.
Thermo Electron
69
Calibration Menu
By selecting Calibration from the Edit Menu, you can call up the
Calibration Menu (Figure 3.14). This section tells you how to enter
your calibration parameters and how each is used by other
autosampler menus.
Number of Levels
0
Samples/Calibration
0
--------------------------------------------------------Injections/Level
1
Injection Volume
10
Type
Reuse same vials
Bracket samples
No
Figure 3.14 The Calibration Menu
Number of Levels
Specify the number of calibration levels (0-9). The autosampler
automatically copies this value to the Calib 1Vial 1 (of X) field, in the
Samples Menu, where X equals the number of levels. (Refer to
page 78 for more information.) A value of zero in the Number of
Levels field indicates that no calibration injections will be made for
the specified sample set.
Samples/Calibration
From this field you can specify the calibration interval (the number of
sample vials to be injected between each recalibration).
Injections/Level
In this field you specify the number of injections (1-99) from each
calibration level (0-9).
Injection Volume
The Injection Volume field in the Calibration Menu allows you to
specify an injection volume for your calibration vials that is different
from the sample injection volume you specified in the Injection
Menu.
Similar to the Injection Volume field in the Injection Menu, this value
depends upon the injection method you select in the More Menu's
Injection Type field. As with the sample injection volume, you can
choose between 0.1-10.0 µL and 1-200 µL if you are using PushLoop
injection. (See page 74.)
NOTE: This field is not displayed if you have selected Full from the More
Menu's Injection Type field or if you are using an AS1000 or AS100.
Using variable-volume injection, you can choose either Pull or
PushLoop injection mode from the More Menu. These methods are
described on page 74. If you use pull-loop injection, you can select a
calibration-vial injection volume from 1-200 µL. If you use PushLoop
70
Thermo Electron
injection, you can specify a calibration-vial injection volume from 110.0 µL or from 1-1500 µL, depending upon which Push type you
selected in the Injection Type field. You should limit the injection
volume to approximately one-half the sample loop size.
If you're using full-loop injection, you can use the [+] and [-] keys to
select one of the following values: 5, 10, 20, 50, 100, 200, 500, and
1000 µL. We recommend that you enter your sample-loop volume as
your calibration-vial injection volume.
NOTE: If you want to determine optimum syringe and sample loop size, see
page 94.
Type
This field allows you to specify one of two types of patterns: 1) Reuse
same vials or 2) Intersperse vials.
Reuse same vials programs the autosampler to use the same
calibration vials each time a recalibration is made. Thus, each time a
calibration is required, the autosampler goes to the first calibrationvial position specified in the Sample Menu's and injects all the
calibration vials. It then injects the number of samples indicated by
the Calibration Menu's Samples/calibration value.
NOTE: If you have selected the Reuse vial calibration type, do not place
calibration vials between sample vials. Instead, place them before the
sample vials in the same tray or in a different location.
NOTE: When you reuse your calibration vials, you may place them in any
positions in the sample tray(s). However, if you are doing multiple-level
calibrations, your calibration vials must be placed sequentially in the tray.
Intersperse vials programs the autosampler to use each calibration
vial only once. You must manually intersperse your calibration vials
between your sample vials within the sample set. The autosampler
makes injections in tray-placement order. If you've specified a
different injection volume for calibration vials versus sample vials,
the autosampler uses the Samples/Calibration field value to determine
whether the next vial is a calibration or a sample vial. It determines
the location of the first calibration vial from the value entered in the
Samples Menu's Calib 1(of X) field. Figure 3.15 illustrates the vial
order used by the two different calibration routines.
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71
Injection
C
C
S
S
C
C
S
S
Total
Reuse
A01
A02
A03
A04
A01
A02
A05
A06
6
Intersperse Order
A01
A02
A03
A04
A05
A06
A07
A08
8
Figure 3.15 The different injection orders and total number of vials
used by the Reuse and Intersperse calibration routines
Bracket Samples
This field lets you specify whether or not you want to automatically
end the sample set with a recalibration. If you select Yes, the
autosampler processes the last sample and then repeats the calibration
series specified for the sample set. If you select No, the autosampler
simply stops and returns home after the last sample is injected.
Timed Events Menu
Accessed from the Edit Menu, the Timed Events (external events)
Menu (Figure 3.16) allows you to create a time program to turn on/off
external devices (such as column-switching valves and fraction
collectors) at preset times during your run. Pins 9-12 of the
autosampler's 12-pin terminal are used for Timed Events. In this
section, we'll show you how to create a time program. Refer to
Chapter 1 for connecting external devices to your autosampler.
Time
TF1
TF2
TF3
TF4
0.00
Lo
Lo
Lo
Lo
--------------------------------------------------------0.50
Hi
Lo
Lo
Lo
2.00
Lo
Hi
Lo
Lo
Figure 3.16 The Timed Events Menu
The Timed Events Menu holds a maximum of nine lines, each of
which sets Hi/Lo one or more timed event outputs at the time
specified. For example, in Figure 3.17 through Figure 3.19, Time
Function 1 (TF1) is set Hi (activated) at 0.5 minutes into the run; at 2
minutes into the run, Time Function 2 (TF2) is set Hi while Time
Function 1 is set Lo (deactivated). By changing the Time value in
each line, you can add, delete, or insert menu lines.
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Thermo Electron
Adding a Line
To add a line to the end of your program:
1. Press [∨] to move the cursor to the blank line after the last
displayed line.
2. Press the [+] key to copy the last existing line with a new time
incremented by one minute.
3. Use the arrow keys and the [+] and [-] keys to change the time
and set the time functions' states (Hi/Lo).
Deleting a Line
To remove a line:
1. Move the cursor to the Time field in the line to be deleted.
2. Hold down the [-] key until the value is zero (0.00).
3. Press the [-] key once to erase the line.
If you are deleting a line from the middle of a timed events program,
the blank line remains until you move the cursor to another line.
NOTE: If you delete both of the visible lines on the display, both lines
disappear, but the cursor remains. Press [∧] or [∨] to move to the
remaining lines in the program.
Inserting a Line
To insert a line between two existing lines:
1. Move the cursor to the blank line at the end of the program.
2. Enter a time that falls between two existing times.
3. Move the cursor off the new line. The autosampler
automatically inserts the new line.
4. Then move the cursor to the newly inserted line(s) and set the
Timed Events Outputs' states as desired.
Figure 3.17 though Figure 3.19 illustrate this process.
Time
TF1
TF2
TF3
TF4
0.00
Lo
Lo
Lo
Lo
--------------------------------------------------------1.00
Lo
Lo
Hi
Lo
2.00
Lo
Hi
Lo
Lo
Figure 3.17 The original timed events program
Time
TF1
TF2
TF3
TF4
0.00
Lo
Lo
Lo
Lo
--------------------------------------------------------1.00
Lo
Lo
Hi
2.00
Lo
Hi
Lo
Lo
Lo
0.50
Lo
Hi
Lo
Lo
Figure 3.18 The original program with an added line -- unsorted
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73
Time
TF1
TF2
TF3
TF4
0.00
Lo
Lo
Lo
Lo
--------------------------------------------------------0.50
Lo
Hi
Lo
Lo
1.00
Lo
Lo
Hi
Lo
2.00
Lo
Hi
Lo
Lo
Figure 3.19 The timed events program with an added line -- sorted
NOTE: The Run-time clock stops and resets at the completion of the Cycle
Time. Time lines longer than the Cycle Time value will never be activated.
Sample Prep
(AS3000/AS3500
only)
Once you've entered your external events parameters, the autosampler
returns you to the Edit Menu with the cursor in the /Sample Prep/
field (if installed). If you did not purchase the Sample Preparation
Option, this field won't appear in the Edit Menu. Instead, the
autosampler will return you to the Edit Menu with the cursor in the
/More/ field. See Chapter 4 for detailed information on the Sample
Preparation Option.
More Menu
The More Menu (Figure 3.20 and Figure 3.21) allows you to enter the
remainder of your run parameters, each of which is discussed below.
Equilibration Time
0.0
Gradient Delay 0.00
--------------------------------------------------------Viscosity
Normal
Prep Viscosity Normal
Flush Volume
400
Needle Height
2.0
Figure 3.20 The More Menu (AS1000 and AS100)
Equilibration Time
Gradient Delay
0.0
0.00
--------------------------------------------------------Viscosity
Normal
Prep Viscosity
Normal
Flush Volume
Injection Type
Injection Range
Needle Height
400
Push
.1-10.0
2.0
Figure 3.21 The More Menu (AS3000 and AS300)
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Thermo Electron
Equilibration Time
In this field, specify the length of time (in minutes) required for the
column or LC system to equilibrate. The equilibration time allows
time for the column to equilibrate prior to the first injection of a new
sample set. It is only in effect before the first injection. The default
value of 0.0 is a good starting value for many applications.
Gradient Delay
The gradient delay field is used to specify the amount of time
required to allow the mobile phase to travel from the pump's
proportioning valve to the column. This field is used only if you are
running a gradient. To determine the gradient delay value, you'll
need to calculate the precolumn volume, which is best done during
installation. Refer to Chapter 4 for a procedure for calculating your
precolumn volume and gradient delay value.
Viscosity
Specify the sample viscosity (Normal, Medium or Viscous). The
viscosity setting affects the sample syringe draw-rate after the sample
loop fills. A higher viscosity setting automatically decreases the
syringe draw-rate to give a more viscous sample more time to be
drawn into and to travel through the sample loop.
HINT: Use the Normal setting for running most applications.
Prep Viscosity (AS3000/AS3500 only with sample prep)
Specify the viscosity (Normal, Medium or Viscous) of the samplepreparation sample. The viscosity setting affects the preparation
syringe (2.5 mL) draw-rate. A higher viscosity setting automatically
decreases the syringe draw-rate and increases the injection delay to
give a more viscous sample more time to be drawn into and to travel
through the sample lines. This also reduces cavitation caused by lowboiling point solvents.
NOTE: This field appears only if the Sample Preparation Option is
installed.
HINT: When performing normal-phase chromatography, the viscous setting
maximizes performance.
Flush Volume
Specify the flush volume to be used for flushing between injections.
The acceptable range is 200 - 5000 µL. The default is 400 µL.
Injection Type (AS3000, AS3500 and AS300 only)
With the AS3000, you can choose from four different injection
methods: 1) full-loop, 2) pull-loop 3) Thermo Finnigan’s PushLoop
injection using the high (Push Hi) injection volume range (1-200 µL)
and 4) PushLoop injection using the low (Push Lo) injection volume
range (0.1-10 µL). The advantages of each type are briefly described
here. Refer to “Theory of Operation” page 56 for a description of
how each method operates and for the excess sample volume required
for injection.
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75
The optimum injection method depends upon the amount of sample
you have and the degree of precision desired.
Full: Full- or fixed-loop injection is useful when you want maximum
precision and have unlimited sample. The maximum injection
volume is 1000 µL and the range from 5-1000 µL.
NOTE: If you select full-loop injection, the Injection Menu's Injection
Volume field changes to Loop Size and the Calibration Menu's Injection
Volume field does not appear.
Pull: Pull-loop injection (traditional variable-volume injection) is
useful when sample conservation is important and when large
injection volumes (>100 µL) are required.
Push: Thermo Finnigan’s PushLoop injection is a variable-volume
method that gives the precision of fixed-loop injection with limited
amounts of sample. This method is most effective for smaller
injection volumes (< 10 µL).
Injection Range
The Injection Range field displays the sample- and calibrationinjection volume range, depending upon the Injection Type that you
selected. For full-loop injection, the range is 5-1000 µL. For pullloop injection, the Injection Range field is automatically filled in with
the 1-1500-microliter range. For PushLoop injection, the ranges are
1) 0.1-10.0 microliters (Push Lo) or 2) 1-200 microliters (Push Hi),
depending upon your sample-loop size.
Needle Height
The Needle Height field enables you to control the distance between
the needle-tip and the bottom of the vial (0.0-20.0 mm). The default
value is 2.0 mm.
NOTES: With this value, you are actually controlling the vial's height
relative to the needle tip. The vial moves; the needle is stationary. If you are
using a standard, 65 mil septa in the vial cap, keep needle height below 50
mm to maximize injection precision.
SAVE YOUR FILE
Your file is saved automatically. Once you have filled in all of the
Edit Menu's parameters, you have finished creating your edit file.
Exiting the Edit Menu automatically saves your edit file but does not
load it. For more about file management, see page 91.
NOTE: Pressing [RUN] after creating your file will not start your run. You
must first add it to the sample queue. For more information, see "Build
Your Sample Queue" below.
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Thermo Electron
LOAD YOUR FILE
The Load function is useful when you want to run a single sample set
manually, for example to turn on your oven or tray heater/cooler.
To load your file, press /FILES/, /Load/. The Load Menu appears:
Load File 1:
FILENAME
Figure 3.22 Load File field
For AS3000, AS3500, and AS300 only: If you want to load the last
file you edited, use the [+] and [-] keys to select its file number and
press [ENTER]. If, instead, you want to load a different file, select
the correct file number and then press [ENTER]. The message **File
Loaded** will be displayed. Once a file is loaded, it becomes your
run file.
The Load function is used to run a single sample set; however, if you
want to run a group of sample sets, this is done using the sample
queue, by pressing [RUN]. Once [RUN] is pressed, the autosampler
automatically loads each file into the queue before running it. You
don't need to manually load each file. For more information on
running your samples from the sample queue, see "Build Your
Sample Queue" below.
BUILD YOUR
SAMPLE QUEUE
The sample queue is the list of sample sets and associated files to be
run. Building your sample queue requires adding each item to the
queue from the Samples Menu and then arranging those items in the
desired run order from the Queue Menu. If you run a group of files
from the sample queue, each file is automatically loaded before it is
run. These procedures are described below.
Adding Items to the
Sample Queue
To add an item to the queue, press [SAMPLES] to access the Samples
Menu. Then enter each of the values shown in the Samples Menu
(Figure 3.23 and Figure 3.24):
HINT: Since the method downloads most of the values automatically, the
only values that require editing are those in the File Number and Vial
location fields.
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77
Sample Set
1
File 1: VITAMINS
--------------------------------------------------------Loop Size
100
Injections/Sample
Cycle Time
1
0.1
Calib Vial 1 (of 1)
A01
First Sample Vial:
A02
Number of Samples
1
Add to Queue?
(If calibration is used)
Yes
Figure 3.23 The Samples Menu (AS1000 and AS100)
Sample Set
1
File 1: VITAMINS
--------------------------------------------------------Injection Volume
100
Injections/Sample
1
Cycle Time
0.1
Calib Vial 1 (of 1)
A01
First Sample Vial:
A02
Number of Samples
1
Add to Queue?
(If calibration is used)
Yes
Figure 3.24 The Samples Menu (AS3000 and AS300)
Sample Set
Select the desired sample set you want to add to the queue. Each
sample set can be used only one time in the sample queue. You can
load a maximum of 39 sets into the sample queue, plus one priority
set.
File
Specify the number of the file to be applied to the specified sample
set. The file's parameters will determine the sequence of Sample Set
prompts.
Injection Volume, Injections/Sample, and Cycle Time
These values are copied from the Injection Menu of the associated
file.
NOTE: The injection volume field does not appear in the AS1000 or
AS100 Samples Menu.
Calib Vial 1 (of X)
This value is copied from the Calibration Menu of the associated file.
Specify the location of the first vial in the calibration series. "X" is an
indicator of the number of calibration levels specified in the file.
NOTE: The vial position automatically wraps. For example, if the first of
three calibration vials is placed in C38, the first sample vial will be A01 for
instruments without tray temperature control.
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Thermo Electron
First Sample Vial
Specify the first sample-vial location.
Number of Samples
Enter the number of sample vials in the sample set.
Add to Queue
Select Yes or No and press [ENTER] to add the selected sample set to
the queue. Each set can be used only one time in the sample queue.
Once you have filled in the Samples Menu for the first sample set,
pressing [SAMPLES] again returns you to the top of the menu so that
you can add the next sample set.
Ordering Sets in the
Sample Queue
You add items into the queue from the Samples Menu, which you
access by pressing [Samples]. You can reorder them in the Queue
Menu (Figure 3.25). You can reorder any sets not currently running.
Order
File Name
Set
1
1:TEST
3
--------------------------------------------------------2
2:SAMPLE1
4
3
3:AA1
5
Figure 3.25 The Queue Menu with no priority-vial sample sets in the
queue
Changing the Order
of Sets Within the
Queue
To change the order of items in the queue, move the cursor to the line
where the change in order should occur and increment/decrement to
the new Order value. Refer to the example in Figure 3.26. To run
sample set 5 before sample set 3, move the cursor to the order column
of the sample-set 5 line, and decrement the Order value to 1
(Figure 3.26).
Order
File Name
Set
--------------------------------------------------------1
1:TEST
3
2
2:SAMPLE1
4
3->1
3:AA1
5
Figure 3.26 Changing the order of sets in the queue
The lines rearrange themselves after you exit the field (Figure 3.27).
Thermo Electron
79
Order
File Name
Set
1
3:AA1
5
--------------------------------------------------------2
1:TEST
3
3
2:SAMPLE1
4
Figure 3.27 The Queue Menu after the Order has been changed
NOTE: Remember, Order 1 can only be changed if the sample set is not
active. (When you press [Status] the QStatus column should read IDLE).
Deleting Sets
from the Queue
To delete an item from the sample queue, move the cursor to the line
containing the item to be deleted, decrement the Order number to 0
and then blank. Moving the cursor to another line deletes the blank
line from the display.
Managing a
Priority Set
Within the Queue
A priority set is inserted into the active set and its samples run before
other samples in the active set. It must be assigned to the active file.
It appears in the Queue Menu with a P in the Set field (Figure 3.28).
You can't assign or change its priority status from the Queue Menu.
A priority set will automatically be deleted from the queue upon
completion of the active set or when the queue (QStatus) returns to its
IDLE state (if you press [STOP]). For more information on running
priority vials, see page 88.
Order
File Name
Set
1
1:SAMPLE1
P
--------------------------------------------------------1
1:SAMPLE1
4
2
3:AA1
5
Figure 3.28 A sample queue that contains a priority set
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Thermo Electron
ENTER YOUR
OTHER LC
PARAMETERS
Prepare your other LC modules for a run and enter the run parameters
for your pump, detector and data-output device (recorder, integrator,
or data system). Before you start your run:
•
Initialize your pump and degas your solvents as required for
your application.
•
Be sure your detector is stabilized. Don't start your analysis
until you've achieved a stable baseline.
•
Set up your data-output device.
For more information on connecting your autosampler to other LC
modules, see Chapter 1.
LOAD
SAMPLE TRAYS
Place your calibration and sample vials into the sample trays included
in your Accessory Kit. Trays A, B, and C are arranged in the
autosampler from left to right. Each tray contains 40 (35, with Tray
Temperature Control) vials arranged in eight (seven) rows of five
(Figure 3.29). The first vial position is A01; the last is C40. The
position labels are on the left of each position.
NOTE: If you purchased the Tray Temperature Control Option, each
sample tray contains 35 vials.
AS-Z002\DT
NOTE: Be sure to use a 40-60 mil (1.0-1.5 mm) rubber or silicone
septum with a 5 mil (0.1 mm) Teflon® liner on one side.
Figure 3.29 An autosampler tray
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81
Place the calibration vials, and samples into the trays. After your
vials are in place, install sample trays A, B, and C as follows:
1. Hold the tray handle, tilting the back end down (Figure 3.30).
2. Insert the tray's "runners" into the slots in position A at the rear
of the tub.
3. Lower the front of the tray into place and press down firmly to
seat the tray.
HINT: Pull the handle back toward you until the tray snaps into
place. Try moving each tray from side to side to be sure that it's
seated completely.
4. Insert the other two sample trays into positions B and C.
NOTE: Be sure that each tray is properly seated. Each tray's upper
front edge should fit under the upper lip of the tub. If a tray is not
completely seated, the autosampler will not be able to correctly pick
up or return vials to that sample tray.
Sample Tray Handle
(front)
A
B
Tub
Tray Positions
Runner
Slots
Figure 3.30 Installing a sample vial tray
82
Thermo Electron
AS-Z003E\FM
C
Starting Your Run
Once you have created and loaded your file, added your sample set(s)
to the sample queue, and installed your sample tray(s), you're ready to
start your run.
Press [RUN] to start your run. If there are sample sets in the sample
queue, the sample queue's "RUN" Status is displayed on the Status
Screen (Figure 3.31).
If the queue is empty, the autosampler prompts you to access the
Samples Menu so that you can add sample sets to the queue. After
you have added sets, press [RUN] to start your run.
MONITORING
THE RUN STATUS
When [RUN] is pressed, the autosampler begins processing vials once
the oven, heater/mixer and/or tray set temperatures have been
reached, the equilibration delay time (entered in the More Menu) has
elapsed, the door is closed, and the pump is ready. During the run,
the Status Screen is displayed (Figure 3.31).
NOTE: You can change the "Ready Requirements" from the Options, Ready
Participation Menu. See "Ready Participation" on page 31 in Chapter 1.
QStatus
Set
#Samples
RUN
1
1 /
1
--------------------------------------------------------Status Vial
Vol
#Inj
0.10
1000
1 / 2
A01-S
File 1:
Oven
TrayTemp
ON:80
ON:4
Figure 3.31 The Status Screen
The status screen gives you four kinds of information (described
below):
•
Autosampler queue status
•
Injection status
•
Oven and tray temperatures
•
Sample preparation status (if the Sample Preparation Option is
installed)
NOTE: The sample preparation status screen is discussed in Chapter 4.
Thermo Electron
83
You can't edit any information on the Status screen; however, editing
during a run can be done from the Samples Menu (below).
Sample Queue
information
Information on the sample queue is presented in the first two lines of
the Status screen. The specific fields are described here.
QStatus
This field displays the status of the queue (IDLE, PAUSE, RUN,
EQUIL, ABORT. These conditions are described here.
IDLE indicates that no sample sets are currently being processed in
the queue.
NOTE: The IDLE status doesn't tell you whether or not there are sets
loaded into the queue. To check the contents of the sample queue, press
[MENU] and select QUEUE.
PAUSE indicates that a sample set is loaded, but that processing has
been suspended. If the queue is paused (the [STOP] key has been
pressed once), you can either resume processing with the current or
subsequent vial, or abort processing of the current sample set. For
more information, see "Stopping a Run" on page 89.
RUN indicates that a sample set in the queue is being processed.
queue (see also Sample queue)states ran
EQUIL indicates that the autosampler is waiting for the equilibration
delay (temperature zone and initial conditions) to be reached before
beginning the processing of the current sample set. This status will be
displayed until the oven and tray reach the set temperatures entered in
the Edit Menu and/or until the Equilibration Time entered in the More
Menu has elapsed.
ABORT indicates that the processing of the current sample set has
been stopped (the [STOP] key has been pressed and ABORT has been
selected from the Abort/Continue Menu) or that a system error has
occurred. Abort is displayed until the autosampler has returned the
active vial to the sample tray and flushed all the lines. For more
information about Abort, Continue, and Pause, see "Stopping a Run"
on page 89.
Set
This field displays the number of the sample set currently being
processed (1-39, P). If the queue is not running (the QStatus field
reads other than RUN or PAUSE), the set field is blank.
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Thermo Electron
#Samples
This field displays the number of the sample currently being
processed and the total number of samples in the set: current sample
number/total of samples. For example, if the autosampler is
processing sample 50 in a set of 120 samples, the #Samples field will
read 50 / 120.
NOTE: The total does not indicate the total number of vials or total number
of injections in the set, -- it indicates the total number of sample vials in the
set. Calibration vials are not included on this screen. This way, you'll have
a more exact status of the run progress if you've chosen to reuse your
calibration vials.
Injection Status
Pressing [∨] displays the next two status screens that contain
information on the injection in progress.
NOTE: The Status Screen is updated every two seconds.
Status
This field displays the progress of the current injection. A time in
minutes in the Status field indicates that the queue is running (RUN)
and that the current sample has been injected. This field displays the
current run time (the time elapsed since the injection). If an injection
has not occurred, the autosampler displays one of the following
messages in the Status field:
RDY indicates that the autosampler is ready for an injection (all
temperature zones and gradient conditions have been reached), but
that no injection is currently in progress.
NOTRDY indicates that the autosampler is not ready for an injection.
NOTRDY may indicate that the temperature zones are not stabilized,
the door is opened, or the Pump Ready contact closure is in the wrong
state.
GRAD indicates that the autosampler is waiting for the set gradient
conditions to be reached (for the gradient delay time set in the More
Menu to elapse.)
HOLD indicates that an injection hold is in effect as determined by
the Inject Hold input or as set in the Input Polarity Menu.
ABORT indicates that the current injection has been stopped in one of
two ways: 1) [STOP] has been pressed to stop the Queue and
ABORT selected from the Abort/Continue Menu, or 2) an error
condition has occurred. During the ABORT sequence, the
autosampler returns the current vial to the sample tray and flushes the
lines.
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85
DOOR indicates that the door is open. The autosampler automatically
moves the arm to the home position to allow easy access to the
sample tray. The arm automatically resumes where it left off when
the door is closed.
Vial
This field indicates the position and type of vial (S for sample; C for
calibration vial) currently being injected. If no injection is in progress
(the Status is "NOTRDY"), this field is blank.
Vol
This field displays the injection volume for the current vial. If the
current vial is a sample vial, this field indicates the Injection Volume
value entered in the Injection Menu. If the current vial is a calibration
vial, this field displays the Injection Volume value entered in the
Calibration Menu.
#Inj
This field displays a fraction that indicates the current injection
number over the total number of injections requested for the current
vial: current injection number/injection per vial. For example, if the
autosampler is performing the third of five injections the #Inj field
will read 3 / 5.
File #
Pressing [∨] displays the number and name of the file assigned to the
sample set currently being processed.
Temperature Status
Pressing [∨] accesses Oven/Tray Temperature Status screen which
displays the actual—not the set temperatures of the oven and tray, if
the Column Oven and Tray Temperature Control Options are
installed. The temperatures are displayed even if these options are
turned off (not set). "OFF" indicates that an option is installed but not
turned on in the current file's Injection Menu (Figure 3.32).
Oven
TrayTemp
OFF:75 RDY:18
Figure 3.32 The Temperature Status screen with the oven turned off
The temperature field remains blank if one of these options is not
present (Figure 3.33). This screen does not appear if neither option is
installed.
Oven
TrayTemp
RDY:75
Figure 3.33 The Temperature status screen without the Tray
Temperature Control Option installed
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Thermo Electron
HINT: To check the set temperature for either the oven or the tray (or other
parameters), you can return to the Edit Menu and view the information for
the appropriate file as long as you have not loaded that file into the Queue.
Therefore, you may wish to copy your file before loading it. That way you
can edit the copy if you want to check any experimental parameters after
your run file is loaded.
Changing the Run in Progress
Once a run has started, you may wish to change the injection volume,
cycle time, calibration interval, or set assignments. You make
changes to the run currently in progress (the run file) from the
Samples Menu. Figure 3.24 and Figure 3.35 list the changeable
parameters in bold-face type and indicates when those changes take
effect. With the exception of the Cycle Time and Calib 1(of X)
values, changes made from the Samples Menu take effect on the next
sample injection. Changes to the Cycle Time take effect on the
current sample injection; changes to the Calib 1(of X) field take effect
on the next recalibration. Samples Menu changes remain in effect
until the next sample set.
Sample Set
File 1:
1
VITAMINS
--------------------------------------------------------Next sample injection
Injections/Sample
1
Cycle Time
Current recalibration
0.1
Calib 1 (of 3) A02
Reuse vials only
First Sample Vial
A05
Cannot change
Number of Samples
1
Next sample injection
Set
2
Next sample set
Figure 3.34 The Samples Menu (AS1000 and AS100)
Sample Set
File 1:
1
VITAMINS
--------------------------------------------------------Injection Volume
5.0
Next sample injection
Injections/Sample
1
Next sample injection
Cycle Time
0.1
Current recalibration
Calib 1 (of 3) A02
Reuse vials only
First Sample Vial
A05
Cannot change
Number of Samples
1
Next sample injection
Set
2
Next sample set
Figure 3.35 The Samples Menu (AS3000 and AS300)
NOTE: Changing one vial location does not affect other vial locations.
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87
Priority sample sets
A priority sample set is one which can be inserted into the currently
active sample set. It shares the same file and resources as specified
for the currently active set. Priority sets can be added at any time.
Priority sample sets are like other sample sets with several important
exceptions:
1. You assign their priority set number (P) in the Samples Menu
(not in the Queue Menu).
2. Priority sets must be assigned to the currently active file.
3. You can't change the order of priority sample sets once they are
loaded into the sample queue.
4. If you want to run priority sample sets, you must select Reuse
same vials as your calibration routine type. (If you select
Intersperse vials, the autosampler won't "know" whether a
subsequent vial is a calibration vial or a sample vial and
therefore won't be able to resume the calibration routine after
the priority sample set is processed.)
5. Since a priority set is inserted into the currently active set, the
file cannot be edited or changed between the currently active
set and the priority set.
To insert a priority sample set into the sample queue, use the
following steps (Figure 3.36):
1. Press [SAMPLES].
2. Use the [+] and [-] keys to change the Set field value to P.
3. Fill in the other file parameters as you would for any other
sample set.
4. Add the priority set to the queue.
Sample Set 1:
P
File 1: VITAMINS
--------------------------------------------------------Injection Volume
Injections/Sample
Cycle Time
First Sample Vial
Number of Samples
Add to Queue?
10
1
10
C23
1
Yes
Figure 3.36 The Samples Menu with a priority sample set
NOTE: The door is equipped with a safety interlock that automatically
moves the autosampler's arm to the "home" position to allow you to add
samples to the tray during a run. Once the door is closed, the arm returns
to its position and resumes the operation in progress before the door was
opened. However, you can disable the door interlock if you wish. Press
[MENU], /OPTIONS/, Ready Participation, move to Door Interlock
(page 31), and use the [+] and [-] keys to select DISABLE.
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Thermo Electron
The autosampler assigns a P (priority) order to the priority set as
shown in the Queue Menu (Figure 3.37):
Order
File Name
Set
1
1:VITAMINS
P
--------------------------------------------------------1
1:VITAMINS
1
2
2:BARBITUA
2
3
3:EXPER 1
3
Figure 3.37 The Queue Menu with one priority sample set
STOPPING A RUN
The [STOP] key has several functions depending upon the
instrument's status when the [STOP] key is pressed. Refer to
Figure 3.38 as you read about the STOP/ABORT sequence.
Queue is running
[STOP]
(User Abort)
STOP requested
Select ABORT SAMPLE/CONTINUE
System
Error
ABORT
CONTINUE
Returns vial to tray
and flushes lines
STATUS goes to ABORT
Pauses the queue
Resumes queue
with current vial
[STOP]
Stops the queue
QSTATUS goes to IDLE
[RUN]
Resumes queue
with current vial
[RUN]
AS-Z015E/DG
Restarts queue from
the beginning
Figure 3.38 The Queue Stop/Abort Sequence
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89
If the queue is in RUN when you press the [STOP] key, the
autosampler suspends its current operation and waits for you to make
a selection to ABORT (Figure 3.39) or CONTINUE (Figure 3.40):
Stop requested.
Select
ABORT Sample
Figure 3.39 The Abort prompt
Stop requested.
Select
CONTINUE
Figure 3.40 The Continue prompt
If you select Continue (by pressing the [+] key), the autosampler
returns the queue to the ACTIVE state and resumes the motor activity
(as if you had not pressed the [STOP] key).
If you select Abort or if a system error occurs, the autosampler:
•
Aborts the processing of the current sample
•
Returns the active vial to the vial tray
•
Flushes the lines
•
Puts the queue in the PAUSE state (Figure 3.41)
QStatus
Set
#Samples
PAUSE
1
1 /
1
Figure 3.41 The Queue Status line with the autosampler in the
PAUSE state
When the queue is PAUSED, you can either:
1. Press [RUN] to resume processing of the current vial or
2. Press [STOP] to stop the queue (the QStatus goes to IDLE) and
then press [RUN] to restart processing from the beginning of
the queue.
This [STOP]/Abort sequence allows you several opportunities to stop
and resume or restart the run as needed.
The ABORT/CONTINUE Menu and QPAUSE states let you remedy
problems that could adversely influence the processing or data
received from the current or subsequent vials (depleted solvent
supply, etc.) and resume the run without loss of previous data.
The ABORT, [STOP], [RUN] choice allows you to stop the current
run and remedy problems that could have influenced the processing
or data received from all prior injections (incorrect file assigned to the
set, incorrect solvent concentration, etc.). This second option
minimizes loss of resources (and time!).
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Thermo Electron
File Management
You've learned how to edit and load files from the Files Menu. You
can also copy, delete, protect files from editing, and lock files to
prevent changes to the run file during a run.
SAVING FILES
As you know, the autosampler automatically saves your file when
you exit the Edit Menu. If a power failure occurs while you are
editing a file, all current changes will be lost. However, if a power
failure occurs after you exit the Edit Menu, your changes will be
saved.
COPYING FILES
(AS1000, AS3000,
AND AS3500 ONLY)
To copy a file, press [MENU], and select /FILES/, /Copy/ to display
the Copy Menu (Figure 3.42).
Copy File
1:
To File 2:
Figure 3.42 The Copy Menu
Enter the number of the file to be copied in the Copy File field. Enter
the number of the file you wish to copy to in the To field. When you
press [ENTER], the message **File Copied** appears briefly and
you are returned to the Files Menu.
DELETING FILES
Deleting a file restores all parameters to their default values. To
delete a file, press [MENU], select /FILES/, and /Delete/. The Delete
Menu appears (Figure 3.43).
Delete File
1:
Figure 3.43 The Delete Menu
Enter the number of the file you wish to delete and press [ENTER].
The message **File Deleted** appears briefly and you are returned to
the Files Menu.
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91
PROTECTING FILES
(AS1000, AS3000,
AND AS3500 ONLY)
You can protect a file from being edited or deleted by using the file
protection feature in the OPTIONS Menu. Because this feature
enables you to control the changes to an original run file, you can
easily incorporate your autosampler files into your standard goodlaboratory-practice (GLP) protocols. To display the Configurations
Menu (Figure 3.44), press [MENU], select /OPTIONS/,
/Configuration/.
Sample Syringe
250
Prep Installed Yes
--------------------------------------------------------Oven Installed Yes
TrayTemp Installed
Yes
Key Repeat Rate
Medium
File Name
Protect
1: VITAMINS
On
2: BARBITUA
Off
3: EXPER 3
Off
4: STAT Off
Mode
Stand Alone
Solv Viscosity Normal
Figure 3.44 The Configurations Menu
To protect a file, move the cursor down to the Protect field in the line
that contains the file to be secured and press the [+]/[-] keys to turn
ON the protect function. For example, File 1 VITAMINS in
Figure 3.45 cannot be edited until the protect function is turned off.
Attempting to edit a protected file displays the following message:
*File Protected*
No Editing Allowed
Figure 3.45 The confirmation message to prevent editing of a
protected file
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Thermo Electron
Changes to realtime editing
If a file is protected, you will not be able to change the injection
volume, number of injections per vial, or cycle time for the run in
progress. These lines will not appear on the Samples Menu
(Figure 3.46).
Sample Set 1
File 1:
VITAMINS
First Sample Vial
A01
Number of Samples 10
Set Already in Queue
Figure 3.46 The Samples Menu for a protected file
Needle Height
For some applications, such as liquid-liquid extractions and lowinjection volumes, you may need to change or optimize the "needle
height." On SpectraSYSTEM autosamplers, the needle doesn't move.
Instead, the bottom of the vial moves relative to the needle tip. The
"needle height" is the approximate distance between the needle tip
and the bottom of the vial (0 - 20 mm).
The More Menu's default setting of 2 mm (Figure 3.47) is sufficient
for most applications and accommodates a wide variety of vials.
However, in cases where you have very limited sample volumes,
further optimization may be required.
NOTE: The thickness of the bottom of the vial varies considerably between
vial types (1.8 mL vials, microvials, and vials with micro-inserts, etc.) and
materials (glass, plastic, etc.).
Equilibration Time
Gradient Delay
0.0
0.00
--------------------------------------------------------Viscosity
Flush Volume
Injection Type
Injection Range
Needle Height
Medium
250.0
Push
1-1500
2.0
Figure 3.47 The More Menu (Needle Height)
NOTE: Operating the autosampler with a needle height different
from the default height may bend the needle. Do not adjust the needle
height without a spare needle available.
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93
Use the following procedure to optimize the needle height for your
sample vials:
1. Place 200 - 300 µL water in a vial in position A01.
2. Create an autosampler file that will make two 200 µL
injections (see Chapter 3 for more information).
a. Set the Injection Menu's Injection Volume field equal to
200.
b. Set the Injection Menu's Injections/Sample to 2.
3. Run the file and observe the amount of fluid remaining in the
vial. The fluid height indicates the position of the needle tip
from the vial's bottom.
4. Adjust the More Menu's Needle Height value until the needle
tip is located approximately 1 - 2 mm from the bottom of the
vial.
5. Repeat steps 1 - 4 to confirm the needle height.
Optimization
Injection Volume
This section describes how to select the sample loop and syringe size
to obtain maximum reproducibility when performing large-volume
injections.
The maximum injection volume depends upon:
•
The volume of the syringe currently installed as designated
under /OPTION/Configuration/Syringe Size/;
•
The Injection Mode selected under /FILE/More/Injection Type;
and
•
The volume of the installed sample loop.
By changing the sample syringe and/or the sample loop, you can set
up your autosampler to deliver injection volumes larger than can be
delivered by the 250 µL syringe shipped with the instrument. Use the
following steps to prepare your instrument for large-volume
injections:
1. Determine the correct sample loop size for the injection volume
desired. (See Table 3.1.)
2. Use the injection volume and algorithms outlined below to
determine the required syringe size.
The following algorithms define the amount of sample needed
for your injection volume:
PushLoop
Pull Loop
Full Loop
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Thermo Electron
(Injection Volume + 15 µL)
(Injection Volume + 1.1 µL)
(Injection Volume x 1.33) + 70 µL
3. Change your sample loop and syringe. (Refer to Chapter 5 for
instructions.)
4. Enter the syringe size in the /FILE/Configuration/Syringe Size
field.
5. Enter the Injection Mode in the /OPTIONS/More/Injection
Type/ field.
6. Enter your injection volume in the Injection Menu.
NOTE: If the Full-loop mode is chosen, the maximum value depends
only on Loop Size (assuming that the syringe size is larger than the
loop size). (Loops come only in the sizes included in the Parts and
Accessories list located at the front of your Reference Manual).
NOTE: For Pull and PushLoop injection, to ensure maximum
reproducibility, we recommend that you do not inject more than half
of your loop volume.
Table 3.1 Recommended Injection Size For Selected Sample Loop Sizes*
Injection Volume ( in µL)
Injection Mode
PushLoop
Pull
Full
Sample Loop Size (µL)
5
2.5
2.5
5
10
5.0
5
10
20
10
10
20
50
25
25
50
100
50
50
100
200
100
100
200
500
200
250
500
1000
200
500
1000
These recommended volumes will provide the best chromatographic performance. Maximum
injection sizes will be determined by the particular system being used.
The syringe size needed will be the total volume determined by these
algorithms. For example, for a 500 µL full-loop injection (using a
1000 µL loop), the Total Volume (TV) needed is
TV = (500 µL x 1.33) + 70 µL
TV = 735 µL
Therefore you will need a 1000 µL syringe since there are no 735 µL
syringes available. (See Table 3.2.)
Table 3.2 Maximum Injection Size for Syringe Size
Syringe Size (in µL)
Injection Type
250
500
1000
Push Hi
200
200
200
Pull
212
461
956
Full
100
200
500
Thermo Electron
2500
200
1500
1000
5000
200
1500
1000
95
SOLVENT
VISCOSITY
You can adjust the syringe speed (draw rate) for your solvent viscosity
from the Solv Viscosity field in the Configurations Menu (Figure 3.48).
Sample Syringe Size
250
Prep Syringe Size
2500
--------------------------------------------------------Prep Installed
Yes
Oven Installed
Yes
Tray Temp Installed
Yes
Key Repeat Rate
Medium
File
Protect
1:
Off
2:
Off
3:
Off
4:
Off
Solv Viscosity
Normal
Figure 3.48 Configurations Menu -- Solv Viscosity field
Using the [+]/[-] keys you can choose among three different syringe
speeds: 1) Normal (fastest), 2) Medium, and 3) Viscous (slowest).
"Normal" lets you optimize your cycle time and is optimal for most
applications. With either very viscous solvents or normal-phase (low
viscosity) solvents, you may wish to use the "Viscous" setting to slow
down the syringe rate to prevent cavitation and nonreproducible
injection volumes.
Special Commands: Commands Menu
The autosampler has several built-in commands that allow you to
manually manipulate some of the autosampler's components (the arm,
injector valve, etc.) outside of a run. These commands, useful at
times during maintenance and troubleshooting procedures, are
accessible from the Commands Menu (Figure 3.49).
Flush Sample Syringe
Flush Prep Syringe
Initialize Hardware
Injector
Arm
Figure 3.49 The Commands Menu
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Thermo Electron
FLUSH SAMPLE
SYRINGE
This command lets you manually flush the 250 µL sample syringe.
You used this command when you primed your autosampler. It can
also be useful for maintenance and troubleshooting. To access this
test:
1. Fill your flush solvent bottle, if you have not already done so.
2. Press [MENU] and select /COMMANDS/, (Figure 3.49).
3. Select /Flush Sample Syringe/ to bring up the menu shown in
Figure 3.50.
4. Use the [+] and [-] keys to select the desired flush volume in
microliters. Then press [ENTER] to initiate the flush
operation.
Flush 5000 µL
Figure 3.50 Flush Syringe Menu
The autosampler automatically flushes the sample syringe and flush
lines with the specified volume of flush solvent and returns the
syringe to its starting position. During the flush sequence a
confirmation message is displayed.
Flush Prep Syringe
(AS3000 and
AS3500 only)
If you purchased the Sample Preparation Option, you can also flush
the large (2.5 mL) sample preparation (prep) syringe and external
solvent lines manually as follows:
1. Fill each solvent bottle with an appropriate solvent.
2. Press [MENU] and select /COMMANDS/.
3. Select Flush Prep Syringe (Figure 3.51).
4. Use the [+] and [-] keys to select the desired flush volume in
microliters. Then move the cursor to the Solv field and select
the desired solvent (S-1, S-4, Btl). Press [ENTER] to initiate
the flush sequence.
Flush 5000 µL Solv Btl
Figure 3.51 Flush Prep Menu
The autosampler automatically flushes the selected solvent line with
the specified volume of flush solvent and then returns the prep
syringe to its starting position.
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97
INITIALIZE
HARDWARE
The Initialize Hardware command allows you to manually control the
autosampler's syringes and injector valve. This command brings the
syringe(s) to the initial position(s) and removes any backlash.
INJECTOR
The Injector commands allows you to manually switch the injector
valve between the inject and fill positions. Selecting /Injector/
displays the menu shown in Figure 3.52.
Rotate Injection Valve INJECT
Figure 3.52 The Injector Menu
Pressing the [+] or [-] key and then pressing [ENTER] changes the
injector valve's position.
ARM
The Arm Menu contains two routines for returning the XYZ arm to
its home position and for aligning the arm. Figure 3.53 shows the
Arm Menu. The two routines are described below.
Home Arm
Align Arm
Clear Mixer
Figure 3.53 The Arm Menu
Home Arm
Pressing [ENTER] with the cursor in this field moves the arm from its
present position to its home location to the left of the rear-left corner
of the tub (Figure 3.54).
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Thermo Electron
Sample Tray Handle
(front)
A
B
Tub
Tray Positions
Runner
Slots
AS-Z003E\FM
C
Figure 3.54 The arm in home position
Align Arm
This routine moves the arm to the vial position requested
(Figure 3.55). It is used primarily by Thermo Electron service
engineers to properly adjust the arm's x-y-z reference points. If you
suspect that the arm's alignment is incorrect, call your
Thermo Electron Service representative.
Align Arm
A01
Figure 3.55 Align Arm Menu
NOTE: The arm "senses" its position relative to its home position.
Therefore, if you request the arm to go to position A01, and it goes to
position A06, the system configuration may incorrectly indicate that the Tray
Temperature Control Option is installed. Check the system configuration in
the OPTIONS Configurations Menu against the installed options.
Clear Mixer
This routine is used to remove a vial from the mixer if a power failure
occurs during a mix step in a sample preparation procedure. The arm
picks up the vial from the mixer and returns to the home position.
You must manually return to the vial to its proper location in the
sample tray.
NOTE: This routine appears on the Arm Menu only with the Sample
Preparation Option installed.
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99
4
Advanced Operations
Introduction
This chapter describes the Sample Preparation Option and function in
the AS3000 and AS3500 autosamplers as well as gradient delay
programming, and built-in diagnostics available on all
SPECTRASYSTEM autosamplers.
This chapter is organized into three parts:
•
Sample Preparation beginning on page 101
•
Gradient Delay beginning on page 127
•
Diagnostics beginning on page 129
Sample Preparation
This section tells you how to use the optional sample preparation
system to automate sample treatment, heating, and mixing. In this
section you will learn:
•
The terminology used in the sample preparation templates
•
How the sample preparation system works
•
How to use the built-in templates
•
How to create your own customized file
•
How to use the Edit and Samples Menus and Status Screen
with the Sample Preparation Option installed
•
How to run an autosampler file that uses automated sample
preparation
Maintenance and troubleshooting procedures for the sample
preparation system are included in Chapter 5, Maintenance, and
Appendix A, Troubleshooting, respectively.
BEFORE YOU
START
In presenting the material in this chapter, we assume that you have
installed your sample preparation bottles and lines as described in
Chapter 1, that you have read and used the procedures described in
Chapter 3, and that you are comfortable with the routine operations of
your autosampler.
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101
OVERVIEW
Preparing to run a sample preparation file is similar to preparing to
run any other autosampler file. First, you prepare your chemicals and
fill the solvent bottle(s). Then you enter your file information into the
Injection, Calibration, Timed Events, and More Menus as always.
Next you enter your sample preparation parameters into one of the
four built-in sample preparation menus (templates) accessible from
the Edit Menu's Sample Prep field (Figure 4.1). Your sample
preparation parameters are stored in the same file as your other run
parameters. You load your file, enter your sample set information
(build your sample queue) and start your run as we discussed in
Chapter 3.
The Overlap Enable field at the end of each template allows you to
program the autosampler to begin preparing the next sample vial
while the current injection vial is still eluting.
The steps for running a sample preparation file are listed on the
Sample Preparation reference card in the front of this manual.
Edit File
1
File Name
--------------------------------------------------------Injection
Calibration
Timed Events
Sample Prep
More
Figure 4.1 The Edit Menu
TERMINOLOGY
Templates
Each of the four sample preparation menus is a built-in procedure
format called a template. Each template contains a different number
of steps arranged in a preset and unchangeable order. Each step
carries out a specific autosampler operation or group of operations.
Some steps transfer liquid between vials or dispense solvents from
any of the four external solvents. Others control the operation of the
heater/mixer (included in the Sample Preparation Option) or the
position of the hook to adjust the distance between the needle tip and
the bottom of the vial.
By using the template default values, you can run one of the four
built-in procedures:
1) Simple dilution
2) DABS-CL derivatization
3) Rainbow dilution
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Thermo Electron
4) Linear dilution
Or, you can enter new values and create your own customized
procedures. The autosampler automatically prepares your sample(s)
according to the steps specified in the template.
Dilution Cycles
Within each template, the numbered steps are grouped into dilution
cycles. For example, steps 1 - 6 in Figure 4.2 comprise the dilution
cycle for Template 1.
Dilution cycles differ across templates in the number of steps and
types of operations they perform. You can program the autosampler
to repeat dilution cycles up to four times within a template to carry
out more complex procedures. We'll see how this works in the
section titled "Template Descriptions" on page 112. But first we'll
discuss what happens in the simple dilution cycle used in Template 1
because its steps are used in all four templates. Once you understand
how these steps control the autosampler's operation, you'll be able to
apply the same principles to Templates 2-4.
Dilution Cycles
1
1- Load 500 µL Solv S-1
2- Pickup
100 µL Sample
3- Pickup
0 µL Sample
4- PickLG
0 µL Sample
5- Add 500 µL to Sam+1
6- Mix for 0.5 minute
Overlap Enable:
No
This Procedure requires
that 1 Extra Vial be
placed immediately after
each Sample Vial.
Figure 4.2 Template 1
Your autosampler's simple dilution cycle is easily understood by
comparing it to a manual dilution procedure. In a manual dilution,
you draw a specific amount of sample into a pipette and expel a
measured volume into a volumetric flask. Next you dilute to final
volume with a desired amount of solvent and mix your
sample/solvent mixture for a specific amount of time.
In the manual dilution we've just discussed, solvent and sample are
added to the target container one at a time. In contrast, the
autosampler first draws all of the resources for a given dilution into
its solvent-holding loop and sample/needle lines. An air bubble is
drawn before and after each resource to isolate it in the loop. Once all
of the resources have been drawn into the lines, the autosampler
Thermo Electron
103
delivers them to the target vial simultaneously. The vortex mixer
then mixes the target vial for the specified amount of time.
All of the templates use this dilution routine. The next section
discusses how the individual template steps carry out this general
dilution procedure.
HOW DILUTION
CYCLES WORK
Delivery lines transport liquid and air through the sample preparation
system as programmed in the template steps. Each step influences the
operation of one or more of the six major components in the sample
preparation system:
1) Prep syringe
2) Sample syringe with 6-port flush valve
3) Solvent holding loop
4) Six-port solvent selection valve with your four solvent bottles
5) Heater/vortex mixer
6) Needle
Refer to Figure 4.3 and keep these components in mind as you learn
the functions of each step in a simple dilution cycle.
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Thermo Electron
Solvent
Selector
Valve
6
1
2
Selector-Flush
Tube
5
3
Solvent
Holding
Loop
4
Flush
Solvent
Prep-Flush
Tube
1
6
2
5
Injection
Tower
4
Sample
Loop
2
1
3
6
4
5
To
Pump/Oven
Sample
Syringe
To
Column
Flush-Inject
Tube
Sample Syringe
Valve
(6-pos.)
Prep
Syringe
KEY
Buffer
Pickup
Pickup
PickLg
Flush Solvent
AS-Z010E/DT
Injector
Valve
Prep
Syringe
Valve
3
Figure 4.3 The sample preparation dilution process
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105
The Simple Dilution
Cycle
Before the dilution cycle begins, the autosampler's lines contain flush
solvent. The simple dilution cycle is made of five different steps:
Load, Pickup, PickLG, Add, and Mix. Each of these steps is
described below.
NOTE: Some of these steps are repeated or skipped in the default methods.
We'll see how this works in the sections that describe each template's
dilution cycle.
Load
The Load step flushes the lines with the selected solvent in a six-step
process:
1) The 6-port solvent-selection valve rotates to the air position to
draw an air bubble into the solvent holding loop.
2) The solvent selection valve then rotates to the selected solvent
position (S-1, S-2, S-3, or S-4).
3) The preparation (prep) syringe draws the specified amount of
solvent (0-1500 µL) plus a constant excess amount into the
solvent holding loop. The excess solvent will be used in step 6
to flush the dead volume from the solvent-selection valve to the
needle tip. The air bubble drawn into the solvent-holding loop
in step 1 isolates the selected solvent (this step) from the flush
solvent already present in the lines.
4) The solvent-selection valve moves back to the air position and
the prep syringe draws another air bubble.
5) The solvent-selection valve rotates to the sample/needle line.
6) The prep syringe moves forward and expels solvent, thus
flushing the dead volume with the selected solvent and
expelling the second air bubble leaving only the selected
solvent behind.
Pickup
The Pickup step causes the sample syringe to withdraw the specified
volume of liquid from the source vial. The source can be the vial in
the Sample (active) position, the vials in the Reagent A, B, or C
locations (anywhere in the trays), or vials in the three positions
adjacent to the Sample vial: Sam+1, Sam+2, or Sam+3. For
example, if the Sample vial position is A01, Sam+1 would be A02,
Sam+2 would be A03, etc. The following operations occur during
Pickup:
1) The hook moves to the selected vial position and picks up the
vial.
2) The autosampler's arm brings the vial to the sample tower and
pushes the funnel out of the way.
106
Thermo Electron
3) The sample syringe draws an air bubble to isolate the sample or
reagent from the selected solvent already present in the solvent
holding loop (from the Load step).
4) The vial is raised in the sample tower. Sample is then drawn
into the sample/needle line.
5) The vial is lowered and the sample syringe draws an air bubble
to isolate the sample slug and to prevent sample loss.
6) The hook returns the vial to the sample tray.
This process is repeated if a second Pickup step is used.
NOTE: Each pickup step allows you to pick up a maximum of 100 µL. If
you want to pick up more than 100 µL, use either the second pickup step or
the PickLg step.
PickLg
The PickLg (Pickup Large) is the same as Pickup except that it uses
the solvent syringe to transport larger volumes (0 - 1000 µL).
Add
During the Add step two events happen:
1) The hook moves to the designated "target" vial position
(Sample, Sam+1, Sam+2, or Sam+3) and transports the vial
into the sample tower.
2) The prep syringe expels the desired volume (0 - 1000 µL) of
the liquids drawn into the solvent-holding loop and
sample/needle lines, during the Load, Pickup, and PickLG
steps, into the "target" vial.
NOTE: Because the Add step uses the prep syringe, the maximum
volume that can be added is the sum of the Load volume and the
PickLG volume. That is, you can't add (deliver to the target vial)
more than you've withdrawn into the large (prep) syringe in the Load
and PickLg steps.
Mix
The final step in the simple dilution cycle is the Mix step. During this
step the target vial is carried to the mixer and vortex mixed for the
specified length of time (0.0 to 9.9 minutes). In templates 1 and 2,
the vial is returned to the sample tray after the Mix step. In templates
3 and 4, you have the option of doing an extraction operation before
returning the vial to the tray.
After each dilution cycle, the autosampler automatically flushes its
lines and syringes with flush solvent. This operation (not shown in
the templates) returns the instrument to a "known" state. Any residual
sample, reagents, or solvents from a previous cycle is flushed from
the lines so that the instrument is ready for the next cycle or injection.
NOTE: If you are using immiscible solvents, be sure to flush the
preparation syringe (prep flush) between solvents to minimize contact
between the immiscible solvents in the autosampler's lines.
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107
TEMPLATE
PROGRAMMING
RULES
Just as there were rules for creating regular files, there are rules that
you need to remember when creating sample preparation procedures.
These nine rules are also included on your Sample Preparation Quick
Reference Card for future reference.
NOTE: You may wish to read through these guidelines briefly and then
refer back to them as you read the section titled, "Template Descriptions."
1. Extra vials are placed in the three vial positions subsequent to
the Sample vial position (Sam+1, Sam+2, Sam+3). For
example, if the Sample position is A02, Sam+1 is A03, Sam+2
is A04, etc.
2. It is impossible to add, delete, or change the order of steps
within a template, but you can skip any step (except an Add
step) by setting its volume parameter to 0 µL.
3. Specify one dilution cycle for each delivery of chemicals to, or
for each operation carried out on, the target vial.
4. The desired final volume is specified in the Add step.
5. To perform an operation on, but not make a delivery to, the
target vial, set the Add step's volume parameter to 0. This
approach is useful, for example, when you want to mix or heat
the contents of a vial.
6. The volume specified in the Add step in a given dilution cycle
must be less than or equal to the sum of the PickLG and the
LOAD volumes.
7. The Add step makes deliveries to the target vial on a last-infirst-out basis.
8. Leave the Overlap Enable function turned on (Yes) if sample
preparation for the next vial to be injected can begin while the
current vial is still running.
9. The autosampler injects only from the last vial referenced in
the method template's last Add step.
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SAMPLE
PREPARATION MENU
DIFFERENCES
Before we describe each template in detail, we'll discuss how to do a
sample preparation run.
Performing a sample preparation run is essentially the same as doing
a run without sample preparation. You create and load your file,
build your sample queue, and start your run just as you did in
Chapter 3. However, with the Sample Preparation Option installed,
the Edit and Samples Menus, and Status Screens are expanded to
allow you to enter and monitor your sample preparation parameters.
The "new" lines that are dedicated to sample preparation in each of
these menus and screens are discussed below.
Edit Menu
With the Sample Preparation Option installed, the Edit Menu has the
Sample Prep field that you saw in Figure 4.1.
Sample Preparation
Menu
Selecting /Sample Prep/ from the Edit Menu accesses the Sample
Prep Menu from which you can use the [+] and [-] keys to select one
of the four sample preparation templates (Figure 4.4):
Use Template
Description
1
Dilution
(The Templates start here)
Figure 4.4 The Sample Prep Menu
Samples Menu
The Samples Menu (Figure 4.5) has several additional lines with the
Sample Preparation Option installed. These new lines reflect the
number of reagent and extra vials used by the method.
Reagent Vials (Reag-A, B, and C)
If reagents (A, B, and C) are used in the sample preparation method, a
line is added on the Samples Menu for each reagent used by the
method to allow you to edit its vial location. If one reagent is used,
only the Reag-A appears; if two reagents are used, the Reag-B line
appears. Figure 4.5 shows the Samples Menu when all three reagents
are used.
NOTE: Reagent vials can be placed in any vial position in any of the vial
trays. They don't have to be in proximity to the sample vial.
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109
Sample Set
1
File 1:
Injection Volume
10
Injections/Sample
1
Cycle Time
Reag-A C23
(If injection is used)
0.1
[If reagent vial(s) is (are) used]
Reag-B C24
Reag-C C25
Calib Vial 1 (of X)
A01
First Sample Vial
A04
Number of Samples
1
(If calibration is used)
File requires n extra
vials after each sample
Add to Queue?
Yes
Figure 4.5 The Samples Menu with reagents, calibration, and extra
vials used in the sample preparation method
File requires n extra vials after each sample
If extra target vials are used after each sample vial, the two lines
indicating the number of extra vials required are copied from the
sample preparation template into the last two lines of the Samples
Menu.
The "n" represents the number of extra (empty) vials placed after the
Sample vial (the Sam+1, Sam+2, and Sam+3 vials). An extra vial
(one in which sample is mixed with solvent, where a reaction takes
place), or a pickup (PickLg) is made. "n" is selected in the Add steps
of each template. Figure 4.5 illustrates the Samples Menu for a file
that uses calibration with the Sample Preparation Option installed.
NOTE: The screen actually displays a number (1, 2, or 3) in the "n" field.
We've represented this value as "n" because it changes depending on
whether you've chosen Sam+1, Sam+2, or Sam+3 during your run.
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Changes to real-time editing
As described in Chapter 3, you can make changes to the run file for
the run in progress from the Samples Menu. Table 4.1 summarizes
when Samples Menu changes to the run in progress take effect.
Table 4.1 When Changes Affect the Run in Progress
File #
Can't edit
Injection Volume
Next injection*
Injections/Sample
Next injection
Cycle Time
Current injection
ReagentA(B,C)
Next sample
Reagent A-C
Next sample
Calib Vial 1 of (x)
Next recalibration
First Sample Vial
Can't edit
Number of Samples
Next injection
*Changes to this field affect sample vials only, not calibration
vials.
Protecting files from real-time editing
As described in Chapter 3, a file can be protected from real-time
editing by turning on the Protect function in the Configurations
Menu. File Protect prevents you from editing the injection volume,
injections per sample and cycle time and eliminates some lines from
the Samples Menu for a protected file that uses sample preparation
just as it did for a regular file. Figure 4.6 shows the Samples Menu
for a protected file that uses sample preparation and calibration.
Sample Set
1
File 1: VITAMINS
Calib Vial 1 (of 3)
A01
First Sample Vial
A04
Number of Samples
10
Already in Queue
Figure 4.6 The Samples Menu for a Protected File that uses sample
preparation and calibration
Status Screen
With the Sample Preparation Option installed, an additional Status
Screen is included to give you information on the current template
step, the heater/mixer status and the resource locations for the active
file. Figure 4.7 shows the Status Screen for a file that contains a
sample preparation procedure and that uses several reagents and
calibration vials.
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111
QStatus
Set
#Sample
RUN
1
5
/ 120
--------------------------------------------------------Status Vial
Vol
#Inj
NRDY
1000
1 / 2
A01
File 1:VITAMINS
Oven
TrayTemp
RDY:80
Prep
RDY:4
Vial
PickLG A01
Htr/Mix (only with sample preparation)
RDY:35
Figure 4.7 The Status Screen for a file with sample preparation
Prep
This line tells you which template step is in progress (Load, Pick_Up,
Pick_Lg, Add, Mix, Wait, or Remove). For example, Figure 4.7
indicates that even though the sample queue is running (RUN), since
a Mix step is in progress, the autosampler is not ready (NRDY) to
inject.
Vial
This field indicates the position of the vial currently undergoing
sample preparation.
Htr/Mix
This field indicates the status (OFF, RDY, and NRDY) of the
heater/mixer and its actual temperature, if turned on and set in the
Heater field in templates 2 and 4. If not in use, this field reads "OFF"
followed by the actual temperature.
NOTE: The temperature is always displayed whether or not the
heater/mixer is in use.
TEMPLATE
DESCRIPTIONS
You are now familiar with the basic template steps and the additional
menu selections you have to make when using automated sample
preparation. In this next section we'll describe the specifics of each
template.
All four templates are based on the liquid transport and delivery
process described in the section titled "The Simple Dilution Cycle"
above. In fact, Template 1 (simple dilution) contains only the steps
described above (Figure 4.2). If your protocol has more than two
dilutions, simply specify the number of dilution cycles you need in
the line before step 1. The autosampler will automatically replicate
the template's dilution cycle a total of four times.
Each template will be discussed in two ways: with a boxed figure
showing how the display looks, and with a line-by-line description of
each step. You can use the line-by-line description to assist you in
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Thermo Electron
creating a customized method file for your own applications. Each
example uses the template's default values.
Template 1: Dilution
Template 1 (Figure 4.8) is a dilution program that you can use to
perform a 1:5 v/v dilution procedure.
Use Template
Description
1
Dilution
# of Dilution Cycles
1
1- Load 500 µL Solv S-1
2- Pickup
100 µL Sample
3- Pickup
0 µL Sample
4- PickLG
0 µL Sample
5- Add 500 µL to Sam+1
6- Mix for 0.5 minute
Overlap Enable:
Yes
This procedure requires
that 1 Extra Vial(s) be
placed immediately
after each Sample Vial.
Figure 4.8 Template 1: Dilution
Line-by-Line Description of Template 1
Use Template 1
Template 1 is selected.
Description
Template 1 is the Dilution Template.
Dilution
# of Dilution Cycles 1
One dilution cycle requested for delivery to Sample vial.
1- Load 500 µL Solv S-1
The prep syringe draws 500 µL Solvent 1 (buffer) into solvent
holding loop
2- Pickup 100 µL Sample
The sample syringe draws 100 µL sample into the sample lines.
3- Pickup 0 µL Sample
Skip this step; the sample syringe is not needed.
4- PickLG 0 µL Sample
Skip this step; the prep syringe not needed.
5- Add 500 µL to Sam+1
Dilute 100 µL sample to 500 µL and deliver to Sam+1.
6- Mix for 0.5 minute
Mix sample and buffer for 0.5 minute.
Overlap Enable Yes
Turn on the Overlap Enable function to begin preparation after the
last injection and flush of the previous vial (sample or calibration).
This procedure requires
that 1 Extra Vial(s) be
placed immediately
after each Sample Vial.
One Extra (empty or extra) vial is required since mixing takes place
in the Sam+1 vial position.
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113
How the simple
dilution steps work
together
Before continuing to the other three templates, it is important to
understand how the Load, Pickup, PickLg, and Add steps affect the
contents of the Add step target vial. Remember that the Add step
makes deliveries to the target vial on a last-in-first-out basis.
Consider the following examples to see how this works.
Use Template
Description
1
Dilution
# of Dilution Cycles
1
1- Load 1000 µL Solv S-1
2- Pickup
0 µL Sample
3- Pickup
0 µL Sample
4- PickLG
0 µL Sample
5- Add 1000 µL to Sam+1
6- Mix for 0.5 minute
Overlap Enable:
Yes
This procedure requires
that 1 Extra Vial(s) be
placed immediately
after each Sample Vial.
Figure 4.9 Template 1: Dilution Cycle with no Pickup or PickLg
In Figure 4.9, 1000 µL of solvent is loaded into the holding loop and
delivered into the target vial since no other reagents are drawn into
the holding loop in the Pickup or Picklg steps.
In contrast, in Figure 4.10, 1000 µL of solvent and 100 µL sample are
loaded into the holding loop. The Add step delivers the last 1000 µL
of liquid drawn: 100 µL sample from the first Pickup step but only
900 µL solvent from the Load step. The Add step designates the
desired final target-vial volume.
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Use Template
Description
1
Dilution
# of Dilution Cycles
1
1- Load 1000 µL Solv S-1
2- Pickup
100 µL ReagA
3- Pickup
0 µL Sample
4- PickLG
0 µL Sample
5- Add 1000 µL to Sam+1
6- Mix for 0.5 minute
Overlap Enable:
Yes
This procedure requires
that 1 Extra Vial(s) be
placed immediately
after each Sample Vial.
Figure 4.10 Template 1: Dilution Cycle showing a 1:10 dilution
Template 2: DABS-CL
Template 2 (Figure 4.11) is a program to perform various chemical
reactions. The default method is specifically designed to perform a
(Dimethylamino)azobenzenesulfonyl chloride (DABS-Cl)
derivatization procedure. This template contains the same steps as
Template 1, plus three new steps described here.
Heater Temp
This line allows you to turn the heater on and off and set the
temperature from 30 - 100 °C.
Heat
In this field you can specify a heating time from 0.0-99.9 minutes.
Mix
This field allows you to remix the reagents for a specified amount of
time (0.0 - 9.9 minutes) after heating. Figure 4.11 shows the first
Template 2 dilution cycle.
Use Template 2
Description
DABS-CL
# of Dilution Cycles
2
Heater: On
Temp: 65 C
1- Load 200 µL
Solv S-1
2- Pickup 100 µL Reag-A
3- Pickup
40 µL Sample
4- PickLG
0 µL Sample
5- Add 140 µL to Sam+1
6- Mix for 0.3 minute
7- Heat for 4.5 minute
8- Mix for 0.3 minute
Overlap Enable
Yes
Figure 4.11 The first dilution cycle of Template 2
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115
The default method's values are those used to perform a simple
DABS-Cl derivatization of amino acids. For the exact reagents,
samples, and concentrations used for this procedure, please refer to
Thermo Finnigan’s Application Note 2: Automated DABSYL
Derivatization.
Line-by-line description of Template 2
Use Template 2
Template 2 selected.
Description DABS-CL
Template 2 is DABS-Cl derivatization.
# of Dilution Cycles
Heater: On
2
Temp: 65
Two dilution cycles is default value: one dilution cycle per delivery
to Extra vial (Sam +1).
Turn on heater; set the temperature.
1- Load 200 µL Solv S-1
Draw buffer into the solvent-holding loop.
2- Pickup 100 µL Reag-A
Sample syringe draws Reag-A into sample/needle line (e.g. DABSCl).
3- Pickup 40 µL Sample
Sample syringe draws 40 µL Sample into sample/needle line (e.g.
Amino Acids).
4- PickLG 0 µL Sample
Skip step.
5- Add 140 µL to Sam+1
Deliver Reag-A and Sample into first extra vial (e.g. DABS-Cl).
6- Mix for 0.3 minute
Mix contents of the first Extra vial for 0.3 minutes to bring vial to
temperature.
7- Heat for 4.5 minute
Heat vial for 4.5 minutes.
8- Mix for 0.3 minute
Mix reaction vial for 0.3 minute.
Repeat Dilution Steps
9- Load 1000 µL Solv S-1
Draw buffer into the solvent-holding loop.
10- Pickup 0 µL Sample
Skip this step.
11- Pickup 0 µL Sample
Skip this step.
12- PickLG 0 µL Sample
Skip this step.
13- Add 860 µL to Sam+1
Deliver 860 µL Solv S-1 into reaction vial (e.g. acetate buffer).
14- Mix for 0.3 minute
Mix reaction vial for 0.3 minutes.
15- Heat for 0.0 minute
Skip this step.
16- Mix for 0.0 minute
Skip this step.
Overlap Enable
Turn on Overlap Enable function.
Yes
This procedure requires
that 1 Extra Vial(s) be
placed immediately
after each Sample Vial.
The protocol requires one reaction vial for vortex mixing Reag-A,
sample, and Solv S-1.
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Thermo Electron
Template 3:
RAINBOW
Template 3 (Rainbow) allows you to do a more complex dilution
process or an extraction process and is the same as Template 1
(dilution) with three additional steps. Wait and Remove steps are
added after the Mix step at the end of each dilution cycle. The FlushP
step has been added after the last dilution cycle before the Overlap
Enable step. These new steps are described here.
NOTE: The default values in Template 3 are designed to illustrate a
dilution procedure that creates a calibration series with two components
(Reagents A and B) and four calibration levels. For this illustration, the
resource locations are as follows: ReagA contains your first standard,
ReagB contains your second standard, and Sample, Sam + 1-3 are empty.
Wait
The Wait step programs a reaction time for the vial contents or to
allow sufficient time for the liquid layer in an extraction to separate.
Wait pauses the autosampler for the specified amount of time (0.0 99.9 minutes).
Remove
The Remove step positions the bottom of the vial at the desired
distance (0.0 - 20.0 mm) from the needle tip, withdraws a desired
volume of solvent (0 - 1500 µL), and delivers it to waste.
FlushP
As you know, the autosampler automatically flushes the lines and
syringes with flush solvent at the end of each dilution cycle. (This
step is not shown on your templates.) The FlushP step allows you to
fill the autosampler's lines with any of the four solvents (or buffers) in
preparation for the sample injection. You can select any of the four
external solvents (S-1, S-2, S-3, S-4), or you can flush the lines with
flush solvent by selecting Solv Btl. After flushing the lines, FlushP
returns the prep syringe to its initial position in preparation for the
next injection. Figure 4.12 illustrates the last dilution cycle for
Template 3 in order to include the FlushP step.
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117
Repeat Dilution Steps
25- Load
1000 µL
Solv
S-1
26- Pickup 0 µL Sample
27- Pickup 30 µL Reagent A
28- PickLG 0 µL Sample
29- Add 1000 µL to Sam+3
30- Mix for 0.5 minute
31- Wait for 0.0 minute
32- Remove 0 µL to
waste at 2.0 mm height
FlushP
0 µL
Solv
S-1
Overlap Enable No
This procedure requires
that 3 Extra Vial(s) be
placed immediately
after each Sample Vial.
Figure 4.12 The last dilution cycle of Template 3
Line-by-line description of Template 3
Use Template 3
Template 3 selected.
Description Rainbow
Template 3 is Rainbow Dilution example.
# of Dilution Cycles 4
Specify a dilution cycle for the reagent(s)/solvent delivery to each
of the four calibration vials.
1- Load 1000 µL Solv S-1
Draw solvent into the solvent holding loop.
2- Pickup 30 µL Reag-B
Sample syringe draws Reagent B into the sample/needle line.
3- Pickup 0 µL Reag-A
Skip step (no Reagent A is used in the first calibration vial).
4- PickLG 0 µL Sample
Skip this step.
5- Add 1000 µL to Sample
Dilute 30 µL Reagent B to 1000 µL with solvent S-1 and deliver to
sample vial.
6- Mix for 0.5 minute
Mix contents of sample vial for 0.5 minutes (first calibration level).
7- Wait for 0.0 minute
Skip this step.
8- Remove 0 µL to
waste at 2.0mm height
Skip this step.
Repeat Dilution Steps
9- Load 1000 µL S-1.
Flush lines and solvent loop with the desired volume of solvent
(S-1).
10- Pickup 20 µL Reag-B
Sample syringe withdraw 20 µL Reagent B into sample/needle line.
11- Pickup 10 µL Reag-A
Sample syringe withdraw 10 µL Reagent A into sample/needle line.
12- PickLG 0 µL Sample
Skip this step.
13- Add 1000 µL to Sam+1
Dilute 20 µL Reagent B and 10 µL Reagent A to 1 mL with solvent
S-1 and deliver to first Extra vial (second calibration level).
14-Mix for 0.5 minute
Mix contents of first extra for 0.5 minutes.
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Thermo Electron
15- Wait for 0.0 minute
Skip this step.
16- Remove 0 µL to waste at
2.0mm height
Skip this step.
17- Load 1000 µL Solv S-1
Draw solvent into the solvent holding loop.
18- Pickup 10 µL Reag-B
Sample syringe draw 10 µL Reagent B into the sample/needle line.
19- Pickup 20 µL Reag-A
Sample syringe draw 20 µL Reagent A into the sample/needle line.
20- PickLG 0 µL Sample
Skip this step.
21- Add 1000 µL to Sam+2
Dilute 10 µL Reagent B and 20 µL Reagent A to 1 µL with solvent
S-1 and deliver to second Extra vial (third calibration level).
22- Mix for 0.5 minute
Mix contents of second Extra for 0.5 minutes.
23- Wait for 0.0 minute
Skip this step.
24- Remove 0 µL to
waste at 2.0mm height
Skip this step.
Repeat Dilution Steps
25- Load 1000 µL S-1.
Flush lines and solvent loop with the desired volume of solvent
(S-1).
26- Pickup 0 µL Reag-B
Skip this step (no Reagent B needed in the last calibration level).
27- Pickup 30 µL Reag-A
Sample syringe withdraw 30 µL Reagent A into sample/needle line.
28- PickLG 0 µL Sample
Skip this step.
29- Add 1000 µL to Sam+3
Dilute 30 µL Reagent A to 1 mL with solvent S-1 and deliver to
third Extra vial (fourth calibration level).
30-Mix for 0.5 minute
Mix contents of third extra for 0.5 minutes.
31- Wait for 0.0 minute
Skip this step.
32- Remove 0 µL to waste at
2.0mm height
Skip this step.
FlushP
0 µL Solv S-1
Overlap Enable
No
This procedure requires
that 3 Extra Vial(s) be
placed immediately
after each Sample Vial.
Skip this step.
Turn off the Overlap Enable function.
One extra vial is required for each standard level. The sample vial
contains the first level; levels 2-4 are in positions Sam+1, 2, and 3,
respectively.
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119
Template 4:
LINEAR
Template 4 template is also named for its default method; it allows
you to do linear dilutions. This template (Figure 4.13) contains the
heating and mixing steps added to Template 2 and the Wait, Remove
and FlushP steps used in Template 3. Figure 4.13 shows the last
dilution cycle of Template 4.
Use Template 4
Description Linear
# of Dilution Cycles
Heater: On
3
Temp: 30 C
21- Load 1000 µL Solv S-1
22- Pickup 5 µL Sample
23- Pickup
0 µL Sample
24- PickLG
0 µL Sample
25- Add 1000 µL to Sam+1
26- Mix for 0.3 minute
27- Heat for 0.0 minute
28- Mix for 0.0 minute
29- Wait for 0.0 minute
30- Remove 0 µL to
waste at 2.0 mm height
FlushP
0 µL Solv S-1
Overlap Enable
No
This procedure requires
that 3 Extra Vial(s) be
placed immediately
after each Sample Vial.
Figure 4.13 The last dilution cycle of Template 4: Linear
This template can be used to perform any function or application
performed by any of the other templates. The default values can be
used to perform a linear dilution procedure, for example, to prepare
calibration vials. If you use the default values, your diluent is Solvent
S-1, Sam+1 will hold your Level 1 standard, Sam+2 will hold your
Level 2 standard, and Sam+3 will hold your Level 3 standard.
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Thermo Electron
Line-by-line description of Template 4
Use Template 4
Template 4 is selected.
Description
Template 4 does linear dilution.
Linear
Dilution Cycles
Heater: On
Temp:
3
30 C
One cycle per delivery to a target vial.
Turn on heat; set temperature.
1- Load 1000 µL Solv S-1
Load 1000 µL buffer into solvent holding loop.
2- Pickup 5 µL Sample
Sample syringe draw sample (for the Level 1 standard) into the
sample/needle line.
3- Pickup 0 µL Sample
Skip this step.
4- PickLG 0 µL Sample
Skip step.
5- Add 1000 µL to Sam+1
Deliver buffer and sample to the first Extra vial (Level 1 standard).
6- Mix for 0.3 minute
Mix the first Extra vial for 0.3 minutes.
7- Heat for 0.0 minute
Skip step.
8- Mix for 0.0 minute
Skip step.
9- Wait for 0.0 minute
Skip step.
10- Remove 0 µL to
waste at 2.0mm height
Skip step.
Repeat Dilution Steps
Begin dilution cycle 2.
11- Load 1000 µL Solv S-1
Draw 1000 µL buffer into the solvent holding loop.
12- Pickup 50 µL Sample
Sample syringe draw sample (for the Level 2 standard) into
sample/needle line.
13- Pickup 0 µL Sample
Skip step.
14- PickLG 0 µL Sample
Skip step.
15- Add 1000 µL to Sam+2
Add buffer and sample to second Extra vial (Level 2 calibration
vial).
16- Mix for 0.3 minute
Mix the Level 2 calibration vial.
17- Heat for 0.0 minute
Skip step.
18- Mix for 0.0 minute
Skip step.
19- Wait for 0.0 minute
Skip step.
20- Remove 0 µL to
waste at 2.0mm height
Skip step.
Repeat Dilution Steps
Begin dilution cycle 3.
21- Load 1000 µL Solv S-1
Load 1000 µL buffer into the solvent holding loop.
22- Pickup 100 µL Sample
Sample syringe draws 100 µL sample into the sample/needle line.
23- Pickup 0 µL Sample
Skip step.
24- PickLG 0 µL Sample
Skip step.
25- Add 1000 µL to Sam+3
Deliver sample and buffer for the Level 3 calibration vial into the
third Extra vial.
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121
26- Mix for 0.3 minute
Mix Level 3 calibration vial for 0.3 minute.
27- Heat for 0.0 minute
Skip step.
28- Mix for 0.0 minute
Skip step.
29- Wait for 0.0 minute
Skip step.
30- Remove 0 µL to
waste at 2.0mm height
Skip step.
FlushP
0 µL Solv S-1
Overlap Enable
Flush lines with selected solvent.
Do not begin sample preparation on next set until current set has
completed injection.
No
You need an extra vial for each calibration vial.
This procedure requires
that 3 Extra Vial(s) be
placed immediately
after each Sample Vial.
STARTING A SAMPLE
PREPARATION RUN
Once you have filled in all of the parameters in one of the four sample
preparation templates, you're ready to enter the rest of your
parameters into the Edit Menu. Before you start your run, you need
to:
•
Enter your Timed Events Menu parameters
•
Enter your More Menu parameters
•
Load your file
•
Press [RUN]
PREPARING AND RUNNING
YOUR SAMPLES
AUTOMATICALLY
A Simple Dilution
Run
If you are doing a simple dilution, you can put your concentrated
sample, diluent, reagent (if used) and extra vial in one sample set.
The autosampler will automatically inject the last vial used during
sample preparation, in this case, the contents of your extra vial.
Figure 4.14 illustrates this example. For this example, vial A01 is a
reagent (Reag-A), vial A02 contains the sample concentrate, and vial
A03 is the extra vial.
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Thermo Electron
Sample Set
File 1:
1
EXAMPLE
Injection Volume
10
Injections/Sample
1
Cycle Time
0.1
Reag-A
A01
First Sample Vial
A02
Number of Samples
File requires 1 extra
vial(s) after each sample
Add to Queue?
Yes
Figure 4.14 An example of a simple dilution run using sample
preparation and injection in the same sample set
NOTE: In Figure 4.14, the vial in the First Sample Vial position (A01) is
your first preparation vial, not your first injection vial.
Preparing
Calibration Vials
and Injecting Them
If you want to prepare several calibration vials and then inject them,
it's easiest to set up two sample sets. The first set will be your sample
preparation set without injections; the second set will be for your
actual run with injections.
For the first set, you set Injections/Sample to 0 in the Edit and
Samples Menus. For the second set, you set Injections/Sample to the
desired number of sample injections in both menus. Figure 4.15
illustrates the Samples Menu for the preparation set; Figure 4.16
illustrates the Samples Menu for the injection set.
Samples Set
File 1:
1
PREP
Injection Volume
10
Injections/Sample
0
Cycle Time
10.0
First Sample Vial
A01
Number of Samples
10
Add to Queue
Yes
Figure 4.15 The Samples Menu for the Prep Set
NOTE: You can decrement the Injections/Sample field to 0 only if you have
first selected a sample preparation template.
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Samples Set
File 2:
2
INJECT
Injection Volume
10
Injections/Sample
5
Cycle Time
10.0
First Sample Vial
A01
Number of Samples
10
Add to Queue
Yes
Figure 4.16 The Samples Menu for the Injection Set
NOTE: If you enter your sample preparation and other run parameters,
load your file and then start your run, the autosampler will prepare your
samples as you would expect, but inject only the vial used by the last Add
step in the sample preparation template.
STOPPING A RUN
Similar to operation without sample preparation, the [STOP] key has
several functions depending upon the instrument's status when the
[STOP] key is pressed. However, the [STOP] sequence discussed in
Chapter 3 is slightly different when sample preparation is used. Refer
to Figure 4.17 as you read about the STOP/ABORT sequence for
sample preparation.
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Queue is running
[STOP]
(User Abort)
STOP requested
Select ABORT SAMPLE/CONTINUE
CONTINUE
Stops the queue
QSTATUS goes to IDLE
Resumes current
activity with
current vial
[RUN]
Restarts queue from
the beginning
AS-Z016E/DG
System
Error
ABORT
Figure 4.17 The Queue Stop/Abort Sequence
If the queue is in RUN when you press the [STOP] key, the
autosampler suspends its current operation and waits for you to make
a selection from the ABORT/Continue Menu as described in
Chapter 3.
If you press [+] to select Continue, the autosampler returns the queue
to the ACTIVE state and resumes the motor activity (as if you had not
pressed the [STOP] key), just as it did without sample preparation.
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125
If you select ABORT or if a system error occurs, the autosampler:
•
Aborts the processing of the current sample
•
Returns the active vial to the vial tray
•
Flushes the lines (both syringes)
•
Puts the queue in the IDLE state (not PAUSE)
You can then press [RUN] to restart processing from the beginning of
the queue.
This [STOP] sequence allows you to resume the run from the
ABORT/CONTINUE menu. But, if you press [STOP] and select
ABORT, you can only restart (not resume) the queue from the
beginning.
CUSTOMIZING THE
TEMPLATES
Guidelines
In our examples so far, the contents of the vials in the template
positions have matched the autosampler's names for those positions.
For example, we have placed the sample in the Sample position and
stock standards in the Reagent A, B, and C positions. We have also
used the vial positions in a sequential order. However, the
autosampler does not limit you to this approach. In customizing the
templates to your own applications, the following more general rules
may help:
1. In each dilution cycle, you have seven vial positions to work
with: three source-vial positions (Reagents A, B, C) and four
vial positions that may be either source or target vials (Sample,
Sample + 1, Sample + 2, and Sample + 3).
2. You can arrange your Pickups and Add vials in any order you
wish within a dilution cycle as long as you remember that the
autosampler locates the extra vials (Sample + 1 - 3) in
relationship to the vial you've assigned to the Sample position.
For example, you could Pickup from the Sam + 3 vial and Add
to the Sample vial. (But, you couldn't Add to the ReagentA
vial since, according to rule 1, it cannot be a target vial.)
3. You can use the vial positions in any order you wish within a
dilution cycle. For example, even though the autosampler
cannot skip Sample vial positions, you can program it to
prepare interspersed calibration vials by Adding to the Sam + 2
position, without using the Sam + 1 position in any of the steps
in a given dilution cycle.
HINT: In loading your tray, if you want your first calibration vial to
be A01, then you can use rule 3 and the autosampler's "wrap" feature
by assigning the First Sample Vial as C39 (Sam + 2 will then be
A01). However, you will still need to create two sample sets: one to
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Thermo Electron
prepare and the other to run your calibration and sample vials.
Gradient Delay
Gradient delay is a user-defined value that delays the injection of a
sample. This delay is the amount of time required for a change in the
mobile phase to travel from the gradient proportioning valve to the
injection valve. The volume of mobile phase contained between the
two valves is also referred to as the precolumn volume (PV).
Although not required for normal autosampler operation, a gradient
delay is recommended when using large volume pre-filters or low
flow rates (<0.5 mL/min). The gradient delay time can be found
under the FILE/Edit/More Menu. You then use the [+] or [-] keys to
enter the desired time in minutes (Figure 4.18).
The following procedure is used to calculate the gradient delay time
for your HPLC system. (See Figure 4.19.)
1. Inject an unretained sample into your LC system with a column
in-line and using 100% A as the mobile phase (e.g., MeOH).
2. Change to 100% B (e.g., 0.1% Acetone in MeOH) as fast as
possible (0.1 minutes for SpectraSYSTEM pumps).
3. Subtract the retention time (Rt) of the unretained peak from the
retention time of the start of the baseline shift (T2-T1).
4. Subtract the time required to switch mobile phases from
100% A to 100% B.
5. The resulting time is the gradient delay time.
EXAMPLE
The following example illustrates how to apply this procedure.
An unretained peak has a retention time of 3.0 min. with a baseline
shift at 4.5 min. and a complete mobile-phase switch time of 0.1 min.
The gradient delay is calculated as follows:
Gradient Delay
= (T2-T1) - 0.1 min
= (4.5 - 3.0) - 0.1
= 1.4 min
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127
Equilibration Time
0.0
Gradient Delay
0.00
--------------------------------------------------------Viscosity
Medium
Flush Volume
250.0
Injection Type
Push
Injection Range
1-1500
Needle Height
2.0
Figure 4.18 The More Menu (Gradient Delay)
Pump to Detector
Injector
to
Detector
Pump
to
Injector
Baseline Shift
Inject
T1
T2
Time (in minutes)
Figure 4.19 The delay time calculation using a hypothetical chromatogram
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AS-Z012E/DG
Unretained Peak
Diagnostics (Tests Menu)
The SpectraSYSTEM autosamplers have built-in diagnostic routines
accessible from the Tests Menu. These tests enable you to access the
operation of your autosampler. Before describing each of these tests,
we'll describe how the diagnostic tests are run and how to interpret
the error screen information, should a test fail.
All of the test screens are arranged similarly. A test is initiated by
pressing [ENTER]. While a test is in progress, the name of the test is
displayed along with three dots (Figure 4.20).
Test External Contacts:
...
Figure 4.20 The In-progress Screen
INTERPRETING THE
RESULTS
When the test is complete, either a PASS or FAIL result is displayed.
Should a test fail, an error screen similar to that shown in Figure 4.21
will appear that contains the following information:
•
Test identification
•
Fault code (diagnostic error)
•
Module identification
•
[RUN]/[STOP]
Connector:
ERROR #3
Syringe1
[RUN]/[STOP]
Figure 4.21 An example of an error screen
NOTE: The error number in this screen is just an example to show you the
screen layout.
Test Identification
The test identification is the name of the test being run.
Fault Code
The fault code is the error number associated with the specific failed
result. Document and report the fault code to your service
representative to receive further assistance in diagnosing and solving
the problem.
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129
Module
Identification
This field displays the specific autosampler board or assembly where
the problem occurred.
[RUN]/[STOP]
This field prompts you to press [RUN] to repeat the test or to press
[STOP] to exit the current test and return to the Tests Menu.
TEST
DESCRIPTIONS
All diagnostics and built-in assessment routines are accessible from
the Tests Menu (Figure 4.22). Each of the routines is described here.
Software Version
Display Input States
Display Output States
ROM Test
RAM Test
Hardware Test
Test External Contacts
Test Comm Port
Initialize EEPROM
Figure 4.22 Tests Menu
Software Version
Selecting this field displays the current software version number,
x.yy.zz.
Display Input States
This menu (Figure 4.23) shows the current states of the TTL-input
device signals (Hi or Lo). This information can be used to check the
level of the Pump Ready and Injection Hold input lines from other LC
system modules. You can use this information to diagnose situations
where the autosampler is waiting for ready or waiting to inject. Make
sure the Input Polarity is set to the proper level for each of these
signals. Pressing the up- or down-arrow keys returns you to the Test
Menu's Display Input States field should you wish to repeat the test.
NOTE: You can change the active polarity of a specific input signal from
the OPTIONS Input Polarity Menu.
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Pump Ready
Inj Hold
Hi
Hi
Figure 4.23 Display Inputs States Menu
Display Output
States
This menu shows the current states of the TTL-output device signals
(Hi or Lo). This information is used when hardwiring your
autosampler to other LC system modules. Pressing the up- or downarrow keys returns you to the Test Menu's Display Output States field
should you wish to repeat the test.
NOTE: You can change the active polarity of a specific input signal from
the OPTIONS Input Polarity Menu. This menu shows the states of the TTL
output device signals (Hi or Lo) as shown in Figure 4.24.
Display Output States
AS Rdy Inject Grad
Stop
Lo
Lo
Lo
Lo
Figure 4.24 Display Output States Menu
Pressing the up- or down-arrow keys returns you to the Test Menu's
Display Output States field should you wish to repeat the test.
Display Timed
Events
This menu (Figure 4.25) displays the current states of the timed
events outputs. This information is used when hardwiring your
autosampler to other LC system modules. Refer to Appendix A
"System Communication" for more information.
TF1
TF2
TF3
TF4
Lo
Lo
Lo
Lo
Figure 4.25 Display Timed Events Menu
RAM Test
This test (Figure 4.26) writes to and then reads the contents of
memory. If the write information is the same as the read information,
the test is successful. You may wish to run this test once a month to
verify proper software operation.
Short Memory Test:
Figure 4.26 RAM Test Menu
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131
If successful, the autosampler returns a PASS result (Figure 4.27). If
unsuccessful, the autosampler returns a "FAIL" result and displays an
error message. Document the test results and error message, and call
your Thermo Electron service representative.
Short Memory Test: PASS
Figure 4.27 Successful RAM Test
Press [STOP] to return to the Tests Menu.
ROM Test
It evaluates check-sums on the contents of the instrument's ROM and
compares the results to known values.
If successful, the autosampler returns a PASS result. If unsuccessful,
the autosampler displays the number of the failed chip(s) along with
the regular error screen information (Figure 4.28). If the test fails,
document the test results (if failed) and call your Thermo Electron
service representative.
ROM Test:
Error #: U78, U79
CPU
[RUN]/[STOP]
Figure 4.28 Failed ROM Test
Hardware Test
This is the most important autosampler diagnostic. It checks all of the
cable connections, sensors, motors, circuitry, and attached
components.
Before running this test, be sure that the XYZ arm is in the home
position (at the left rear of the sample compartment), or false errors
may be reported. You can use the Home Arm command in the
Commands Menu (see "Special Commands" on page 96) if the arm is
not at home.
If the Hardware Test fails, the screen displays the test identification,
fault code, applicable module or connection, and the [RUN]/[STOP]
fields. For example, the message "J10/J40/J45" in Figure 4.29
indicates that further troubleshooting is required. Document the
screen information and report it to your Thermo Electron service
representative when you call for assistance.
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Connector:
ERROR #3
J10/J40/J45
[RUN]/[STOP]
Figure 4.29 An example of an error screen
Pressing [STOP] returns you to the Tests Menu.
Most of the failed Hardware Test results require that you contact your
service representative for assistance. However, there are several
messages that indicate problems that you can most likely resolve
without a service call. These are described here.
Opt Config
This message indicates an incorrect configuration. Check the
Configurations Menu (under OPTIONS) and verify that the options
field entries match your installed options.
Arm,*
This message indicates that the XYZ arm was not in its "home"
position when the Hardware Test was initiated. Select
/COMMANDS/, /Arm/, /Home arm/.
Test External
Contacts
This test activates all of the external contacts and tests for proper
activation. It should not be run while the external contacts are wired
to other equipment, or the test will report false errors. Before running
this test, disconnect the 12-pin connector block from the rear of the
instrument. After the test is complete, plug the connector back in to
resume normal instrument operation.
Pressing [ENTER] with the cursor in the Test External Contacts field
displays a warning prompt (Figure 4.30).
Disconnect all external contacts
CONTINUE
Figure 4.30 The External Devices Menu
From this menu, you can use the [+] and [-] keys to select
CONTINUE or ABORT. Selecting Continue starts the test.
Selecting ABORT discontinues the test and returns you to the Test
External Contacts field of the Tests Menu. If ABORT is displayed
and you want to resume to test, press the left- or right-arrow keys.
The display will again show CONTINUE.
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133
Test COMM Port
A loop-back connector is required to run this test used to assesses the
operation of the communications port located on the instrument's rear
panel. This test sends a signal to the communications port and
expects a signal in return. A successful result is reported as a PASS;
an unsuccessful result is indicated by the regular error screen.
NOTE: If you wish to perform this test, contact your Thermo Electron Field
Service Engineer for the procedure and necessary parts information.
Initialize EEPROM
The Electronically Erasable Programmable Read Only Memory
(EEPROM) stores all files, user-set variable values, and system
configuration information when the power is turned off.
Reinitializing the EEPROM restores the default values to all files and
erases the system configuration information. Pressing [ENTER] in
this field displays the message shown in Figure 4.31:
Reset all system values
YES
Figure 4.31 The Initialize EEPROM Menu
If you want to exit this routine before running it, you can:
•
Press the [+] [-] keys to select No
•
Press [MENU] to call up the Main Menu
•
Press [Status] to display the Status Screen
•
Press the up-arrow key to return to the Tests Menu.
NOTE: This routine erases any current file values and returns all
field values to their default values. Initialize EEPROM also erases all
configuration information. After running this routine, be sure to reset
all installed options to "YES" in the Configurations Menu.
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5
Required Maintenance
Introduction
This chapter contains the maintenance schedule and procedures for
your autosampler.
SPECIFIC HAZARDS
For your own safety, please familiarize yourself with and follow these
safety precautions when using this instrument:
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.
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135
Maintenance Schedule
Your autosampler requires only a few simple maintenance procedures
to keep it in optimal working condition. Table 5.1 contains the
Maintenance Schedule for the autosampler. The yearly items should
be performed only by a Thermo Electron service representative or
other qualified person.
Table 5.1 Maintenance Schedule
Frequency
Procedure
Performed By
Daily
Empty flush-solvent tray
User
Monthly
Inspect the sample needle
module.
User
Check the solvent tubing
and connections for leaks.
User
Run a standard sample.
User
Clean the drop catch
User
Replace the flush-solvent
inlet filter.
User
Replace needle assembly
User
Replace the injector
valve's rotor seal
User
Semi-annually
Annually
Clean and lubricate the
syringe drive mechanism.
Service
Clean and lubricate the
XYZ arm.
Service
Verify the XYZ arm
alignment.
Service
NOTES: Maintenance of the autosampler is the responsibility of the user.
Routine maintenance is not provided under warranty; however, planned
maintenance contracts are generally available. Please contact your local
representative if you are interested in purchasing a planned maintenance
contract.
This chapter contains recommended maintenance intervals based
upon operation of the autosampler under typical conditions.
Operation of the autosampler under certain harsh conditions
(corrosive solvents, extreme temperatures, etc.) may require more
frequent maintenance than indicated in this chapter.
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Procedures
EMPTYING THE
FLUSH-SOLVENT
TRAY
For normal operation, you may need to empty the tray at least once or
twice a day. To empty the tray:
1. Be sure the autosampler is in idle status.
2. Carefully slide the tray out without spilling the collected
solvent.
3. Empty the tray, rinse it off with water, and slide it back into
place under the black elbowed, barbed piece of tubing.
CAUTION! Wear skin and eye protection and dispose of hazardous
waste according to local, state, and federal regulations.
NOTE: Be sure that the level of the liquid in the waste container is
below the level of the instrument or waste will collect in the lines and
could backflush into the instrument.
REPLACING THE
ROTOR SEAL
Replacing the rotor seal includes the following procedures:
•
Removing the injector valve
•
Disassembling the valve
•
Cleaning the stator
•
Reassembling and replacing the valve
NOTE: If you are only replacing the rotor seal, you don't have to remove
the injector valve from the instrument; however, it's easier. If you wish to
replace the rotor seal with the valve installed, proceed to the section
entitled, "Disassembling the Injector Valve" on page 140.
Removing and
reinstalling the
Injector Valve
To disassemble the injector valve (Figure 5.1):
1. Using a 1/4" open-end wrench, disconnect tubing from ports 2,
3, 5, and 6.
2. If you have the Column Oven Option and are using the 100 µL
loop shipped with your instrument, you remove the sample
loop. Otherwise, proceed to step 3.
Thermo Electron
137
3. If you have purchased the column oven, loosen and remove the
two Phillips-head screws that secure the injector valve collar to
the front of the oven. Then remove the collar.
4. Remove the two Phillips screws that secure the valve mounting
plate.
5. Carefully pull the valve out of the instrument.
6. Reverse these steps to reinstall the valve.
2
3
1
4
6
5
RUN
STATUS
MENU
STOP
ENTER
SAMPLES
SpectraSYSTEM
AS3000
AS-Z007/FM
WARNING
Figure 5.1 The injector valve
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Thermo Electron
NOTE: When reinstalling the bracket/valve assembly, be sure to
orient the bracket with the "locating-hole" corner in the lower right
as shown in Figure 5.2. You should be able to see the pin through the
locating hole.
Mounting
Screw
Locating
Hole
AS-Z019E/DT
Mounting
Screw
Figure 5.2 The correct injector valve bracket orientation
Disassembling the
Injector Valve
NOTE: This repair must be done in a clean environment. Component
cleanliness will affect the useful life of your injector valve.
NOTE: Do not activate the valve mechanism with the injector valve removed
from the instrument or valve mechanism damage could occur.
To disassemble the injector valve:
1. Follow the procedures above for removing the injector valve.
2. Using an Allen wrench, remove the 9/64" hex cap screws from
the top of the valve (Figure 5.3).
3. Carefully lift the stator from the valve body and set it onto a
clean surface.
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139
Valve Shaft
Rotor Seal
Valve Body
Hex Cap Screws
Stator
Figure 5.3 The injector valve's components
NOTE: Be careful not to drop the two rotation stop pins that protrude from
the valve base (Figure 5.4).
4. Remove the valve body.
5. Before removing the worn rotor seal, note the following on the
rotor seal (Figure 5.4):
140
•
the three circular port grooves on one side of the rotor (the
other side is smooth)
•
the small locating notch on the rotor's rim (not the V
stamped on the seal).
Thermo Electron
AS-Z021E\DT
Valve Base
3 Circular
Port Grooves
Rotor Seal
Vertical
Stop Pins
Notch
Horizontal
Locating Pin
Valve Body
AS-Z018E\DT
Valve Base
Figure 5.4 The injector valve with valve body removed and rotor seal
exposed (NOTE: The mounting bracket is not shown.)
6. Remove the old rotor seal. A thin, flat-bladed tool may be
needed.
Cleaning the Stator
Clean the stator as follows:
1. Inspect the stator to determine if it requires cleaning.
2. If it is dirty or greasy, swab it with HPLC-grade methanol. If
more stringent cleaning is required, use a sonicator.
3. Inspect the remaining valve components for contamination.
Clean as necessary.
NOTE: If the stator is scratched, replace it. Scratches can damage
the rotor seal and cause valve leaks.
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141
Installing the
Rotor Seal
To install the rotor seal:
1. Install the new rotor seal in exactly the same orientation as the
old one. Be sure that the rim notch is in line with the
horizontal locating pin and that the circular port grooves are
visible.
2. Install the valve body.
3. Install the stator onto the valve body carefully, observing that
the two locating pins in the valve body (Figure 5.4) line up
with the two holes on the bottom side of the stator assembly
(Figure 5.5).
NOTE: Be sure to hold the valve shaft so that the horizontal locating
pin touches the left stop pin as you reassemble the valve. This puts
the valve into the INJECT mode (the same mode as when removed
from the instrument). If the pins are not in the correct orientation, the
valve shaft may not be in the proper orientation for reinsertion into
the instrument.
Locating Hole
Screw Hole
(3X)
Stator
Face
Locating
Hole
Figure 5.5 The bottom of the stator assembly
NOTE: These locating pins orient the stator assembly in one of two
positions, but only one position aligns the mounting holes in the valve body
with the screw holes in the stator.
4. Insert the three cap screws so that each one just contacts the
stator.
NOTE: There should be a uniform gap between the stator and the valve
body to maintain the necessary pressure for proper valve operation and to
avoid valve binding. Do not over-tighten the valve.
5. Tighten each cap screw an equal amount until all setscrews are
in equal contact with the valve body.
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AS-Z017E\DT
Setscrew
(3X)
REPLACING THE
FLUSH SOLVENT
INLET FILTER
Properly filtering your flush solvent extends the life of your injector
valve and column. Replace the inlet filter as follows:
1. Be sure the autosampler is in the IDLE mode.
2. Remove the flush-solvent bottle cap and lift out the inlet
tubing.
NOTE: Be sure to follow all of the safety precautions for each
solvent as prescribed by the manufacturer's MSDS.
3. Put on a new filter cartridge and put the line back into the
bottle. Tighten the solvent cap.
4. Flush the inlet filter and lines with an appropriate volume of
solvent (~5 µL).
SAMPLE NEEDLE
Module
The sample-needle module resides in the sample tower and is
responsible for piercing the vial septum and removing sample from
the vial. It easily snaps out of the needle mount and is very easy to
maintain.
Removal
Use the following steps to remove the sample needle module:
1. Remove the sample needle fitting from port 2 of the injection
valve using a 1/4" open-end wrench.
2. Carefully (to avoid damage to the sample tubing) release the
sample needle module from the tower by grasping the module
edges and pulling towards the rear of the autosampler.
3. Pass the needle fitting through the autosampler's front bulkhead
and remove the needle module from the autosampler.
NOTE: If you are replacing (not inspecting) the needle, you may wish to cut
the used needle to make it easier to pass the fitting through the bulkhead.
Cleaning
To clean the sample needle module:
1. Inspect the needle module.
2. Rinse in water or sonicate if contamination is present.
3. If internal plugging is suspected, pass a cleaning wire through
the needle and flush the needle with high flow from an LC
pump.
NOTE: The sample needle module is glued during manufacture and
cannot be disassembled. Attempts to do so will damage the module.
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143
Installation
Replace the sample needle module every six months. The needle
module is easy to replace because it is designed to snap into place.
1. Remove the new module from the replacement kit.
2. Pass the valve end of the needle through the hole on the
underside of the column compartment.
3. Snap the needle assembly into the mounting brackets on the
backside of the sample tower (Figure 5.6).
4. Place the tube fitting and ferrule onto the needle and install it
into port 2 of the injection valve (Figure 5.1).
Drop Catch
The drop catch (Figure 5.6) removes the last drop of sample from the
end of the needle after injection. It must be flushed monthly as
follows:
1. Remove the needle module assembly as described above, but
leave the drop catch installed.
2. Flush the drop catch with water, an organic solvent, an acid or
base (as appropriate for the sample), and once more with water.
Then dry it with a clean tissue.
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Needle Module Assembly
Front Bulkhead
Drop-Catch
Location
Needle
Sample
Tub
Injection
Tower
AS-Z020E\DT
THIS
SIDE
OUT
Figure 5.6 Replacing the sample needle module
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145
SYRINGE
REPLACEMENT
The section describes how to replace the sample (250 µL) syringe and
the large (2.5 mL) sample preparation syringe.
250 µL Sample
Syringe
You can replace the 250 µL sample syringe without removing the
syringe drive assembly, but you must move the syringe drive about
two inches out by interrupting a flush cycle. Replace the sample
syringe as follows:
1. From the Main Menu, choose /Commands/, and /Flush Sample
Syringe/.
2. When the plunger is lowered, turn off the power.
3. Hold the silver base of the plunger to prevent it from turning,
and loosen the knurled syringe set screw under the syringe's
plunger.
HINT: You may need to insert a screwdriver into the slot in the set screw to
get the screw started and then lower the plunger.
NOTE: The syringe set screw is "captive" and won't fall down into the
instrument's chassis.
4. To loosen the top of the syringe, hold the metal tip on the top
of the syringe barrel, unscrew the used syringe barrel and lift it
out of the instrument.
NOTE: Don't try to loosen the syringe by holding the barrel. The syringe
could break between the glass barrel and the metal tip.
HINT: Push up on the syringe plunger while lifting it out of place.
HINT: Install the syringe in its fully expelled position to prevent an air
bubble in the syringe barrel.
5. Reverse steps 4 and 5 to install your new syringe.
2.5 mL Preparation
(prep) Syringe
If you have the Sample Preparation Option, you can position the large
(2.5 mL) syringe's plunger in the down position by interrupting a
flush cycle, as follows:
1. Be sure the autosampler is in the IDLE mode.
2. Move the XYZ arm out of your way by pressing [MENU],
Commands, /Arm/, /Align Arm/, and selecting position A01.
Then press [ENTER].
3. To start the flush cycle, from the Main Menu, choose
Commands, and Flush Prep Syringe.
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Thermo Electron
4. When the plunger has lowered, turn off the power.
5. Hold the plunger to prevent it from turning, and loosen the
knurled syringe set screw under the syringe's plunger.
HINT: You may need to insert a screwdriver into the slot in the set screw to
get the screw started.
NOTE: The syringe set screw is "captive" and won't fall down into the
instrument's chassis.
6. To loosen the top of the syringe, hold the metal tip on the top
of the syringe barrel, unscrew the used syringe barrel and lift it
out of the instrument.
NOTE: Don't try to loosen the syringe by holding the barrel. The
syringe could break between the glass barrel and the metal tip.
HINT: Push up on the syringe plunger while lifting it out of place.
7. Reverse steps 5 and 6 to install your new syringe.
HINT: When reinserting the bottom set screw, you may need to pull the
plunger down slightly so that the end of the screw will reach the bottom of
the plunger.
SAMPLE LOOP
A number of different size sample loops are available for the
SpectraSYSTEM and SpectraSERIES autosamplers. Refer to the list
of spare parts and consumables in the Appendix C of your manual.
To change the sample loop:
1. Be sure that the autosampler is in the IDLE mode.
2. Using a open-end wrench, loosen the fittings at ports 1 and 4,
and pull out the two ends of the sample loop.
3. Insert the ends of the new sample loop in the same orientation
as the old one and tighten the fittings.
NOTE: If the Sample Preparation Option is installed, to ensure maximum
reproducibility, do not perform small-volume injections with the large
(preparation) syringe.
FUSES
Instrument power is supplied by two T5A fuses housed in the fuse
compartment on the lower left corner as you are facing the
instrument's rear panel (Figure 5.7).
Thermo Electron
147
6-Port Solvent-Selection
Valve
COMM Port
12-Pin Terminal
1
6
3
4
2
Bottle-Bracket
Screws
5
TERMINAL CONFIGURATION
Fuse Compartment
AC Power
Figure 5.7 The autosampler's rear panel
148
Thermo Electron
Air-Supply Port
AS-Z004E\DT
WARNING
To replace the fuses:
1. Use a flat blade to press down on the plastic tab at the top edge
of the fuse compartment and pull out the fuse holder.
2. Pull both fuses out of the holder and discard. Place the new
fuses into the holder with the metal end visible.
3. Snap the fuse holder back into place.
Maintenance Options
SHUTDOWN
If the autosampler is not used for a period of time (after completing
an overnight or over-the-weekend run), you can set it to turn off
automatically. You enter the shutdown parameters in the OPTIONS
Shutdown Menu (Figure 5.8).
Configurations
Ready Participation
Input Polarity
Output Polarity
Shutdown
Maintenance Log
Communications
Figure 5.8 The Options Menu
Figure 5.9 illustrates the autosampler's Shutdown Menu.
Delay Time
Off
Oven Shutoff
Yes (Only if oven installed)
TrayTemp Shutoff
No (Only if tray temperature control installed)
Heater Shutoff
No (Only if sample prep installed)
Pump Shutoff
Yes
Flush Volume
400
Prep Flush Volume
1000 (Only if sample prep installed)
Figure 5.9 The Shutdown Menu
Delay Time
In this field you specify the desired amount of time that the
autosampler should wait before initiating the shutdown procedure
(Off, 5, 10, 20, 30, 45, 60, 90, 120, 240, 480 minutes).
Oven Shutoff
Use the [+] and [-] keys to select Yes or No. Selecting Yes causes the
autosampler to automatically turn off the oven after the Shutoff Delay
time has elapsed.
NOTE: If you are using the Shutdown function with methods that call for
heating and cooling, be sure to allow a long enough delay time (not less
than 10 minutes) for thermal equilibrium to occur.
Thermo Electron
149
TrayTemp Shutoff
Use the [+] and [-] keys to select Yes or No. Selecting Yes causes the
autosampler to automatically turn off the Tray Temperature Control
Option after the Shutoff Delay time has elapsed. You may or may not
wish to turn off tray temperature control depending upon your sample
requirements.
Heater Shutoff
Use the [+] and [-] keys to select Yes or No. Selecting Yes causes the
autosampler to automatically turn off the heater after the Delay Time
has elapsed. This field appears only if the Sample Preparation Option
is installed.
Pump Shutoff
Use the [+] and [-] keys to select Yes or No. Selecting Yes causes the
autosampler to automatically activate the Pump Stop output after the
Delay Time has elapsed. Turning off the pump retains your solvent
supply and prevents air from being pumped into the lines should the
solvent reservoirs become depleted.
Flush Volume
This field allows you to flush the lines using the 250 µL sample
syringe. Specify the flush volume in microliters (200 - 5000). The
default value is 400 µL.
Prep Flush Volume;
This field allows you flush the lines using the 2.5 mL sample
preparation syringe. Specify the flush volume in microliters
(0 - 6500). This field appears only if the Sample Preparation Option
is installed.
MAINTENANCE LOG
The Maintenance Log Menu (Figure 5.10), accessible from the
OPTIONS Menu, monitors the instrument's maintenance status and
counts and displays the number of activations for each of the
autosampler's routine maintenance parts listed in the menu.
Total System Cycles
Injector Valve
Needle Wear
Sample Syringe
Prep Syringe
Figure 5.10 The Maintenance Log Menu
Instrument Maintenance Status
The Maintenance Log Menu provides a convenient way to keep track
of autosampler maintenance. When you first access the Maintenance
Log Menu from the OPTIONS Menu, the "Maintenance Required"
confirmation message is displayed if instrument maintenance is
required. This message is displayed if any of the #Cycles field values
150
Thermo Electron
in any of the Maintenance Log menus is greater than the number of
cycles entered in the Due fields. See below for more information on
these fields.
No message is displayed if maintenance is not required. Pressing the
down-arrow key clears the confirmation message and returns you to
the Maintenance Log Menu.
Total System Cycles Menu
The Total System Cycles Menu (Figure 5.11) displays the total
number of instrument cycles in the Total Cycles field. This field
value is set to zero at the factory and cannot be reset. The Note field
is a six-character, alphanumeric, user-programmable field. You can
use the [+]/[-] keys to enter the last service date or other comments
regarding the maintenance of your instrument.
#Cycles
35
Note
Feb 93
Figure 5.11 The Total System Cycles Menu with the Note field used
to indicate the next maintenance due date
Injector Valve Menu
With the exception of the Total System Cycles Menu, all of the
Maintenance Log menus are identical to the Injector Valve Menu
(Figure 5.12). Therefore, only the Injector Valve Menu will be
described in detail in this chapter.
#Cycles
202
Due
Note
24000
021593
--------------------------------------------------------Reset #Cycles
YES
Figure 5.12 The Injector Valve Menu
#Cycles: This field displays the number of activations of the specific
autosampler part since the last time this field (counter) was reset
(0 - 999,999). The #Cycles value for each of the Maintenance Log
menus is included in the menu descriptions below.
Due: This is a six-character numeric field where you enter the
number of cycles before maintenance is due. If the #Cycles field
value is larger than the Due field value, the "Maintenance Due"
message is displayed when you access the Maintenance Log Menu
from the Options Menu. The default maintenance Due field value is
12,000 (approximately once a year). The default values for the other
Maintenance Log menus are included in the menu descriptions below.
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151
NOTE: The Due field default values for all of the Maintenance Log menus
are estimates only. The actual values appropriate for your instrument will
vary with the type of applications and the specific conditions being used
with your LC. If the "Maintenance Required" message is displayed, parts
replacement is recommended. However, you may wish to increase the Due
value and experiment with extending the time before maintenance is
performed. Conversely, if instrument performance indicates, you may wish
to decrease the Due field value for some of the Maintenance Log menus.
Note: Similar to the Total System Cycles Menu, this is a sixcharacter, alphanumeric, user-programmable field. We recommend
that you use the [+]/[-] keys to enter the last service date in this field.
However, you can also use this field to document contact information
or other comments regarding the maintenance of the specific
autosampler part. Reset #Cycles: Pressing the down-arrow key
displays the Reset #Cycles field used to reset the #Cycles counter for
each autosampler part. To reset the counter to 0 after maintenance is
complete, press the [+]/[-] keys to select YES, and press [ENTER].
The confirmation message ** Counter Reset ** is displayed. Most
likely, your service representative will reset the #Cycles field value
when the instrument is serviced.
Needle Wear Menu
The #Cycles field in this menu is incremented each time the needle
pierces a vial. The Due field's default value is 10,000, which
corresponds to approximately every six months.
Sample and Prep Syringe Menus
The #Cycles field in both of these menus displays each full syringe
stroke. However, the instrument tracks the actual distance of linear
travel, thus compensating for different injection (stroke) volumes.
The default Due field value for both menus is 24,000, which
corresponds to approximately once a year.
152
Thermo Electron
A
Troubleshooting
Introduction
This appendix provides information to help you troubleshoot possible
autosampler problems. We have included the following:
•
Table 1.1: Troubleshooting guide to general chromatographic
problems
•
Table 1.2: Troubleshooting guide to autosampler-specific
chromatographic problems
•
Table 1.3: Troubleshooting guide to autosampler hardware
problems
•
Display Messages
•
Information on testing tubing restrictions
If, after reviewing the troubleshooting table and diagnostics included
in this chapter, you would like additional assistance in
troubleshooting a particular problem, don't hesitate to contact the
Thermo Electron Technical Support Group or your service
representative. The telephone numbers are found on the back cover
of this manual.
General Chromatography Troubleshooting
This section focuses on general chromatographic problems and is
designed to help you define the particular instrument or condition
causing the problem. Once you have ascertained the cause, use the
ordered outline to fix the problem or refer to the individual
SpectraSYSTEM instrument's troubleshooting guide. Each of these
problems can be detected by monitoring the chromatography running
conditions or by observing the chromatogram.
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153
Table A.1 General LC Troubleshooting Table
Symptom
Cause/Remedy
1. No flow.
a) Check mobile phase connections
b) Check for leaks
c) Check pump troubleshooting guide
2. High back pressure.
a) Check flow rate and system/column
specifications
b) Check for tubing or column blockage
c) Check pump troubleshooting guide
3. Unstable baseline or
drift.
a) System/column not equilibrated; allow
more time
b) Check detector troubleshooting guide
c) Check pump troubleshooting guide
4. Baseline noise.
a) Check for air bubbles in system, degas
solvents
b) Check for system/solvent contamination
c) Check pump troubleshooting guide
d) Check your chromatography data system
troubleshooting guides
5. No peaks.
a) Check detector and data system
connections
b) Check autosampler troubleshooting
guide
c) Check sample retention with
chromatographic conditions
6. Contaminating/ghost
peaks.
a) Clean system and column
b) Check autosampler troubleshooting
guide
c) Check pump troubleshooting guide
154
Thermo Electron
Table A.1 General LC Troubleshooting Table, continued
7. Poor peak shape.
a) Check system for leaks
b) Check fittings and tubing lengths
c) Check column performance
d) Check autosampler troubleshooting
guide
e) Check pump troubleshooting guide
f) Check detector troubleshooting guide
8. Poor retention time
reproducibility.
a) Check system for leaks and bubbles
b) System/column not equilibrated, allow
more time
c) Check column performance
d) Check pump troubleshooting guide
e) Check autosampler troubleshooting
guide
f) Check data system troubleshooting
guide
9. Poor peak area
reproducibility.
a) Check column performance
b) Check autosampler troubleshooting
guide
c) Check data system troubleshooting
guide
10. Non-integrated or
too many peaks.
a) Check integrator or data system
troubleshooting guide
11. No instrument of
device control.
a) Check cable connections
b) Check system configuration
c) Check individual instrument
troubleshooting guide
d) Check integrator, ISM, or data system
troubleshooting guide
For more detailed chromatographic troubleshooting, refer to any
HPLC troubleshooting reference book or call your local sales or
service representative.
Thermo Electron
155
Table A.2 Autosampler-specific Chromatographic Problems
Symptom
Possible Problem
Remedy
1) Baseline drift when the
injector valve changes
from INJECT to FILL.
a) Lack of thermal stability in
the column oven
compartment, detector, or
mobile phase.
a) Eliminate drafts around the column
compartment. Wait until the column
oven has reached thermal
equilibrium. If the problem persists,
contact your local Thermo Electron
service representative.
2) Shifting retention times.
a) Temperature variations in the
lab.
a) Stabilize temperature.
b) Column-heater problems.
b) Call your Thermo Electron service
representative.
a) Inconsistent sample injection
volumes.
a) Check for a partially blocked injector
valve and observe the pressure. If
the problem persists, replace the
injection valve (refer to Chapter 5).
b) Leaks between the injector
valve and the column
causing inconsistent samplevolume injection.
b) Tighten/remake fitting.
c) Air bubbles aspirated into the
lines along with sample.
c) Replenish/prepare fresh sample and
repeat the injection.
3) Sensitivity changes.
c') Slow down syringe speed.
156
d) Incomplete sample mixing
during sample preparation.
d) Check mix time (Sample Preparation
Option, only). Thaw frozen samples
completely prior to sample
preparation to prevent sample
"layering."
e) Sample adsorption to the
walls of the injector valve or
sample loop.
e) Change the mobile phase
concentration.
Thermo Electron
Table A.2 Autosampler-specific Chromatographic Problems, continued
Symptom
3) Sensitivity changes,
continued
4) Irregular peak shapes.
5) Band broadening.
6) Smaller than expected
peak heights.
Possible Problem
Remedy
f) Sample solvent incompatible
with the mobile phase.
f) Change solvent, use mobile phase if
possible.
g) Sample insoluble in solvent.
g) Be sure sample solvent is lower
elution strength than mobile phase.
h) Air in syringe/sample lines.
h) Flush sample lines.
a) The volume of sample
injected exceeds the
column's capacity.
a) Decrease the volume injected or
dilute the sample.
b) The strength of the solvent
used to dilute the sample
exceeds the initial
concentration of the mobile
phase.
b) If appropriate for your application,
ensure that the flush solvent is of the
same composition as the solvent used
to dissolve the sample and that it
never exceeds the initial composition
of the mobile phase.
c) Degradation of sample
component(s).
c) Prepare fresh sample and repeat
injection.
a) Fitting problem at the
injector-column connection.
a) Inspect for damage, mismatch,
incorrect assembly.
b) Tubing ID too large.
b) Use 0.010-inch ID tubing.
a) Small injection due to
wrong-size syringe.
a) Check syringe size.
b) Wrong injection.
b) Check injection volume value in file.
c) Air in lines.
c) Flush.
d) Misadjusted syringe-drive
mechanism.
d) Call your Thermo Electron service
representative.
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157
Table A.2 Autosampler-specific Chromatographic Problems, continued
Symptom
7) Peaks during a blank
injection (from a
previous injection).
8) No peaks.
9) Incorrect precision and/or
accuracy problem on a
sample of known
content-random error.
10) Irreproducibility of peak
areas.
158
Possible Problem
Remedy
a) Sample carryover due to
residual sample in the sample
loop.
a) Insufficient flush volume. Increase
Injection Volume in Edit Menu; add
a blank vial between sample vials; or
use a stronger flush solvent.
b) Sample carryover due to
incorrectly assembled
sample-loop fittings (adding
dead volume that can contain
residual sample).
b) Check and remake fittings.
c) Sample too concentrated.
c) Dilute sample.
a) Plugged or bent needle.
a) Clean/replace needle. Tighten
fittings and syringe.
b) Leaking flush valve fitting.
b) Tighten fitting.
c) Plugged or leaking tubing.
c) Replace faulty tubing.
a) Air leak.
a) Tighten all fittings and repeat the
run.
b) Worn out syringe.
b) Replace.
a) Plugged or bent needle.
a) Clean/replace needle. Tighten
fittings and syringe.
b) Leaking flush valve fitting.
b) Tighten fitting.
c) Plugged or leaking tubing.
c) Replace faulty tubing.
d) Sample viscosity too low or
too high.
d) Change viscosity setting in the
FILE/Edit/More Menu.
e) Solvent viscosity too low or
too high.
e) Change Solv Viscosity setting in the
OPTIONS/Configurations Menu.
Thermo Electron
Table A.3 Autosampler Hardware Problems
Symptom
Possible Problem
Remedy
Startup
1) Nothing works when
instrument is turned on.
a) Power cord unplugged or
faulty.
a) Plug in/replace cord.
b) Blown fuse.
b) Replace fuse.
c) Defective power switch,
driver, transformer.
c) Call your local Thermo Electron
service representative.
2) Fan is running but
display doesn't work.
a) Display component(s)
failure.
a) Call your local Thermo Electron
service representative.
3) No response from keypad
but display works.
a) Defective keypad.
a) Call your Thermo Electron service
representative.
4) Column oven and/or tray
temperature control won't
turn on.
a) File isn't loaded.
a) Load file.
5) No RS-232-C
communications.
a) Further troubleshooting
required.
a) Call your Thermo Electron service
representative.
6) EEPROM fails to retain
memory.
a) Reinitialize the EEPROM.
a) If unsuccessful, may be a defective
PCB. Call your Thermo Electron
service representative.
7) External devices (timed
events) don't work.
a) Wrong settings.
a) Check Chapter 1 for proper pinouts.
b) Interface equipment problem.
b) Refer to respective instrument
manuals.
Communications
Thermo Electron
159
Table A.3 Autosampler Hardware Problems, continued
Symptom
Possible Problem
Remedy
Vials
8) Vial stuck in tower.
a) Needle bent.
a) Replace needle module (see
Chapter 5).
9) Vial stuck in mixer.
a) Run Clear Mixer routine in
the Commands Menu.
a) See page 99.
10) Instrument can't locate
vial.
a) Incorrect vial position
entered into Samples Menu.
a) Check Samples Menu.
b) TrayTemp Option incorrectly
selected in Configurations
Menu. (Instruments with
tray temperature control have
fewer tray rows.)
b) Check configuration.
c) Sample incorrectly placed in
sample tray.
c) Check tray arrangement.
d) Arm misaligned.
d) Home arm. See "Special
Commands" on page 96.
e) Arm movement restricted.
e) Remove restriction and home arm.
11) Vials don't fit in tray.
a) Incorrect vials.
a) Vial diameter should be 0.44 0.47 inches.
12) Broken vials.
a) Tray not aligned or not
seated properly.
a) Seat tray completely in tray chamber
and pull handle back toward you.
13) Torn septa.
a) Bent needle.
a) Replace needle.
160
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Table A.3 Autosampler Hardware Problems, continued
Symptom
Possible Problem
Remedy
Injector
14) Injector valve won't
switch.
a) Slipping injector drive.
a) Call your Thermo Electron service
representative.
15) Injector valve continues
to rotate until fault is
generated.
a) Loose or faulty sensor
cable.
a) Call your Thermo Electron service
representative.
16) Injector valve does not
rotate to the proper
position.
a) Valve requires excessive
torque.
a) Call your Thermo Electron service
representative.
17) Motor fails to stop after
syringe is fully returned.
a) Possible defective sensor.
a) Call your Thermo Electron service
representative.
18) Sample loop won't fill.
a) Blocked injector valve.
a) Follow procedures on page 143 to
isolate blockage and flush. Or take
apart valve and sonicate. DO NOT
clean valve with a cleaning wire or you
could damage it.
b) Blocked sample loop.
b) Backflush loop. If problem persists,
replace the loop.
c) Worn syringe.
c) Substitute a "known-good" syringe and
determine if symptoms persist.
d) Blocked needle.
d) Clean with a cleaning wire.
a) Too much valve seal
pressure against valve.
a) Decrease LC system pressure.
b) Particulate matter in valve.
b) Use an in-line filter before the injector
valve.
c) Inadequately filtered
sample.
c) Filter with a 0.5 µm filter before
injection.
19) Rapid valve-seal wear.
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161
Table A.3 Autosampler Hardware Problems, continued
Symptom
Possible Problem
Remedy
Injector, continued
20) Rapid valve-seal wear,
continued
d) Buffer crystallization.
d) Don't allow buffers to stand in system.
Run a flush sequence from the
Commands Menu when the system is
not in use.
21) Injection from incorrect
vial.
a) Incorrect vial position
entered in Samples Menu.
a) Check Samples Menu.
b) Sample placed in
incorrect position in vial
tray.
b) Check tray arrangement.
a) Error in file (calibration
vials placement/sample
preparation resource
placement).
a) Check file (calibration/sample
preparation template).
a) Incorrect vials.
a) See list of recommended replacement
parts in the front of the manual.
b) Alignment problem.
b) Call Thermo Electron.
c) Arm sticking.
c) Call your Thermo Electron service
representative.
a) Multiple/incorrect septa.
a) Verify a single septum of type
recommended.
22) Samples injected
incorrectly in a
pattern.
Needle
23) Needle bent or broken.
24) Needle plugging.
162
Thermo Electron
Table A.3 Autosampler Hardware Problems, continued
Symptom
Possible Problem
Remedy
Syringe
25) Loud buzzing from
syringe drive.
a) Defective limit switch.
a) Call your Thermo Electron service
representative.
26) Flush valve won't
activate.
a) Defective
valve/coupler/motor.
a) Call your Thermo Electron service
representative.
27) Flush valve actuated, but
syringe can't draw flush
solvent.
a) Plugged inlet filter in
solvent reservoir.
a) Replace inlet filter (Chapter 5).
a) XYZ arm motor stepping
failure.
a) Call your Thermo Electron service
representative.
29) Leakage during loop
filling.
a) Blocked loop or waste line
or valve not deflected far
enough.
a) Flush sample loop or replace; check
and flush waste line. Check the
sensor.
30) Cross-port leakage.
a) Worn rotor seal.
a) Replace according to instructions in
Chapter 5.
31) Liquid on vial caps.
a) Leaky flush valve.
a) Replace flush valve.
b) Leaky injector valve.
b) Replace rotor seal (Chapter 5).
Arm
28) Hook doesn't lift vial.
Leaks
Thermo Electron
163
Table A.3 Autosampler Hardware Problems
Possible Problem
Remedy
32) An automatic sequence is
interrupted, or instrument
will not start.
a) Column-oven, heater, door
interlock(s) activated.
a) Check the Ready Participation Menu.
33) In Stand Alone mode,
Status Screen indicates a
run in progress, but the
instrument is locked up.
a) Configuration is incorrectly
set to SpectraNET mode.
a) In the Configurations Menu, set Mode
to Stand Alone.
34) Contaminated sample.
a) Dirty vials.
a) Replace vials.
b) Improper septa made of
silicone or other rubber-like
material.
b) Use recommended septa.
NOTE: Thermo Electron septa are
made of Teflon-faced silicone.
a) Block between column and
autosampler.
a) Crack each fitting and observe
pressure. See instructions in
Appendix A "Testing for Tubing
Restrictions" on page 169.
Symptom
Other
35) Elevated system
pressures.
164
Thermo Electron
Display Messages
As we mentioned in Chapter 1, there are three different kinds of
messages that can appear on your autosampler's display: user
messages, confirmation messages, and error messages. This section
describes the possible conditions that generate these messages.
USER MESSAGES
User messages are displayed as a result of an incorrect operation on
the part of the user.
You can't issue a command from a menu or press a key while the
autosampler is in the middle of a previously requested operation.
Autosampler Busy
Autosampler Busy
Loading Allowed
No
You can't load a sample set while the queue is running.
Can not add Priority
Set during final
calibration
You cannot add a priority set during bracketing (after all the sample vials
have been injected).
Can not add Priority
Set at this time
You cannot add a priority set to other than set 1. You cannot add a priority
set unless the file's Calibration Menu calls for interspersed calibration vials
(you must select reuse calibration vials in order to add priority sets to the
queue).
File in Queue
Cannot be Copied to
You cannot copy to a file assigned to a sample set in the queue.
File in Queue
Cannot be Deleted
You cannot delete a file assigned to a sample set in the queue.
File in Queue
Cannot be Edited
You cannot edit a file assigned to a sample set in the queue from the Edit
Menu.
NOTE: You can edit the run-file copy of the active file from the Samples
Menu.
File Protected
Cannot Be Copied To
You can't copy to a file that is protected (Protect is set to On) in the
Configurations Menu.
Use SAMPLES key to Add
Samples to Queue
You can't start a run by pressing the [RUN] key when the queue is empty.
You must first load or add sample set(s) to the queue.
Thermo Electron
165
CONFIRMATION MESSAGES
Confirmation messages inform you that the autosampler is completing
a requested operation or command. These messages disappear when
the operation is complete.
Aligning Arm
The Align Arm command is in progress.
Clearing Mixer
The Clear Mixer command is in progress.
File Copied
The requested file has been copied.
File Deleted
The requested file has been deleted.
File Loaded
The requested file has been loaded into the queue.
Flush Samp Syringe
The Flush Sample Syringe command is in progress.
Flush Prep Syringe
The Flush Prep Syringe command (page 97) is in progress.
Homing Arm
The Home Arm command is in progress.
Initializing
The Initialize Hardware command (page 98) is in progress.
Set Added to Queue
The requested sample set has been added to the sample queue.
ERROR MESSAGES
There are three kinds of errors: 1) hardware errors, 2) system errors,
and 3) diagnostics errors. Hardware and system errors are discussed
below. Diagnostic errors are discussed in the diagnostics section on
page 129.
HARDWARE
ERRORS
Hardware errors occur during the normal use and operation of the
instrument. Most of the time you will be able to clear them. For
some, you may need to call your Thermo Electron service
representative. Pressing any key clears the error message. Then take
the steps suggested for each message to resolve the problem.
Arm Failed to Home on
X, Y, Z
The XYZ arm could not make the final X-, Y-, or Z-axis motion required
return to the home position. Check for a dislodged vial, tray, or other
obstruction. Run the Initialize Hardware routine. If the error persists,
contact your Thermo Electron service representative.
Arm Jam on X, (Y, Z)
Axis
The XYZ arm is obstructed on its left-to-right (X) axis; its front-to-back (Y)
axis; or its vertical (Z) axis. Check for a dislodged vial, tray, or other
obstruction. Run the Initialize Hardware routine. If the error persists,
contact your Thermo Electron service representative.
166
Thermo Electron
Flush Valve Misaligned
The flush valve did not turn all the way to the required position. Perform the
Initialize Hardware routine. If the error persists, run the Test Connectors
routine. If the error persists, contact your Thermo Electron service
representative.
Hardware Failure
A problem occurred with the valves or syringes. Run the Initialize
Hardware diagnostic on page 98 and/or the Injector Test (page 98). If you
are unable to clear the error, contact your Thermo Electron service
representative.
Hardware Not Homed
The solvent valve was requested to move to a certain position, but the
hardware was not initialized properly. Or, on powerup, the solvent valve
could not home properly. Run the Initialize Hardware diagnostic on page
98. If you are unable to clear the error, contact your Thermo Electron
service representative.
Hardware Timeout
A requested mechanical operation was not completed in the allotted time.
The problem could be with the arm, valves, or syringe(s). Run the Initialize
Hardware routine on page 98. If the problem persists, try manually flushing
the syringe(s) using the Flush Sample (page 97) and Flush Prep Syringe
(page 97) commands. If the problem persists, contact your Thermo Electron
service representative.
Injector Valve Failure
The injector valve failed to move (between the INJECT and FILL positions,
or vice versa) for the requested operation. Issue the Injector command from
the Commands Menu. If the error persists, call your Thermo Electron
service representative.
Inject Valve
Misaligned
The injector valve is positioned between the INJECT and FILL positions.
Run the Injector test (page 98), and call your Thermo Electron service
representative if the error persists.
IValve Move Is
Impossible
The injector valve cannot move to the requested position from its current
position. Run the move the injector valve to another position using the
Injector command, and call your service representative if the error persists.
IValve Move Not
Detected
The system did not detect a requested movement to the required position.
Check for a loose connection to the injector. Try to manually move the
injector to another position. If the error persists, call your service
representative .
Inject Valve Overshoot
The injector valve moved past the desired position. Run the Injector test
(page 98), and call your Thermo Electron service representative if the error
persists.
Injector Valve
Undershoot
The injector valve failed to move far enough to the desired position. Run
the Injector test (page 98), and call your Thermo Electron service
representative if the error persists.
Parameter Out of Range
A value too large or too small was entered. For example, an injection
volume too large for the installed syringe was entered. Check the installed
syringe/sample loop and versus the edit file parameters.
Thermo Electron
167
Solvent Valve
Misaligned
The 6-port solvent selection valve failed to turn far enough to reach the next
required port or turned past the next required port. Issue the Initialize
Hardware command on page 98. If the error persists, issue the Flush Prep
Syringe (page 97) using various external solvent positions (S-1, S-2, S-3,
S-4). This message appears only on instruments with the Sample
Preparation Option installed.
SValve Not Present
A Load or Flush Prep Syringe command was issued without the Sample
Preparation Option installed. The Configurations Menu incorrectly indicates
that the Sample Preparation Option is installed (Sample Prep Yes). Check
the Configurations Menu under OPTIONS. If the problem persists, call your
Thermo Electron service representative.
Syr 1(2) Not Enough to
Expel
One of the syringe(s) is not retracted far enough to expel the requested
volume in the ADD step (sample preparation syringe) or flush operation
(both syringes). Run the Initialize Hardware test (page 98), and restart the
run (sample preparation) or the flush cycle.
Syringe Failure
A syringe failed to moved to the appropriate position required for an
operation. Run the Initialize Hardware test (page 98). If the error repeats,
call your Thermo Electron service representative.
Syringe Overfill
A volume has been requested for injection or ADD that is too large for the
size of the installed syringe. Check the method and syringe volume(s).
TrayTemp Failure
The Tray Temperature Control Option was unable to reach the desired set
point and has turned itself off returning the instrument to a READY state.
Any method loaded will be executed without the Tray Temperature Control
Option. Pressing any key clears the error. This error indicates a problem
with the Tray Temperature Control Option. Call your Thermo Electron
service representative for assistance.
Unexpected Vial in
Hook
This message appears if a vial is found in the hook where no vial is expected
for the requested operation. To clear the error, remove the vial and restart or
resume the requested operation.
Vial Not Found in Hook
This message appears if a vial is not found in the hook when a vial is
expected. Verify that a vial exists in the requested tray location, check the
file and vial assignments in the Samples Menu, and check to see if a vial is
lodged in the sample tower. (If so, the needle may be bent.)
168
Thermo Electron
SYSTEM ERRORS
System errors occur very rarely and abort normal operation. Since
they are not usually problems you can correct, we will list some of
them, but not describe them in detail. Should you see any of the
following messages or other messages not described in this section,
document them, and contact your Thermo Electron service
representative for assistance.
Invalid Current Location
Invalid Command
Not in GET/RET Sequence
Z Must be Up/No Vial
Move Z Not Allowed
Get LocNum: Bad Row, Col
Zup Only in Serv Block
Invalid Target Location
SValve Extra Interrupt
Bad Vial Number
Unexpected Syr Event
Testing for Tubing Restrictions
If there is a restriction in the solvent lines, you will probably hear the
a loud grinding sound from the syringe-drive mechanism due to
increased resistance to syringe travel.
NOTE: Before beginning this procedure, be sure to set up a beaker or other
container to catch the solvent.
To isolate a block to a section of tubing:
1. Place the Autosampler injection valve into the FILL position
(press [MENU], COMMANDS, Injector Test, and use the [+]
and [-] keys to select /Fill/).
2. Disconnect the injection valve-to-flush valve tubing at the
injection valve. Initiate a flush sequence (press [MENU],
/COMMANDS/, /Flush Sample Syringe/, page 97 or /Flush
Prep Syringe/, page 97. Solvent will flow freely on the return
stroke of the plunger if there are no restrictions present. Flow
restriction is unlikely at this point due to the large internal
tubing diameter.
3. Reconnect the tubing.
4. Disconnect the needle and check the flow.
5. If no restriction is found, reattach all of the tubing, and flush
the syringe to verify that the restriction still exists.
6. If a restriction is still present, remove and clean the sample
needle as described in Chapter 5, page 143.
Thermo Electron
169
B
Glossary
Introduction
We have included a glossary to define certain technical terms used
throughout the manual's text. These terms should be consistent with
standard definitions used throughout the analytical industry, and are
added here as a quick reference only.
A
active set/vial
The sample set or vial currently being processed. The active vial refers to
the vial currently being injected or currently undergoing sample preparation.
Add
1) A step used in sample preparation used to deliver all Pickup volumes and
the Load volume to the target vial. 2) The process of putting a sample set
into the sample queue.
B
backlash
The slack in the syringe and its drive mechanism.
C
calibration interval
The number of sample vials to be injected between calibration-vial
injection(s).
calibration level
The number of different concentration levels used in the calibration series.
calibration vial
A vial that contains a known concentration of the compound(s) of interest; a
standard.
cycle time
The time between autosampler injections.
Thermo Electron
171
D
dead volume
The volume from the solvent-selection valve to the autosampler needle tip.
decrement
Press the [-] key to decrease the value of a parameter.
diluent
The solvent or buffer used to dilute a sample. On the autosampler, the
volume of diluent is specified in the Load step of all of the sample
preparation templates.
dilution cycle
A preset sequence used in the sample preparation templates that transfers
solvents, sample, and reagents from the solvent bottle(s) or source vial(s) to
the target vial(s).
E
extra vials
In sample preparation, an empty or extra reagent vial where samples and
solvents are mixed; an empty reaction vial. It is always placed in
relationship to the sample vial (Sam+1, Sam+2, Sam+3).
F
file/method
"File" and "method" are used interchangeably to describe the autosampler
run parameters. However, the word "file" is often used in a more specific
sense to mean the autosampler files in which the parameters are stored.
flush solvent
The solvent used to wash the internal tubing, syringes, and sample needle
after an injection. This solvent should be of sufficient strength to easily
remove the small amounts of sample that can cling to the tubing walls.
full-loop injection
A technique that injects a sample volume equal to the loop volume.
G
guard column
A small column placed just before the separation column. The guard
column is used to remove contaminants that might otherwise damage the
main column or interfere with the separation.
H
holding loop
The isolated section of connecting line used in sample preparation to hold
measured amounts of sample, reagents, and solvents until they are delivered
into a vial.
home
The position to the left of the rear-left corner of the sample tub where the
XYZ arm resides when it is not transporting a vial.
172
Thermo Electron
hook
The attachment on the end of the pod that moves a vial up and down (along
the Z axis). The hook lifts the vial from the sample tray and holds it during
transport to the sample tower for injection.
I
increment
Press the [+] key to increase the value of a parameter.
initialization
The process that takes place between dilution cycles that brings the
autosampler to a "known" state. During this process the lines and syringes
are automatically flushed with flush solvent. Any residual resources present
from a previous dilution cycle are flushed out of the lines to prepare the
autosampler for the next injection.
J-L
linear dilution
A dilution sequence in which resource A (buffer) is added to resource B, C,
and D.
loading a file
Loading a file copies the original edit file into the run-file location to be run.
The original edit file remains.
M
method
A group of parameters applied to one or more vials.
mobile phase
In HPLC, the liquid used to move the sample through the column.
N-O
open-collector outputs
An output, usually a transistor, that has no connection to the collector.
P
PickLg
A step used in the sample preparation templates that retracts the prep syringe
to draw a large volume of solvent or buffer into the holding loop.
Pickup
A step used in the sample preparation templates that retracts the sample
syringe to draw a small volume of sample or reagent into the holding loop.
prep syringe
The large 2.5 mL sample preparation syringe used in to transfer large
volumes of liquid (usually buffer or solvents).
pod
The autosampler component attached to the XYZ arm that contains the hook.
Thermo Electron
173
precision
The degree to which results are reproducible (analysis of the same sample
yields similar results).
prime
The process that flushes lines with flush solvent, etc. Priming is done during
installation and whenever a syringe is replaced.
PCB
A printed circuit board.
pull-loop injection
Variable-loop injection.
pull-up resistor
Resistors used to pull inputs or nonactive open-collector outputs to "Hi"
states.
Q
queue
One or more vial sequences/sets subjected to one or more methods run in a
series that use(s) the same column and solvents. The sample queue lists the
order and file name for each sample set to be processed during a given run.
R
reagent transfer step
In a sample preparation program, a step used to deliver solvents, reagents, or
samples from one location (vial or solvent bottle) to another (target or
sample vial).
real-time editing
The ability to edit the run file for a run currently in progress. On the
SpectraSYSTEM autosamplers, this function is provided by the [SAMPLES]
key; on other SpectraSYSTEM instruments, this function is accessed by
pressing the [STATUS] key.
recalibration
The reinjection of the calibration vial(s) after the injection of each series of
sample vials within a sample set.
recalibration interval
The number of sample vials injected between each calibration series
injection. The value in the Samples/Calibration field.
resource
One of the samples, calibration vials, reagents, or solvents used in an
autosampler run.
routine
The type of order in which calibration vials are arranged within a sample set.
Calibration vials may be reused throughout the set (Reuse same vials) or
they may be interspersed (Intersperse) among the sample vials in the sample
tray.
run file
The copy of the edit file that has been loaded and the file that is currently
running. From the Samples Menu, changes can be made to the run file
during a run, but those changes can't be saved until after the run has
completed. Also, changes can't be copied to a file that is in use (assigned to
a sample set in the queue).
174
Thermo Electron
S
sample needle
The needle that pierces the septum of the vial cap and through which vial
contents are drawn into the holding loop.
sample syringe
The 250 µL syringe. In sample preparation, this syringe is used in sample
preparation to transfer small volumes of liquid (usually samples or reagents).
sample tower
The autosampler location and component where injection takes place.
sample tub
The autosampler compartment that holds the sample trays.
sample tray
One of the three racks that holds the sample vials in position in the
autosampler.
sampling valve
The valve through which sample is injected onto the column.
sample vial
The container that holds the sample in the sample tray. Sample vials must
have a 12 mm diameter.
set
A group of samples vials and associated calibration vials run with a
particular method.
solvent-selection valve
The 6-port valve that regulates the flow of solvents and air into the sample
holding loop. This valve allows the flow of waste out of the loop.
solvent reservoir
One of the four bottles on the back of the autosampler that contains the
external solvents used in sample preparation. The solvent in the first
position usually contains the buffer.
standard
A vial that contains a known concentration of sample. Peaks generated from
this vial are used to plot the calibration curve used to determine the
concentration of the compound of interest. (See also calibration vial.)
syringe refill and loopisolation valve
The valve in the sample preparation system that regulates the flow of liquid
from the syringes.
T-V
template
In sample preparation, one of the four file formats that contain a unique,
preset, unchangeable sequence of steps used to carry out a series of sample
preparation procedures. The four types of templates include simple dilution,
heating, extraction, and heating and extraction.
W-X
Thermo Electron
175
Wait
A step used in the sample preparation templates that pauses all autosampler
activity. This step may be used to allow a mixture to settle or to react.
X
In the Calibration Menu's Calib 1 (of X) field, X indicates the number of
calibration levels in the calibration sequence. X is not visible on the screen;
it is used for discussion purposes only.
XYZ arm
Also referred to as the "arm," this component carries the sample vials to and
from the sample tray and to the various workstations (sample tower, heater
mixer) in the autosampler.
Y-Z
Z
176
In the sample preparation templates, the variable used to represent the
number of clean (extra) vials subsequent to the sample vial. Z equals 1, 2,
or 3.
Thermo Electron
C
Kits and Parts Lists
Shown below is a list of spare parts, consumables, and kits available
from Thermo Electron for use with your SpectraSYSTEM
autosampler.
Clear Vials and Vial Kits
Shell Vials
A4946-010
A4948-010
Shell Vials, 100/package
Shell Vial Kit
Standard Vials
A4951-010
A4952-010
A4953-010
A4954-010
Standard Vials, 100/package
Standard Vial Septa, 100/package
Standard Vial Caps, 100/package
Standard Vial Kit, 100/package
Crimp Vials/Caps
A4955-010
A4956-010
A4957-010
Crimp Caps, 100/package
Crimp Vials, 100/package, 1.8 mL
Crimp Vial Kit, 100/package
Poly Kits
A4958-010
A4959-010
Polypropylene Kits, 600 mL, 100/package
Polypropylene Kits, 100 mL, 100/package
Inserts
A4981-010
A4960-010
A4961-010
Spring for Insert, 100/package
Insert, 50 mL, 100/package
Insert, 100 mL, 100/package
Amber Vials and Vial Kits
Crimp Vials
A4964-010
A4965-010
Amber Crimp Vials, 100/package
Amber Vial Crimp Kit, 100/package
Standard Vials
A4966-010
A4967-010
Standard Amber Vials, 100/package
Standard Amber Vial Kit, 100/package
PFTE Liners/Septa
A4968-010
A4969-010
A4970-010
PTFE Liners, 100/package
PTFE Vial Caps/Septa, 100/package
Standard Vial Station
Sample Trays
A3695-010
A3696-010
Sample Trays
A3695-010
Sample Trays (3), 40-vial capacity
Sample Trays (3), 35-vial capacity
(For autosamplers with Tray Temperature Control.)
Sample Trays (3), 40-vial capacity
Thermo Electron
177
A3696-010
Cables and Adapters
A3981-010
A3663-010
Sample Trays (3), 35-vial capacity
(For autosamplers with the Peltier cooling option/upgrade.)
Autosampler-to-integrator Cable/Adapter
BCD Communications Option
(This option requires field installation by Thermo Electron unless purchased
simultaneously with a SpectraSYSTEM autosampler.)
Standard Autosamplers
178
A3566-010
Injector Valve Assembly for the AS1000 autosamplers, 20 mL, SS
(Assembly includes the complete autosampler valve assembly.)
A3566-030
Injector Valve Assembly for the AS3000, 100 mL, SS
(Assembly includes the complete autosampler valve assembly.)
A4101-010
Standard Maintenance Kit
(Kit includes everything necessary to maintain the instrument for one year: inlet filter,
needle replacement kit, a valve rotor seal, syringe, lubricant, cleaner and an LC test
sample.)
A4051-010
Standard Fittings Kit
(Kit includes stainless steel fittings and tubing used in a SpectraSYSTEM LC system.)
A4719-010
Autosampler Needle Replacement Kit
(Kit includes the standard, stainless-steel, deflected-point replacement needle.)
A4719-020
Autosampler Needle Replacement Kit
(Kit includes the standard, inert, deflected-point replacement needle.)
Thermo Electron
Standard Autosamplers, continued
Standard Sample Loops and Syringes
3302-0230
Rheodyne Sample Loop, 20 µL, SS
3302-0010
Rheodyne Sample Loop, 50 µL, SS
3302-0020
Rheodyne Sample Loop, 100 µL, SS
3302-0220
Rheodyne Sample Loop, 200 µL, SS
3302-0030
Rheodyne Sample Loop, 500 µL, SS
3302-0040
Rheodyne Sample Loop, 1 mL, SS
A3588-020
A3588-010
A3587-020
A4094-010
Test Mixes
A4991-010
A5135-010
Screw-tip Syringe, 250 µL, (standard and prep #1 drive)
Screw-tip Syringe, 500 µL
Screw-tip Syringe, 2.5 mL, (prep #2 drive)
Solvent Inlet Filter Cartridge (pkg. of four)
Autosampler Test Mix
Autosampler Dilution Test Mix
Inert/Biocompatible Autosamplers
A3566-020
Injector Valve Assembly
(Assembly includes the complete autosampler valve assembly.)
A4102-010
Inert/Biocompatible Maintenance Kit
(Kit includes the inert/biocompatible versions of everything in the Standard Maintenance Kit.)
A4061-010
Inert/Biocompatible Fittings Kit
(Kit includes PEEK fittings and tubing used in an inert/biocompatible SpectraSYSTEM LC system.)
A3564-020
Inert/Biocompatible Autosampler Needle Replacement Kit
Inert/Biocompatible Sample Loops and Syringes
A4169-010
A4169-020
A4169-030
A4169-040
A4169-050
A4169-060
A4169-070
Rheodyne Sample Loop, 20 µL
Rheodyne Sample Loop, 50 µL
Rheodyne Sample Loop, 100 µL
Rheodyne Sample Loop, 200 µL
Rheodyne Sample Loop, 500 µL
Rheodyne Sample Loop, 1 mL
Rheodyne Sample Loop, 2 mL
A3588-020
A3588-010
A3587-020
A4094-010
Screw-tip Syringe, 250 µL
Screw-tip Syringe, 500 µL
Screw-tip Syringe, 2.5 mL, (prep #2 drive)
Solvent Inlet Filter Cartridge (pkg. of 4)
Test Mixes
A4991-010
A5135-010
Autosampler Test Mix
Autosampler Dilution Test Mix
Upgrade Kits
Upgrade kits are available for SpectraSYSTEM and SpectraSERIES stainless steel and inert/biocompatible autosamplers. Contact
your local Thermo Electron Representative for details.
Thermo Electron
179
INDEX
KEYS
[+] and [-] keys, 41, 44
[<] key, 44
[>] key, 44
[ENTER] key, 44
[MENU] key, 44
[RUN] key, 43
[SAMPLES] key, 43, 52
[STATUS] key, 43
[STOP] key, 43
[∧] key, 44
[∨] key, 44
A
ABORT Sample, 90
AC Power, 17
Active
sample, 85
vial, 86
Add step
calculating the volume for, 108
defined, 171
described, 107
Add to Queue, 79
Air
input line, location, 10
supply pressure, heater/mixer for, 10
supply requirements, 10
Aligning arm message, 166
alphabetical entries, increasing and decreasing, 41, 44
alphanumeric entries, 44
analytical LC preheated solvent volume, 13
Arm
Arm Menu, 98
see also XYZ arm:, 176
arrow keys, 41, 44, 48
AS1000 (see also Autosampler)
Loop size field, 67, 71
More Menu, 74
AS3000 (see also Autosampler)
full-loop injection, 67
More Menu, 74
Samples Menu, figure of, 87
variable-loop injection, see pull-loop injection:, 71
asterisks, 46
Autosampler
busy message, 165
components, 56
configurations for, 31
detector connection to, 16
front panel illustration, 42
front view illustration, 49
installation
advanced, 21
basic, 6
modes described, 43
operation
advanced, procedures for, 101
basic, procedures for, 55
theory of, 56
priming
2.5-milliliter syringe, 19
250 microliter syringe, 18
pump connection to
with column oven, 12
rules for operation, 41
shutting down, 149
SpectraNET operation, configuration for, 20
Stand Alone operation, configuration for, 21
turning on the, 50
B
backlash, defined, 171
BCD communication, 30
Blank key, 43
Bracket samples field, 72
Brackets, reason for, xix
C
cables and adapters, list of, 178
Calib Vial 1 (of X) field
Calibration Menu in, 70
effect on vial injection order, 71
Samples Menu in, 78
Calibration
interval, defined, 70, 171
level, defined, 171
levels, number, 70
parameters, entering, 70
routines, 72
vial(s)
defined, 171
injections per level, 70
number of injections per, 70
tray positions, 71
Calibration Menu, 70
Calibrators, see Calibration vials:, 71
Can Not Add Priority Set messages, 165
capitalization, standard use of, xx
Carryover, test for, 34
Thermo Electron
index - i
cautions, defined, xiv, 135
Changing the run file, 87
checklist, startup, 2
Clearing Mixer message, 166
column
switching valves, 72
Column
cover, removing the, 13
equilibration
specifying time for, 75
installation
with column oven, 15
without column oven, 15
oven
operation described, 63
ready participation
interlock
disabling the, 30
turning on the, 52
oven field, 68
temperature
setting the, 68
COMM port test, 134
Commands
Flush Sample Syringe, 146
Software Version, 130
Test COMM Port, 134
Test External Contacts, 133
Commands Menu, 18, 45, 96
Communication
BCD, 28
modes, 20
standalone, 20
to older/non-SpectraSYSTEM products, 28
Communication, autosampler
with Thermo Finnigan integrators, 29
Communications Menu
default parameters for, 26
failure to appear, reason for, 26
Communications parameters
normal operation settings for, 26
Concentration levels, standards, 70
Configuration
SpectraNET operation, 20
stand alone operation, 21
Configurations Menu, 92
Confirmation messages, 46, 166
Continue? message, 90
conventions used in manual
brackets, xix
capitalization, xx
caution, xxi
hint, xxi
icons, xxi
note, xxi
slashes, xix
standard words, xx
two-line display, xix
Copy Menu, 91
Copying files, 91
index - ii
Cover
front, removing the, 13
top, see Door interlock:, 30
Creating a file, 51, 65
CSA, see separate Safety Section:, 41
Current vial, information on, 86
cursor
movement, 41
square
blinking, 41
triangular, 41
customer support
Australasia and Asia, vii
Europe, vi
North America, v
Cycle time
defined, 171
field
Injection Menu in, 67
Samples Menu in, 78
D
dead volume, defined, 172
decrement, defined, 172
Default
method, description, 53
delay
gradient, setup, 127
Delay Time, 149
Delete Menu, 91
Deleting
files, 91
sets from the queue, 80
Diagnostics
COMM port, 134
Connector Test, 132
described, 129
RAM Test, 131
ROM Test, 132
diluent, defined, 172
Dilution cycle, 172
described, 103
determining the number of, 108
process, figure, 104
Steps
Add, 107
FlushP, 117
Heat, 115
Load, 106
Mix, 107, 115
PickLG, 107
Pickup, 107
Remove, 117
Wait, 117
Templates, see Templates:, 113
Display
contrast, 31
Inputs States Menu, 131
Output States Menu, 131
Thermo Electron
Display Timed Events Menu, 131
Door
column compartment
removing the, 13
interlock
Configurations Menu in, 92
disabling the, 30
turning off the, 30
down-arrow (¯ key, right side of display), 42
Draw rate
Prep syringe, 75
sample syringe, 75
Drop catch
cleaning the, 144
Due field, 151
E
Edit
file
definition, 77
field, 66
saving an, 76
parameters during a run, 44
Menu, 109
Edit Menu, 66, 102
electrical requirements
instrument, 4
eluants, degassing, ix
Empty vials, number of, 110
ENTER key, 44
Enter, as a standard word, xiii
Entering your parameters
from the Edit Menu, 51
from the Samples Menu, 50
Equilibration
Time field, 75
time, beginning, 52
Error messages, 46
Europe
customer support, iv
technical support, iv
exclamation points, 46
external
events (see also Timed events), 72
solvent lines, plumbing, 9
Contacts Tests, 133
Devices Menu, 133
devices, autosampler control, 72
Extra vials
defined, 172
locations of, 108
Extraction
process, 117
template used for, see Rainbow template, 117
F
FCC, see separate Safety Section
Fields
#Cycles, 151
#Inj, 86
#Samples, 85
Add to queue, 79
Bracket samples, 72
Calib vial 1 (of X)
Samples Menu in, 78
Calibration Menu in, 70
Column
oven, 68
COMMANDS, illustration, 45
Cycle Time
Injection Menu in, 67
Samples Menu in, 78
Due, 151
Edit File, 66
Equilibration Time, 75
File, 78
File #, 86
FILES, 45
File requires n Extra vials, 110
First Sample Vial, 79
Flush volume, 75, 150
Gradient delay, 75
Heater shutoff, 150
Htr/Mix, 112
Injection
Type, 75, 76
volume
calibration vials, 70
sample vials, 67
Injections
/level, 70
/vial, 67
Needle Height, 76
Note, 151, 152
Number of Samples, 79
OPTIONS, illustration, 45
Oven TrayTemp, 86
Overlap Enable, 102
Prep, 112
Flush Volume, 150
Viscosity, 75
Protect, 92
Pump shutoff, 150
QStatus, 84
QUEUE
illustration, 45
Reset #Cycles, 152
Sample Set, 78
Samples/calibration, 70
Set, 84
Status, 85
Total Cycles, 151
Thermo Electron
index - iii
Vial
Status Screen, 86
Status Prep Screen, 112
Viscosity, 75
File
assigning a to a sample set, 86
copying a, 91
creating a, 65
defined, 172
Deleted message, 91
deleting a, 91
Edit, 76
field
Samples Menu, 78
in Queue messages, 165
management, 91
Protect, 92
requires n extra vials field, 110
restricting editing of a, 92
running a
with sample preparation, 102
saving a, 76, 91
File name
Edit Menu in, 66
field, Queue Menu in, 79
requirements, 66
File Protected Cannot Be Copied To message, 165
File# field, 86
Files Menu, 45
Fill position, injector valve, 59
filter, flush solvent inlet, replacing, 143
First Sample Vial field, 79
Fixed-loop
AS1000, see Autosampler or AS1000:, 71
injection volume, entering the, 71
injection, see Full-loop injection:, 67
Flush
bottle bracket, installation, 7
bottle, installation, 9
solvent, defined, 172
cycle, described, 108
Prep Menu, 97
prep syringe, 19, 147
sample syringe, 18
Syringe Menu, 18, 97
Volume field, 75, 150
FlushP, step described, 117
fraction collectors, autosampler use with, 72
front cover, removing the, 12
front panel
connections, 6
illustration of, 42
Full-loop injection, 56
defined, 172
figure, 59
Fuses, replacing, 147
index - iv
G
Good laboratory practices, see separate Safety Section
gradient
delay, 75
Gradient
start output, 23
Ground, 23
grounding, 41
guard column, defined, 172
H
Hardware Test, 132
Hardwire
connections, SpectraSYSTEM modules for, 25
procedures, described, 21
hardwire connections, SpectraSYSTEM modules, 25
Heat step, described, 115
Heater
ready interlock, turning off the, 30
Shutoff field, 150
Heater/mixer, 64
actual temperature, 112
air pressure for, 10
operation described, 64
Heating and extraction template
description, 120
Heating template
description, 115
hints
defined, xiv
holding loop, defined, 172
home position, described, 18
hook, defined, 173
hot surface
defined, xiv, 135
Htr/Mix field, 112
I
I/O states, testing the, 133
ICONS, xxi
increment, defined, 173
initialization, defined, 173
Initialize EEPROM Menu, 134
Inject
hold input, 24
out output, 24
position, injector valve, 59
Injection
Menu, 66
methods
full-loop, 56
figure, 59
pull-loop, 57
figure, 60
PushLoop, 60
Thermo Electron
figure, 61, 62
parameters, entering, 66
Range field, 76
single, 67
status, 85
Type
advantages of each, 75
field, 75
valve, positions, 59
Injection reproducibility, test for, 32
Injection volume
Calibration Menu, 70
calibration vials, 70
field, 67
fixed-loop, 67
Injection Menu, 67
optimization of, 94
sample vials, 67
Samples Menu, 78
Injections
/level field, 70
/vial field, 67
flushing between, 75
multiple, 67
injector
valve bracket orientation, 139
injector valve
disassembling, 140
reinstalling, 137
Input Polarity Menu, 27
Input signals
described, 23
states, 131
Installation
basic, 6
connections
front panel, 6
external solvents, 10
flush solvent, 9
overview, 6
sample tray, 16, 82
Instrument
control, 42
dimensions, 4
operation
hazards, 135
integrators, sending vial information to, 29
Interlocks
Col Oven, 30
Door, 30
Heater, 30
Intersperse vials, 71
K
keypad, 43
illustration, 42
moving around, 41
keys, 43
[+] and [-], 41, 44
[<], 44
[>], 44
[ENTER], 44
[MENU], 44
[RUN], 43
[SAMPLES], 43
[STATUS], 43
[STOP], 43
[∧], 44
[∨] arrow, 44
arrow, 44
blank, 43
Keys
Samples, 77
STOP, 89, 124
Kits
vial, 177
L
laboratory safety, see separate Safety Section
LCD, see Display:, 31
linear dilution, 173
Line-by-line descriptions, templates
Template 1 (dilution), 113
Template 2 (DABS-CL), 116
Template 3 (Rainbow), 118
Load
Menu, 77
step, described, 106
Loading
a file, 173
a sample tray, 17, 82
Loop size
AS3000, 67
AS1000, 67
Loop-back cable, 134
low-dead-volume union, 13
M
Main Menu, 45
illustration, 45
menus accessible from, 45
Maintenance Log Menu, 150
maintenance schedule, 136
Managing priority sets within the queue, 80
Manual, conventions used in, xii
Menu
key, 44, 48
Menus
Calibration, 70
figure, 70
Commands, 19, 45
Configurations, 92
Copy, 91
Delete, 91
Display
Thermo Electron
index - v
Inputs States, 131
Outputs States, 131
Timed Events, 131
display conventions, 45
Edit, 51, 66
External devices, 133
Flush Prep, 20, 97
Flush syringe, 19, 97
Hardware Test, 132
Initialize EEPROM, 134
Injection, 66
Injector Valve, 151
Input Polarity, 28
Load, 77
Maintenance Log, 150
More, 74, 127
Needle Wear, 152
Options, 45
Output polarity, 28
Prep Syringe, 152
Queue, 45, 79
RAM Test, 131
Ready Participation, 31
ROM Test, 132
Sample Preparation, 109
Sample Syringe, 152
Samples
described, 43
without sample preparation, 77
Shutdown, 149
Software Version, 130
Status (see Status Screen):, 109
Test COMM Port, 134
Test External Contacts, 133
Tests, 130
Timed Events, 72
Total System Cycles, 151
Menus and Screens, general description of
Commands Menu, 45
Files Menu, 45
Main Menu, 45
Options Menu, 45
Queue Menu, 45
Status Screen, 45
Messages, 46
Autosampler busy, 165
Aligning arm, 166
Can Not Add Priority Set messages, 165
Clearing Mixer, 166
File
Protected Cannot Be Copied To, 165
File Deleted, 91
File in Queue, 165
Use Samples Key to Add Samples to Queue, 165
user, 46
method, 172, 173
Mix step described, 107, 115
mobile phase, 173
Monitoring the run, 83
More Menu, 127
Mounting a column
index - vi
with column oven, 15
without column oven, 15
moving within and between menus, 44
N
narrow-bore configuration, 55
narrow-bore preheated solvent volume, 13
needle (see sample needle module), 143
Needle height (see also vial height)
adjusting, 93
Needle height (see also vial height):, 76
needle, sample, 143
Normal-phase solvents
syringe speed adjustment for, 96
North America
customer support, iii
ordering replaceable parts, iii
technical support, iii
Note field, 151, 152
notes
defined, xxi
Number of
Levels field, 70
Samples field, 79
numerical entries, 44
increasing and decreasing, 41, 44
O
on/off switch, 50
open-collector outputs, 173
Optimization
injection volume, 94
Injection volume of, 94
Options Menu, 45
Options, available for autosampler, 63
ordering replaceable parts
North America, iii
Ordering sets in the sample queue, 79
output
Signals
described, 24
Output
Polarity Menu, 28
Signals
states, 131
Oven
Shutoff field, 149
Temperature field, 86
Overlap Enable field, 102, 108
Thermo Electron
P
P
defined, 80
queue menu in, 89
PCB, defined, 174
Performance Tests
reproducibility, 32
sample carryover, 34
PickLg step
defined, 173
described, 107
Pickup step
defined, 173
described, 107
Pin assignments, 23
+5VDC, 23
A/S ready, 24
gradient start, 23
ground, 23
inject hold, 24
inject out, 24
pump ready, 23
pump stop, 23
timed outputs, 24
pinouts, recommended configuration, 24
plumbing
external solvents
with bottle caps, 10
without bottle caps, 9
flush solvent, 9
procedures, autosampler for, 7
pod, defined, 173
Polarity
input, 28
output, 28
setting the, 27
power requirements, instrument, 4
Practice example, running the, 47
precision, defined, 174
preheat tube, connection to, 13
preheated solvent volume, 13
Prep
field, 112
Flush Volume, 150
Sample, viscosity, 75
Viscosity field, 75
Prep syringe
defined, 173
used in priming, 19, 97
Prep Syringe Menu, 152
Preparation (see also Sample Preparation)
sample vial, 48
solvents, 48
Syringe
priming, 19
viscosity adjustment for, 75
preset choices, selecting from, 44
prime, defined, 174
priming
250 microliter syringe, 18, 97
prep syringe (sample preparation), 19, 97
Priority sample sets
calibration routine for, 88
within a queue, 88
Protect
field, 92
files, 92
pull-loop injection
defined, 174
figure, 60
pull-up resistor, defined, 174
pump
outlet line connection
with oven, 13
Pump
autosampler connection installation
with column oven, 13
without column oven, 12
outlet line connection
without oven, 13
Ready input, 24
Shutoff field, 150
stop output, 24
Push Lo
Injection Type field in, 75
PushLoop injection, 60
figure, 61, 62
Q
Queue (see also Sample queue)
defined, 174
building, 77
deleting sets from, 80
managing priority sets in, 80
ordering sets in, 79
Queue Menu, 45, 79
states
abort, 84
equil, 84
PAUSE, 84
run, 84
status, 84, 86
R
Rainbow template, 117
RAM Test
Menu, 131
Reaction vials, see Vials, extra, 109
Ready Participation Menu, 30
Reagent
A, B, C, 109
transfer step, defined, 174
Vials field, 109
Rear panel
Thermo Electron
index - vii
figure, 8
rear panel, autosampler, 148
recalibration
defined, 174
interval, 70
reinstalling injector valve, 137
Remixing step, described, 115
Remove step, described, 117
Reordering sample sets in queue, 79
reproducibility
dilution, 37
Reproducibility, test for, 32
Reset #Cycles field, 152
resource, defined, 174
Reuse same vials, 71
reviewing parameters during a run, 44
ROM Test, 132
rotor seal
installing, 141
rotor seal, removing/reinstalling, 137
routine, defined, 174
Run
in progress
changes to sample preparation, 111
preparation
sample vials of without sample preparation, 64
calibration vials using Sample Preparation
Option, 123
preparing other modules for a, 81
starting a, 53
starting a, 83
status of a, 86
steps for setting up a, 47
without sample preparation
in progress, changing a, 43, 93
run file
definition, 174
RUN key functions, 43
Running
a dilution with sample preparation, 122
a file
with sample preparation, 102
without sample preparation, 83
the practice sample, 51
S
Safety
certification, see separate Safety Section
general precautions, see separate Safety Section
Safety precautions
electrical shock, 135
fuses, 135
power cord, 135
Safety regulations (Safety Section), vii
Sample (to be run)
filtering your, 65
injection volume, entering your, 67
practice, running the, 51
index - viii
preparation
automated, see Sample Preparation:, 65
manual, 65
Set field, 78
viscosity, 75
vial currently being processed, 85
Sample carryover, test for, 34
Sample compartment cover
see Door interlock:, 30
sample loop
installation, 15, 147
sizes offered, 31
Sample loop vs. sample size, table for, 95
sample loops and syringes, list, 179
sample needle
defined, 175
module
cleaning the, 144
installing the, 144
removing the, 143
module:, 143
Needle Wear Menu, 152
Sample preparation
dilution cycle in, 103
operation, 64
overview, 102
program running a, 109
simple dilution cycle described (see also Template 1),
105
steps in (see also Dilution cycle, steps in):, 106
template in, 102
Templates (see Templates):, 106
template step, determining the current, 112
terminology, 102
Sample queue (see also queue)
adding items to a, 77
arranging sets in a, 79
definition of a, 77
deleting items from a, 80
Sample set
adding to sample queue, 79
automatic recalibration, 72
changing the order in queue, 79
defined, 43, 64
number of
levels in, 70
vials in, 79
priority, 80, 88
Sample syringe
defined, 175
priming the, 19
rate, viscosity adjustment for, 75
Sample Syringe Menu, 152
Sample to be run
preparation, automatic. See Sample Preparation
sample tower, 175
Sample tray
defined, 175
installing a, 16, 82
installing a, 16, 82
Thermo Electron
temperature control
operation, 64
turning on the (see also Tray Temperature), 52
vial placement in, 16, 71, 81
sample tub, defined, 175
Sample vial(s)
adding during a run, 30
calibration between, 70
defined, 175
SAMPLES key, 43
Samples Menu, 43, 77, 87
affected by other menus, 70
calibration and sample preparation method, 110
entering parameters from the, 48
figure, 77
Figure of, 87
number of empty vials, 110
real-time editing from, 44
with sample preparation, 109, 110
Samples/calibration field, 70
sampling valve, defined, 175
Saving
files, 91
schedule, maintenance, 136
Screens
Oven TrayTemp, 86
Status, 45, 83, 111
Screw-top vials, 17
scrolling through choices, 44
select, xx
self-diagnostics, 17
Septa, 16
set, defined, 175
Shutdown Menu, 149
Signal levels
input, 27
output, 28
Simple dilution
template, description, 113
Site requirements, 4
slashes, reason for, xix
solvent
disposal, ix
preparation, ix
requirements, ix
reservoir, defined, 175
selection valve, 175
solvent tray
waste collection, 10
Solvent viscosity
optimization for, 96
Solvents
disposal of wastes, see separate Safety Section:, 41
external, with sample preparation, installation, 9
incompatible, procedures for using, 107
space requirements, 4
Specific hazards, 135
SpectraNET operation, configuration for, 21
Stand Alone operation, configuration for, 22
standard
defined, 175
Standard
injections per level, 70
levels, 70
positions in tray, 71
standard words, xx
Starting a run, 53, 83
stator
cleaning, 141
Status
Injection Status, 85
instrument, 86
priority status, sample sets, 80
STATUS key, 43
Status Screen, 84
injector status
illustration, 45
oven/tray temperature status
illustration, 46
overview, 45
QStatus, 80
queue status
illustration, 45
with sample preparation, 111
without sample preparation, 83
Status states
ABORT, 85
GRAD, 85
HOLD, 85
IDLE, 84
NOTRDY, 85
PAUSE, 84
READY, 85
Step (see also Dilution cycle, steps)
template, skipping a, 108
templates in:, 108
STOP key, 43
functions, 89, 124
Stopping a run, 89, 124
Syringe
large syringe (see Prep syringe):, 173
refill and loop-isolation valve, defined, 175
sample (see Sample Syringe), 175
see also Prep Syringe:, 173
Syringe speed
adjusting the, 96
System
equilibration, specifying time for, 75
T
technical support, v
Australasia and Asia, vii
Europe, vi
North America, v
Template
defined, 102, 175
number of dilution cycles allowed in, 112
Thermo Electron
index - ix
Templates
general descriptions
Template 1 (dilution), 113
Template 2 (DABS-Cl), 115
Template 3 (Rainbow), 117
Template 4 (linear), 120
rules for programming, 108
steps in (see Dilution cycle, steps in):, 113
Test
COMM Port, 134
External Contacts, 133
Tests Menu, 130
text, conventions, xix
TF (time functions), 72
TF1-TF4
programming, 72
Theory of operation, autosampler, 56
Time
between injections, calculating the, 67
functions (see Timed events outputs):, 72
Status Screen in, 86
timed
outputs, 24
Timed Events
Menu, 72
outputs
external devices for, 72
states, 131
programs
adding lines to, 73
deleting lines from, 73
inserting lines into, 73
Total Cycles field, 151
Total System Cycles Menu, 151
Tray temperature (see also sample tray temperature
control)
turning on the, 52
Tray Temperature Control
Shutoff, 150
troubleshooting
autosampler chromatography problems, 156
autosampler hardware problems, 159
LC, general, 154
Tubing restrictions, testing for, 169
TÜV/GS certification, see separate Safety Section:, 41
U
unpacking procedures
index - x
instrument, 5
Use Samples Key to Add Samples to Queue message,
165
User messages
defined, 165
described, 46
V
Variable-loop injection (see also Pull-loop injection):, 57,
60
VDE, see separate Safety Section:, 41
Vial
bottom thicknesses, 93
Extra, 110
field
Prep Screen in, 112
Status Screen in, 86
height, controlling the, 76
location and Injection Report, 30
placement
in calibration routines, 71
screw-top, 16
trays, installing the, 16, 82
Vial kits, 177
Viscosity field, 75
Volume
flush between injections, 75
injection (see Injection volume):, 75
vortex mixer, 64
W
Wait step
defined, 175
described, 117
Warnings, xxi
X
X, defined, 175
XYZ arm, defined, 176
Z
Z, defined, 176
Thermo Electron
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