Accela Autosampler Hardware Manual Rev.G

Accela Autosampler Hardware Manual Rev.G
Accela Autosampler
Hardware Manual
60357-97000 Revision G
March 2011
© 2011 Thermo Fisher Scientific Inc. All rights reserved.
Accela, ChromQuest, and Xcalibur are registered trademarks of Thermo Fisher Scientific Inc. in the United
States.
Tri-Flow is a trademark of Sherwin-Williams Company. PEEK is a trademark of Victrex PLC. VIEWseal and
POWERseal are trademarks of Greiner Bio One.
The following are registered trademarks in the United States and other countries: Teflon is a registered
trademark of E. I. du Pont de Nemours and Company. Rheodyne is a registered trademark of Rheodyne, LLC.
Valco is a registered trademark of Valco Instruments Co. Inc. and VICI AG.
HyperTerminal is a registered trademark of Hilgraeve, Inc. in the United States.
Thermo Fisher Scientific Inc. provides this document to its customers with a product purchase to use in the
product operation. This document is copyright protected and any reproduction of the whole or any part of this
document is strictly prohibited, except with the written authorization of Thermo Fisher Scientific Inc.
The contents of this document are subject to change without notice. All technical information in this
document is for reference purposes only. System configurations and specifications in this document supersede
all previous information received by the purchaser.
Thermo Fisher Scientific Inc. makes no representations that this document is complete, accurate or errorfree and assumes no responsibility and will not be liable for any errors, omissions, damage or loss that might
result from any use of this document, even if the information in the document is followed properly.
This document is not part of any sales contract between Thermo Fisher Scientific Inc. and a purchaser. This
document shall in no way govern or modify any Terms and Conditions of Sale, which Terms and Conditions of
Sale shall govern all conflicting information between the two documents.
Software versions: Xcalibur 2.0.x or later data system, ChromQuest 4.2 or later data system,
Thermo Foundation 1.0.x or later and Xcalibur 2.1.x or later
Revision history: Revision A, September 2006; Revision B, January 2007; Revision C, March 2008;
Revision D, February 2009; Revision E, October 2009; Revision F, April 2010; Revision G, March 2011
For Research Use Only. Not for use in diagnostic procedures.
Regulatory Compliance
Thermo Fisher Scientific performs complete testing and evaluation of its products to ensure full compliance with
applicable domestic and international regulations. When the system is delivered to you, it meets all pertinent
electromagnetic compatibility (EMC) and safety standards as described in the next section or sections by product name.
Changes that you make to your system might void compliance with one or more of these EMC and safety standards.
Changes to your system include replacing a part or adding components, options, or peripherals not specifically
authorized and qualified by Thermo Fisher Scientific. To ensure continued compliance with EMC and safety standards,
replacement parts and additional components, options, and peripherals must be ordered from Thermo Fisher Scientific
or one of its authorized representatives.
Accela Autosampler
EMC Directive 89/336/EEC amended by 92/31/EEC and 93/68/EEC
EMC compliance has been evaluated by TUV Rheinland of North America, Inc.
EN 61326
1997; A1, 1998; A2, 2001; A3, 2003
EN 61000-4-4
1995; A1, 2000; A2, 2001
EN 61000-3-2
2000
EN 61000-4-5
2001
EN 61000-3-3
1995; A1, 2001
EN 61000-4-6
2003
EN 61000-4-2
2001
EN 61000-4-8
2001
EN 61000-4-3
2002
EN 61000-4-11
2001
FCC Class A, CFR 47 Part 15 Subpart B: 2005
Low Voltage Safety Compliance
Low Voltage Safety Compliance has been evaluated by TUV Rheinland of North America, Inc.
This device complies with Low Voltage Directive 73/23/EEC and harmonized standard EN 61010-1:2001,
IEC 61010-1:2002, UL 61010 A-1:2004, CAN/CSA 22.2 61010-1:2004.
FCC Compliance Statement
THIS DEVICE COMPLIES WITH PART 15 OF THE FCC RULES. OPERATION IS SUBJECT TO
THE FOLLOWING TWO CONDITIONS: (1) THIS DEVICE MAY NOT CAUSE HARMFUL
INTERFERENCE, AND (2) THIS DEVICE MUST ACCEPT ANY INTERFERENCE RECEIVED,
INCLUDING INTERFERENCE THAT MAY CAUSE UNDESIRED OPERATION.
CAUTION Read and understand the various precautionary notes, signs, and symbols contained inside
this manual pertaining to the safe use and operation of this product before using the device.
Notice on Lifting and Handling of
Thermo Scientific Instruments
For your safety, and in compliance with international regulations, the physical handling of this Thermo Fisher Scientific
instrument requires a team effort to lift and/or move the instrument. This instrument is too heavy and/or bulky for one
person alone to handle safely.
Notice on the Proper Use of
Thermo Scientific Instruments
In compliance with international regulations: Use of this instrument in a manner not specified by Thermo Fisher
Scientific could impair any protection provided by the instrument.
Notice on the Susceptibility
to Electromagnetic Transmissions
Your instrument is designed to work in a controlled electromagnetic environment. Do not use radio frequency
transmitters, such as mobile phones, in close proximity to the instrument.
For manufacturing location, see the label on the instrument.
WEEE Compliance
This product is required to comply with the European Union’s Waste Electrical & Electronic Equipment (WEEE)
Directive 2002/96/EC. It is marked with the following symbol:
Thermo Fisher Scientific has contracted with one or more recycling or disposal companies in each European Union
(EU) Member State, and these companies should dispose of or recycle this product. See www.thermo.com/
WEEERoHS for further information on Thermo Fisher Scientific’s compliance with these Directives and the
recyclers in your country.
WEEE Konformität
Dieses Produkt muss die EU Waste Electrical & Electronic Equipment (WEEE) Richtlinie 2002/96/EC erfüllen.
Das Produkt ist durch folgendes Symbol gekennzeichnet:
Thermo Fisher Scientific hat Vereinbarungen mit Verwertungs-/Entsorgungsfirmen in allen EU-Mitgliedsstaaten
getroffen, damit dieses Produkt durch diese Firmen wiederverwertet oder entsorgt werden kann. Mehr Information
über die Einhaltung dieser Anweisungen durch Thermo Fisher Scientific, über die Verwerter, und weitere Hinweise,
die nützlich sind, um die Produkte zu identifizieren, die unter diese RoHS Anweisung fallen, finden sie unter
www.thermo.com/WEEERoHS.
Conformité DEEE
Ce produit doit être conforme à la directive européenne (2002/96/EC) des Déchets d'Equipements Electriques et
Electroniques (DEEE). Il est marqué par le symbole suivant:
Thermo Fisher Scientific s'est associé avec une ou plusieurs compagnies de recyclage dans chaque état membre de
l’union européenne et ce produit devrait être collecté ou recyclé par celles-ci. Davantage d'informations sur la
conformité de Thermo Fisher Scientific à ces directives, les recycleurs dans votre pays et les informations sur les
produits Thermo Fisher Scientific qui peuvent aider la détection des substances sujettes à la directive RoHS sont
disponibles sur www.thermo.com/WEEERoHS.
CAUTION Symbol
CAUTION
VORSICHT
ATTENTION
PRECAUCION
AVVERTENZA
Electric Shock: This instrument uses
high voltages that can cause personal
injury. Before servicing, shut down the
instrument and disconnect the instrument
from line power. Keep the top cover on
while operating the instrument. Do not
remove protective covers from PCBs.
Elektroschock: In diesem Gerät werden
Hochspannungen verwendet, die
Verletzungen verursachen können. Vor
Wartungsarbeiten muß das Gerät
abgeschaltet und vom Netz getrennt
werden. Betreiben Sie Wartungsarbeiten
nicht mit abgenommenem Deckel. Nehmen
Sie die Schutzabdeckung von Leiterplatten
nicht ab.
Choc électrique: L’instrument utilise des
tensions capables d’infliger des blessures
corporelles. L’instrument doit être arrêté et
débranché de la source de courant avant
tout intervention. Ne pas utiliser
l’instrument sans son couvercle. Ne pas
enlever les étuis protecteurs des cartes de
circuits imprimés.
Descarga eléctrica: Este instrumento
utiliza altas tensiones, capaces de
producir lesiones personales. Antes de
dar servicio de mantenimiento al
instrumento, éste debera apagarse y
desconectarse de la línea de alimentacion
eléctrica. No opere el instrumento sin sus
cubiertas exteriores quitadas. No remueva
las cubiertas protectoras de las tarjetas
de circuito impreso.
Shock da folgorazione. L’apparecchio è
alimentato da corrente ad alta tensione
che puo provocare lesioni fisiche. Prima di
effettuare qualsiasi intervento di
manutenzione occorre spegnere ed isolare
l’apparecchio dalla linea elettrica. Non
attivare lo strumento senza lo schermo
superiore. Non togliere i coperchi a
protezione dalle schede di circuito
stampato (PCB).
Chemical: This instrument might contain
hazardous chemicals. Wear gloves when
handling toxic, carcinogenic, mutagenic,
or corrosive or irritant chemicals. Use
approved containers and proper
procedures to dispose waste oil.
Chemikalien: Dieses Gerät kann
gefährliche Chemikalien enthalten. Tragen
Sie Schutzhandschuhe beim Umgang mit
toxischen, karzinogenen, mutagenen oder
ätzenden/reizenden Chemikalien.
Entsorgen Sie verbrauchtes Öl
entsprechend den Vorschriften in den
vorgeschriebenen Behältern.
Chimique: Des produits chimiques
dangereux peuvent se trouver dans
l’instrument. Portez des gants pour
manipuler tous produits chimiques
toxiques, cancérigènes, mutagènes, ou
corrosifs/irritants. Utiliser des récipients
et des procédures homologuées pour se
débarrasser des déchets d’huile.
Química: El instrumento puede contener
productos quimicos peligrosos. Utilice
guantes al manejar productos quimicos
tóxicos, carcinogenos, mutagenos o
corrosivos/irritantes. Utilice recipientes y
procedimientos aprobados para
deshacerse del aceite usado.
Prodotti chimici. Possibile presenza di
sostanze chimiche pericolose
nell’apparecchio. Indossare dei guanti per
maneggiare prodotti chimici tossici,
cancerogeni, mutageni, o
corrosivi/irritanti. Utilizzare contenitori
aprovo e seguire la procedura indicata per
lo smaltimento dei residui di olio.
Heat: Before servicing the instrument,
allow any heated components to cool.
Hitze: Warten Sie erhitzte Komponenten
erst nachdem diese sich abgekühlt haben.
Haute Temperature: Permettre aux
composants chauffés de refroidir avant
tout intervention.
Altas temperaturas: Permita que lop
componentes se enfríen, ante de efectuar
servicio de mantenimiento.
Calore. Attendere che i componenti
riscaldati si raffreddino prima di
effetturare l’intervento di manutenzione.
Fire: Use care when operating the system
in the presence of flammable gases.
Feuer: Beachten Sie die einschlägigen
VorsichtsmaBnahmen, wenn Sie das
System in Gegenwart von entzündbaren
Gasen betreiben.
Incendie: Agir avec précaution lors de
l’utilisation du système en présence de
gaz inflammables.
Fuego: Tenga cuidado al operar el
sistema en presencia de gases
inflamables.
Incendio. Adottare le dovute precauzioni
quando si usa il sistema in presenza di gas
infiammabili.
Eye Hazard: Eye damage could occur
from splattered chemicals or flying
particles. Wear safety glasses when
handling chemicals or servicing the
instrument.
Verletzungsgefahr der Augen:
Verspritzte Chemikalien oder kleine
Partikel können Augenverletzungen
verursachen. Tragen Sie beim Umgang mit
Chemikalien oder bei der Wartung des
Gerätes eine Schutzbrille.
Danger pour les yeux: Des projections
chimiques, liquides, ou solides peuvent
être dangereuses pour les yeux. Porter des
lunettes de protection lors de toute
manipulation de produit chimique ou pour
toute intervention sur l’instrument.
Peligro par los ojos: Las salicaduras de
productos químicos o particulas que
salten bruscamente pueden causar
lesiones en los ojos. Utilice anteojos
protectores al mnipular productos
químicos o al darle servicio de
mantenimiento al instrumento.
Pericolo per la vista. Gli schizzi di
prodotti chimici o delle particelle presenti
nell’aria potrebbero causare danni alla
vista. Indossare occhiali protettivi quando
si maneggiano prodotti chimici o si
effettuano interventi di manutenzione
sull’apparecchio.
General Hazard: A hazard is present that
is not included in the above categories.
Also, this symbol appears on the
instrument to refer the user to instructions
in this manual.
Allgemeine Gefahr: Es besteht eine
weitere Gefahr, die nicht in den
vorstehenden Kategorien beschrieben ist.
Dieses Symbol wird im Handbuch
auBerdem dazu verwendet, um den
Benutzer auf Anweisungen hinzuweisen.
Danger général: Indique la présence
d’un risque n’appartenant pas aux
catégories citées plus haut. Ce symbole
figure également sur l’instrument pour
renvoyer l’utilisateur aux instructions du
présent manuel.
Peligro general: Significa que existe un
peligro no incluido en las categorias
anteriores. Este simbolo también se utiliza
en el instrumento par referir al usuario a
las instrucciones contenidas en este
manual.
Pericolo generico. Pericolo non
compreso tra le precedenti categorie.
Questo simbolo è utilizzato inoltre
sull’apparecchio per segnalare all’utente
di consultare le istruzioni descritte nel
presente manuale.
When the safety of a procedure is
questionable, contact your local Technical
Support organization for Thermo Fisher
Scientific San Jose Products.
Wenn Sie sich über die Sicherheit eines
Verfahrens im unklaren sind, setzen Sie
sich, bevor Sie fortfahren, mit Ihrer
lokalen technischen
Unterstützungsorganisation für Thermo
Fisher Scientific San Jose Produkte in
Verbindung.
Si la sûreté d’une procédure est
incertaine, avant de continuer, contacter
le plus proche Service Clientèle pour les
produits de Thermo Fisher Scientific San
Jose.
Cuando la certidumbre acerca de un
procedimiento sea dudosa, antes de
proseguir, pongase en contacto con la
Oficina de Asistencia Tecnica local para
los productos de Thermo Fisher Scientific
San Jose.
Quando e in dubbio la misura di sicurezza
per una procedura, prima di continuare, si
prega di mettersi in contatto con il
Servizio di Assistenza Tecnica locale per i
prodotti di Thermo Fisher Scientific San
Jose.
CAUTION Symbol
CAUTION
Electric Shock: This instrument uses
high voltages that can cause personal
injury. Before servicing, shut down the
instrument and disconnect the instrument
from line power. Keep the top cover on
while operating the instrument. Do not
remove protective covers from PCBs.
Chemical: This instrument might contain
hazardous chemicals. Wear gloves when
handling toxic, carcinogenic, mutagenic,
or corrosive or irritant chemicals. Use
approved containers and proper
procedures to dispose waste oil.
Heat: Before servicing the instrument,
allow any heated components to cool.
Fire: Use care when operating the system
in the presence of flammable gases.
Eye Hazard: Eye damage could occur
from splattered chemicals or flying
particles. Wear safety glasses when
handling chemicals or servicing the
instrument.
General Hazard: A hazard is present that
is not included in the above categories.
Also, this symbol appears on the
instrument to refer the user to instructions
in this manual.
When the safety of a procedure is
questionable, contact your local Technical
Support organization for Thermo Fisher
Scientific San Jose Products.
C
Contents
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .xiii
Related Documentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .xiii
Finding the Manuals. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .xiv
Accessing the Data System Help for the Autosampler . . . . . . . . . . . . . . . . . . . . xv
Safety and Special Notices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .xvi
Special Precautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xvii
Good Laboratory Practices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xx
Contacting Us . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xxii
Thermo Scientific
Chapter 1
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1
Autosampler Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Tray Compartment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Injection System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Column Oven and Tray Compartment Heater/Cooler . . . . . . . . . . . . . . . . . 15
Status LEDs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Injection Modes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
No Waste Injection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Partial Loop Injection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Full Loop Injection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Injection Sequence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Getting Ready for an Injection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Drawing Transport Solvent (Partial Loop and No Waste Injections) . . . . . . . 23
Drawing Sample . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
Pushing Transport Solvent (Partial Loop and No Waste Injections) . . . . . . . 26
Loading Sample into the Sample Loop . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
Injecting Sample onto the Column . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Post-Injection Events. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
Autosampler Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
Automated Sample Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Specifications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
Chapter 2
Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33
Software Compatibility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
Installation Checklist. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
Unpacking and Inspecting the Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Accela Autosampler Hardware Manual
ix
Contents
Making Initial Instrument Preparations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Making the Back Panel Connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Connecting the System Interconnect Cable . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Connecting the Ethernet Cable . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Checking the Unit ID Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Connecting the Autosampler to Line Power . . . . . . . . . . . . . . . . . . . . . . . . . 39
Connecting the Solvent Lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
Connecting the Wash Bottle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
Connecting the Needle Tubing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
Connecting the Autosampler to the Pump. . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Powering On the Autosampler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Calibrating the Accela Autosampler. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
Changing the Sample Loop . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
x
Chapter 3
Sample Loading . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .51
Standard Tray Type. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
Installing the Standard Tray Type . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
Moving the XYZ Arm to the Back of the Tray Compartment . . . . . . . . . . . . 53
Carrier Trays for Microplates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
Important Precautions for Sample Loading . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Recommended Vials, Microplates, and Microplate Covers . . . . . . . . . . . . . . . . 57
Recommended Vials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Recommended Microplates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Recommended Microplate Covers. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
Chapter 4
Routine Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .61
Maintenance Schedule. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
Replacing the Rotor in the Valco Injection Valve . . . . . . . . . . . . . . . . . . . . . . . 63
Removing the Injection Valve from the Autosampler . . . . . . . . . . . . . . . . . . 64
Disassembling the Injection Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
Cleaning the Stator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
Installing a New Rotor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
Reinstalling the Injection Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
Replacing the Inlet Filter for the Wash Bottle Solvent Line . . . . . . . . . . . . . . . . 71
Replacing the Transfer Tubing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
Maintaining the Syringe Drive Assembly. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
Troubleshooting a Blockage in the Injection System . . . . . . . . . . . . . . . . . . . . . 79
Testing the Needle Tubing and the Syringe Valve for Blockage. . . . . . . . . . . 81
Testing the Needle for Blockage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82
Testing the Transfer Tube for Blockage. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
Cleaning the Injection Valve or the Sample Loop . . . . . . . . . . . . . . . . . . . . . 83
Accela Autosampler Hardware Manual
Thermo Scientific
Contents
Replacing the Needle Tubing Assembly. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85
Setting the Needle to the Removal Position. . . . . . . . . . . . . . . . . . . . . . . . . . 85
Removing the Needle Tubing Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86
Installing the Needle Tubing Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
Setting the XYZ Arm to the Home Position . . . . . . . . . . . . . . . . . . . . . . . . . 91
Replacing the Needle Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
Changing the Syringe or Replacing the Inner Plunger . . . . . . . . . . . . . . . . . . . . 93
Setting the Syringe to the Removal Position . . . . . . . . . . . . . . . . . . . . . . . . . 93
Removing the Syringe from the Autosampler . . . . . . . . . . . . . . . . . . . . . . . . 93
Replacing the Inner Plunger of the Syringe . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Installing the Syringe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Setting the Syringe to the Home Position . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
Clearing a Plugged Heat Exchanger. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96
Replacing a Fuse . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97
Maintenance Cycles. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98
Setting the Maintenance Cycles from the Xcalibur Data System . . . . . . . . . . 98
Setting the Maintenance Cycles from the ChromQuest Data System . . . . . . 99
Chapter 5
Diagnostics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .103
Power Up Diagnostics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103
Error Log in the Xcalibur Data System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105
Error Log in the ChromQuest Data System . . . . . . . . . . . . . . . . . . . . . . . . . . 107
Autosampler Validation Information. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107
Chapter 6
Routine Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .109
General Chromatography Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . 110
Autosampler-Specific Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
Chromatography Problems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 112
Hardware Problems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116
Common Operator Errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120
Insufficient Wash Solvent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120
Damaged Needle Tubing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120
Incorrect Syringe Type . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120
Inappropriate Injection Volume . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
Incorrect Dead Volume . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
Overtightened Transfer Tube or Wash Tube Fittings . . . . . . . . . . . . . . . . . 121
Determining the Transfer Tubing Volume . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
Appendix A Consumables and Service Parts. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .125
Consumables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
Service Parts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
Accela AS Accessory Kit. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
Accela System Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
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Contents
Appendix B Firmware Updates. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .131
Preparing the Autosampler for a Firmware Download . . . . . . . . . . . . . . . . . . . 131
Downloading Firmware to the Autosampler . . . . . . . . . . . . . . . . . . . . . . . . . . 133
Appendix C Quick Reference Guides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .135
Replacing the Rotor Seal in the Rheodyne Valve . . . . . . . . . . . . . . . . . . . . . . . 137
Replacing the Rotor in the Valco Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
Using the Xcalibur Direct Control Commands . . . . . . . . . . . . . . . . . . . . . . . . 141
Using the ChromQuest Direct Commands . . . . . . . . . . . . . . . . . . . . . . . . . . . 143
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .145
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Accela Autosampler Hardware Manual
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P
Preface
This manual describes the features, installation, and maintenance of the Accela™ Autosampler.
This manual also describes portions of the Xcalibur™ or ChromQuest™ data systems required
to perform diagnostics and maintenance procedures for the autosampler.
Contents
• Related Documentation
• Finding the Manuals
• Accessing the Data System Help for the Autosampler
• Safety and Special Notices
• Special Precautions
• Good Laboratory Practices
• Contacting Us
Related Documentation
In addition to this guide, Thermo Fisher Scientific provides the following documentation as
PDF files for the Accela family of LC instruments:
• Accela Preinstallation Requirements Guide
• Accela Getting Connected Guide
• Accela User Guide for LC Devices (formerly known as the Accela User Guide for
Xcalibur 2.1.x)
• Accela User Guide for the ChromQuest Data System
• Help from within the data system
Thermo Scientific
Accela Autosampler Hardware Manual
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Preface
Finding the Manuals
For the ChromQuest data system, you can find the manuals on the ChromQuest software
installation CD.
For Thermo Scientific mass spectrometry applications, such as the Xcalibur data system, you
can find the manuals (provided as PDF files) on the data system computer.
• To access the manuals from LC Devices 2.1.0 or 2.2.0, choose Start > All Programs >
Xcalibur > Manuals > LC Devices > Accela.
• To access the manuals for LC Devices 2.2.1 or later, from the data system computer,
choose Start > All Programs > Thermo Instruments > Manuals > LC Devices > Accela.
.
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Preface
Accessing the Data System Help for the Autosampler
Your data system provides context-sensitive Help for these autosampler topics:
• Instrument configuration parameters for the autosampler
• Injection cycle and temperature control parameters for an acquisition method
(instrument method in the Xcalibur data system or method in the ChromQuest data
system)
• Calibration options for the autosampler
• Direct controls for the autosampler
 To open context-sensitive Help
Do one of the following:
• Press F1.
• Click the Help button, if available.
• For the ChromQuest data system, click the Help icon,
Thermo Scientific
.
Accela Autosampler Hardware Manual
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Preface
Safety and Special Notices
Make sure you follow the precautionary statements presented in this guide. The safety and
other special notices appear in boxes.
Safety and special notices include the following:
CAUTION Highlights hazards to humans, property, or the environment. Each CAUTION
notice is accompanied by an appropriate CAUTION symbol.
CAUTION Alerts you to the presence of high voltage and to the potential injury that could
occur from electrical shock were you to come in contact with a specific instrument area or
component. It also tells you how to avoid contact with the high-voltage areas in your
instrument.
CAUTION Alerts you to the presence of hot surfaces and to the potential injury that could
occur were you to come in contact with a specific instrument area.
CAUTION Highlights a heavy lifting hazard.
IMPORTANT Highlights information necessary to prevent damage to software, loss of
data, or invalid test results; or might contain information that is critical for optimal
performance of the system.
Note Highlights information of general interest.
Tip Highlights helpful information that can make a task easier.
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Preface
Special Precautions
Follow these precautions when handling the autosampler:
• Loading the Autosampler Tray Compartment
• Connecting the Waste Line
• Lifting and Carrying the Autosampler
• Avoiding Contact with Hot Surfaces
Loading the Autosampler Tray Compartment
Do not place tall objects taller than 4.6 cm (1.8 in.) in the tray compartment. Placing tall
objects in the tray compartment can damage the autosampler arm as it moves within the tray
compartment to the sample vial and well locations specified in the injection sequence.
CAUTION Placing tall objects in the tray compartment can damage the autosampler’s
XYZ arm.
Connecting the Waste Line
Ensure that the waste line connected to the drainage manifold is routed to a waste bottle as
shown below.
ACCELA
PDA Detector
Power
Communication
Run
Lamps
Power
Communication
Run
Temperature
Power
Communication
Run
Degas
ACCELA
Autosampler
ACCELA
Pump
Waste
bottle
Wall outlet #1
115 or 230 Vac
Secondary
containment
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Preface
Lifting and Carrying the Autosampler
The autosampler, at 30 kg (66 lbs) and with a height, width, and depth of 37× 38× 51 cm
(14.5 × 15 × 20 in.), is too heavy and too bulky for one person alone to handle safely. In
addition, any jarring or uneven movement can cause misalignment of the autosampler’s
XYZ arm.
CAUTION For your safety and to avoid instrument damage, take care when handling the
autosampler. Lifting or moving the autosampler requires a team effort.
Accela Autosampler
z-axis
height
37 cm
(14.5 in.)
x-axis
width
38 cm
(15 in.)
y-axis
depth
51 cm
(20 in.)
View of the XYZ arm
x-axis
lead screw
XYZ arm
y-axis
stepper
motor
x-axis
stepper
motor
y-axis
lead screw
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Preface
Avoiding Contact with Hot Surfaces
The autosampler has two controlled temperature zones: the tray compartment and
the column oven compartment. Because the column oven can reach temperatures as high as
95 °C (203 °F), ensure that the oven is at room temperature before installing or removing the
LC column and before connecting the pump outlet to the inlet of the autosampler heat
exchanger.
CAUTION To prevent personal injury, before installing the LC column or connecting the
pump to the autosampler, make sure that the column oven compartment is at room
temperature.
THERMO
LC column outlet
connection
LC column inlet
connection
Heat exchanger inlet
connection
20
µL
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Preface
Good Laboratory Practices
To obtain optimal performance from your LC system and to prevent personal injury or injury
to the environment, do the following:
• Keep good records.
• Read the manufacturers’ Material Safety Data Sheets (MSDSs) for the chemicals being
used in your laboratory.
• Remove particulate matter from your samples before you inject them into the liquid
chromatograph.
• Use HPLC-grade solvents.
• Connect the drainage tubes from the pump, autosampler, and detector to an appropriate
waste receptacle. Dispose of solvents as specified by local regulations.
Keep Good Records
To help identify and isolate problems with either your equipment or your methodology, for
best results, keep good records of all system conditions (for example, % RSDs on retention
times and peak areas, and peak shape and resolution). At a minimum, keep a chromatogram
of a typical sample and standard mixture, well documented with system conditions, for future
reference. Careful comparison of retention times, peak shapes, peak sensitivity, and baseline
noise can provide valuable clues to identifying and solving future problems.
Chemical Toxicity
Although the large volume of toxic and flammable solvents used and stored in laboratories can
be quite dangerous, do not ignore the potential hazards posed by your samples. Take special
care to read and follow all precautions that ensure proper ventilation, storage, handling, and
disposal of both solvents and samples. Become familiar with the toxicity data and potential
hazards associated with all chemicals by referring to the manufacturers’ Material Safety Data
Sheets.
Sample Preparation
Always consider the solubility of your sample in the solvent/mobile phase. Sample
precipitation can plug the column, tubing, or flow cell causing flow restriction. This
obstruction can result in irreparable damage to the system. You can remove particulate matter
by filtering the samples through 0.45 or 0.2 μm (or less) filters.
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Preface
Solvent Requirements
Many chemical manufacturers provide a line of high-purity or HPLC-grade reagents that are
free of chemical impurities. Typically, HPLC-grade solvents do not require filtration. You can
significantly prolong the life and effectiveness of the inlet filters, check valves and seals,
injector, and column by routine filtration of other types of solvents through a 0.45 or 0.2 μm
(or less) fluorocarbon filter before placing them in the solvent reservoir bottles.
Choose a mobile phase that is compatible with the sample and column you have selected for
your separation. Remember that some solvents are corrosive to stainless steel.
Solvent Disposal
Make sure you have a solvent waste container or other kind of drain system available at or
below the benchtop level. Most solvents have special disposal requirements and should not be
disposed of directly down a drain. Follow all governmental regulations when disposing of any
chemical.
High-Pressure Systems and Leaks
LC systems operate at high pressures. Because liquids are not highly compressible, they do not
store much energy. Accordingly, the high pressures in an LC system pose little immediate
danger. However, correct any leak that occurs as soon as possible. Always wear eye and skin
protection when operating or maintaining an LC system. Always shut down the system and
return it to atmospheric pressure before attempting any maintenance.
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Preface
Contacting Us
There are several ways to contact Thermo Fisher Scientific for the information you need.
 To contact Technical Support
Phone
800-532-4752
Fax
561-688-8736
E-mail
[email protected]
Knowledge base
www.thermokb.com
Find software updates and utilities to download at mssupport.thermo.com.
 To contact Customer Service for ordering information
Phone
800-532-4752
Fax
561-688-8731
E-mail
[email protected]
Web site
www.thermo.com/ms
 To get local contact information for sales or service
Go to www.thermoscientific.com/wps/portal/ts/contactus.
 To copy manuals from the Internet
Go to mssupport.thermo.com, agree to the Terms and Conditions, and then click
Customer Manuals in the left margin of the window.
 To suggest changes to documentation or to Help
• Fill out a reader survey online at www.surveymonkey.com/s/PQM6P62
–or–
Click the link below.
• Send an e-mail message to the Technical Publications Editor at
[email protected]
xxii
Accela Autosampler Hardware Manual
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1
Introduction
The Accela Autosampler is a member of the Accela family of ultra-high-performance liquid
chromatography instruments. The Accela Autosampler (see Figure 1) holds up to
200 standard vials, three 96-well plates, or three 384-well plates; contains a built-in column
oven (5 to 95 °C); provides tray/sample temperature control (0 to 60 °C); and is capable of
performing automated sample preparation routines.
Contents
• Autosampler Components
• Status LEDs
• Injection Modes
• Injection Sequence
• Autosampler Operation
• Automated Sample Preparation
• Specifications
Figure 1.
Accela Autosampler
Status LEDs
On/Off power switch
Thermo Scientific
Accela Autosampler Hardware Manual
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1
Introduction
Autosampler Components
Autosampler Components
The main components of the Accela Autosampler are as follows:
• Tray Compartment
• Injection System
• Column Oven and Tray Compartment Heater/Cooler
Tray Compartment
Two types of sample trays are supplied with the Accela Autosampler: a conventional sample
tray and a microwell carrier. The tray compartment can hold up to five conventional sample
trays or one microwell carrier. The tray compartment also holds up to four 16 mL capacity
reservoir vials that can hold solvent, reagent, or diluent. The reservoir vials are located behind
the wash station and are designated RV1, RV2, RV3, and RV4. Figure 2 shows an empty tray
compartment.
Figure 2.
Tray compartment
XYZ arm
mechanism
A
Injection port
B
C
D
E
From the left to the right side of the tray compartment, the five conventional sample trays are
designated A, B, C, D, and E. Each conventional sample tray holds up to 40 standard 1.8 mL
vials for a total capacity of 200 samples. Overlays allow the sample trays to accommodate
different vial sizes.
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1 Introduction
Autosampler Components
The microwell carrier can hold up to three low-density microplates or up to three
high-density microplates. From the front to the back of the tray compartment, the
microplates are designated A, B, and C.
The low-density microplate contains 96 wells in an 8 × 12 configuration, designated A1
through H12, for a total capacity of 288 samples. Each well can hold up to 250 μL of sample.
The high-density microplate contains 384 wells in a 16 × 24 configuration, designated A1
through P24, for a total capacity of 1152 samples. Each well can hold up to 50 μL of sample.
The tray compartment door contains a magnetic switch. The magnet is located in the door
and the switch is attached to the chassis. When you open the door, the switch signals the
autosampler that the door is open. If you turn on the verification option for the door position
when you configure the autosampler, the XYZ arm automatically moves to the back of the
tray compartment when you open the door so that you can remove trays or replace vials.
Opening the tray compartment door while the autosampler is making an injection does not
interrupt the current run. The XYZ arm moves to the back of the compartment after the
injection is complete and the sequence pauses. Closing the autosampler door allows the
sequence to resume.
Injection System
Figure 3 shows the injection system of the Accela Autosampler. The major components of the
injection system are as follows:
• Wash Bottle and Wash Bottle Tube
• XYZ Arm Mechanism
• Needle Tubing Assembly and Needle Assembly
• Syringe Valve
• Syringe
• Wash Station
• Injection Port of the Autosampler and Transfer Tube
• Injection Valve
• Sample Loop
You can order the replaceable components of the injection system by Thermo Scientific part
number. “Consumables” on page 126 lists the part numbers.
CAUTION Because the XYZ arm moves to the sample position, do not place objects taller
than 4.5 cm (1.8 in.) in the tray compartment.
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1
Introduction
Autosampler Components
Figure 3.
Accela Autosampler injection components
Syringe
valve
Needle tubing
assembly
Wash
tube
Concentric
syringe
Wash
bottle
Reservoir
vial
Wash
station
Injection port of
autosampler
Standard
1.8 mL vial
Dead
volume
Inject
position
Transfer tube
To
column
1
2
6
3
Wash
tube
Mobile phase
Wash solvent
Sample
4
Accela Autosampler Hardware Manual
4
5
Injection valve
From
pump
Thermo Scientific
1 Introduction
Autosampler Components
Wash Bottle and Wash Bottle Tube
The wash bottle rests in the solvent platform on the top of the Accela stack. The wash bottle
tube connects the wash bottle to the syringe valve. Both of the direct commands, Flush
(from bottle) and Wash Needle (from bottle), draw solvent from the wash bottle. If you let the
wash bottle run dry, the syringe barrel and the wash bottle tube fill with air. When this
happens, the autosampler cannot draw sample into the needle tubing during an injection
sequence.
IMPORTANT Check the volume of solvent in the wash bottle before performing a
sequence of injections. Letting the wash bottle run dry compromises the performance of
the autosampler.
XYZ Arm Mechanism
The XYZ arm mechanism moves the needle along the X-Y plane to the requested vial or well
location. After it positions the needle above the vial or well, the XYZ arm lowers the needle
along the z axis to the requested needle height. The syringe plunger descends, drawing sample
into the needle tubing. The XYZ arm mechanism moves back to the home position, above the
injection port of the autosampler, and then lowers the needle into the injection port. The
syringe plunger ascends, expelling the sample from the needle tubing.
Do not move the XYZ arm manually. Instead, use the commands and options listed in
Table 1 to control the position of the XYZ arm. For information on using the data system
commands, see “Using the Xcalibur Direct Control Commands” on page 141 or “Using the
ChromQuest Direct Commands” on page 143.
Table 1. Commands and options that control the position of the XYZ arm
Action
Data system
Command or option
Make the XYZ arm automatically move
to the back of the tray compartment when
you open the tray door.
Xcalibur
Select the Verify Door Is Closed check box when
you specify the instrument configuration options.
ChromQuest
Select the Verify Door Is Closed check box when
you specify the instrument configuration options.
Xcalibur
Use the Position Arm to Access Tray direct
command.
ChromQuest
Use the Position Arm to Access Tray direct control
command.
Xcalibur
Use the Set Arm to Home Position direct
command.
ChromQuest
Use the Go to Home direct control command.
Xcalibur
Use the Remove Needle direct command.
ChromQuest
Use the Needle Removal direct control command.
Move the XYZ arm to the back of the
tray compartment.
Move the XYZ arm to its home position
above the injection port and behind the
wash station.
Move the XYZ arm to the center front of
the tray compartment, allowing easy access
to the needle.
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1
Introduction
Autosampler Components
Needle Tubing Assembly and Needle Assembly
Figure 4 shows the needle assembly and the needle tubing assembly.
The needle assembly consists of a blunt-tip needle, a latch nut, a flag, a compression spring,
and an externally threaded fitting that connects to the needle tube assembly.
The needle tube assembly connects the solvent path between the needle and the syringe valve
and consists of low-pressure tubing, an internally threaded fitting that connects to the needle
assembly fitting, a sleeve, a black or red PVC cap, a needle tube guide that attaches to the
x-axis positioning frame, and an externally threaded fitting with a flangeless ferrule that
connects to the right side of the syringe valve.
Figure 4.
Needle assembly and needle tubing assembly
Needle assembly
Latch nut
Compression spring
Flag
Needle tubing assembly
Externally threaded nut and flangeless
ferrule that connect to the right side of
the syringe valve
10-32
fitting
Low-pressure tubing
Blunt-tip
needle
PEEK fitting
(that connects to the
needle assembly)
Sleeve
Two-pronged
needle guide
Black or red
PVC cap
The needle mount on the XYZ arm holds the needle assembly. To prevent the needle tubing
from interfering with the movement of the XYZ arm, the following items secure the needle
tubing to the autosampler (see Figure 5):
• A bracket on the XYZ arm
• A guide on the x-axis positioning frame
• A bracket on the left wall of the tray compartment
• A PVC cap that holds the tubing below the metal runner for the x-axis positioning frame
• A bracket on the inner wall of the tray compartment door
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1 Introduction
Autosampler Components
Figure 5.
Needle tubing assembly connections
Bracket on left wall of the tray compartment
(alternate view of the x-axis positioning frame at the
back of the tray compartment)
Needle tubing guide
connected to the x-axis
positioning frame
Bracket
on the XYZ arm
Bracket
on the inside wall of the
tray compartment door
Connection to the right side
of the syringe valve
Black or red
PVC cap
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1
Introduction
Autosampler Components
The standard needle tubing assembly holds approximately 560 μL of liquid. During an
injection, the syringe draws sample into the needle tubing, not into the syringe barrel. If your
autosampler is configured with a 2500 μL standard syringe, and you want to make partial
loop injections greater than 500 μL or full loop injections greater than 150 μL, you must add
additional needle tubing to your autosampler.
The tubing extension, which the autosampler needs to make large volume injections, holds up
to 1 mL of liquid. Adding this additional tubing to your autosampler allows the withdrawal of
up to 1500 μL from a sample location or a reservoir vial. To attach this additional tubing to
your autosampler, detach the needle tubing assembly from the syringe valve. Use the Teflon™
connector to attach the additional tubing to the needle tubing assembly. Attach the other end
of the additional tubing to the syringe valve.
Syringe Valve
The syringe valve is a 2-position rotary valve (see Figure 6). In the wash bottle position, the
downward movement of the syringe plunger draws wash solvent from the wash bottle into the
syringe barrel. In the needle position, the downward movement of the syringe plunger draws
liquid from a sample vial or a reservoir vial into the needle tubing. As the syringe plunger
moves upward, it pushes liquid out of the needle tubing.
Figure 6.
Two-position rotary syringe valve
To
needle tubing
assembly
From
wash bottle
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Accela Autosampler Hardware Manual
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1 Introduction
Autosampler Components
Syringe
The Accela Autosampler uses a syringe with two concentric barrels to perform liquid transfers
(see Figure 7). The autosampler uses the inner barrel plunger to draw and deliver sample
amounts equal to or less than the nominal size of the syringe and the outer barrel plunger to
draw and expel volumes up to 560 μL.
Figure 7.
Dual concentric syringe
Outer barrel
Inner plunger
Inner barrel
The inner plunger wears more rapidly than the rest of the syringe. The lifespan of the inner
plunger is approximately 50000 injections. Replace the inner plunger if you notice liquid
leaking from the bottom of the syringe. For ordering information, see page 126.
Dual concentric syringes are available in 100, 250, and 500 μL sizes. In addition, a 2500 μL
standard syringe is available. The size of the syringe determines the injection volume range.
Table 2 lists the minimum and maximum injection volumes for each syringe size.
Table 2. Allowable injection volumes based on the syringe size
Injection volume (μL)
a
Thermo Scientific
Syringe size (μL)
Minimum
Maximum
(No waste)
Maximum
(Partial loop)
Maximum
(Full loop)
100
0.1
20
20
20
250
0.1
100
100
100
500
0.1
250
250
250
2500
0.1
1000
1000
1000a
Requires an extension to the standard needle tubing
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Autosampler Components
Wash Station
The wash station is a waste cup containing an inner sleeve slightly larger than the needle
(see Figure 8). Low-pressure tubing connects the waste cup to the waste bottle. To wash the
outside of the needle, the XYZ arm moves the needle to the wash station and lowers the
needle into the inner sleeve of the waste cup. The syringe draws solvent from the wash bottle
or requested reservoir vial, and then expels the solvent through the needle. As it fills the waste
cup, the expelled solvent flows up over the exterior surface of the needle, and then out the
waste tubing to the waste bottle.
Figure 8.
Wash station
Dual concentric
syringe
Waste cup
Needle
Sleeve
To
waste
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Autosampler Components
Injection Port of the Autosampler and Transfer Tube
A transfer tube connects the injection port (see Figure 9) to the injection valve (see Figure 10).
The transfer tubing has an inner diameter of 0.012 in. One end of the tubing has a flanged
stainless steel fitting that connects to the injection port. The other end of the tubing has a
PEEK™ fitting that connects to port 2 of the injection valve (see Figure 11).
IMPORTANT
1. The label attached to the transfer tube assembly specifies the internal volume of the
tubing. Because the autosampler uses this value for its injection algorithm, you must
type this value in the Dead Volume box when you configure the Accela Autosampler.
If you do not enter the value listed on the label, your data might be compromised.
2. You can constrict the transfer tube by overtightening the fitting to port 2 of the
injection valve. Lack of injection precision is a symptom of a constricted transfer tube.
Figure 9.
Injection port of the Accela Autosampler
Injection port of
autosampler
Transfer tubing
Figure 10. Transfer tubing connections
Calibrated
volume of the
transfer tubing
21µL
Injection valve
Waste
tubing
Thermo Scientific
Transfer
tubing
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Autosampler Components
Injection Valve
The injection valve is a six-port, two-position valve that introduces sample onto the column
by way of the sample loop (see Figure 11).
Figure 11. Six-port rotary injection valve
Connection from pump to
heat exchanger
Volume label
20
µL
Port 3
Connection to
waste
Port 2
Connection to
transfer tube
2
Port 1
1
3
Port 4
6
4
5
Port 6
Connection to
column
Port 5
Return from
heat exchanger
Sample loop
There are two positions for the injection valve: fill (load) and inject. Three passages are open
in each position.
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Autosampler Components
In the fill (load) position (see Figure 12), the three passages connect the following ports:
• The injection port of the injection valve (port 2) is connected to the front of the
sample loop (port 1).
• The back of the sample loop (port 4) is connected to the waste line (port 3).
• The temperature-equilibrated mobile phase that has passed through the
heat exchanger (port 5) is connected to the LC column (port 6).
To isolate the sample loop from the mobile phase stream, the autosampler rotates the valve to
the load position (see Figure 12). While the mobile phase stream bypasses the loop, the
syringe pushes sample through the transfer tubing and into the front of the sample loop. As
the sample loop fills with sample, excess solution passes out of the back of the sample loop to
waste.
Figure 12. Fill (load) position for the six-port injection valve
Autosampler
injection port
To
LC column
Heat
exchanger
Transfer
tubing
2
1
6
To
waste
3
4
From
pump
5
Mobile phase
Sample
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Introduction
Autosampler Components
In the inject position (see Figure 13), the three passages connect the following ports:
• The solvent return from the heat exchanger (port 5) is connected to the back of the loop
(port 4).
• The front of the sample loop (port 1) is connected to the column inlet (port 6).
• The injection port of the injection valve (port 2) is connected to the waste line (port 3).
To put the sample loop into the path of the mobile phase stream, the autosampler rotates the
valve to the inject position (see Figure 13). The mobile phase stream enters the sample loop
from the back, backflushing the contents of the sample loop onto the column. Excess sample
left in the transfer tube passes directly to waste. To provide ample rinsing of the sample loop
with mobile phase, the injection valve remains in the inject position during the entire run.
Figure 13. Inject position for the six-port injection valve
Autosampler
injection port
To
column
Heat
exchanger
2
1
6
3
To
waste
4
From
pump
5
Sample moving
toward the LC column
Mobile phase
Sample
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Autosampler Components
Sample Loop
The sample loop is a stainless steel tube with end fittings that holds the sample prior to its
introduction onto the column. The sample loop is an interchangeable part swaged onto
ports 1 and 4 of the six-port injection valve (see Figure 14).
Figure 14. Schematic of the sample loop attached to the injection valve
25 µL
Sample loop
Thermo Fisher Scientific ships the Accela Autosampler with a 25 μL sample loop. Sample
loops are available in nominal sizes ranging from 5 to 1000 μL. The accuracy of the sample
loop volume is ± 20%. This means that the actual volume of a 25 μL sample loop is between
20 to 30 μL. For ordering information, see “Consumables” on page 126.
Column Oven and Tray Compartment Heater/Cooler
The built-in column oven controls the temperature of the air surrounding the
chromatographic column. Isothermal temperature control is achieved with a Peltier device.
The Peltier device is a solid-state, heat-transferring assembly used to heat or cool the column
oven. The temperature control range is 5 to 95 °C.
Between the pump outlet and the autosampler injection valve, the mobile phase passes
through a heat exchanger that is located behind the column oven (see Figure 75 on page 96).
As the mobile phase passes through the heat exchanger, it equilibrates to the temperature of
the column oven. The heat exchanger consists of a length of 0.004 in. ID tubing that adds
only 3 μL of gradient delay volume to the system (see Figure 75 on page 96).
The tray temperature control feature provides temperature control of the samples in the range
from 0 to 60 °C. A Peltier device maintains the tray temperature.
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Introduction
Status LEDs
Status LEDs
Four status light-emitting diodes (LEDs) labeled Power, Communication, Run, and
Temperature are located on the front of the autosampler’s tray compartment door
(see Figure 15). Table 3 lists the states of the LEDs.
Figure 15. Status LEDs
Power
Communication
Run
Temperature
Note To control the temperature of the column oven compartment and the tray
compartment, select the Wait for Temperature Ready check box when you add the Accela
Autosampler to the data system instrument configuration.
Table 3. Status LED states
LED
State
Meaning
Power
Green
The autosampler power is on.
Comm
Amber
Communication with the data system has not
been established.
Green
Communication with the data system has been
established.
Flashing Amber
An error condition, such as an XYZ arm jam or
initialization startup error, has occurred.
Green
The autosampler is in the Ready state.
Flashing Green
An injection or timed event is in progress.
Amber
A temperature change within the column oven
or tray temperature zones is in progress.
Green
The column oven and tray temperature zones
are in equilibrium at the set temperature.
Run
Tempa
a
16
The Temp LED remains green if you do not select the Wait for Temperature Ready check box when you add the
Accela Autosampler to the instrument configuration.
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Introduction
Injection Modes
Injection Modes
The Accela Autosampler can perform the following injection modes:
• No Waste Injection
• Partial Loop Injection
• Full Loop Injection
The optimum injection mode depends on the amount of sample that you have and the degree
of precision that your application requires.
No Waste Injection
In the no waste injection mode, the autosampler withdraws only the exact amount of sample
requested from the sample vial. Of the three injection modes, the no waste injection mode
uses the least amount of sample, but it is also the least precise. Use this injection mode to
conserve sample.
IMPORTANT
For no waste injections, do the following:
• Use a sample loop that is at least 5 μL larger than the injection volume. Because the
accuracy of the nominal size is ± 20%, use an estimate of 80% for the actual size. For
example, use 20 μL as an estimate for the actual volume of a 25 μL loop, and inject no
more than 15 μL with this loop size.
• Consider matching the chemistry of the sample matrix, the flush solution, and the
mobile phase. For no waste injections, the autosampler loads approximately 2 μL of
flush solvent and 3 μL of air into the sample loop (see Figure 16), regardless of the
requested injection volume.
• Inject at least 1.0 μL of sample.
Approximately 0.25 μL of the sample is lost as it travels from the injection port through the
transfer tubing and into the injection valve. Because of this loss, inject at least 1.0 μL of
sample with the no waste injection mode.
The quantity of lost sample depends on the syringe rate. Decreasing the syringe rate decreases
the sample loss. Thermo Fisher Scientific recommends a syringe rate no greater than 4 μL/s
for the no waste injection mode.
In addition to the sample, the no waste injection mode loads approximately 2 μL of flush
solvent and 3 μL of air into the sample loop (see Figure 16). Because these values are
independent of the injection volume, the chemistry of the flush solution can affect your
chromatographic results. For best chromatographic results, consider matching the chemistry
of the sample matrix, the flush solution, and the mobile phase.
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Introduction
Injection Modes
Figure 16. No waste injection mode – load position
To
column
1
2
6
3
Mobile phase
Flush solvent
Sample
4
5
Air bubbles
From
pump
Note The two 1.5 μL air bubbles isolate the sample from the flush solvent.
Partial Loop Injection
In the partial loop injection mode, the autosampler withdraws 22 μL of excess sample from
the vial in addition to the requested injection volume. The autosampler expels approximately
one-half of the excess volume to waste before it meters the center of the sample plug into the
front of the sample loop. It expels the second portion of excess sample to waste after the
mobile phase stream backflushes the contents of the sample loop onto the column.
Because the autosampler meters sample into the sample loop, the partial loop injection mode
is less precise but more accurate than the full loop injection mode. You can inject variable
amounts of sample, ranging from a minimum of 0.1 μL to a practical maximum of one-half
the volume of your sample loop. The laminar flow of fluid within the stainless steel sample
loop causes this maximum volume limitation.
IMPORTANT To make precise partial loop injections, limit the maximum injection
volume to less than half the nominal sample loop size. Because the accuracy of the
nominal volume of the sample loop is ± 20% (which means that the actual volume of the
standard 25 μL sample loop is between 20 μL and 30 μL), for best results, limit the
maximum injection volume to 10 μL for the standard sample loop.
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Introduction
Injection Modes
Full Loop Injection
In the full loop injection mode, the autosampler withdraws a sample volume sufficient to
overfill the loop (according to the equation given on page 19). Because the actual injection
volume is determined by the size of the loop, not the metering action of the stepper motor, a
full loop injection is very reproducible. However, because the full loop injection mode
completely fills the sample loop, you cannot inject variable amounts of sample.
Use the full loop injection mode when you want maximum precision and have unlimited
sample. However, if you want to change the injection volume, you must replace the attached
sample loop with one of a different size. The available sample loop sizes are listed in
“Consumables” on page 126.
Note You enter the size of the sample loop when you configure the Accela Autosampler.
In the full loop injection mode, the autosampler withdraws a large excess of solution from the
sample vial according to the following equation:
Amount withdrawn = 3 × IV + DV + 7.5 μL
Where:
IV = requested injection volume
DV = Volume of the transfer tube + volume of the injection port of the autosampler and
the rotor slot (1.6 μL)
This equation is valid until the volume exceeds the maximum capacity of the syringe. If you
request a 100 μL full loop injection and your autosampler has a 250 μL syringe, the
autosampler withdraws 265 μL of sample solution from the vial, which is the maximum
capacity of the syringe. A sample volume equal to the actual volume of the sample loop is
injected onto the column. Table 4 lists the maximum capacity for each syringe.
Table 4. Maximum volume withdrawn for full loop injections
a
Thermo Scientific
Nominal syringe size
Maximum volume withdrawn
Concentric 100 μL
101 μL
Concentric 250 μL
265 μL
Concentric 500 μL
535 μL
Standard 2500 μL
1544 μLa
The data system limits the maximum amount withdrawn to 1544 μL.
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Injection Sequence
Injection Sequence
The primary sequence of events for the full loop injection mode is as follows:
1. Getting ready for an injection
2. Drawing sample from the vial
3. Loading sample into the loop
4. Injecting sample
5. Post-injection events
The full loop injection mode draws enough sample to overfill both the transfer tube and the
sample loop. The no waste injection mode, however, draws only the injection volume
requested, and the partial loop injection mode draws only the injection amount requested
plus an additional 22 μL.
As a result, unlike the full loop mode, both the partial loop and no waste injection modes
require additional solvent to push the sample all the way through the transfer tubing to the
injection valve before the sample is loaded into the loop. The syringe draws this extra
transport solvent from the wash bottle and uses it to compensate for the volume held by the
injection port of the autosampler, the transfer tube, and the rotor slot of the injection valve.
Before the sample is loaded into the loop, the syringe uses the transport solvent to push the
sample to the injection valve.
The primary sequence of events for both the partial loop injection mode and the no waste
injection mode is as follows:
1. Getting Ready for an Injection
2. Drawing Transport Solvent (Partial Loop and No Waste Injections)
3. Drawing Sample
4. Pushing Transport Solvent (Partial Loop and No Waste Injections)
5. Loading Sample into the Sample Loop
6. Injecting Sample onto the Column
7. Post-Injection Events
Table 5 outlines the sequential events that occur during an injection.
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Injection Sequence
Table 5. Injection sequence
#
Full loop
#
Partial loop
No waste
1
Get ready for injection
1
Get ready for injection
Get ready for injection
Switch injector to inject position
Switch injector to inject position
Draw transport solvent
Draw transport solvent
Switch syringe valve to wash
solvent position
Switch syringe valve to wash
solvent position
Draw wash solvent
(transport volume = total
dead volume + 7.5 μL)
Draw wash solvent
(transport volume = total
dead volume)
Draw Sample
Draw Sample
Move XYZ arm to vial
Move XYZ arm to vial
Move XYZ arm to vial
Switch syringe valve to needle
position
Switch syringe valve to needle
position
Switch syringe valve to needle
position
Draw 3 μL air bubble
Draw 3 μL air bubble
Draw 1.5 μL air bubble
Draw sample
[(3 × IV) + DV + 7.5 μL]
Draw sample equal to
Inject Volume (IV) = 22 μL
Draw sample equal to
Inject Volume (IV)
Draw 3 μL air bubble
Draw 3 μL air bubble
Draw 1.5 μL air bubble
Push Transport Solvent
Push Transport Solvent
Move XYZ arm to injection port
of autosampler
Move XYZ arm to injection port
of autosampler
Push the transport volume into
the transfer tube
Push the transport volume
(minus 1 μL) into the transfer
tube
Load Sample into Loop
Load Sample into Loop
Switch the injector to fill position
Switch the injector to fill position
Meter the sample plug into the
loop
Meter the sample plug + air
bubbles + 2 μL wash solvent into
the loop
N/A
2
Switch injector to fill position
2
Draw Sample
N/A
3
4
Move XYZ arm to injection
port of autosampler
3
Load Sample into Loop
5
Overfill the sample loop
4
Switch the injector to inject
position
6
Switch the injector to inject
position
Switch the injector to inject
position
5
Post-Injection Events
7
Post-Injection Events
Post-Injection Events
Perform optional flush/wash
Perform optional flush/wash
Perform optional flush/wash
Home syringe
Home syringe
Home syringe
Prepare for next injection
Prepare for next injection
Prepare for next injection
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Injection Sequence
Getting Ready for an Injection
Before an injection sequence begins, the autosampler initializes the syringe plunger to the
Ready position (see Figure 17).
• If the syringe plunger is in the Home Side of Ready position, the autosampler switches
the syringe valve to the solvent bottle position, and then returns the syringe plunger
downward to the Ready position at flush speed.
• If the syringe plunger is below the Ready position, the autosampler switches the syringe
valve to the needle position, the XYZ arm moves the needle to the wash station, and the
syringe plunger moves upward to the Ready position at flush speed.
Figure 17. Syringe, syringe valve, and needle position prior to injection (Ready position)
From
wash bottle
Home
Needle tube
Ready
Wash station
Injection port
To
waste
To
injection valve
Wash solvent
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Injection Sequence
Drawing Transport Solvent (Partial Loop and No Waste Injections)
To draw transport solvent, the syringe valve switches to the wash bottle position. The amount
of solvent that the syringe draws from the wash bottle, referred to as the transport volume,
depends on both the injection mode and the configured value for the dead volume.
IMPORTANT A transfer tube connects the injection port of the autosampler to port 2 of
the injection valve. The transfer tube is labeled with its factory calibrated volume. When
you configure the autosampler, you must type the label value in the Dead Volume box of
the Accela Configuration dialog box.
In the no waste injection mode, the syringe draws only enough solvent to compensate for the
total dead volume of the system according to the following equation:
Transport solvent volume = Total dead volume
Where:
Total Dead Volume = Transfer tube volume + volume of the injection port of the
autosampler and the rotor slot of the injection valve (1.6 μL)
In the partial loop injection mode, the syringe draws enough solvent from the wash bottle to
compensate for the total dead volume of the system plus an additional 7.5 μL of solvent (see
Figure 18).
Figure 18. Drawing wash solvent from the wash bottle
Two-way
syringe valve
Needle tubing
assembly
Wash
bottle
Wash
tubing
Wash solvent
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Introduction
Injection Sequence
Drawing Sample
The autosampler draws sample from the sample vial or well as follows:
Note For the full loop injection mode, the injection valve rotates to the load position. For
both the no waste injection mode and the partial loop injection mode, the injection valve
remains in the inject position.
1. The XYZ arm moves to the sample vial.
2. The syringe valve switches to the needle position.
3. The inner syringe plunger descends, drawing an air bubble into the tip of the needle.
Depending on the injection mode, the autosampler draws the following volume of air (see
Figure 19):
• For partial loop and full loop injections, the syringe draws 3 μL of air.
• For no waste injections, the syringe draws 1.5 μL of air.
Figure 19. Drawing the first air bubble
Needle
tubing
Wash
bottle
Air bubble
Wash solvent
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Injection Sequence
4. The XYZ arm lowers the needle tip into the sample vial to the depth requested (see
Figure 20).
Figure 20. Needle withdrawing sample from a standard vial
Needle height
from bottom
Bottom
of vial
2 mm
Wash solvent
Sample
Note The autosampler lowers the needle tip to the position requested in the Needle
Height From Bottom box of the Accela Autosampler page of the Instrument Setup
dialog box. The allowable range is 0.1 to 18 mm from the bottom of the vial.
5. The inner syringe plunger descends further, drawing sample at syringe sample speed. The
volume of sample that the autosampler draws from the vial depends on the requested
injection volume and the injection mode as follows:
• For no waste injections, the autosampler draws only the requested injection volume.
• For partial loop injections, the autosampler draws the requested injection volume
plus an additional 22 μL.
6. The XYZ arm raises the needle out of the sample vial (see Figure 21). The inner syringe
plunger descends further, drawing a second air bubble into the needle. The sample plug is
now isolated between the two air bubbles.
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Injection Sequence
Figure 21. Drawing a second air bubble, which isolates the sample plug
Wash
bottle
Air
bubbles
Wash solvent
Sample
Pushing Transport Solvent (Partial Loop and No Waste Injections)
The XYZ arm moves to the home position, and then lowers the needle into the injection port
of the autosampler. The inner syringe plunger moves upward, pushing the sample plug
followed by the transport solvent into the transfer tube.
The actions that occur during the pushing transport solvent step depend on the injection
mode.
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Injection Sequence
Pushing Transport Solvent Step for No Waste Injections
The needle expels 1 μL less than the transport volume into the transfer tube. The sample plug
falls just short of the entry to the injection port (port 2) of the injection valve (see Figure 22).
Figure 22. No Waste injection—Pushing transport solvent step
Injection port of
autosampler
Air bubbles
To
column
1
2
6
3
5
4
Mobile phase
Flush solvent
Sample
From
pump
Waste line
Pushing Transport Solvent Step for Partial Loop Injections
The needle expels the transport volume, which is equal to the dead volume between the
injection port of the autosampler and the injection valve plus an excess of 7.5 μL, into the
transfer tube. This pushes 4.5 μL of the sample plug plus the first 3 μL bubble through the
injection valve and out to waste (see Figure 23).
Figure 23. Partial Loop Injection—Pushing transport solvent step
Injection port of
autosampler
Air bubbles
To
column
1
2
6
3
Mobile phase
Flush solvent
Sample
Thermo Scientific
4
Waste line
5
From
pump
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Injection Sequence
Loading Sample into the Sample Loop
The injection valve switches to the fill position for both the partial loop injection mode and
the no waste injection modes. For a full loop injection, the injection valve switches to the fill
position at the beginning of the injection sequence. The syringe then pushes the sample into
the sample loop. The Accela Autosampler issues a momentary Gradient Start signal.
The actions that occur during the sample loading step depend on the injection mode.
Sample Loading for No Waste Injections
The syringe meters the requested sample volume plus 5 μL into the sample loop. This pushes
the sample plug, the two bracketing air bubbles, and approximately 2 μL of wash solvent into
the sample loop (see Figure 24).
Figure 24. No Waste injection—Loading the sample loop
Air bubbles
From
injection port
To
column
1
2
6
3
5
4
Mobile phase
Wash solvent
Sample
From
pump
Waste line
Sample Loading for Partial Loop Injections
The syringe meters the requested sample volume into the sample loop. This leaves
approximately 16 μL of excess sample plus the second 3 μL air bubble in the transfer line
(see Figure 25).
Figure 25. Partial Loop injection—Loading the sample loop
From
injection port
To
column
1
2
6
3
Air bubbles
Mobile phase
Wash solvent
Sample
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To waste
5
From
pump
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1 Introduction
Injection Sequence
Sample Loading for Full Loop Injections
The needle expels the sample into the sample loop. As the sample loop becomes overfilled,
excess sample exits through the waste line (see Figure 26).
Figure 26. Full Loop injection—Loading the sample loop
From
injection port
To
column
1
2
6
3
Air bubbles
Mobile phase
Wash solvent
Sample
5
4
From
pump
To waste
Injecting Sample onto the Column
The autosampler backflushes the contents of the sample loop onto the column as follows:
1. The injection valve switches to the inject position.
2. Mobile phase enters the sample loop from the back, sweeping the contents of the sample
loop onto the column (see Figure 27).
3. The autosampler issues a momentary Inject Out signal.
Figure 27. Partial Loop injection—Injecting sample
From
injection port
To
column
1
2
6
3
Mobile phase
Wash solvent
Sample
Thermo Scientific
4
To waste
5
From
pump
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Autosampler Operation
Post-Injection Events
At the end of an injection, the autosampler performs the following operations:
1. The autosampler performs an optional flush, wash, or both as specified in the instrument
method.
• Flush (optional): During an internal flush, the autosampler lowers the needle into
the injection port, and then dispenses a specified volume of solvent through the
needle and transfer line out to waste. You can select the solvent wash bottle or a
reservoir vial as the flush solvent source. The internal flush removes excess or residual
sample from the injection flow path. It does not flush the sample loop.
• Wash (optional): During an external wash, the autosampler lowers the needle into
the wash station, and then dispenses a specified volume of solvent through the needle.
As the wash station overfills, the wash solvent flows up over the exterior of the needle,
and then out to waste. You can select the solvent wash bottle or a reservoir vial as the
wash solvent source.
2. The autosampler moves the syringe plunger to the Ready position, and then switches the
syringe valve to point to the wash bottle.
3. The autosampler prepares for the next injection by moving the XYZ arm to the Home
position, located above the injection port of the autosampler.
Autosampler Operation
You have full control of the autosampler from the Xcalibur and ChromQuest data systems
and limited control of the autosampler from other Thermo Scientific mass spectrometry
applications. The power switch is the only manual control. The data system Help, the Accela
User Guide for LC Devices, and the Accela User Guide for the ChromQuest Data System provide
information on how to operate the autosampler.
Note You can create instrument methods to control the Accela Autosampler from several
Thermo Scientific mass spectrometry applications; however, you can calibrate the Accela
Autosampler only from the ChromQuest or Xcalibur data system.
The Help and the manuals (provided as PDF files) include information on how to do the
following:
• Calibrate the well bottom position for custom vials or wellplates (see “Calibrating the
Accela Autosampler” on page 48).
• Perform direct commands to flush the syringe, move the XYZ arm (see “XYZ Arm
Mechanism” on page 5), inject sample, and so on.
• Create an instrument method for data acquisition.
For information on accessing the manuals, see “Related Documentation” on page xiii.
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Automated Sample Preparation
Automated Sample Preparation
With the automated sample preparation feature, you can build a pretreatment routine that the
autosampler performs before it performs an injection. The pretreatment routine can consist of
multiple tasks including transferring liquid between sample vials, transferring liquid from one
of the four reservoir vials to a sample vial, mixing, and more.
The autosampler performs each task by drawing liquid into or expelling liquid from the
needle tubing. It uses the inner plunger of the concentric syringe for liquid transfers that
require a high degree of precision, such as a transfer of liquid between sample vials. It uses the
outer plunger of the concentric syringe for liquid transfers that require less precision, such as
sample mixing, which is performed using an aspirate and expunge technique.
For a complete list of sample preparation tasks that the autosampler can perform and a set of
rules for creating sample preparation routines, refer to the Help provided with the data
system.
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Introduction
Specifications
Specifications
These are specifications for the Accela Autosampler.
Sample capacity:
• 200 vials in five removable trays
• 288 samples in three 96-well microplates
• 1152 samples in three 384-well microplates
Standard vial:
• 1.8 mL volume
Injection precision:
• < 1.0% RSD at 5 μL and up
Partial loop injection volume:
• 10 μL with standard 25 μL sample loop and 250 μL
syringe
• Up to 1000 μL injection volume with optional loops
and syringes
Fixed loop (full loop)
injection volume:
• 25 μL with standard loop and 250 μL syringe
• Up to 1000 μL injection volume with optional loops
and syringes
Sample carryover:
• < 0.1%
• 6 mL flush volumes, user-specified for internal and
external needle wash cycles
Minimum sample volume:
• 1 μL can be injected from 5 μL
Needle height control:
• Programmable in 0.1 mm increments
• Active vial bottom search selectable on/off
Column oven:
• Programmable in 1 °C increments from 5 to 95 °C
Tray temperature control:
• Programmable in 1 °C increments from 0 to 60 °C
• Typical vial temperature from 4 to 50 °C
Reagent vials:
• Four 16 mL vials can be specified for flush/wash or
dilution functions
Remote inputs:
• Pump ready, inject hold
Remote outputs:
• Pump stop, autosampler ready, inject, gradient start,
four timed events
Ambient environment:
• 10 to 40 °C, 5 to 95% relative humidity,
non-condensing
Dimensions:
• 37 × 36 × 50 cm (h×w×d)
• 14.5 × 15 × 20 in. (h×w×d)
32
Weight:
• 30 kg (66 lbs)
See “Lifting and Carrying the Autosampler” on
page xviii.
Power requirements:
• 100/120/220/240 Vac nominal; 50/60 Hz, 550 VA
Product certification:
• CSA, TÜV/GS, FCC (EMI), VDE (EMI)
Accela Autosampler Hardware Manual
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2
Installation
To install the Accela Autosampler as part of an Accela UHPLC system, use the “Installation
Checklist” on page 34 and follow the procedures in this chapter. Make a copy of the checklist
and fill it out when the installation is complete. Include the completed checklist in your
maintenance records.
“Changing the Sample Loop” on page 49 describes how to change the sample loop, a piece of
stainless steel tubing with two end fittings connected to the injection valve. For full loop
(fixed loop) injections, the sample loop size determines the injection volume.
Note Install the Accela Autosampler in the sequence presented in the Installation
Checklist.
Contents
• Software Compatibility
• Installation Checklist
• Unpacking and Inspecting the Instrument
• Making Initial Instrument Preparations
• Making the Back Panel Connections
• Connecting the Solvent Lines
• Powering On the Autosampler
• Calibrating the Accela Autosampler
• Changing the Sample Loop
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2
Installation
Software Compatibility
Software Compatibility
You have full control of the Accela Autosampler from the ChromQuest chromatography data
system and the Xcalibur mass spectrometry application and limited control from several other
Thermo Scientific mass spectrometry applications. The following data systems provide
compatible firmware and a firmware upgrade utility (see “Downloading Firmware to the
Autosampler” on page 133):
• For the ChromQuest 4.2 data system, you can find the Accela device drivers on the
Accela Device Driver Add-on CD.
• For the ChromQuest 5.0 data system, you can find the Accela device drivers on the
ChromQuest CD.
• For the Xcalibur data system, you can find the Accela device drivers on the
LC Devices DVD.
Installation Checklist
The following installation summarizes the steps that you must complete to properly install the
Accela Autosampler.

Unpack and inspect your instrument (page 35).

Read the safety notices (front of manual).

Make the initial instrument preparations (page 35).

Make the initial back panel connections (page 36).

Power on the autosampler for the first time (page 47).

Install the Xcalibur 2.0 (or later) data system or the ChromQuest 4.2 (or later) data
system and connect remote communication outputs, as required for external devices
that are not controlled by the data system.

Add the Accela Autosampler to the data system instrument configuration. For
instructions on specifying the configuration options for the Accela Autosampler, refer to
the Accela User Guide for LC Devices, the Accela User Guide for the ChromQuest Data
System, or the Help provided by the data system.
This Accela Autosampler was installed by:
(Name)
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(Date)
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2 Installation
Unpacking and Inspecting the Instrument
Unpacking and Inspecting the Instrument
Carefully remove the autosampler from the shipping container and inspect both the
autosampler and packing for any signs of damage. If you find any damage, save the shipping
materials and immediately contact the shipping company.
The shipping container contains the Accela Autosampler and an accessory kit. Carefully check
to make sure you received all the items listed on the packing list. Ensure that the needle
tubing assembly is attached to the XYZ arm and the right side of the syringe valve, a 250 μL
dual-concentric syringe is attached to the syringe drive mount, and a 25 μL sample loop is
attached to the injection valve.
If any items are missing, contact your Thermo Fisher Scientific representative immediately.
To connect the autosampler to the other instruments in the Accela stack, you must have the
Accela System Kit (see “Accela System Kit” on page 129). This kit is provided with orders for
instruments sold together with either the Xcalibur or the ChromQuest data system or as a
separate option.
Making Initial Instrument Preparations
Place the autosampler on a benchtop. Allow at least 15 cm (6 in.) of space between the back
panel of the autosampler and any wall or obstruction. This provides clearance for the
back-panel connectors and cool air flow.
Installing the autosampler requires these tools:
• 5/16 in. open-end wrench
• Narrow flat-head screwdriver
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Installation
Making the Back Panel Connections
Making the Back Panel Connections
To operate the Accela Autosampler, connect the power, Ethernet, and system interconnect
cable to the back panel (see Figure 28) of the autosampler, and check the unit ID setting.
Figure 28. Back panel of the Accela Autosampler
r
Rotary switches
(unit ID)
Ethernet port
Interconnect cable
connection
Power receptacle
To make the back panel connections, follow these procedures:
• Connecting the System Interconnect Cable
• Connecting the Ethernet Cable
• Checking the Unit ID Settings
• Connecting the Autosampler to Line Power
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Making the Back Panel Connections
Connecting the System Interconnect Cable
The system interconnect cable that synchronizes the timing of the Accela devices is in the
Accela System Kit. The interconnect cable has seven combicon connectors: three have a blue
DETECTOR label; two have a yellow PUMP label; one has a small A/S tag on its adjacent
cable; and one has a small M/S tag on its adjacent cable (see Figure 29).
Figure 29. System interconnect cable with seven combicon connectors
DETECTOR
DETECTOR
DETECTOR
M/S
PUMP
Autosampler
connector
PUMP
A/S
 To connect the interconnect cable
1. Plug the AS connector into the left 8-pin socket on the back panel of the Accela
Autosampler (see Figure 30).
2. Attach one of the pump connectors to the Accela pump (Accela Pump, Accela 600 Pump,
or Accela 1250 Pump).
3. If your LC system contains an Accela detector, attach one of the detector connectors to it.
Refer to the Accela Getting Connected Guide for instructions on how to connect the system
interconnect cable to a Thermo Scientific MS detector.
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Installation
Making the Back Panel Connections
Figure 30. System interconnect cable connections
Connector labeled
DETECTOR
Connector labeled
DETECTOR
9
9
7 8
ENET
7 8
GND B
CHB - 1V
CHB - 100mV
CHB - 10mV
GND A
CHA - 1V
CHA - 100mV
CHA - 10mV
EVENT
GND
ZERO
READY
RUN
NC
+5V @ 150mA
GND
Accela PDA Detector
–or–
Accela UV/Vis Detector
1 2 3 4 5 6 7 8
9 10 11 12 13 14 15 16
NC
CHASSIS GND
INTEGRATOR INTEGRATOR +
NC
CHASSIS GND
RECORDER RECORDER +
UNIT ID
PURGE
GND
ZERO
READY
RUN
NC
+5V @ 150mA
GND
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
5 6
RS232
4
5 6
0 1
2 3
4
0 1
2 3
–or–
ENET
Accela RI Detector
Connector with cable
labeled A/S
9
9
7 8
7 8
9 10 11 12 13 14 15 16
5 6
1 2 3 4 5 6 7 8
4
5 6
0 1
2 3
4
0 1
2 3
A/S READY
INJECT OUT
INJECT HOLD
PUMP READY
GRAD START
PUMP STOP
+5V @ 150mA
GND
GND
GND
TF 1
TF 2
TF 3
TF 4
+5V @ 150mA
GND
UNIT ID
ENET
Accela Autosampler
System
interconnect
cable
Connector labeled
PUMP
Connector labeled
PUMP
!
RS232
1 2 3 4 5 6 7 8
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Accela Autosampler Hardware Manual
RS232
1 2 3 4 5 6 7 8
USB
RESERVED
GND
INJECT HOLD
PUMP READY
GRAD. START
RESERVED
5V
GND
PRESSURE
GND
INJECT HOLD
PUMP READY
GRAD. START
PUMP STOP
5V
GND
Accela Pump
USB
DO NOT USE
TOGETHER
WITH
D-SUB25
DIGITAL I/O
–or–
Accela 600 or 1250 Pump
Thermo Scientific
2 Installation
Making the Back Panel Connections
Connecting the Ethernet Cable
Connect the Ethernet port on the back panel of the Accela Autosampler to the Ethernet
switch using the 7 ft length, CAT-5 Ethernet cable with ferrite supplied in the accessory kit.
Checking the Unit ID Settings
The Accela Autosampler is shipped with the unit ID preset using the two rotary switches
located on the back panel (see Figure 31). The unit ID must correspond with the stack
number that you specify in configuring the Accela Autosampler.
Figure 31. Rotary switches set to 01
9
5 6
7 8
9
4
5 6
0 1
2 3
4
0 1
2 3
7 8
The Communication page (see Figure 84 on page 123) of the Accela Autosampler
Configuration dialog box in the Thermo Foundation Instrument Configuration application
contains one box for the unit ID value. The default is 1, and the range is 1 to 99.
The Accela AS Configuration dialog box (see Figure 85 on page 124) in the ChromQuest
data system contains two boxes for the unit ID. The default value is 01 and the range is
01 to 99. The value of 0 or 00, respectively, is reserved for special service functions.
Connecting the Autosampler to Line Power
The Accela Autosampler is shipped with one of these power cords.
Destination
Plug type
Voltage rating
Current rating
P/N
United Kingdom
BS 1363
250 Vac
5A
6003-0810
Switzerland
SEV 1011
250 Vac
10 A
6003-0620
China and Europe
CEE 7/7
250 Vac
10 A
6003-0330
United States
and Canada
NEMA 5-15P
125 Vac
10 A
6003-0160
Local codes in your area might require installing another type of plug and receptacle. The
Thermo Fisher Scientific field engineer for your country will provide the appropriate power
plugs.
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2
Installation
Connecting the Solvent Lines
 To connect the autosampler to line power
1. Ensure that the power switch on the front of the unit is in the Off position (released or
out position).
2. Check the fuses (see “Replacing a Fuse” on page 97).
3. Connect the plug to the power receptacle (see Figure 28 on page 36) on the back panel of
the autosampler.
4. Connect the other end of the power cord to line power.
When you are ready to power on the autosampler, go to “Powering On the Autosampler” on
page 47.
Connecting the Solvent Lines
Before you can operate the LC system, connect the wash bottle that is contained in the solvent
platform to the left side of the syringe valve by using the wash bottle tubing, connect the
needle tubing to the right side of the syringe valve, and connect the autosampler to the pump.
To connect the solvent lines, follow these procedures:
• Connecting the Wash Bottle
• Connecting the Needle Tubing
• Connecting the Autosampler to the Pump
IMPORTANT
If the wash bottle tubing runs dry during a sequence of injections, the
autosampler makes “blank” injections.
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2 Installation
Connecting the Solvent Lines
Connecting the Wash Bottle
The Wash Bottle Kit supplied in the Accela System Kit contains a 1 liter bottle and a tubing
assembly (see Figure 32).
Figure 32. Wash bottle tubing assembly
Bottle cap assembly
Teflon filter
PEEK fitting
 To connect the wash bottle
1. Fill the wash bottle with an appropriate solvent, and then place it in the solvent platform
on the top of the system stack.
Note The solvent platform holds the 1 L wash bottle and four 1 L solvent reservoir
bottles. In case of breakage, the solvent platform can hold up to 7 L and acts as a
secondary container for the liquid in the solvent bottles.
2. Insert the Teflon filter into the bottom of the bottle and screw on the cap.
3. Connect the PEEK fitting to the left side of the syringe valve (see Figure 33).
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Installation
Connecting the Solvent Lines
Figure 33. Wash tubing and needle tubing connections
Wash tubing
fitting
Syringe
valve
Needle tubing
fitting
Connecting the Needle Tubing
The Accela Autosampler is shipped with the needle tubing assembly connected and routed
through the appropriate brackets. The internally threaded fitting of the needle tubing
assembly is connected to the needle assembly and the externally threaded fitting is connected
to the syringe valve. The needle tubing is routed through a bracket on the XYZ arm, a bracket
on the left wall of the tray compartment, and a bracket on the tray compartment door. In
addition, the two-pronged needle tubing guide is connected to the x-axis positioning frame,
and the PVC cap (red or black) is pushed below the metal runner for the x-axis positioning
frame.
Before you operate the autosampler, make sure that the needle tubing is routed through the
appropriate brackets.
CAUTION The needle tubing is easily damaged. Before you operate the autosampler, make
sure that the needle tubing is routed through the support brackets, that the needle tubing
guide is connected to the x-axis positioning frame, and that the PVC cap is pushed below
the metal runner for the x-axis positioning frame.
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2 Installation
Connecting the Solvent Lines
Connecting the Autosampler to the Pump
After you set up the Accela stack on the laboratory benchtop, connect the pump outlet to the
autosampler inlet (heat exchanger inlet).
CAUTION To prevent personal injury, before connecting the pump to the autosampler,
make sure that the column oven compartment is at room temperature.
CAUTION To prevent personal injury caused by skin contact with hazardous solvents,
ensure that the pump is off before you connect it to the autosampler.
 To connect the autosampler heat exchanger to the Accela pump outlet
1. Connect a length of high-pressure tubing to the Accela pump outlet as follows:
a. Using a three-piece compression fitting with a 0.45 in. nut (see Figure 34), connect a
length of 0.005 in. ID × 1/16 in. OD tubing to the pump outlet.
The tubing length depends on the pump model:
• For the Accela Pump, connect a 20 cm (8 in.) length of precut tubing to the
outlet of the dynamic mixer.
• For the Accela 600 Pump or the Accela 1250 Pump, connect a 26 cm (10 in.)
length of precut tubing to the outlet of the placeholder module or the optional
dynamic mixer.
Tip To identify the tubing ID, the precut tubing has a color-coded band:
Thermo Scientific
• Red = 0.005 in.
• Black = 0.007 in.
• Blue = 0.010 in.
• Yellow = 0.020 in.
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Installation
Connecting the Solvent Lines
Figure 34 shows the 0.45 in. length nut and the front and back ferrules for
1/16 in. OD tubing.
The 0.45 in. nut has a longer threaded portion than the 0.53 in. nut
(0.295 in. > 0.225 in.). The longer threaded portion makes the 0.45 in. nut suitable
for use with the deep ports of the dynamic mixer and the placeholder module.
Figure 34. High-pressure fitting with a front and back ferrule and a 0.45 in. nut
0.45 in.
0.295 in.
b. Using a 0.25 in. open-ended wrench, tighten the nut.
2. Route the tubing through the access slot in the autosampler drip tray.
3. Connect the other end of the tubing to the heat exchanger inlet as follows:
a. Using a three-piece compression fitting with a 0.53 in. nut, connect the other end of
the tubing to the heat exchanger inlet.
Figure 35 shows the 0.53 in. length nut and the front and back ferrules for
1/16 in. OD tubing.
Figure 35. High-pressure fitting with a front and back ferrule and a 0.53 in. nut
0.53 in.
0.225 in.
b. Using a 0.25 in. open-ended wrench, tighten the nut.
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2 Installation
Connecting the Solvent Lines
Figure 36 through Figure 38 show the high-pressure connection between the autosampler and
the pump.
Figure 36. Connection between the last version of the Accela Pump and the autosampler
Connection to the
heat exchanger inlet
Access slot in the
autosampler drip tray
Dynamic mixer
PU R GE
Inline filter holder
PULSE
D A MPENER
F L USH
Figure 37. Connection between the Accela 600 Pump or the Accela 1250 Pump and the
autosampler
20
Connection to the
heat exchanger inlet
µL
3
4
2
5
1
6
Access slot in the
autosampler drip tray
Dynamic
mixer
–or–
Placeholder
module
Static mixer
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Installation
Connecting the Solvent Lines
Figure 38. Connection between the autosampler and an earlier version of the Accela Pump
without a dynamic mixer
PURGE
PULSE
DAMPENER
FLUSH
Tubing,
10 cm length
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Accela Autosampler Hardware Manual
Filter holder
Tubing,
20 cm length
Thermo Scientific
2 Installation
Powering On the Autosampler
Powering On the Autosampler
After installing solvent lines and connecting the autosampler to line power, you can power on
the autosampler.
 To turn on the power to the autosampler
1. Ensure that the power switch at the front of the unit is in the Off position (released or out
position).
2. Attach the power cord to the power entry module on the back panel of the autosampler
and connect it to the power source.
3. Turn the power on by pushing the power button in to engage it.
4. Make sure that the power indicator briefly lights amber and then turns solid green. If it
does not light at all, see Chapter 6, “Routine Troubleshooting.” If it stays amber or
flashes amber or green, turn the power off for several seconds, and then turn it back on
again. Call your Thermo Fisher Scientific service representative if you require assistance.
These events occur during the autosampler startup sequence:
1. The Power LED turns solid green.
2. If the autosampler is communicating with the data system computer, the Com LED turns
from amber to green.
3. The Run LED and Temperature LED turn solid green.
4. The autosampler initializes the syringe, injection valve, and XYZ arm positions as follows:
a. Switches the two-way syringe valve to the flush bottle position.
b. Switches the injection valve to the inject position.
c. Moves the syringe to the home (empty) position.
d. Moves the syringe to the ready position.
e. Switches the two-way syringe valve to the needle position.
5. If the autosampler is communicating with the data system computer, the Com LED turns
from amber to solid green.
6. Every time you close and reopen the data system, the autosampler initializes its hardware
components (see step 4).
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2
Installation
Calibrating the Accela Autosampler
Calibrating the Accela Autosampler
The Accela Autosampler is factory calibrated. The data system provides calibration options for
the following:
• Column oven temperature
• Tray compartment temperature
• XYZ arm position
• Depth of custom vials or microplate wells
Only a Thermo Fisher Scientific service representative can perform the first three calibration
options.
Before you use any of the three custom tray options, perform the calibration that determines
the z-axis distance that the needle must travel to reach the bottom of your custom vial or well
plate.
 To access the well bottom distance calibration option from the Xcalibur data system
1. From the Roadmap view, click the Instrument Setup icon.
The Instrument Setup window appears.
2. Click the Accela AS icon in the viewbar.
The Instrument Setup view for the Accela Autosampler appears.
3. From the Accela AS menu, choose Calibration.
The Calibration dialog box appears.
4. Select the Well Bottom Distance option.
 To access the well bottom distance calibration option from the ChromQuest data
system
1. From the Instrument window, choose Control > Instrument Status.
2. Click the Accela AS tab.
3. Click Diagnostics.
4. Click the Calibration tab.
5. Select the Bottom Distance Calibration option.
For instructions on how to perform this calibration, refer to the data system Help.
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Changing the Sample Loop
Changing the Sample Loop
Several sample loop sizes are available for the Accela Autosampler (see Table 6).
Table 6. Sample loops
Description
Part number
5 μL
00109-99-00023
10 μL
00109-99-00024
20 μL
00109-99-00025
25 μL
00109-99-00026
50 μL
00109-99-00027
100 μL
00109-99-00028
500 μL
00109-99-00029
1000 μL
00109-99-00030
 To change the sample loop
1. Make sure that the autosampler is in Idle mode.
2. Using an open-end wrench, loosen the fittings at ports 1 and 4, and gently pull out the
two ends of the sample loop. If you want to reuse the sample loop, label the end fittings
with the port number (1 or 4) that the swaged fitting was connected to.
3. Insert the ends of the new sample loop in the same orientation as the old one.
4. Firmly press one end of the sample loop into port 1 of the injection valve as you
hand-tighten the nut.
5. Firmly press the other end of the sample loop into port 4 of the injection valve as you
hand-tighten the nut.
6. Using a 1/4 in. open-end wrench, gently tighten the fittings by an additional 3/4 turn.
Tip To connect tubing with a two-piece stainless steel fitting to a receiving port, press the
tubing against the bottom of the port as you hand-tighten the fitting. Then tighten the
fitting by an additional 3/4 turn with a wrench. Once you use the wrench to tighten the
fitting, the ferrule portion of the two-piece fitting is permanently connected to the tubing.
To connect tubing with a pre-swaged fitting to a receiving port, press the tubing against
the bottom of the port as you hand-tighten the fitting. Then tighten the fitting by an
additional 1/4 to 1/2 turn with a wrench.
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3
Sample Loading
Before you load your samples into the autosampler, make sure that your samples are
completely soluble in the mobile phase and that you have filtered your samples and solvents
through a 0.5 μm filter. These techniques minimize sample precipitation in the lines and
remove particulate matter that could obstruct the flow through the autosampler injector or
the column. In addition, make sure that the vial caps are securely fastened to the vials.
After you finish preparing your samples, you are ready to load them into the Accela
Autosampler. The Accela AS Accessory Kit (see “Accela AS Accessory Kit” on page 128)
contains trays that hold standard vials, inserts that hold 100 μL vials, and carrier plates that
hold 96- and 384-microwell plates.
Contents
• Standard Tray Type
• Carrier Trays for Microplates
• Important Precautions for Sample Loading
• Recommended Vials, Microplates, and Microplate Covers
Standard Tray Type
The Accela Autosampler Accessory Kit contains five standard trays. Trays A, B, C, D, and E
are arranged in the autosampler from left to right. Each tray holds 40 standard, 1.8 mL vials
arranged in two rows of 20. The accessory kit also contains a supply of standard, 1.8 mL vials,
silicone and Teflon septa, and screw-top vial caps. You can order additional vials from
Thermo Fisher Scientific.
The part numbers of the replaceable components in the accessory kit are listed in “Accela AS
Accessory Kit” on page 128.
IMPORTANT
If you are using screw cap vials, the caps must be firmly tightened. If the
caps are not firmly tightened, the needle might push the septum into the bottom of the
vial.
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Sample Loading
Standard Tray Type
To install the standard tray type, follow these procedures:
• Installing the Standard Tray Type
• Moving the XYZ Arm to the Back of the Tray Compartment
Installing the Standard Tray Type
 To install the sample trays into the tray compartment of the autosampler
1. Open the left door of the autosampler.
• If you selected the Verify Door Is Closed check box when you configured the
autosampler, the XYZ arm moves to the back of the tray compartment.
• If the XYZ arm does not move to the back of the tray compartment, perform the
appropriate direct command from the data system.
2. Hold the tray handle, tilting the back end of the tray down.
3. Insert the tray runner into the slot at the back of the tray compartment (see Figure 39).
4. Lower the front of the tray into place and press down firmly to seat the tray.
Figure 39. Tray compartment and standard tray
Slot for tray
Tray runner
R4
R3
R2
Standard
1.8 mL vial
R1
Tray handle
A
B
C
D
E
IMPORTANT Be sure that each tray is properly seated. If a tray is not completely seated,
the autosampler will not operate correctly.
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Sample Loading
Standard Tray Type
IMPORTANT To trigger the vial sensor, you must position custom vials in the tray so
that the top of the vial reaches the minimum height of 1.55 in. Custom vials that fall
below this minimum height do not trigger the vial sensor. When an acquisition sequence
incurs a vial that is below the minimum height, the sequence halts and the following
message appears: Vial Not Found.
Note When you open the autosampler door, the XYZ arm automatically moves to the
back of the autosampler, allowing you to add samples to the tray during a run. After the
door is closed, the arm returns to its position and resumes the operation that was in
progress before the door was opened. You can enable this door interlock feature in the
Configuration menu for the autosampler.
The accessory kit also contains a tray adapter for 100 μL vials and a tray adapter plate for
microcentrifuge tubes.
 To use the tray adapter
Insert the tray adapter into the standard tray, and then secure it with the 1/4 in. Phillips
head screw from the accessory kit.
Moving the XYZ Arm to the Back of the Tray Compartment
If you did not select the Verify Door Is Closed check box when you configured the
autosampler, the XYZ arm does not move to the back of the tray compartment when you
open the tray compartment door. From the data system, use a direct command to move the
XYZ arm.
 To move the XYZ arm to the back of the tray compartment
• For the Xcalibur data system, open the Direct Control dialog box for the autosampler
(see “Using the Xcalibur Direct Control Commands” on page 141). Select Position
Arm to Access Tray, and then click Apply.
• For the ChromQuest data system, open the Direct Controls page for the autosampler
(see “Using the ChromQuest Direct Commands” on page 143). Select Position Arm
to Access Tray, and then click Submit.
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Sample Loading
Carrier Trays for Microplates
Carrier Trays for Microplates
In addition to the standard tray option, the accessory kit contains a short microwell plate
carrier and a tall, solid microwell plate carrier. These carrier trays hold up to three microplates.
To use PCR plates, you can order a cooling adapter from Thermo Fisher Scientific.
Use the carrier trays as follows:
• For either 1 mL or 2 mL deep well plates, use the short microwell carrier.
• For PCR plates, use the short microwell carrier in combination with the cooling adapter.
• For standard depth 96- or 384-well plates, use the tall, solid microwell carrier.
Note If your samples require low temperature cooling, use the cooling adapter and the
PCR plates instead of the standard 96-well microplates.
Figure 40 and Figure 41 show the short microwell plate carrier and the tall, solid microwell
plate carrier, respectively.
Figure 40. Short microwell plate carrier
Figure 41. Tall, solid microwell plate carrier
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Carrier Trays for Microplates
 To install the microplates onto the carrier tray
1. If this is an option in your data system, determine the microplate orientation.
With the Xcalibur and ChromQuest data systems, you can select the A1 top-left or A1
bottom-right orientation.
2. Hold the plates in the appropriate orientation as you snap them into the carrier slots.
Figure 42 shows a carrier with three 96-well plates in the A1 top-left orientation.
Figure 42. Carrier with three 96-well plates in the A1 top-left orientation
Carrier runner
The A1 well is in
the top-left corner.
Plate C
Plate B
Plate A
Carrier handle
 To load the carrier tray into the autosampler tray compartment
1. Holding the carrier handle, insert the carrier runner into the slot in the back of the tray
compartment tub.
Tip Use two hands to hold the tall, solid microwell carrier, as it weighs approximately
2.6 kg (5.8 lbs).
2. Lower the front of the carrier tray into place.
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Sample Loading
Important Precautions for Sample Loading
Important Precautions for Sample Loading
Because the needle mechanism moves to the vial or well during an injection sequence,
punctures the vial septum or microwell plate cover, and then draws sample from a specified
depth, follow these precautions when you load samples into the autosampler tray
compartment:
• Check the height of custom trays before you place them in the tray compartment.
Objects taller than 4.5 cm (1.8 in.) can stall the autosampler arm. You must check the
height of custom trays before you place them in the tray compartment.
• Use only recommended vials, vial caps, microplates, and microplate covers.
Thermo Fisher Scientific has tested the vials, vial caps, microplates, and microplate covers
listed in the next section, “Recommended Vials, Microplates, and Microplate Covers,” for
use with the Accela Autosampler. Because the tip of the needle is blunt, the durometer
(material hardness) of the vial cap septum is extremely important, as the needle might not
properly pierce septa that are not recommended.
• Ensure that you screw on the vial caps tightly.
If the vial cap is loose, the needle can dislodge the septum during an injection. The needle
can also dislodge the septum if it pierces the edge of the septum, rather than its center. If
you suspect the needle is out of alignment, call your local Thermo Fisher Scientific service
engineer to calibrate its alignment.
• Calibrate the depth of custom vials or microwell plates.
If you are using one of the three custom tray configurations, you must perform a
calibration that determines the distance between the tip of the needle and the bottom of
your vials or wells. Performing this calibration ensures that the needle tip descends to the
height specified in your instrument method as it withdraws sample from the vial or well.
Perform this calibration whenever you change the custom tray type configuration or use a
different type of custom vial or microplate.
For instructions on how to perform bottom distance calibration, refer to the data system
Help for the autosampler or the Accela user guide or getting started guide for your data
system.
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Recommended Vials, Microplates, and Microplate Covers
Recommended Vials, Microplates, and Microplate Covers
Thermo Fisher Scientific has tested and recommends the following vials, microplates, and
microplate covers for use in your Accela Autosampler.
Recommended Vials
Table 7 lists the recommended sample vials.
Table 7. Approved vials
Description
Qty/pkg
Part number
1.8 mL vial with septa and screw cap
100
A4954-010
1.8 mL amber vial with septa and screw cap
100
A4965-010
Shell vial with cap
100
A4948-010
Shell vial with 100 μL insert and cap
100
60053-62008
1.8 mL vial with cross-slit septa and screw cap
100
A5400-010
Recommended Microplates
The microplate carriers hold three low-density 96-well or three high-density 384-well
microplates.
Low-Density Microplates
All 96-well microplates that meet the dimensions of the Society for Biomolecular Screening
and that have a maximum height of 0.80 cm (0.32 in.) are suitable. Polypropylene is the
preferred material when working with organic samples.
Table 8 lists the 96-well microplates that Thermo Fisher Scientific approves for use with the
Accela Autosampler. The supplier is SUN-SRi, a subsidiary of Thermo Fisher Scientific Inc.
Table 8. Recommended 96-well microplates
Thermo Scientific
Description
Qty/pkg
Part number
96-well, U-bottom, Polypropylene,
working well volume 250 μL
10
500 833
100
500 811
96-well, V-bottom, Polypropylene,
working well volume 190 μL
10
500 835
100
500 813
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Sample Loading
Recommended Vials, Microplates, and Microplate Covers
High-Density Microplates
All 384-well microplates that meet the dimensions of the Society for Biomolecular Screening
and that have a maximum height of 0.32 in. are suitable. Polypropylene is the preferred
material when working with organic samples.
Table 9 lists the 384-well microplates that Thermo Fisher Scientific approves for use with the
Accela Autosampler.
Table 9. Recommended 384-well microplates
Description
Qty/pkg
Part number
384-well, U-bottom, Polypropylene,
working well volume 90 μL
10
500 841
100
500 819
Recommended Microplate Covers
Table 10 lists the covers for 96-well microplates that Thermo Fisher Scientific approves for use
with the Accela Autosampler.
Table 10. Recommended 96-well microplate covers
Supplier
catalog number
Description
Qty/pkg
Source
Micromat II/PS Round Pink
Material: Teflon-coated silicone
rubber
10/pkg
Thermo Fisher
Scientific
400 079
Micromat CLR 96-well PS 7 mm
Material: clear silicone rubber
10/pkg
Thermo Fisher
Scientific
300 005
Clear heat-seal film
100/pkg
Thermo Fisher
Scientific
Matrix
Technologies Inc.™
AB-0685
VIEWseal™ dry film
adhesive-backed tape
100/pkg
Greiner Bio-One
676070
Table 11 lists the 384-well microplate covers that Thermo Fisher Scientific approves for
use with the Accela Autosampler.
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Recommended Vials, Microplates, and Microplate Covers
Table 11. Recommended 384-well microplate covers
Supplier
catalog number
Description
Qty/pkg
Source
Clear heat-seal film
100/pkg
Thermo Fisher
Scientific
Matrix Technologies
AB-0685
VIEWseal™ dry film adhesive
backed tape
100/pkg
Greiner Bio-One
676070
For more information about the microplates, heat-seal covers, and sealing equipment supplied
by Thermo Fisher Scientific, click the following icon:
Microplate
Heat Sealing
Instruments
and
Accessories
CAUTION Avoid using the following covers with your Accela Autosampler:
• Parafilm “M”, manufactured by American National Can, or similar material
• POWERseal™, manufactured by Greiner Bio One, or similar material
• Adhesive sealing tapes
Parafilm “M” and POWERseal covers leave small pieces of material on the needle that
eventually clog the injection port. Adhesive sealing tapes leave an adhesive film on the
needle causing system contamination. The adhesive builds up on the needle and in the
injection port, and eventually the needle becomes completely clogged.
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Routine Maintenance
This chapter contains information on the maintenance record keeping options provided by
your Thermo Scientific data system, the routine maintenance procedures necessary to
optimize the performance of the autosampler, and the suggested maintenance schedule.
Contents
• Maintenance Schedule
• Replacing the Rotor in the Valco Injection Valve
• Replacing the Inlet Filter for the Wash Bottle Solvent Line
• Replacing the Transfer Tubing
• Maintaining the Syringe Drive Assembly
• Troubleshooting a Blockage in the Injection System
• Replacing the Needle Tubing Assembly
• Replacing the Needle Assembly
• Changing the Syringe or Replacing the Inner Plunger
• Clearing a Plugged Heat Exchanger
• Replacing a Fuse
• Maintenance Cycles
For your own safety and to protect the autosampler from damage, follow the safety
precautions in this manual when performing maintenance on the autosampler.
CAUTION To avoid electrical shock, do not remove the top cover or remove parts beyond
the descriptions in the operation portions or in the maintenance topics of this manual.
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Routine Maintenance
Maintenance Schedule
Maintenance Schedule
The Accela Autosampler requires only a few simple maintenance procedures to keep it in
optimum working condition. Table 12 contains the maintenance schedule for the
autosampler. A Thermo Fisher Scientific service representative or other qualified person must
perform the last procedure, the alignment of the XYZ arm.
To make the maintenance of your autosampler easier, the data system keeps track of how
many injections, injection valve cycles, needle cycles, and syringe cycles the autosampler has
performed. The data system alerts you to perform maintenance when the number of cycles
exceeds a preset value. For information on setting up the tracking system, see “Maintenance
Cycles” on page 98.
.
Note You are responsible for maintaining your autosampler. Routine maintenance does
not fall under Thermo Fisher Scientific warranty; however, planned maintenance
contracts are generally available. Contact your local representative if you are interested in
purchasing a planned maintenance contract.
Table 12. Maintenance procedures and schedule
Frequency
Procedure
Performed by
Monthly
Checking the solvent tubing connections for leaks
User
Monthly
Running a standard sample
User
Every 15 000 to
20 000 injections
Replacing the rotor seal (Rheodyne™ injection
valve)
User
Replacing the rotor (Valco™ injection valve)
Semiannually
Replacing the flush solvent inlet filter
User
As necessary
Replacing the injection port
User
As necessary
Clearing a plugged line or needle
User
As necessary
Changing the syringe
User
As necessary
Changing the sample loop
User
As necessary
Replacing a fuse
User
Semiannually
Cleaning and lubricating the syringe drive
mechanism
User
Annually
Cleaning and lubricating the XYZ arm
Field service
Annually
Verifying the XYZ arm alignment
Field service
Note Table 12 lists recommended maintenance intervals based on operating the
autosampler under typical conditions. Operating the autosampler under certain harsh
conditions (corrosive solvents, extreme temperatures, and so on) might require more
frequent maintenance than indicated in the table.
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Replacing the Rotor in the Valco Injection Valve
Replacing the Rotor in the Valco Injection Valve
The rotor forms a high-pressure seal with the stator. Replace the rotor when you notice a leak
between either the stator and rotor or the adjacent ports of the injection valve, or when you
notice a decrease in injection precision.
This section describes how to replace the rotor for the autosampler’s Valco injection valve. For
information on replacing the rotor seal for the Rheodyne injection valve that shipped with
earlier versions of the autosampler, see “Replacing the Rotor Seal in the Rheodyne Valve” on
page 137.
CAUTION Before you remove the injection valve from the autosampler, turn off the power
to the autosampler and unplug the autosampler from line power.
Replacing the rotor requires these tools.
Tool
Use
1/4 in. open-end wrench
To remove the tubing from injection valve ports 5 and 6
#2 Phillips screwdriver
To disconnect the injection valve from the autosampler
9/64 in. hex wrench
To disassemble the injection valve
IMPORTANT Because component cleanliness affects the useful life of the injection
valve, replace the rotor in a clean environment.
To replace the rotor, follow these steps:
1. Removing the Injection Valve from the Autosampler
2. Disassembling the Injection Valve
3. Cleaning the Stator
4. Installing a New Rotor
5. Reinstalling the Injection Valve
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Replacing the Rotor in the Valco Injection Valve
Removing the Injection Valve from the Autosampler
Before you remove the injection valve from the autosampler, prepare a clean environment
where you can disassemble the valve.
 To remove the injection valve from the autosampler
1. Disconnect the solvent lines (see Figure 43) connected to the valve as follows:
• Disconnect the waste tubing from port 3 of the injection valve.
• Disconnect the transfer tubing from port 2 of the injection valve.
• Using a 1/4 in. open-end wrench, disconnect the 1/16 in. OD high-pressure tubing
from ports 5 and 6.
IMPORTANT You do not need to remove the sample loop from the injection valve
to disassemble the injection valve. Once the sample loop has been swaged onto the
injection valve, the fittings are no longer interchangeable. If you do remove the
sample loop from the injection valve, take note of which fitting was connected to
port 1 and which fitting was connected to port 4. When you reinstall the sample loop,
connect the fittings to their respective ports.
Figure 43. Injection valve solvent line connections
20
µL
Waste line
fitting
Transfer tube
fitting
2
1
3
6
4
5
Sample loop
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Replacing the Rotor in the Valco Injection Valve
2. Using a #2 Phillips screwdriver, remove the two screws that secure the injection valve to
the autosampler (see Figure 44).
Figure 44. Injection valve secured to the autosampler with two screws
Screws that secure the injection valve
20
µL
2
Return from
heat exchanger
1
3
6
4
5
3. Slide the injection valve forward out of the autosampler.
Figure 45 shows the drive shaft on the back side of the injection valve.
Figure 45. Injection valve with a view of the drive shaft on the back side of the valve
Drive shaft
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Replacing the Rotor in the Valco Injection Valve
Disassembling the Injection Valve
Leave the sample loop connected to the stator as you disassemble the injection valve.
 To disassemble the injection valve
1. Using a 9/64 in. L-hex wrench, remove the two socket head cap screws that secure the
stator to the driver (see Figure 46).
CAUTION The polished (sealing) surface of the stator contains six ports that excess
handling can easily damage. As you remove the stator from the injection valve, avoid
touching this polished surface, and never place the polished surface face down on a
hard surface.
Figure 46. Injection valve (exploded view)
Driver
Rotor
Stator
Socket head
cap screws
2. Gently pry the rotor away from the driver (see Figure 46).
3. Examine the sealing surface of the rotor for scratches. If scratches are visible, replace the
rotor.
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Replacing the Rotor in the Valco Injection Valve
Cleaning the Stator
Before you replace the rotor, clean the stator.
 To clean the stator
1. Inspect the stator to determine if it requires cleaning.
2. If the stator is dirty, swab it with HPLC-grade methanol. If more stringent cleaning is
required, use a sonicator.
3. Inspect the remaining valve components for contamination. Clean them as necessary.
IMPORTANT If the stator is scratched, replace the valve. Scratches can damage the
rotor and cause cross-port leaks.
Installing a New Rotor
The rotor has two sides. The side containing the engraved flow passages (see Figure 47) faces
the stator.
Figure 47. Engraved flow passages in the Valco rotor
Engraved flow passages
 To install a new rotor
1. Replace the rotor in the driver, making sure that the rotor sealing surface with its
engraved flow passages is facing out.
The pattern is asymmetrical to prevent improper placement.
2. Replace the stator.
3. Insert the two socket head screws.
a. Using a 9/64 in. L-Hex wrench, tighten each screw until you feel resistance
(approximately fingertight).
b. Tighten each screw by 1/8 turn, and then tighten each screw again, until the stator is
secured to the driver. Take care not to overtighten the screws.
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Routine Maintenance
Replacing the Rotor in the Valco Injection Valve
Reinstalling the Injection Valve
 To reinstall the injection valve
1. Align the drive shaft (see Figure 45) and the two prongs of the coupling unit with the
three holes in the injection valve receptacle located in the lower portion of the column
oven compartment. Then insert the injection valve into the receptacle (see Figure 48).
Figure 48. Injection valve receptacle in the lower portion of the column oven compartment
Transfer tubing
20
µL
Waste
tubing
Injection valve
receptacle
Pump connection
Return from
heat exchanger
2. Align the holes in the mounting plate of the injection valve with the small holes next to
the injection valve receptacle.
3. Using the two screws that you removed in step 1 on page 64, secure the injection valve to
the autosampler with a #2 Phillips screwdriver.
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Replacing the Rotor in the Valco Injection Valve
4. Reconnect the solvent lines to the injection valve as follows (see Figure 48):
• Hand-tighten the transfer tube fitting to port 2 and the waste tube fitting to port 3.
Never use a wrench to tighten these fittings.
CAUTION Never use a wrench to tighten the transfer tube fitting.
Over-tightening the fitting can do the following:
• Restrict the end of the tubing, which leads to less sample reaching the sample
loop and results in smaller than expected peak areas or blank injections. A
restriction in the end of the tubing can also cause the syringe to make a
grinding sound during an injection sequence.
• Strip the threads on the fitting, which leads to solvent leaks.
• Hand-tighten the waste tube fitting to port 3.
• Press the end of the return tubing from the heat exchanger to the bottom of port 5 as
you hand-tighten the high-pressure fitting. Then use a wrench to tighten the fitting
by an additional 1/4 turn.
• Using high-pressure tubing and two high-pressure fittings, reconnect the LC column
to port 6 of the injection valve.
Note To reconnect tubing with a pre-swaged stainless steel fitting to a valve port,
hand-tighten the fitting as you press the tubing against the bottom of the port,
and then use a wrench to tighten the fitting by an additional 1/4 to 1/2 turn.
To connect tubing with a two-piece stainless steel fitting to a mating port, press
the tubing against the bottom of the port as you hand-tighten the fitting. Then
tighten the fitting by an additional 3/4 turn with a wrench. Once you use the
wrench to tighten the fitting, the ferrule portion of the two-piece fitting is
permanently connected to the tubing.
• If you removed the sample loop, connect the appropriate ends of the sample loop to
ports 1 and 4.
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Routine Maintenance
Replacing the Rotor in the Valco Injection Valve
Figure 49. Schematic of the tubing connections to the injection valve
2
Transfer tube low-pressure fitting
(hand-tighten only)
1
3
Sample loop fitting
(tighten with 1/4 in. wrench)
Waste-tube
low-pressure fitting
(hand-tighten only)
2 1
3
6
4 5
6
4
Sample loop fitting
(tighten with 1/4 in.
wrench)
5
LC column connection
high-pressure fitting
(tighten with 1/4 in. wrench)
Return from heat exchanger
connection
high-pressure fitting
5. Turn on the pump flow and check for leaks.
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Replacing the Inlet Filter for the Wash Bottle Solvent Line
Replacing the Inlet Filter for the Wash Bottle Solvent Line
Properly filtering the flush solvent extends the life of the injection valve and the LC column.
 To replace the inlet filter
1. Be sure the autosampler is in Idle mode.
2. Remove the flush-solvent bottle cap and lift out the inlet tubing.
3. Remove the old inlet filter.
CAUTION Be sure to follow all of the safety precautions for each solvent, as prescribed
by the MSDS.
4. Install a new filter, and then put the line back into the bottle. Tighten the solvent cap.
5. Flush the inlet filter and lines with an appropriate volume of solvent.
Replacing the Transfer Tubing
The transfer tubing that connects the injection port of the autosampler to port 2 of the
injection valve can become worn, damaged, or plugged with use.
Replace the transfer tubing if it becomes plugged or if your injections become less
reproducible (which can indicate of a worn or damaged transfer tube). In addition, replace the
transfer tubing when you replace the needle.
The transfer tubing assembly consists of two fittings and a 9 in. length of 0.012 in. ID tubing
(see Figure 50). The stainless steel nut connects to the autosampler injection port and the
green PEEK fitting connects to port 2 of the injection valve. The label near the PEEK fitting
end of the tubing specifies the internal volume of the tubing.
Figure 50. Transfer tubing assembly
18 μL
Stainless steel nut
18μL
Internal volume
of the tubing
Thermo Scientific
PEEK
fitting
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Routine Maintenance
Replacing the Transfer Tubing
Tip Because the flanged end of the transfer tubing (see Figure 51) conforms to the shape
of the needle, it is good practice to replace the transfer tubing when you replace the
needle. The reverse is not true. The tip of the needle does not conform to the shape of the
transfer tube fitting, so you do not need to replace the needle when you replace the
transfer tubing.
Figure 51. Transfer tube nut and flanged end of the transfer tubing
Flanged tubing
(that conforms to the
shape of the needle)
Transfer tube nut
 To replace the transfer tubing
1. To remove the aluminum retention clip (see Figure 52) from the wash station housing,
pull it forward and then upward.
Figure 52. Retention clip for the injection port
Retention clip
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Replacing the Transfer Tubing
2. Pull the injection port out from the wash station (see Figure 53).
Figure 53. Injection port seated in the wash station
Injection port
Spring
3. Unscrew the transfer tube nut from the injection port fitting (see Figure 54) and from
port 2 of the injection valve, and then pull the tubing to the left through the hole in the
wall between the tray compartment and the oven compartment (see Figure 10 on
page 11). Discard the used transfer tube.
Figure 54. Injection port connected to the transfer tube nut
Injection port
Transfer tube nut
4. Screw the injection port onto the nut of the new transfer tubing.
5. Set the injection port into the spring (see Figure 55). Then insert it into the port in front
of the wash station.
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Routine Maintenance
Replacing the Transfer Tubing
Figure 55. Transfer tubing, spring, and injection port
Injection port
Assembled
Transfer tube fitting
(metal)
Spring
6. Reinstall the retention clip (see Figure 56) as follows:
a. Align the retention clip with the slot located in front of the wash station.
b. Using the retention clip to push the injection port down, align the circular cutout in
the retention clip over the alignment screw.
c. Insert the end of the retention clip into the wash station slot as far as it will go.
Figure 56. Retention clip installation
Slot in wash station
Injection port
Alignment screw
Retention clip
installed
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Maintaining the Syringe Drive Assembly
7. Route the other end of the transfer tube through the hole in the wall between the tray and
column oven compartments (see Figure 10 on page 11).
8. Taking care to avoid overtightening, hand-tighten the fitting to port 2 of the injection
valve. Never use a wrench to tighten this fitting.
CAUTION Never use a wrench to tighten the transfer tube fitting. Overtightening
constricts the transfer tube, causing poor injection precision or blank injections.
Eventually, the damaged transfer tube becomes completely plugged.
IMPORTANT You must modify the instrument configuration by entering the new
value for the internal volume of the transfer tubing. For instructions on updating the
instrument configuration for the autosampler, refer to the data system Help.
9. Modify the instrument configuration for the internal volume of the new transfer tube.
For information on specifying the volume of the transfer tubing, refer to the data
system Help.
Maintaining the Syringe Drive Assembly
If you have an older version of the Accela Autosampler with an uncoated, stainless steel lead
screw, as a part of routine maintenance, lubricate the lead screw once every six months or as
needed. If the syringe drive skips steps and does not return to the home position after you
perform a Flush Syringe direct command, lubricate the lead screw before you continue
operation.
Note The syringe drive assembly in the current version of the Accela Autosampler does
not require routine maintenance. The lead screw is coated with black Teflon and the drive
nut is made of plastic, so there is minimal friction between these moving parts.
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Routine Maintenance
Maintaining the Syringe Drive Assembly
Figure 57 shows the uncoated, stainless steel lead screw in an older version of the Accela
Autosampler.
Figure 57. Syringe drive assembly with an uncoated, stainless steel lead screw
Lead screw
Lubricating the lead screw requires these tools and materials:
Tools and materials
Flat-head screwdriver
Tri-Flow™ lubricant (For ordering information, see Lubricant in the list of “Consumables”
on page 126.)
 To clean and lubricate the lead screw of the syringe drive assembly
1. Remove the syringe drive assembly from the autosampler as follows:
a. Remove the wash bottle from the solvent platform and place it on the countertop
next to the Accela LC stack.
b. Disconnect the needle tubing from the right side of the syringe valve.
c. Disconnect the wash tubing from the left side of the syringe valve.
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Maintaining the Syringe Drive Assembly
d. Remove the two thumbscrews that connect the syringe drive assembly to the inner
side of the tray compartment door (see Figure 58).
e. To disconnect the cable that is routed through the back of the syringe drive assembly
and connected to the P2 receptacle on the inner side of the tray compartment door,
press the connector tab (see Figure 59) and pull the connector out of the receptacle.
Figure 58. Syringe drive assembly disconnected from the inner side of the door
P2 receptacle
Thumbscrews
Alignment
holes
Tray compartment door
Figure 59. Syringe drive assembly cable
Connector tab
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Routine Maintenance
Maintaining the Syringe Drive Assembly
Note You can lubricate the lead screw without removing the base plate from the back
of the syringe drive assembly.
2. Lubricate the lead screw as follows:
a. Through the cutout in the front of the syringe drive assembly, apply 2 to 3 drops of
Tri-Flow lubricant to the top part of the lead screw.
b. Using a flat head screwdriver, turn the screw at the bottom of the motor
(see Figure 60) counterclockwise until the arm is at the top position.
Figure 60. Screw at the bottom of the syringe drive motor
Screw
c. Apply lubricant to the bottom part of the lead screw.
3. Reconnect the syringe drive assembly to the autosampler as follows:
a. Plug the syringe drive assembly cable into the P2 receptacle on the inner side of the
tray compartment door.
b. Realign the syringe drive assembly with the alignment holes on the inner side of the
tray compartment door (see Figure 58 on page 77).
c. Using the two thumbscrews that you removed in step 1d on page 77, secure the
syringe drive assembly to the door.
4. Set the syringe to its home position (see “Setting the Syringe to the Home Position” on
page 95).
5. To ensure that the lead screw has been properly lubricated, set up the autosampler as
follows and watch the movement of the syringe as the autosampler flushes the syringe:
a. Fill the wash bottle with HPLC-grade water.
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Troubleshooting a Blockage in the Injection System
b. Perform a Flush Syringe direct command using these values:
Parameter
Value
Reservoir
Flush Bottle (Xcalibur data system or Tune window)
Bottle (ChromQuest data system)
Volume (μL)
6000
Flush Speed (μL/sec)
100
If the lead screw has been properly lubricated, it does not stall as the autosampler
flushes water through the system at a speed of 100 μL/sec.
6. Refill the wash bottle with an appropriate solvent mixture for your applications.
Troubleshooting a Blockage in the Injection System
A blocked tubing or a plugged needle can cause the syringe drive to stall. A stalled syringe
drive makes a grinding sound because the syringe motor must overcome a greater force to
move the plunger.
If the injection system has a plug (as indicated by an abnormally loud grinding sound made
by the syringe drive), determine where the plug is located by systematically checking the
following sections of the injection system (see Figure 61):
• Section 1: Needle tubing assembly
• Section 2: Needle assembly
• Section 3: Transfer tubing
• Section 4: Injection valve and sample loop
Tip The transfer tube, needle, and small ports in the stator of the injection valve are the
most commonly plugged components of the injection system. A damaged syringe valve
can also reduce the solvent flow and cause a grinding sound.
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Figure 61. Possible locations of a plug
1 Needle tubing assembly
2 Needle assembly
Wash station
Injection port
3 Transfer tube
1
2
6
Waste line
3
5
4
4
Injection valve and
sample loop
To find and clear a plug, follow these steps until you find the blockage:
1. Testing the Needle Tubing and the Syringe Valve for Blockage
2. Testing the Transfer Tube for Blockage
3. Testing the Transfer Tube for Blockage
4. Cleaning the Injection Valve or the Sample Loop
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Troubleshooting a Blockage in the Injection System
Testing the Needle Tubing and the Syringe Valve for Blockage
 To test the needle tubing for blockage
1. Move the XYZ arm to the front-center of the tray compartment (see “Setting the Needle
to the Removal Position” on page 85).
2. Unscrew the needle tubing assembly from the needle assembly (see Figure 4 on page 6)
and place the needle tubing fitting into a beaker.
3. As you perform a Wash Needle direct command with a 2000 μL volume, observe the
solvent flow and listen for a grinding sound from the syringe drive.
4. Depending on whether solvent flows freely out the end of the needle tubing and you hear
a grinding sound from the syringe drive, do one of the following:
• If the wash solvent flows out of the needle tubing at a normal rate and you do not
hear a grinding sound, the needle tubing is not the source of blockage. Skip step 5
and step 6 and go to the next procedure to test the needle for blockage.
–or–
• If the flow appears to be constricted, go to step 5.
5. Verify that the syringe valve is not the cause of the constricted flow:
a. Unscrew the needle tubing from the right side of the syringe valve.
b. Perform a Wash Needle direct command using a 2000 μL volume, and observe the
solvent flow out of the right side of the syringe valve.
c. Depending on whether solvent flows freely out of the right side of the syringe valve,
do one of the following:
• If solvent flows out of the right side of the syringe valve, replace the needle
tubing, which was the source of the blockage.
• If solvent does not flow out of the right side of the syringe valve, replace the
syringe valve.
6. After you replace the syringe valve or the needle tubing, home the position of the syringe
(see “Setting the Syringe to the Home Position” on page 95).
If the needle tubing was not the blockage source, go to the next procedure, “Testing the
Needle for Blockage.”
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Testing the Needle for Blockage
 To test the needle for blockage
1. Remove the needle from the needle mount on the XYZ arm by pulling the latch nut of
the needle assembly forward, and then pulling the needle up out of the needle mount.
2. Reconnect the needle tubing to the needle.
3. Holding the needle over the beaker, perform a Wash Needle direct command using a
2000 μL volume. Watch how freely solvent flows from the end of the needle, and listen
for a grinding sound from the syringe drive.
4. Depending on whether solvent flows freely out of the end of the needle and you hear a
grinding sound from the syringe drive, do one of the following:
• If the solvent flow is constricted, the needle is constricted.
–or–
• If solvent flows out of the end of the needle at a normal rate and you do not hear a
grinding sound from the syringe drive, the needle is not the source of blockage.
Go to step 6.
5. To remove the needle blockage, run a clean wire through the needle or replace the needle
as described in “Replacing the Needle Tubing Assembly” on page 85. After you replace
the needle assembly, set the syringe to its home position.
6. If the needle is not the source of blockage, reconnect the injection system as follows:
a. Disconnect the needle from the needle tubing.
b. Move the XYZ arm to the front-center of the tray compartment (see “Setting the
Needle to the Removal Position” on page 85).
c. Insert the needle into the needle mount on the XYZ arm, and then push the latch nut
backward.
d. Reconnect the needle tubing to the needle, and then ensure that the needle tubing is
routed through the appropriate brackets (see “Replacing the Needle Tubing
Assembly” on page 85).
If the needle was not the blockage source, go to the next procedure, “Testing the Transfer
Tube for Blockage.”
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Troubleshooting a Blockage in the Injection System
Testing the Transfer Tube for Blockage
 To test the transfer tube for blockage
1. Disconnect the transfer tube from port 2 of the injection valve (see Figure 61 on
page 80).
2. Place the free end of the transfer tube into a beaker to collect solvent.
3. As you perform a Flush Syringe direct command, listen for an abnormally loud grinding
sound.
• If the syringe drive does not stall, then the plug is in the injection valve or the sample
loop. Reconnect the transfer tube to port 2 of the injection valve and go to the next
procedure, “Cleaning the Injection Valve or the Sample Loop.”
• If the solvent flow is constricted through the transfer tube, replace the transfer tube
(see “Replacing the Transfer Tubing” on page 71).
4. After you replace the transfer tube, home the position of the syringe (see “Setting the
Syringe to the Home Position” on page 95).
If the transfer tube was not the blockage source, go to the next procedure, “Cleaning the
Injection Valve or the Sample Loop.”
Cleaning the Injection Valve or the Sample Loop
If the blockage was not in the needle tubing assembly or the transfer tube, it must be in either
the injection valve or the sample loop. Clear the blockage, and then set the syringe to the
home position.
CAUTION Do not clean the injection valve with a cleaning wire, which can damage the
valve.
 To clear the blockage from the sample loop
1. Remove the sample loop from the valve.
2. Clear blockage from the sample loop by pushing a wire through it. Or, connect a new
sample loop to the valve.
3. After you reconnect the sample loop, home the position of the syringe.
 To clear the blockage from the injection valve
1. Remove the sample loop from the valve (see “Changing the Sample Loop” on page 49).
2. Remove the valve from the autosampler (see “Removing the Injection Valve from the
Autosampler” on page 64).
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3. Disassemble the valve (see “Disassembling the Injection Valve” on page 66).
CAUTION The polished (sealing) surface of the stator contains six ports that excess
handling can easily damage. As you remove the stator from the injection valve, avoid
touching this polished surface, and never place the polished surface face down on a
hard surface.
4. Place the stator (with the front face down) and the rotor (Valco injection valve) or rotor
seal (Rheodyne injection valve) side-by-side in a beaker. Fill the beaker with HPLC-grade
methanol or acetonitrile, and then place the beaker in an ultrasonic bath and sonicate for
approximately 10 minutes.
5. Reassemble the valve (see “Installing a New Rotor” on page 67).
6. Reinstall the valve (see “Reinstalling the Injection Valve” on page 68).
7. After you clear the source of the blockage, home the position of the syringe (see “Setting
the Syringe to the Home Position” on page 95).
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Replacing the Needle Tubing Assembly
Replacing the Needle Tubing Assembly
The needle tubing assembly connects the solvent path between the needle and the syringe
valve.
CAUTION The needle tubing is easily damaged. Before you operate the autosampler, take
care to route the needle tubing through the support brackets.
To replace the needle tubing assembly, follow these steps:
1. Setting the Needle to the Removal Position
2. Removing the Needle Tubing Assembly
3. Installing the Needle Tubing Assembly
4. Setting the XYZ Arm to the Home Position
Setting the Needle to the Removal Position
Setting the needle to the removal position moves the XYZ arm to the front-center of the tray
compartment and lowers the needle one inch (see Figure 62).
Figure 62. XYZ arm set to the front center of the tray compartment
XYZ arm
Needle
assembly
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Replacing the Needle Tubing Assembly
 To set the needle to the removal position
Do the following:
• For the Xcalibur data system, open the Direct Control dialog box for the autosampler
(see “Using the Xcalibur Direct Control Commands” on page 141). Select Needle
Removal, and then click Apply.
• For the ChromQuest data system, open the Direct Controls page for the autosampler
(see “Using the ChromQuest Direct Commands” on page 143). Select Position Arm
to Replace Needle, and then click Submit.
To replace the needle tubing assembly, go to the next procedure, “Removing the Needle
Tubing Assembly.”
Removing the Needle Tubing Assembly
 To remove the needle tubing assembly
1. Move the XYZ arm to the front center of the tray compartment (see “Setting the Needle
to the Removal Position” on page 85).
2. Unscrew the needle tubing from the needle assembly.
3. Remove the needle tubing from the bracket on the XYZ arm (see Figure 64 on page 88).
4. Pull the two-pronged needle tubing guide out of the x-axis positioning frame (see
Figure 65 on page 88).
5. Move the XYZ arm to the back of the tray compartment (see “Moving the XYZ Arm to
the Back of the Tray Compartment” on page 53).
6. Remove the needle tubing from the bracket on the left wall of the tray compartment
(see Figure 66 on page 89).
7. Remove the needle tubing from the bracket on the inside of the tray compartment door
(see Figure 68 on page 90).
8. Pull the tubing out of the tray compartment. A black or red PVC cap keeps the tubing
positioned below the metal runner for the x-axis positioning frame.
9. Unscrew the needle tubing from the right side of the two-way syringe valve
(see Figure 69 on page 90).
Go to the next procedure, “Installing the Needle Tubing Assembly.”
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Replacing the Needle Tubing Assembly
Installing the Needle Tubing Assembly
 To install a new needle tubing assembly
1. Connect the internally-threaded fitting of the needle tubing assembly to the needle
assembly fitting (see Figure 63).
2. Verify that the length of tubing between the internally threaded fitting and the black
sleeve is approximately 16.5 cm (6.5 in.) in length (see Figure 63).
Tip When the needle tubing assembly is connected to the needle assembly, the
distance between the needle tubing fitting and the letter E on the tray compartment is
approximately 16.5 cm (6.5 in.).
Figure 63. Appropriate length of tubing between the needle tubing fitting and the needle
tubing sleeve
Needle assembly fitting
Needle tubing
internally threaded fitting
Needle tubing sleeve
3. Route the needle tubing through the bracket on the XYZ arm. Seat the black sleeve in the
middle of the bracket (see Figure 64).
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Figure 64. Needle tubing routed through the bracket on the XYZ arm
Bracket on the XYZ arm
4. Insert the prongs of the needle tubing guide into the alignment holes in the
x-axis positioning frame (see Figure 65).
Figure 65. Needle tubing guide inserted into the alignment holes in the x-axis frame
Needle tubing guide
x-axis positioning frame
5. Move the XYZ arm to the back of the tray compartment (see “Moving the XYZ Arm to
the Back of the Tray Compartment” on page 53).
6. Route the needle tubing through the bracket on the left wall of the tray compartment
(see Figure 66).
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Figure 66. Bracket on the left wall of the tray compartment
Bracket on the left wall of
the tray compartment
7. Push the black or red PVC cap (that is attached to the needle tubing assembly) below the
metal runner for the x-axis positioning frame (see Figure 67).
Figure 67. Black or red PVC cap that keeps the needle tubing below the metal runner of the
x-axis positioning frame
Black or red PVC cap
Metal runner for the x-axis
positioning frame
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Replacing the Needle Tubing Assembly
8. Route the needle tubing through the bracket on the inside of the tray compartment door
(see Figure 68).
Figure 68. Bracket on the inside of the tray compartment door
Needle tubing threaded through
the bracket on inner side of the
tray compartment door
Note The bracket on the tray compartment door of your autosampler might look
different from the bracket shown in Figure 68.
9. Connect the externally threaded fitting of the needle tubing fitting to the right side of the
syringe valve (see Figure 69).
Figure 69. Needle tubing connected to the right side of the syringe valve
Syringe valve
Externally-threaded fitting
Go to the next procedure, “Setting the XYZ Arm to the Home Position.”
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Replacing the Needle Tubing Assembly
Setting the XYZ Arm to the Home Position
To set the XYZ arm to the home position, use the appropriate command from the data
system.
 To set the XYZ arm to the home position
Do the following:
• For the Xcalibur data system, open the Direct Control dialog box for the autosampler
(see “Using the Xcalibur Direct Control Commands” on page 141). Select Set Arm
to Home Position, and then click Apply.
• For the ChromQuest data system, open the Direct Controls page for the autosampler
(see “Using the ChromQuest Direct Commands” on page 143). Select GoTo Home,
and then click Submit.
The home position of the XYZ arm is directly behind the wash station (see Figure 70).
Figure 70. XYZ arm in the home position
XYZ arm in the
home position
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Replacing the Needle Assembly
Replacing the Needle Assembly
The needle assembly is a welded piece of 0.012 in. ID stainless steel tubing with an
externally-threaded fitting, a needle flag, a latch, and a compression spring (see Figure 71).
It slides into the needle mount on the XYZ arm and is secured with the latch.
Figure 71. Needle assembly
10-32 fitting
Latch
Compression
spring
Needle flag
Needle mount
on the XYZ arm
 To replace the needle assembly
1. Move the XYZ arm to the front-center of the tray compartment (see “Setting the Needle
to the Removal Position” on page 85).
2. Remove the needle assembly as follows:
a. Unscrew the needle tubing assembly from the needle assembly.
b. Pull the latch nut of the needle assembly forward.
c. Pull the needle up and out of the needle mount on the XYZ arm.
3. Install a new needle assembly as follows:
a. Slide the needle into the needle mount on the XYZ arm.
b. Turn the latch to the right.
c. Attach the needle tubing assembly to the needle assembly.
4. Home the position of the XYZ arm (see “Setting the XYZ Arm to the Home Position” on
page 91).
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Changing the Syringe or Replacing the Inner Plunger
Changing the Syringe or Replacing the Inner Plunger
You must change the syringe if a different size is required for your application. For the
dual-concentric syringe, the lifespan of the inner plunger is approximately 50000 injections.
If the inner plunger of the 250 μL syringe wears out—that is, if fluid leaks out of the bottom
of the syringe, you must replace the plunger. For the inner plunger part number, see page 126.
To replace the syringe or the syringe plunger, follow these steps:
1. Setting the Syringe to the Removal Position
2. Removing the Syringe from the Autosampler
3. Replacing the Inner Plunger of the Syringe
4. Installing the Syringe
5. Setting the Syringe to the Home Position
Setting the Syringe to the Removal Position
 To set the syringe to the removal position
Do the following:
• For the Xcalibur data system, open the Direct Control dialog box for the autosampler
(see “Using the Xcalibur Direct Control Commands” on page 141). Select Set
Syringe to Removal Position, and then click Apply.
• For the ChromQuest data system, open the Direct Controls page for the autosampler
(see “Using the ChromQuest Direct Commands” on page 143). Select Position
Syringe for Removal, and then click Submit.
The inner plunger of the dual-concentric syringe and the syringe drive move downward.
Removing the Syringe from the Autosampler
 To remove the syringe
1. Set the syringe to the removal position (see the previous procedure, Setting the Syringe to
the Removal Position).
2. Undo the screw that connects the syringe plunger to the syringe drive (see Figure 72).
3. Loosen and remove the syringe by turning the knurled top of the syringe
counterclockwise (see Figure 72).
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Figure 72. Syringe mounted to the syringe drive assembly
Knurled nut
Screw that connects the inner
plunger to the syringe drive
Replacing the Inner Plunger of the Syringe
 To replace the inner plunger
1. Remove the syringe from the autosampler (see “To set the syringe to the removal position”
on page 93).
2. Pull the worn plunger out of the bottom of the syringe (see Figure 73).
3. Replace the worn plunger with a new replacement inner plunger.
Figure 73. Inner plunger removed from the dual-concentric syringe
Inner plunger
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Changing the Syringe or Replacing the Inner Plunger
Installing the Syringe
 To reinstall the syringe
1. Screw the top of the syringe into the syringe valve.
2. Reconnect the bottom of the inner plunger to the syringe drive.
3. Home the position of the syringe plungers (see the next procedure, Setting the Syringe to
the Home Position).
Setting the Syringe to the Home Position
Figure 74 shows the relative positions of the syringe plungers in the syringe removal position
and the home position.
Figure 74. Relative positions of the syringe plungers
Removal position
Home position
 To set the syringe to the home position
Do the following:
• For the Xcalibur data system, open the Direct Control dialog box for the autosampler
(see “Using the Xcalibur Direct Control Commands” on page 141). Select Set
Syringe to Home Position, and then click Apply.
• For the ChromQuest data system, open the Direct Controls page for the autosampler
(see “Using the ChromQuest Direct Commands” on page 143). Select Position
Syringe to Home, and then click Submit.
The inner plunger moves upward until the nut on the bottom of the plunger meets the
washer on the bottom of the syringe. Then both plungers move upward until they are
1.5 mm (0.06 in.) below the top of the outer syringe barrel.
IMPORTANT If you change the size of the syringe, you must change the syringe
type parameter for the configuration of the Accela Autosampler.
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Clearing a Plugged Heat Exchanger
Clearing a Plugged Heat Exchanger
The heat exchanger is a piece of 0.004 in. ID × 1/32 in. OD stainless steel tubing located
behind the column oven plate (see Figure 75).
To clear a plug from the heat exchanger, backflush the tubing by reversing the solvent flow.
 To backflush the heat exchanger tubing
1. Using a 1/4 in. open-end wrench, detach the tubing connected to both the inlet and the
outlet of the heat exchanger.
2. Connect the tubing from the pump to the port on the right side of the heat exchanger.
3. Set the flow rate from the pump to 1 mL/min.
4. After you clear the plug, reconnect the pump to the inlet of the heat exchanger and
reconnect the outlet of the heat exchanger to port 5 of the injection valve.
Figure 75. Schematic of heat exchanger
Normal direction
of solvent flow
Inlet from
pump
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Routine Maintenance
Replacing a Fuse
Replacing a Fuse
Instrument power is supplied by two 5.0 A, 250 V, TL, 5 OD × 20 mm length fuses. The
fuses are housed in the fuse compartment on the lower right corner as you are facing the back
panel of the autosampler (see Figure 76). If an electrical component in your autosampler stops
working, first check for a blown fuse.
Figure 76. Power entry module
Fuse holder
release tab
Time-lag fuses
Fuse holder
CAUTION Disconnect the power cord before you replace fuses.
 To replace a fuse
1. Turn off the autosampler and disconnect its power cord.
2. 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.
3. Inspect each fuse for a burned or broken wire (which indicates a blown fuse).
4. Pull the blown fuse out of the holder and discard it. Place the new fuse (5.0 A, 250 V, TL,
5 × 20 mm) into the holder with the metal end visible.
CAUTION When you are replacing fuses, use only fuses of the type and current rating
specified. Do not use repaired fuses and do not short-circuit the fuse holder.
5. Snap the fuse holder back into place.
6. Reconnect the power cord and turn on the autosampler.
If the replacement fuse fails in a short time, you have a serious electrical problem. Leave the
blown fuse in place and contact your Thermo Fisher Scientific service representative.
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Maintenance Cycles
Maintenance Cycles
The rotor (Valco injection valve) or rotor seal (Rheodyne injection valve), injection needle,
and inner syringe plunger wear with use. From the data system, you can set the scheduled
maintenance time (SMT) for these autosampler components.
To set the maintenance cycles, follow the procedure for your data system:
• Setting the Maintenance Cycles from the Xcalibur Data System
• Setting the Maintenance Cycles from the ChromQuest Data System
Setting the Maintenance Cycles from the Xcalibur Data System
Each page of the Maintenance Information dialog box contains a Usage Limit box, a Current
Counter readback, a Set New Limit button, and a Zero Counter button. Use each page to set
a new usage limit, view the current counter readback, or zero the current counter reading.
If the counter exceeds the usage limit, the Accela Autosampler cannot start a run until you
perform the scheduled maintenance, or you select the Enable Maintenance Log check box of
the Accela Autosampler Configuration dialog box.
 To set the scheduled maintenance limits from the Xcalibur data system
1. In the Instrument Setup window for the Accela AS, choose Accela AS > Maintenance.
The Maintenance Information dialog box appears (see Figure 77).
Figure 77. Maintenance Information dialog box (Xcalibur data system)
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Maintenance Cycles
2. Depending on the usage limit you want to set, open the appropriate page.
Page
Usage limit
Injections
Maximum number of autosampler injections
Valve Cycles
Maximum number of times that the autosampler can switch the
valve between the fill and inject positions
Needle Cycles
Maximum number of times that the autosampler can insert the
needle into a vial septum, the injection port, or the wash station
Syringe Cycles
Maximum number of times that the autosampler can return the
syringe to the Ready position
3. To change the usage limit, type the new value in the Usage Limit box, and then click
Set New Limit.
4. To rezero the counter, click Zero Counter.
Setting the Maintenance Cycles from the ChromQuest Data System
The Maintenance page in the ChromQuest data system contains four options: Injections,
Valve Cycles, Syringe Cycles, and Needle Cycles. After you select an option, its corresponding
Details box appears on the right side of the Maintenance page. You can use each Details box
to set a usage limit, view the current counter readback, or zero the current counter reading
(see Figure 78).
Figure 78. Maintenance page (ChromQuest data system)
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Maintenance Cycles
If the counter exceeds the usage limit, the Accela Autosampler cannot start a run until you
perform the scheduled maintenance or you clear the Enable Maintenance Log check box of
the Accela Autosampler Configuration dialog box.
 To set up the maintenance cycles from the ChromQuest data system
1. In the online Instrument window, choose Control > Instrument Status.
The Instrument Status window appears.
2. Click the Accela AS tab.
The Accela AS status page appears (see Figure 79).
Figure 79. Accela AS status page (ChromQuest data system)
Diagnostics button
3. Click Diagnostics.
The Diagnostics dialog box appears.
4. Click the Maintenance tab.
The Maintenance page appears (see Figure 78 on page 99).
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Maintenance Cycles
5. Select the option that you want to set:
• To set the usage limit for the number of user-defined injections, select the Injections
option.
• To set the usage limit for the number of user-defined valve cycles (from Fill to Inject),
select the Valve Cycles option.
• To set the usage limit for the number of user-defined syringe cycles where the syringe
is put in the Ready position, select the Syringe Cycles option.
• To set the usage limit for the number of user-defined needle cycles where the needle is
inserted into a septum, injection port, or wash station, select the Needle Cycles
option.
6. To change the usage limit, type the new value in the Usage Limit box, and then click
Submit.
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Diagnostics
Use the Accela Autosampler diagnostics to test the major electronic circuits within the
instrument and to indicate whether the circuits pass or fail the tests. If the instrument
electronics fail, the diagnostics can often locate the problem. However, to distinguish a
hardware failure from an electronic failure, someone who is familiar with system operation
and basic hardware theory must run the diagnostics.
You can use the diagnostics to test many of the Accela Autosampler components. The
Thermo Fisher Scientific service philosophy is to troubleshoot the problem to the lowest part,
assembly, PCB, or module listed in Appendix A, “Consumables and Service Parts.” You can
replace the parts listed in “Consumables” on page 126, and a Thermo Fisher Scientific field
service engineer can replace the parts listed in “Service Parts” on page 127.
Contents
• Power Up Diagnostics
• Error Log in the Xcalibur Data System
• Error Log in the ChromQuest Data System
• Autosampler Validation Information
Power Up Diagnostics
Accela Autosampler power up diagnostics run automatically when you turn on the
autosampler and take about 2 minutes to run. Most of the time required for the power up
diagnostics is occupied testing the RAM.
If there is a memory error of a fatal type, the right three lights of the Status LEDs turn amber.
If this happens, contact Thermo Fisher Scientific Technical Support. The CPU PCB probably
needs to be replaced.
If diagnostics detect any other type of error, the Run Status LED illuminates amber and the
autosampler records the error in its persistent memory log. You can access this error log
through the data system.
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Power Up Diagnostics
Table 13 lists the startup diagnostics error messages. The left column lists the error messages
that your Thermo Scientific field service engineer can view from HyperTerminal™ (or
equivalent application). The right column lists the error messages that might appear in the
autosampler error log.
When the startup test passes, the error log displays this message: No error. No other events
occur.
Table 13. Autosampler startup error messages (Sheet 1 of 2)
HyperTerminal error messages
Error log messages
EC_SDIAGS_BAD_Z_FLAG
The z-axis home flag failed.
EC_SDIAGS_BAD_S_FLAG
The s-axis home flag failed.
EC_SDIAGS_BAD_X_FLAG_INT
The x-axis interrupt failed.
EC_SDIAGS_BAD_Y_FLAG_INT
The y-axis interrupt failed.
EC_SDIAGS_BAD_Z_FLAG_INT
The z-axis interrupt failed.
EC_SDIAGS_BAD_S_FLAG_INT
The s-axis interrupt failed.
EC_SDIAGS_BAD_1_MOTION_TIMER
Motion Timer A failed.
EC_SDIAGS_BAD_2_MOTION_TIMER
Motion Timer B failed.
EC_SDIAGS_BAD_3_MOTION_TIMER
Motion Timer C failed.
EC_SDIAGS_BAD_4_MOTION_TIMER
Motion Timer D failed.
EC_SDIAGS_BAD_Z_BOTTOM_FLAG
The z-bottom flag failed.
EC_SDIAGS_TIMER_SETUP_ERROR
Occurs if an error status is returned by the disable
interrupt call.
EC_SDIAGS_NO_INJ_CABLE
The injector motor cable is not connected.
EC_SDIAGS_BAD_INJ_POS
The injection valve did not change position.
EC_SDIAGS_BAD_SYRINGE_VALVE_INT
The syringe valve interrupt did not occur.
EC_SDIAGS_BAD_FLUSH_VALVE_INT
The flush value interrupt did not occur.
EC_SDIAGS_BAD_CAPACITOR
The capacitor did not charge to the acceptable level.
EC_SDIAGS_X_MOTOR_NOT_PRESENT
The x-axis motor was not detected.
EC_SDIAGS_Y_MOTOR_NOT_PRESENT
The y-axis motor was not detected.
EC_SDIAGS_Z_MOTOR_NOT_PRESENT
The z-axis motor was not detected.
EC_SDIAGS_S_MOTOR_NOT_PRESENT
The s-axis motor was not detected.
EC_SDIAGS_XZ_CONNECTOR_NOT_
CONNECTED
The cable that connects the x- and z-axis motors is not
connected.
EC_SDIAGS_BAD_REG_VALUE
One of the memory registers is not set correctly.
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5 Diagnostics
Error Log in the Xcalibur Data System
Table 13. Autosampler startup error messages (Sheet 2 of 2)
HyperTerminal error messages
Error log messages
EC_SDIAGS_MOTOR_ERROR
The motor diagnostic bits did not change to the
correct state.
EC_SDIAGS_ISR_TIMEOUT
The interrupt did not occur before the interrupt
timeout did.
EC_SDIAGS_METAL_OUTOF_RANGE
The oven or tray metal readings were not within the
specified low and high.
EC_SDIAGS_OVEN_AIR_OUTOF_RANGE
The oven air readings were not within the specified low
and high.
EC_SDIAGS_24V_OUTOF_RANGE
The oven or tray 24 V readings were not within the
specified low and high.
EC_SDIAGS_12V_OUTOF_RANGE
The oven or tray 12 V readings were not within the
specified low and high.
EC_SDIAGS_OVEN_MIRROR_OUTOF_RANGE
The readings for the oven mirror were not within the
specified low and high.
EC_SDIAGS_UNKNOWN_DEVICE
A call was made to the temperature diagnostics with
an unknown device.
EC_SDIAGS_VIAL_MIRROR_OUTOF_RANGE
The readings for the tray mirror were not within the
specified low and high.
EC_SDIAGS_HOME_ERROR
A move to the home flag did not set the home flag bit
correctly.
Error Log in the Xcalibur Data System
The error log in the Xcalibur data system lists the diagnostic error messages. You can access
the error log through the status page of the Information View. The Information View is
normally displayed on the left side of the Home Page window and contains two pages: Status
and Acquisition Queue. If the Home Page window does not display this view, it has been
turned off.
 To open the Information View
From the Home Page window, choose View > Info View or in the toolbar, click
turn the Information View on and off. See Figure 80.
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5
Diagnostics
Error Log in the Xcalibur Data System
Figure 80. Status page of the Information View with the Accela AS selected in the directory
tree
 To access the error log for the Accela Autosampler
1. Click the Accela AS directory in the Status window to activate its status pages.
2. Click the Error log tab to open the Error Log page (see Figure 81).
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5 Diagnostics
Error Log in the ChromQuest Data System
Figure 81. Error log for the Accela Autosampler
3. Click Refresh to display the list of errors.
4. After reviewing the error list, click Clear to erase the list.
Error Log in the ChromQuest Data System
The error log in the ChromQuest data system lists the diagnostic error messages.
 To open the error log
1. From the Instrument window, choose Control > Instrument Status to display the
Instrument Status window.
2. Click the Accela AS tab to display the Accela AS page of the Instrument Status window.
3. Click Diagnostics to display the Diagnostics dialog box.
4. Click the Error log tab to display the error log for the Accela Autosampler.
• To view the error log, click Refresh.
• To clear the error log, click Clear.
Autosampler Validation Information
The Xcalibur data system keeps track of validation information. Use the Validation
Information dialog box to enter the date when the validation occurred and the name of the
person who performed the validation.
 To open the Validation Information dialog box
From the Instrument Setup window, choose Accela AS > Validation.
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Diagnostics
Autosampler Validation Information
The Validation Information dialog box has the following pages:
• Sample Tray Low Temp
• Sample Tray High Temp
• Column Oven Low Temp
• Column Oven High Temp
• Inject Reproducibility
• Dilution Reproducibility
Use these pages to enter the calendar dates when the validation occurred and the initials of the
person who performed the validation. See Figure 82.
Figure 82. Validation Information dialog box
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6
Routine Troubleshooting
This chapter provides information to help you troubleshoot possible autosampler problems.
When troubleshooting, consider the following:
• Did the system fail when you were running samples?
• Did problems occur after you performed maintenance on the instrument?
• Did you change the system configuration, cables, or peripherals just before the problem
occurred?
If the answer is yes to the first question above, the issue might be a hardware failure.
If the answer is yes to one of the latter two questions above, the issue is probably mechanical,
not electrical. Verify that alignment, configurations, and cable connections are correct.
If, after reviewing the troubleshooting table included in this chapter, you would like
additional assistance in troubleshooting a particular problem, contact Thermo Fisher
Scientific Technical Support or your service representative.
Contents
• General Chromatography Troubleshooting
• Autosampler-Specific Troubleshooting
• Common Operator Errors
• Determining the Transfer Tubing Volume
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General Chromatography Troubleshooting
General Chromatography Troubleshooting
Table 14 lists general chromatographic problems. For more information about
chromatographic troubleshooting, refer to any HPLC troubleshooting reference book, or
contact technical support or your service representative.
Table 14. General chromatography troubleshooting table (Sheet 1 of 2)
Symptom
110
Remedy
No flow
• Check the mobile phase connections.
• Check for leaks.
• Refer to the hardware manual for your pump.
High backpressure
• Check the flow rate and system/column
specifications.
• Check for tubing or column blockage.
• Refer to the hardware manual for your pump.
Unstable baseline or drift
• System/column not equilibrated; allow more time.
• Refer to the detector troubleshooting guide.
Baseline noise
• Check for air bubbles in system, degas solvents.
• Check for system/solvent contamination.
• Check the hardware manual for your pump.
No peaks
• Check the detector and data system computer
connections.
• See “Autosampler-Specific Troubleshooting” on
page 111.
• Check sample retention with chromatographic
conditions.
Contamination/ghost peaks
• Clean the system and column.
• See “Autosampler-Specific Troubleshooting” on
page 111.
• Refer to the hardware manual for your pump.
Poor peak shape
•
•
•
•
Poor retention time
reproducibility
•
•
•
•
•
Accela Autosampler Hardware Manual
Check the system for leaks.
Check fittings and tubing lengths.
Check column performance.
See “Autosampler-Specific Troubleshooting” on
page 111.
• Refer to the hardware manual for your pump.
• Refer to the hardware manual for your detector.
Check the system for leaks and bubbles.
System/column not equilibrated; allow more time.
Check column performance.
Check the hardware manual for your pump.
See “Autosampler-Specific Troubleshooting” on
page 111.
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6 Routine Troubleshooting
Autosampler-Specific Troubleshooting
Table 14. General chromatography troubleshooting table (Sheet 2 of 2)
Symptom
Remedy
Poor peak area reproducibility
• Check column performance.
• See “Autosampler-Specific Troubleshooting” on
page 111.
Non-integrated peaks or too
many peaks
• Refer to the hardware manual for each of the modules
in your LC system.
• Refer to the data system manual for information on
setting peak identification and integration.
No instrument or device
control
• Check the cable connections.
• Check the system configuration.
• Refer to the hardware manuals for each of the
modules in your LC system.
Autosampler-Specific Troubleshooting
This section focuses on problems that might occur with the autosampler portion of your
HPLC system. It contains the following topics:
• Chromatography Problems
• Hardware Problems
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Autosampler-Specific Troubleshooting
Chromatography Problems
Table 15 lists potential chromatography problems associated with the autosampler.
Table 15. Chromatography problems (Sheet 1 of 4)
Symptom
Possible problem
Remedy
1. Baseline drift when the
injection 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 Thermo Fisher
Scientific service representative.
2. Shifting retention times
a. Temperature variations in the lab
a. Stabilize the temperature.
b. Column heater problems
b. Contact your Thermo Fisher
Scientific service representative.
a. Inconsistent sample injection
volumes
a. Check for a partially blocked
injection valve and observe the
pressure. If the problem persists, see
“Replacing the Rotor in the Valco
Injection Valve” on page 63.
3. Sensitivity changes
b. Leaks between the injection valve
b. Tighten or change the fitting.
and the column causing inconsistent
sample volume injection
c. Air bubbles aspirated into the lines
along with sample
c. Replenish/prepare fresh sample and
repeat the injection.
Decrease the syringe speed.
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d. Incomplete sample mixing during
sample preparation
d. Thaw frozen samples completely
before sample preparation to
prevent sample "layering."
e. Sample adsorption to the walls of
the injection valve or sample loop
e. Change the mobile phase
composition.
f.
f.
Sample solvent incompatible with
the mobile phase
Change the solvent. Use a different
mobile phase if possible.
g. Sample insoluble in solvent
g. Be sure the sample solvent has a
lower elution strength than the
mobile phase.
h. Air in syringe/sample lines
h. Flush sample lines.
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6 Routine Troubleshooting
Autosampler-Specific Troubleshooting
Table 15. Chromatography problems (Sheet 2 of 4)
Symptom
4. Irregular peak shapes
Possible problem
Remedy
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
its organic content never exceeds the
initial composition of the mobile
phase.
c. Degradation of sample components c. Prepare fresh sample and repeat the
injection.
5. Band broadening
6. Smaller than expected
peak heights
Thermo Scientific
a. Fitting problem at the injection
valve-column connection
a. Inspect for damage, mismatch, or
incorrect assembly.
b. Tubing ID too large
b. Use 0.010 in. ID tubing from the
injection valve to the column and
from the column to the detector.
a. Small injection due to incorrect size a. Check the syringe size in the
syringe in configuration
autosampler device driver
configuration.
b. Incorrect injection volume for the
injection mode
b. Check the injection volume value in
the file. For partial loop injections,
limit the maximum injection
volume to half the sample loop size.
For no waste injections, limit the
maximum injection volume to 5 μL
less than the sample loop size.
c. Incorrect value for the transfer
tubing
c. Check the dead volume value in the
configuration. The dead volume for
the transfer tubing is specified on its
attached label. See Figure 10 on
page 11.
d. Air in lines
d. Perform a Needle Wash direct
command to remove air from the
wash bottle tubing and the syringe.
e. Incorrectly adjusted syringe drive
mechanism
e. Contact your Thermo Fisher
Scientific service representative.
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Autosampler-Specific Troubleshooting
Table 15. Chromatography problems (Sheet 3 of 4)
Symptom
7. Peaks during a blank
injection (from a previous
injection)
8. No peaks
Possible problem
Remedy
a. Sample carryover due to residual
sample in the sample loop
a. Increase flush volume. Increase
injection volume, 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 redo connections.
c. Sample carryover due to a
contaminated needle guide
c. Clean the needle guide.
d. Sample too concentrated
d. Dilute the sample.
a. Empty wash bottle or excessive air
in the wash tubing
a. Fill the wash bottle with solvent.
Remove air from the wash tubing
and the syringe.
b. Plugged or bent needle
b. Clean or replace the needle. Tighten
the fittings and the syringe.
c. Leaking syringe valve fitting at the
wash tubing connection
c. Tighten the fitting.
d. Plugged or leaking tubing
d. Replace faulty tubing.
9. Lack of precision, accuracy, a. Air leak
or both on a sample of
known content-random
b. Worn out syringe
error
a. Tighten all fittings and repeat the
run.
b. Replace the syringe (see “Changing
the Syringe or Replacing the Inner
Plunger” on page 93).
c. Worn out rotor (Valco injection valve) c. Replace the rotor (see “Replacing
the Rotor in the Valco Injection
Valve” on page 63)
d. Worn out rotor seal (Rheodyne
injection valve)
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d. Replace the rotor seal (see “Replacing
the Rotor Seal in the Rheodyne
Valve” on page 137)
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6 Routine Troubleshooting
Autosampler-Specific Troubleshooting
Table 15. Chromatography problems (Sheet 4 of 4)
Symptom
Possible problem
Remedy
10. Irreproducibility of peak
areas
a. Plugged or bent needle
a. Clean/replace needle (see
“Replacing the Needle Assembly”
on page 92). 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.
e. Wash solvent viscosity too low or
too high
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e. Change wash solvent or flush speed.
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Autosampler-Specific Troubleshooting
Hardware Problems
Table 16 lists the potential hardware problems associated with the autosampler.
Table 16. Hardware problems associated with the Accela Autosampler (Sheet 1 of 4)
Symptom
Possible problem
Remedy
a. Power cord unplugged or faulty
a. Plug in or replace the power cord.
b. Blown fuse
b. Replace the fuse (see “Replacing a
Fuse” on page 97).
c. Defective power switch, driver,
transformer
c. Contact your Thermo Fisher
Scientific service representative.
a. Incorrect computer NIC settings
a. Check the address. Use only
TCP/IP protocol.
b. Ethernet hardware problem
b. Check the Ethernet switch and
Ethernet cables.
Startup
1. Nothing works when
instrument is turned on.
Communications
2. No Ethernet
communication
3. External devices are not
triggered.
a. Incorrect timed event settings in the a. Check the timed event settings in
acquisition method
the acquisition method.
b. Interface equipment problem
b. See “Connecting the System
Interconnect Cable” on page 37 or
refer to the Accela Getting Connected
Guide for information on contact
closure connections.
a. Incorrect vial position entered into
sample sequence
a. Check the sample sequence.
Vials
4. Instrument cannot locate
vial.
b. Sample incorrectly placed in sample b. Check the tray arrangement.
tray
5. Vials do not fit in tray.
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c. XYZ arm misaligned
c. Recalibrate the XYZ arm. Refer to
the data system Help.
d. XYZ arm movement restricted
d. Remove restriction and home arm.
See “Important Precautions for
Sample Loading” on page 56.
e. Loose left-hand guide bracket
e. Ensure that the bracket is tightly
attached and level.
a. Incorrect vials
a. Check that the vial diameter is
11 to 12 mm (0.43 to 0.47 in.).
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6 Routine Troubleshooting
Autosampler-Specific Troubleshooting
Table 16. Hardware problems associated with the Accela Autosampler (Sheet 2 of 4)
Symptom
Possible problem
Remedy
6. Torn septum
a. Bent needle
a. Replace the needle (see “Replacing
the Needle Tubing Assembly” on
page 85).
7. Septum pushed into vial
a. Incorrect vial caps
a. Use only recommended vials and
vial caps. See “Recommended Vials,
Microplates, and Microplate
Covers” on page 57.
b. Loose vial caps
b. Ensure that the vial caps are fully
tightened.
a. Slipping injector drive
a. Contact your Thermo Fisher
Scientific service representative.
Injection valve
8. Injection valve will not
switch.
9. Injection valve continues to a. Loose or faulty sensor cable
rotate until fault is
generated.
a. Contact your Thermo Fisher
Scientific service representative.
10. Injection valve does not
rotate to the proper
position.
a. Valve requires excessive torque.
a. Contact your Thermo Fisher
Scientific service representative.
11. Motor fails to stop after
syringe is fully returned.
a. Possible defective sensor
a. Contact your Thermo Fisher
Scientific service representative.
12. Sample loop will not fill.
a. Blocked injection valve
a. Follow “Troubleshooting a Blockage
in the Injection System” on page 79
to isolate blockage. DO NOT clean
valve with a cleaning wire or you
could damage the valve.
13. Valve seal wears rapidly.
a. Too much valve seal pressure against a. Decrease LC system pressure.
valve
Thermo Scientific
b. Particulate matter in valve
b. Use an inline filter before the
injection valve.
c. Inadequately filtered sample
c. Filter with a 0.5 m filter before
injection.
d. Buffer crystallization
d. Do not allow buffers to stand in the
system. Run a flush sequence when
the system is not in use.
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Autosampler-Specific Troubleshooting
Table 16. Hardware problems associated with the Accela Autosampler (Sheet 3 of 4)
Symptom
Possible problem
Remedy
a. Incorrect vials
a. See the list of recommended
replacement parts in Appendix A,
“Consumables and Service Parts.”
b. Alignment problem
b. Contact your Thermo Fisher
Scientific service representative.
c. Arm sticking
c. Contact your Thermo Fisher
Scientific service representative.
a. Multiple/incorrect septum
a. Verify a single septum of the type
recommended (see “Recommended
Vials, Microplates, and Microplate
Covers” on page 57).
b. Worn needle
b. Replace the needle (see “Replacing
the Needle Assembly” on page 92).
a. Crimped needle tubing
a. Replace the needle tubing (see
“Replacing the Needle Tubing
Assembly” on page 85).
a. Plugged lines or needle
a. Clear lines or needle (see
“Troubleshooting a Blockage in the
Injection System” on page 79).
b. Defective limit switch
b. Contact your Thermo Fisher
Scientific service representative.
Needle
14. Needle bent or broken
15. Needle plugging
Needle Tubing
16. Blockage in the needle
tubing
Syringe
17. Loud buzzing from syringe
drive
Transfer tube
18. Lack of injection precision
a. Constricted or plugged transfer tube a. Replace the damaged transfer tube.
and accuracy, lack of precision
Avoid overtightening the fitting to
or blank injections
port 2 of the injection valve (see
“Replacing the Transfer Tubing” on
page 71).
Leaks
19. Leakage during loop filling
118
a. Blocked loop or waste line, or valve
not deflected far enough
Accela Autosampler Hardware Manual
a. Flush or replace the sample loop;
check and flush the waste line.
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6 Routine Troubleshooting
Autosampler-Specific Troubleshooting
Table 16. Hardware problems associated with the Accela Autosampler (Sheet 4 of 4)
Symptom
Possible problem
Remedy
20. Cross-port leakage
a. Worn rotor seal (Rheodyne
injection valve) or rotor (Valco
injection valve
a. Replace the rotor seal (see
“Replacing the Rotor Seal in the
Rheodyne Valve” on page 137).
Replace the rotor (see “Replacing
the Rotor in the Valco Injection
Valve” on page 63).
21. Liquid on vial caps
a. Leaky syringe valve
a. Replace the syringe valve.
b. Leaky injection valve
b. Replace the rotor seal (see
“Replacing the Rotor Seal in the
Rheodyne Valve” on page 137).
Replace the rotor (see “Replacing
the Rotor in the Valco Injection
Valve” on page 63).
a. Dirty vials
a. Replace vials.
b. Improper septum made of silicone
or other rubber-like material
b. Use the recommended septum (see
“Recommended Vials, Microplates,
and Microplate Covers” on
page 57).
a. Block between column and
autosampler
a. Check for blockage and tubing
restrictions.
Other
22. Contaminated sample
23. Elevated system pressures
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Routine Troubleshooting
Common Operator Errors
Common Operator Errors
Operator errors can compromise the performance of the Accela Autosampler.
These are the most common operator errors:
• Letting the wash bottle run dry
• Routing the needle tubing incorrectly
• Selecting an incorrect syringe type in the Accela Configuration Autosampler dialog box
• Requesting an inappropriate injection volume for the sample loop size and the injection
mode
• Entering an incorrect dead volume for the transfer tubing
• Overtightening the transfer tube fitting to port 2 or the wash tube fitting to port 3 of the
autosampler injection valve
Insufficient Wash Solvent
Allowing the wash bottle to run dry disrupts the ability of the syringe to draw liquid into the
needle tubing, which consequently results in blank injections. Fill the wash bottle with
solvent, and remove air from the wash line tubing before running a sequence of injections.
Damaged Needle Tubing
Because the needle tubing is easily damaged, routing the needle tubing correctly is critical to
the performance of the Accela Autosampler. Take care to avoid kinking the tubing as you
route it through the bracket on the XYZ arm.
Incorrect Syringe Type
Because the autosampler cannot detect the syringe type connected to the syringe drive
assembly, confirm that the syringe type in the Accela Configuration Autosampler dialog box
matches the autosampler syringe.
The injection volume algorithm used by the autosampler depends on the syringe type that
you select when you configure the data system. The distance that the inner syringe plunger
must descend to draw the specified injection volume increases as the syringe size decreases. For
example, to draw 10 μL of sample, the inner plunger of a 100 μL syringe must descend farther
than the inner plunger of a 250 μL syringe. If you replace the 250 μL syringe with a 100 μL
syringe, but neglect to modify the data system configuration, the autosampler injects a smaller
than expected injection volume because the syringe plunger does not descend far enough to
draw the requested injection volume.
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Routine Troubleshooting
Common Operator Errors
Inappropriate Injection Volume
For best results, follow these rules:
• For no waste injections, limit the maximum injection volume to 5 μL less than the sample
loop volume. Limit the minimum injection volume to 1 μL of sample.
• For partial loop injections, limit the maximum injection volume to 40 percent of the
nominal sample loop volume.
Incorrect Dead Volume
The dead volume of the 0.012 in. ID transfer tubing is specified on the label attached to the
tubing. Entering an incorrect dead volume when you specify the configuration options for the
autosampler can result in lower than expected peak areas or blank injections.
Overtightened Transfer Tube or Wash Tube Fittings
Avoid overtightening the low-pressure fittings to the injection valve (see Figure 83):
• Overtightening the transfer tube fitting constricts the transfer tube, causing a lack of
injection precision. Eventually, the constricted transfer tube becomes plugged.
• Overtightening the waste tube fitting constricts the waste tube, increasing the
backpressure in the injection system.
Figure 83. Transfer tubing connection to port 2 of the injection valve
Dead volume label
(internal volume of the transfer tube)
20
µL
Hand-tightened
transfer tube fitting
2
1
3
6
4
5
Hand-tightened
waste tube fitting
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Determining the Transfer Tubing Volume
Determining the Transfer Tubing Volume
The Dead volume configuration parameter is critical for the no waste injection mode.
Specifying an incorrect dead volume value can result in lower than expected peak areas or
blank injections. For best results, the specified dead volume must equal the actual volume of
the transfer tubing that connects the autosampler injection port to port 2 of the injection
valve.
 To determine the volume of the transfer tubing
1. From the Xcalibur or ChromQuest data system, set up an acquisition method for an
analyte that you know the chromatographic conditions for.
Use chromatographic conditions that result in a relatively short retention time for the
analyte (to minimize the run time). In the method, select the No Waste Injection option.
You can also use the autosampler test mix (0.5 percent toluene in methanol) that comes
with the autosampler. With the chromatographic conditions listed in Table 17, toluene
elutes at approximately two minutes.
Table 17. Chromatographic conditions for toluene
Parameter
Setting
Mobile phase
80:20 methanol/water
Flow rate
1 mL/min
Column
100 × 4.6 mm, C-18 column, 5 μm particle size, or equivalent
Temperature
Ambient
Wavelength
260 nm, 1 nm bandwidth for the PDA detector
2. Change the value for the Dead Volume autosampler configuration parameter. Start with a
value of 15.
Figure 84 shows the Communication page of the Accela Autosampler Configuration
dialog box for Thermo Foundation Instrument Configuration, and Figure 85 shows the
Accela AS Configuration dialog box in the ChromQuest data system. The default dead
volume value is 17.0. The manufacturer determines the volume of the tubing used to
produce each lot of transfer tubes (see Figure 83 on page 121) and places a label on each
tube.
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6 Routine Troubleshooting
Determining the Transfer Tubing Volume
Figure 84. Communication page (Thermo Foundation Instrument Configuration)
The Stack Address value
must match the unit ID setting
on the back panel of the autosampler.
Default dead volume setting
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Routine Troubleshooting
Determining the Transfer Tubing Volume
Figure 85. Accela AS Configuration dialog box (ChromQuest data system)
The Stack value must match
the unit ID setting on the back panel
of the autosampler.
Default dead volume setting
3. From the Xcalibur or ChromQuest data system, inject 1 μL of sample using the no waste
injection mode.
4. Repeat step 2 and step 3, increasing the dead volume by 1 μL each time until you have
data points from 15 to 23 μL.
5. Plot the peak area versus dead volume. Check for an increase in peak area until it reaches
a plateau.
Due to injection variability, the plateau might not be flat. Select the dead volume where
the maximum is first reached.
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A
Consumables and Service Parts
A consumable item is an item that is expected to wear with use or that comes in contact with
system fluids or samples. As a result, you must periodically replace a consumable item. A
service part is a subassembly of the autosampler. Normally, Thermo Fisher Scientific service
representatives or other qualified personnel replace a service part. This chapter provides a
comprehensive listing of consumables and Accela Autosampler service parts that you can
order and replace.
This chapter also includes an itemized list of the parts in the Accela System Kit. This kit
contains the hardware required to interconnect the system modules and the instrument device
drivers required to operate the modules. The Accela System Kit is not packaged with the
Accela Autosampler. You must order it separately or as part of a system bundled with either
the Xcalibur or ChromQuest data system.
Note To control the Accela Autosampler, you must have the Xcalibur or ChromQuest
data system and the Accela device drivers.
• For the Xcalibur data system, the Accela device drivers are provided on the LC
Devices software CD.
• For the ChromQuest 4.2 data system, the Accela device drivers are provided on the
Accela Add-On Device Drivers CD.
• For the ChromQuest 5.0 data system, the Accela drivers are provided on the
ChromQuest data system CD.
Contents
• Consumables
• Service Parts
• Accela AS Accessory Kit
• Accela System Kit
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Consumables and Service Parts
Consumables
Consumables
These parts are consumables for the Accela Autosampler. The sample loops are compatible
with the Valco injection valve.
Accessory Kit, Accela Autosampler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-62001
Assembly, needle, inert . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-60017
Assembly, needle tubing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-60102
Assembly, syringe valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A3692-010
Assembly, transfer tube, 0.012 in. ID . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-60014
Assembly, wash bottle kit. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-60041
Cooling adapter, 96 well . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-20002
Ferrule, front, Swagelok. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .00101-08-00001
Ferrule, back, Swagelok . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .00101-08-00002
Filter, flush solvent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A4258-010
Grease, silicon/Teflon (for lead screw) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00301-01910
Kit, Maintenance Accela AS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-62002
Kit, Needle Tubing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-62043
Kit, Wash Bottle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-62009
Lubricant, Tri-flow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1611-0030
Nut, compression, long, 10-32, 1/16 in. OD tube . . . . . . . . . . . . . . . . . . .00101-07-00001
Plunger, replacement, concentric, 100 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1123-010
Plunger, replacement, concentric, 250 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1123-020
Plunger, replacement, concentric, 500 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1123-030
Port, needle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-20031
Reservoir vials, 16 mL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00301-07527
Retainer, needle port (injection port). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-10035
Ring, bearing (Rheodyne injection valve) . . . . . . . . . . . . . . . . . . . . . . . . . .00109-99-00022
Rotor, Valco injection valve (also known as a rotor seal) . . . . . . . . . . . . . . .00110-03-00019
Syringe, dual concentric, 100 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1100-010
Syringe, dual concentric, 250 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1100-020
Syringe, dual concentric, 500 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1100-030
Syringe, standard, 2500 μL (with needle tubing extension) . . . . . . . . . . . . . . 60053-62002
Syringe, 2.5 mL. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-60006
Sample loop, 1000 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .00109-99-00030
Sample loop, 5 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .00109-99-00007
Sample loop, 10 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .00109-99-00008
Sample loop, 20 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .00109-99-00009
Sample loop, 25 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .00109-99-00010
Sample loop, 50 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .00109-99-00011
Sample loop, 100 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .00109-99-00012
Sample loop, 500 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .00109-99-00013
Sample loop, 1000 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .00109-99-00014
Seal, rotor (for Rheodyne injection valve . . . . . . . . . . . . . . . . . . . . . . . . . . .00109-99-00021
Stripper, vial . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1034-010
Valve, injection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-60018S
Vial caps, 100 μL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-40009
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Consumables and Service Parts
Service Parts
Consumables (continued)
Tubing assembly, 2.5 mL syringe. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-60005
Vial kit, 1.8 mL w/250 μL insert, 100 per pack. . . . . . . . . . . . . . . . . . . . . . . . 60053-62017
Vial kit, 2 mL, 100 count, includes caps and septa (2 each). . . . . . . . . . . . . . . . . A4954-010
Service Parts
Normally, Thermo Fisher Scientific service representatives or other qualified personnel replace
these service parts:
Assembly, arm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-60015
Assembly, cooler . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-60014
Assembly, cooler fan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1110-010
Assembly, door left . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60057-60011
Assembly, door right . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60057-60015
Assembly, oven . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-60011
Assembly, oven fan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-63006
Assembly, oven sensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F9050-010
Assembly, tray sensor. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F9090-010
Base, pod, z-axis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1033-010
Cable, injector sensor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A3513-010
Cable, door LED. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-63002
Cable, temperature control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1002-010
Cable, assembly, DC oven motor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1017-010
Cable, assembly temperature control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1002-010
Cable, A/C input power . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1079-010
Cable, A/C to options PS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1080-010
Cable, backplane power. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1082-010
Cable, Oven DC power. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1083-010
Cable, cooler DC power . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1083-020
Cable, injector power . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1116-010
Column clamp . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-60012S
Drive, injector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-60010
Drive, syringe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60357-60013
Enclosure for the autosampler chassis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60057-10055
Gasket, oven door . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60057-40003
Kit, Autosampler Calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-62001
Kit, adjustable arm lifting stop bracket . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-62028
PCB, Arm Motor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F9045-010
PCB, CPU spare . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-60049
PCB, Interconnect . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F9040-010
PCB, LED . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60057-61000
PCB, Temperature Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F9020-010
Power supply, main . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4004-0062
Power supply, option . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4004-0663
Switch, door . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A3520-020
Wash station . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60057-40012
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Consumables and Service Parts
Accela AS Accessory Kit
Accela AS Accessory Kit
The Accela AS Accessory Kit includes consumables (such as vials, fittings, and test mixtures)
as well as durable accessories (such as microplates, tray adapters, tools, and a power cord). The
Accela AS Accessory Kit, P/N 60357-62001, contains the following items:
Bottle, 16 mL, includes cap for sample prep (4 each) . . . . . . . . . . . . . . . . . . . 00301-07527
Carrier, microwell, short, 0.32 in. height. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-20004
Handle, microwell carrier . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-20003
Cable, shielded, ethernet, CAT-5, 7 ft length. . . . . . . . . . . . . . . . . . . . . . . . . . 70111-63302
Carrier, microtitre, 1.26 in. height. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-20062
Handle, microtitre carrier . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60065-20061
Clamp, waste tube. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00007-07612
Clip, microwell handle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1072-010
Connector, plug, 8-position, 3.8 mm pitch, minicombicon (2 each) . . . . . . . . 00004-02511
Fuse, 5 A, 250 V, 5 × 20 mm, time lag, (4 each) . . . . . . . . . . . . . . . . . . . . . . . . . 5101-1856
Procedure, microplate carrier assembly instructions. . . . . . . . . . . . . . . . . . . . . 60053-97024
Screw, cross-recessed, 100 degree, flat countersunk head, type 1,
6–32 × 0.375 in. length . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2816-4216
Screw, 4–40 × 1/4 in. length, pan head, stainless steel . . . . . . . . . . . . . . . . . . . . . 2851-1044
Screw, pan head, 4–40 × 1/4 in. length, stainless steel,
for tray adapters (5 each) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7111-0404
Test mixture, pkg., 1 mL, 5 count, for performance qualification . . . . . . . . . . . . A4991-010
Test mixture, pkg., 1 mL, 5 count, dilution, for performance qualification . . . . . A5135-010
Tray adapter for 300 μL vial (5 each). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F1124-010
Tray adapter plate for microcentrifuge tubes (5 each) . . . . . . . . . . . . . . . . . . . . . F1008-020
Tubing, 1 L bottle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60053-60041
Tubing, convoluted, for autosampler solvent waste . . . . . . . . . . . . . . . . . . . . . . . F5034-050
Wrench, combination, 1/4 × 5/16 in. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5401-0400
Vial kit, screw top, 2 mL, 100 count, includes caps and septa (2 each) . . . . . . . . A4954-010
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Consumables and Service Parts
Accela System Kit
Accela System Kit
The Accela System Kit, P/N 60057-60060 revision F, contains the cables, tubing, fittings, and
so on used to interconnect the system modules.
5-port Ethernet switch, 5 port, 10/100 Base-T, 100 - 240 V . . . . . . . . . . .00825-01-00024
Cable, Ethernet with two ferrites. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97355-98006
Cable, system interconnect cable, 7-connector . . . . . . . . . . . . . . . . . . . . . . . . 60053-63034
Ferrule, 0.188 in. length, for 1/16 in. OD tubing,
stainless steel (quantity 12) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00101-18187
Ferrule, Valco zero dead volume for 1/16 in. OD tubing,
10 000 psi rating (quantity 12) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00101-18122
Filter cartridge, UHP, 0.5 μm particle size, Teflon . . . . . . . . . . . . . . . . . . . .00109-99-00020
Marker letter A, clip on for 1/8 in. OD tubing . . . . . . . . . . . . . . . . . . . . . . . . . . 608250001
Marker letter B, clip on for 1/8 in. OD tubing . . . . . . . . . . . . . . . . . . . . . . . . . . 608250002
Marker letter C, clip on for 1/8 in. OD tubing . . . . . . . . . . . . . . . . . . . . . . . . . . 608250003
Marker letter D, clip on for 1/8 in. OD tubing . . . . . . . . . . . . . . . . . . . . . . . . . . 608250004
Nut, compression, 0.45 in. length, for 1/16 in. OD tubing,
10-32 thread, stainless steel (quantity 12) . . . . . . . . . . . . . . . . . . . . . . . . . . . 2522-1880
Solvent bottle, 1 liter (quantity 4) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1413-0430
Solvent bottle caps (outer portion) (quantity 4). . . . . . . . . . . . . . . . . . . . . . . . . . A0343-010
Solvent bottle cap adapter (inner portion) (quantity 4. . . . . . . . . . . . . . . . . . . . . A3191-010
Solvent bottle label set. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A4158-020
Solvent bottle holder, solvent platform . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60057-60014
Tubing, stainless steel, 0.005 in. ID × 1/16 in. OD, 10 cm length . . . . . . . .00301-01-00008
Tubing, stainless steel, 0.005 in. ID × 1/16 in. OD, 20 cm length
(to connect the Accela Pump outlet to the inlet
of the Accela Autosampler heat exchanger) . . . . . . . . . . . . . . . . . . . . . .00301-01-00009
Tubing, stainless steel, 0.005 in. ID × 1/16 in. OD, 26 cm length . . . . . . . .00028-01-00035
Tubing, stainless steel, 0.005 in. ID × 1/16 in. OD, 60 cm length . . . . . . . .00028-01-00039
Vial kit, clear vials with 1.8 mL capacity, black screw-on caps,
100 vials and caps per package (quantity 2) . . . . . . . . . . . . . . . . . . . . . . . . . A4954-010
Instrument control software, LC Devices
(for use with the Xcalibur data system and
several other Thermo Scientific mass spectrometry applications) . . . . . . . 60257-62005
Instrument control software,
Thermo pumps add-on for the ChromQuest 5.0 data system . . . . . . . .CHROM-93034
Instrument control software,
Accela detectors add-on for the ChromQuest 5.0 data system. . . . . . . .CHROM-98036
Instrument control software,
Accela Open Autosampler add-on
for the ChromQuest 5.0 data system . . . . . . . . . . . . . . . . . . . . . . . . . .CHROM-98037
Instrument control software, data system independent
for the Accela Open Autosampler
(PAL loader and object manager) . . . . . . . . . . . . . . . . . . . . . . . . . . . . .CHROM-98035
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Firmware Updates
Periodically, upgrades to the Accela Autosampler firmware might become available. In
anticipation of future upgrades, both the Xcalibur and ChromQuest data systems come with
the Accela Firmware Upgrade utility, which you can use to download firmware files to your
Accela instrument modules. Firmware upgrade files have a .bin extension.
For the Xcalibur 2.0.7 or earlier data system and the Thermo Foundation application, the
current firmware files and the Accela Firmware Upgrade utility are automatically installed to
your personal computer during the installation of the data system.
For the ChromQuest data system, you can access the utility by choosing Start > Programs >
Chromatography > Accela Firmware Upgrade Utility.
Contents
• Preparing the Autosampler for a Firmware Download
• Downloading Firmware to the Autosampler
Preparing the Autosampler for a Firmware Download
 To prepare the autosampler for a firmware download
1. Make sure you have the Accela AS firmware files that you intend to download and the
Accela Firmware Upgrade Utility on your computer.
For the Xcalibur 2.0.7 or earlier data system, these files reside in the following folder:
drive:\Xcalibur\system\Accela Firmware
For the Thermo Foundation 1.0 or later application, these files reside in the following
folder (see Figure 86):
drive:\Thermo\Instruments\LC Devices\Accela Firmware
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Firmware Updates
Preparing the Autosampler for a Firmware Download
Figure 86. Accela Firmware folder installed with the Accela devices on the LC Devices CD
2. Make a note of the current settings of the rotary switches on the back panel of the Accela
Autosampler.
At the conclusion of this procedure, you return the switches to these settings.
3. Make sure that your autosampler is connected to the computer through the standard
Ethernet connection.
4. Turn off the power to the Accela Autosampler.
5. Use a small, flat-head screwdriver to set the rotary switches on the back panel of the
autosampler to 00.
6. Turn the autosampler power on.
The Communication LED blinks amber to confirm that the Accela Autosampler is
configured for downloading the firmware.
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Downloading Firmware to the Autosampler
Downloading Firmware to the Autosampler
 To download firmware to the Accela Autosampler
1. Start the Accela Firmware Upgrade Utility by doing one of the following:
• For the Xcalibur data system, browse to the following directory, and then
double-click Accela Firmware Upgrade Utility.exe file.
drive:\Xcalibur\system\Accela Firmware
• For the ChromQuest data system, from the computer desktop, choose Start >
Programs > Chromatography > Accela Firmware Upgrade Utility.
The Accela Firmware Upgrade Utility window opens (see Figure 87).
Figure 87. Accela Firmware Upgrade Utility window, showing the Accela Autosampler
device selected
CAUTION Avoid interrupting the firmware download process. After you click
Connect, do not turn off the power to the Accela Autosampler Plus or close the Accela
Firmware Upgrade Utility window until the download process is finished. If you
terminate the connection during the download process or lose power to the
autosampler, contact your local Thermo Fisher Scientific representative. The firmware
board will need to be reprogrammed.
Thermo Scientific
Accela Autosampler Hardware Manual
133
B
Firmware Updates
Downloading Firmware to the Autosampler
2. In the Accela Firmware Upgrade Utility window, initiate communication between the
utility and the Accela Autosampler:
a. Under Device Type, select the Accela Autosampler option (see Figure 87).
b. Click Connect.
The status display at the bottom of the Accela Firmware Upgrade Utility window
indicates that a connection has been made.
3. Under File Names, select the appropriate firmware files for the Accela Autosampler:
a. For the ROM box, click Browse and select ROM(108).bin.
The file name appears in the ROM box.
b. For the App1 box, click Browse and select Apps1_V2.15.bin.
The file name appears in the App1 box.
c. For the App2 box, click Browse and select Apps2_V2.15.bin.
The file name appears in the App2 box.
IMPORTANT You must select a ROM file, an APP1, and an APP2 file. If you
do not download these files together, your Accela AS will not work properly when
you return it to normal operation.
4. After you select the appropriate firmware files, click Download and wait for the
download to finish.
The utility notifies you when the download is complete.
Note When the Download Completed Successfully message appears, you can safely
turn off the power to the autosampler and close the Accela Firmware Upgrade Utility.
5. Return the autosampler to normal operation after you finish upgrading the firmware as
follows:
a. Close the Accela Firmware Upgrade Utility to terminate the connection with the
Accela Autosampler.
b. Turn off the power to the Accela Autosampler.
IMPORTANT Make sure to turn the Accela Autosampler power off before
adjusting the rotary switches.
c. Set the rotary switches back to their original settings.
IMPORTANT The Accela Autosampler will not connect to the Xcalibur data
system when the rotary switch address is set to 00.
d. Turn on the Accela Autosampler power to resume normal operation.
134
Accela Autosampler Hardware Manual
Thermo Scientific
C
Quick Reference Guides
This appendix contains the following two-page quick reference guides:
• Replacing the Rotor Seal in the Rheodyne Valve
• Replacing the Rotor in the Valco Valve
• Using the Xcalibur Direct Control Commands
• Using the ChromQuest Direct Commands
Replacing the rotor in the Valco injection valve or the rotor seal in a Rheodyne injection valve
requires these tools and materials:
Thermo Scientific
Tools and materials
Part number
Rotor seal for the Rheodyne injection valve
00109-99-00021
Rotor seal for the Valco injection valve
00110-03-00019
9/64 in. L-hex wrench
N/A
#2 Phillips screwdriver
N/A
1/4 in. open-end wrench
N/A
Accela Autosampler Hardware Manual
135
Replacing the Rotor Seal in the Rheodyne Valve
Rheodyne valves shipped with earlier versions of the autosampler.
 To remove the injection valve from the autosampler
To replace the rotor seal, follow these steps:
•
•
•
•
•
1. Disconnect the fingertight fittings from ports 2 and 3.
2. Using a 1/4 in. open-end wrench, loosen and remove the swaged
fittings from ports 5 and 6.
3. Using a #2 Phillips screwdriver, loosen and remove the two
screws that secure the valve to the autosampler, and then pull the
valve out of the autosampler.
Step 1: Removing the Injection Valve
Step 2: Disassembling the Injection Valve
Step 3: Installing a New Rotor Seal
Step 4: Reassembling the Injection Valve
Step 5: Reinstalling the Injection Valve
µL
Stator ring
Ports 2 and 3
fingertight
fittings
20
The rotor seal forms a high-pressure seal between the stator and the
rotor. Replace the rotor seal when leaking occurs between the stator
and the stator ring or when you notice a decrease in injection
precision.
Stator
Step 1: Removing the Injection Valve
Screw
Pressure
adjustment nut
3
4
2
5
1
6
Ports 5 and 6
swaged
fittings
Screw
Step 2: Disassembling the Injection Valve
 To disassemble the injection valve
Log ID
Coupling unit
The lot ID sticker on the stator ring identifies the week and year
(WWYY) of the manufacture date.
Each valve has a pressure adjustment nut that is factory adjusted to
the optimal pressure setting. Depending on the manufacture date,
you can identify the optimal pressure setting as follows:
• For valves manufactured on week 38 of 2009 or later, the pressure
adjustment nut is fixed at the optimal pressure setting.
• For injection valves manufactured from week 37 of 2008 to
week 37 of 2009, the alignment of the factory-applied, red mark
on the valve body and the dimple on the pressure adjustment nut
corresponds to the optimal pressure setting for the valve.
• For injection valves manufactured before week 37 of 2008, the
factory-applied red marks do not correspond to the optimal
pressure setting. You must make your own marks.
Manufacture date
Alignment
markings
Adjustable
pressure nut
Initial release to week 36 of 2008
No
Yes
Week 37 of 2008 to week 37 of 2009
Yes
Yes
Week 38 of 2009 or later
No
No
Copyright© 2009 Thermo Fisher Scientific Inc.
All rights reserved.
1. Determine the manufacture date of the valve.
2. For valves manufactured before week 37 of 2008, make reference
marks on the pressure adjustment nut and the valve body to
identify the current alignment.
After you replace the rotor seal and reassemble the valve, check
the alignment of these marks to ensure that the position of the
pressure adjustment nut is unchanged.
Drive shaft
Coupling unit
Pressure adjustment nut mark
Valve body mark
CAUTION Do not remove the coupling unit from
the drive shaft of the injection valve. If you do
remove the coupling unit, return the valve to
Thermo Fisher Scientific for realignment.
3. Using a 9/64 in. hex wrench, remove the three socket head cap
screws that secure the stator to the valve.
Stator ring
P/N
7715-00
Log ID
sticker
3
4
2
Socket head
cap screws
1
5
Body locating pin
6
Alignment hole in the
valve body
Drive shaft
stop pin
Valve body
3. To mount the stator onto the stator ring, hold the stator with its
polished surface facing the stator ring. Align the alignment hole
in the stator with the stator locating pin on the stator ring. Then
insert the locating pin into the alignment hole.
CAUTION The polished (sealing) surface of the
stator contains six easily damaged ports. Avoid
touching this polished surface as you remove the
stator from the injection valve. Do not place the
polished surface face down on a hard surface.
25 μL
4. Pull the stator off the valve. Place it on its side on a clean surface.
Alignment
hole in the
stator
Socket head
cap screws (3x)
Stator locating
pin
Stator ring
Polished sealing surface
7715-00
5.
6.
7.
8.
P/N
Alignment hole
4. Insert the three socket head cap screws into the stator.
5. Using a 9/64 in. L-hex wrench, tighten each screw a little at a
time, keeping the stator surface parallel to the stator ring surface.
6. If you have an adjustable torque wrench with a 1/4 in. shaft and a
hex driver attachment, evenly torque the three socket head cap
screws to 20 inch-pounds.
7. For valves manufactured before week 38 of 2009, ensure that the
position of the pressure alignment nut is unchanged.
If the polished surface of the stator is scratched, replace the valve.
Pull the stator ring off the valve.
Pull the rotor seal off the three pins on the drive shaft.
Inspect the valve components for contamination. Clean as
necessary.
Step 3: Installing a New Rotor Seal
 To install a new rotor seal
Hold the rotor seal with the engraved flow passages facing away
from the drive shaft and mount the rotor seal onto the drive
shaft’s three alignment pins. The pins align with the rotor seal in
only one way.
Step 5: Reinstalling the Injection Valve
 To reinstall the injection valve
1. Align the holes in the valve plate with the two small holes to the
left and right of the valve receptacle, and then insert the valve
drive shaft into the valve receptacle.
µ
20
Rotor seal with three grooves
L
Alignment pins (for rotor seal)
Valve receptacle
2. Using a #2 Phillips screwdriver, tighten the screws that secure the
valve to the autosampler.
3. Reconnect the transfer tubing to port 2 and the waste tubing to
port 3. Hand-tighten the fittings.
4. Reconnect the heat exchanger outlet to port 5. Hand-tighten the
fitting, and then tighten the fitting an additional 90 degrees
(1/4 turn) with a 1/4 in. open-end wrench.
5. Start the solvent flow from the LC pump, and check for leaks.
Step 4: Reassembling the Injection Valve
 To reassemble the injection valve
1. Ensure that the drive shaft stop pin is positioned within the slot
in the valve body.
2. To mount the stator ring onto the valve body, align the body
locating pin with the alignment hole in the valve body. Then
insert the body locating pin into the alignment hole.
138
Replacing the Rotor in the Valco Valve
4. Using a #2 Phillips screwdriver, remove the two screws that secure
the injection valve to the autosampler.
20
The rotor forms a high-pressure seal with the stator. Replace the rotor
when you notice a leak between either the stator and the rotor or the
adjacent ports of the injection valve, or when you notice a decrease in
injection precision.
µL
Contents
• Step 1: Removing the Injection Valve
2
• Step 2: Disassembling the Injection Valve
1
3
• Step 3: Cleaning the Stator
• Step 4: Installing the Rotor
Screws that secure
the injection valve
6
4
5
• Step 5: Reinstalling the Injection Valve
Replacing the rotor seal requires these tools.
Tool
Use
5. Slide the injection valve out of the autosampler.
1/4 in. open-end wrench
To remove the tubing from injection
valve ports 5 and 6
Step 2: Disassembling the Injection Valve
9/64 in. hex wrench
To disassemble the injection valve
#2 Phillips screwdriver
To disconnect the valve from the
autosampler
IMPORTANT Because component cleanliness affects the useful
life of the injection valve, replace the rotor seal in a clean
environment.
Step 1: Removing the Injection Valve
 To remove the injection valve
 To disassemble the injection valve
1. Using a 9/64 in. L-hex wrench, remove the two socket head cap
screws that secure the stator to the stator ring and the valve body.
CAUTION The polished (sealing) surface of the
stator contains six ports that excess handling can
easily damage. As you remove the stator from the
injection valve, avoid touching this polished surface,
and never place the polished surface face down on a
hard surface.
Driver
1. Disconnect the waste tubing from port 3 of the injection valve.
2. Disconnect the transfer tubing from port 2 of the injection valve.
3. Using a 1/4 in. open-end wrench, disconnect the 1/16 in. ID
high-pressure tubing from ports 5 and 6.
Stator
Socket head
cap screws
20
Waste
line
Rotor
µL
Transfer
tubing
2
Tubing to LC
column
1
3
6
4
5
© 2010 Thermo Fisher Scientific Inc.
All rights reserved.
Return from
heat
exchanger
2. Gently pry the rotor seal away from the driver.
3. Examine the sealing surface of the rotor seal for scratches. If
scratches are visible, replace the rotor seal.
Step 3: Cleaning the Stator
4. Reconnect the solvent lines as follows:
• Hand-tighten the transfer tube
fitting to port 2 and the waste tube
fitting to port 3. Never use a wrench
to tighten these fittings.
 To clean the stator
1. Inspect the stator to determine if it requires cleaning.
2. If the stator is dirty, swab it with HPLC-grade methanol. If more
stringent cleaning is required, use a sonication bath.
3. Inspect the remaining valve components for contamination.
Clean as necessary.
CAUTION Never use a wrench to tighten the
transfer tube fitting. Over-tightening the fitting can
do the following:
IMPORTANT If the stator is scratched, replace it. Scratches can
damage the rotor seal and cause cross-port leaks.
• Restrict the end of the tubing, which leads to
less sample reaching the sample loop and results
in smaller than expected peak areas or blank
injections. A restriction in the end of the tubing
can also cause the syringe to make a grinding
sound during an injection sequence.
Step 4: Installing the Rotor
 To install a new rotor
1. Replace the rotor in the driver, making sure that the rotor sealing
surface with its engraved flow passages is facing out.
• Strip the threads on the fitting, which leads to
solvent leaks.
The pattern is asymmetrical to prevent improper placement.
• Hand-tighten the waste tube fitting to port 3.
• Press the end of the return tubing from the heat exchanger to
the bottom of port 5 as you hand-tighten the high-pressure
fitting. Then use a wrench to tighten the fitting by an
additional 1/4 turn.
Engraved flow passages
• Using high-pressure tubing and two high-pressure fittings,
reconnect the LC column to port 6 of the injection valve.
Note To reconnect tubing with a pre-swaged stainless steel
fitting to a valve port, hand-tighten the fitting as you press
the tubing against the bottom of the port, and then use a
wrench to tighten the fitting by an additional 1/4 to 1/2 turn.
2. Replace the stator.
3. Insert the two socket head screws.
4. Alternating between the screws, slowly tighten the screws until
both are snug. Do not overtighten them.
To connect tubing with a two-piece stainless steel fitting to a
mating port, press the tubing against the bottom of the port
as you hand-tighten the fitting. Then tighten the fitting by an
additional 3/4 turn with a wrench. Once you use the wrench
to tighten the fitting, the ferrule portion of the two-piece
fitting is permanently connected to the tubing.
Step 5: Reinstalling the Injection Valve
 To reinstall the injection valve
1. Align the drive shaft and the two prongs of the coupling unit with
the three holes in the injection valve port located in the lower
portion of the column oven compartment. Then insert the
injection valve into the receptacle.
5. Turn on the solvent flow from the LC pump and check for leaks.
20
µL
Injection valve
receptacle
2. Align the holes in the steel plate of the injection valve with the
small holes next to the injection valve receptacle.
3. Using the two screws that you removed in step 4 on page 139,
secure the injection valve to the autosampler.
140
Using the Xcalibur Direct Control Commands
The On/Off switch is the only manual control provided with the Accela Autosampler. To perform tasks, such as
moving the XYZ arm to the back of the tray compartment, use the direct control commands in the Instrument
Setup window or the tune application for your Thermo Scientific mass spectrometer.
Direct Control
Commands
Command
Description
Position arm to
access tray
Moves the XYZ arm to the back of the tray compartment so that you can remove
trays from or place trays into the tray compartment.
Note. If the tray compartment door is open and you selected the Verify Door Is
Closed check box when you configured the autosampler, the autosampler does not
execute this command until you close the tray compartment door.
Flush syringe
Flushes the needle tubing and the interior of the needle with flush solvent.
Flush syringe at
Fill position
Flushes the needle tubing, the interior of the needle, and the sample loop with flush
solvent.
Wash needle
Washes the exterior of the needle with solvent.
Needle removal
Sets the needle to the needle removal position.
CAUTION. To avoid damaging the needle, execute this command before you
remove the needle.
Set oven temperature
Sets the temperature of the column oven compartment without downloading an
instrument method. The allowable temperature range is 5 to 95 °C.
Important. Avoid setting the temperature above the boiling point of the mobile
phase.
Set tray temperature
Sets the temperature of the tray compartment without downloading an instrument
method. The allowable temperature range is 0 to 60 °C.
Turn off
oven temperature
Turns off the oven temperature control, allowing the temperature of the column oven
compartment to return to ambient.
Turn off
tray temperature
Turns off the tray temperature control, allowing the temperature of the tray
compartment to return to ambient.
Inject sample
Injects a sample. The available parameters are Vial, Volume, and Injection Mode.
The allowable vial or well entries depend on the tray configuration:
• For conventional trays, the allowable entries are A:01 to E:40.
• For 96-well plates, the allowable entries are A:A1 to C:H12.
• For 384-well plates, the allowable entries are A:A1 to C:P24.
Move needle
Moves the XYZ arm to a specific vial or well location.
Set injector position
Switches the position of the injection valve. The injection valve has two positions: fill
and inject.
Set syringe to
removal position
Sets the syringe to its removal position.
Set syringe to
home position
Sets the syringe to its home position.
Set Arm to
Home Position
Moves the XYZ arm to its home position, which is just above the injection port.
Copyright© 2009 Thermo Fisher Scientific Inc.
All rights reserved.
Applying a
Direct
Command
You can access the direct control commands from the Xcalibur data system or the tune application for your mass
spectrometer.
 To open the Inlet Direct Control dialog box from the tune application
1. In the Tune window, choose Setup > Inlet Direct Control. The Inlet Direct Control dialog box appears with
tabbed pages for each configured LC device.
2. Click the Accela AS tab.
The Accela AS page appears.
 To open the Direct Control dialog box from the Xcalibur Roadmap view
1. Click the Instrument Setup icon.
The Instrument Setup window appears. The view bar on the left side of the window displays the configured
instruments.
2. In the view bar, click the Accela AS icon. The Instrument Setup window for the autosampler appears.
3. From the menu bar, choose Accela AS > Direct Control. The Direct Control dialog box appears.
 To apply a direct command
1. In the Commands list, select a command.
If the command requires additional parameters, these parameters appear below the Commands list. Make the
appropriate entries and selections.
2. To execute the command, click Apply.
142
Using the ChromQuest Direct Commands
The On/Off switch is the only manual control provided with the Accela Autosampler. To perform tasks, such as
moving the XYZ arm to the back of the tray compartment, use the direct commands.
Direct
Commands
Command
Description
Position arm to access tray
Moves the XYZ arm to the back of the tray compartment so that you can remove
trays from or place trays into the tray compartment.
Note. If the tray compartment door is open and you selected the Verify Door Is
Closed check box when you configured the autosampler, the autosampler does
not execute this command until you close the tray compartment door.
Flush Syringe
Flushes the needle tubing and the interior of the needle with flush solvent.
Flush With Injection In Fill
Position
Flushes the needle tubing, the interior of the needle, and the sample loop with
flush solvent.
Set Oven Temperature
Sets the temperature of the column oven compartment without downloading an
instrument method. The allowable temperature range is 5 to 95 °C.
Important. Avoid setting the temperature above the boiling point of the mobile
phase.
Set Tray Temperature
Sets the temperature of the tray compartment without downloading an
instrument method. The allowable temperature range is 0 to 60 °C.
Turn Off
Column Oven
Turns off the oven temperature control, allowing the temperature of the column
oven compartment to return to ambient.
Turn Off
Tray Temperature
Turns off the tray temperature control, allowing the temperature of the tray
compartment to return to ambient.
Temperature Control Upload
Determines if the autosampler has the optional temperature control feature.
Wash Needle
Washes the exterior of the needle with solvent.
Move Needle
Moves the XYZ arm to a specific vial or well location.
Set Injector Position
Switches the position of the injection valve. The injection valve has two positions:
fill and inject.
Initialize Hardware
Performs the following sequence of operations:
1. Switches the two-way syringe valve to the flush bottle position.
2. Homes the syringe.
3. Sets the syringe plunger back to the ready position.
4. Switches the two-way syringe valve back to the needle position.
5. Homes the syringe.
After the autosampler initializes, it goes into the Waiting for Download state.
Position Arm to
Replace Needle
Sets the needle to the needle removal position.
Position Syringe to Home
Sets the syringe to its home position.
Position Syringe for Removal
Sets the syringe to its removal position.
Go to Home
Moves the XYZ arm to its home position, which is just above the injection port.
Copyright© 2009 Thermo Fisher Scientific Inc.
All rights reserved.
CAUTION. To avoid damaging the needle, execute this command before you
remove the needle.
Submitting
a Direct
Command
You can access the direct control commands from the Instrument Status window.
 To submit a direct command
1. In the online Instrument window, choose Control > Instrument Status. The Instrument Status window
appears.
2. Click the Accela AS tab. The status page for the autosampler appears.
3. Click Diagnostics. The Diagnostics dialog box appears with the Direct Controls page displayed.
4. In the Direct Commands list, select a command.
If the command requires additional parameters, these parameters appear below the Commands list.
5. Make the appropriate entries and selections.
6. To execute the command, click Submit.
144
I
Index
A
Accela Firmware Upgrade Utility 133
air bubbles
loaded into the sample loop for no waste injections 17
separating the sample from the transfer solvent in
variable volume injections 24
autosampler
calibration 48
diagnostics 103
installation 35
lifting xviii
loading 51
ordering with Accela System Kit 35
startup sequence 47
troubleshooting 112
updating firmware 133
validation information, Xcalibur data system 107
B
back ferrule, high-pressure fitting 44
back panel connections 36
C
cables
Ethernet, connecting 39
system interconnect, connecting 37
calibration, autosampler 48
carrier trays for microwell plates 54
carryover, sample
specification 32
troubleshooting 114
CAUTIONS
Do not overtighten the transfer tube fitting 75
Do not place tall objects in the tray compartment 3
Do not remove the autosampler’s housing 61
Do not remove the injection valve coupling unit 137
Do not touch the polished surface of the stator 66, 84
chemicals, filtering and preparing 51
chromatography, troubleshooting (table) 110
Thermo Scientific
ChromQuest data system
Accela AS Configuration dialog box 124
control 30
direct commands 143
firmware downloads 133
maintenance cycles, recording autosampler 99
color-coded, precut stainless steel tubing 43
column oven
description 15
temperature range 15
compliance
FCC iv
WEEE v
concentric dual syringe assembly. See syringe
connections
back panel 36
injection valve 70
needle tubing 6
transfer tubing 11
wash bottle 41
covers
for 384-well microplates 59
for 96-well microplates 57
types to avoid 59
D
data system control 30
dead volume, transfer tubing 122
device drivers
compatibility 34
ordering 129
diagnostics
description 103
error messages (table) 104
power up 103
Accela Autosampler Hardware Manual
145
Index: E
E
H
electromagnetic compatibility iv
error log
ChromQuest data system 107
error messages (table) 104
Xcalibur data system 105
errors, common operator 120
Ethernet cables 39
Ethernet switch
connecting 39
ordering 129
external wash, description 30
heat exchanger
clearing a plug 96
description 15
heavy object, lifting and carrying xviii
Help, data system xv
high-pressure tubing 43
homing
syringe position 83
XYZ arm position 5, 91
F
fatal error 103
FCC compliance iv
firmware
downloading, hardware connections for 132
files 131
fittings
high pressure 44
injection port 11
needle assembly 6
needle tube assembly 6
sample loop 49
syringe valve 41
transfer tubing 64, 139
wash tubing 41
waste tubing 64, 139
fixed loop injection, description 19
flush bottle. See wash bottle
flush solvent
effect on no waste injections 17
replacing the inlet filter 71
front ferrule, high-pressure fitting 44
front panel LEDs
names of 16
full loop injection, description 19
fuses
replacing 97
schematic 97
G
Gradient Start signal 28
146
Accela Autosampler Hardware Manual
I
Injection Out signal 29
injection valve
description 12
replacing rotor seal (Rheodyne) 137
rotor (Valco), replacing 63
troubleshooting 117
injections
modes 17
precision of 17, 75
sequence of events during 20
volume errors 121
installation
autosampler, positioning 35
changing the sample loop 49
checklist 34
turning on the power 47
unpacking 35
wash bottle, connecting 41
interconnect cable, ordering 129
internal flush, description 30
K
kits
Accela Accessory 128
Accela System 129
L
lead screw, lubricating 75
leaks, injection valve 63, 139
lifting and carrying the autosampler xviii
linear range of no waste injections 17
loading trays 52
Thermo Scientific
Index: M
M
Q
Maintenance page, ChromQuest data system 99
maintenance procedures
flush solvent inlet filter, replacing 71
fuses, replacing 97
heat exchanger, clearing blockage in 96
injection system blockage, troubleshooting 79
lead screw, lubricating 75
plugged needle, clearing 79
rotor (Valco), replacing 63
schedule 62
syringe drive assembly, mounting 77
syringe, changing 93
transfer tube, replacing 71
manuals, Accela xiv
Matrix Technologies, Web site link 59
microplate covers 59
microwell plates
description 3
recommended 57
quick reference guides
replacing rotor seal (Valco) 139
Using the ChromQuest Direct Commands 143
Using the Xcalibur Direct Control Commands 141
N
needle assembly
clearing 79
description 6
needle port
description 11
replacing 71
needle tubing assembly
attachments 35
connecting 42
description 6
no waste injection, description 17
P
partial loop injection, description 18
placeholder module, pump 45
plugged transfer tube, cause of 75
plungers, syringe 9, 93
power cord 39
power indicator 47
precision, injection-to-injection
no waste injection mode 17
partial loop injection mode 18
troubleshooting 118
with an overtightened transfer tube 75
precut stainless steel tubing 43
pressure adjustment nut, injection valve 137
Pump Ready signal 22
PVC cap on the needle tubing assembly 89
Thermo Scientific
R
rear panel connections 36
recommended microplate covers 58–59
removal position
for the needle 53
for the syringe 93
Rheodyne injection valve 137
rotor, Valco
replacing 63
two-sheet replacement guide 139
S
safety precautions
electrical shock 61
solvents 71
sample carryover. See carryover, syringe
sample loop
changing 49
description 15
sample trays
loading 51
loading (caution) 52
signals
gradient start 28
inject out 29
pump ready 22
solvent lines, connecting
high-pressure tubing between the pump and the
autosampler 43
wash bottle to syringe valve 41
stainless steel tubing 43
startup sequence, autosampler 47
static mixer connections, Accela 600 or 1250 Pump 45
stator (injection valve), scratches (caution) 67
syringe
changing 93
homing 95
maximum capacity 19
removal position 93
standard 9
troubleshooting 118
syringe rate recommended for no waste injections 17
Accela Autosampler Hardware Manual
147
Index: T
syringe valve
description 8
testing performance 81
system interconnect cable, description 37
T
temperature caution xix
Thermo Foundation Instrument Configuration 123
transfer tube
constriction of 75
description 11
determining the dead volume of 122
replacing 71
tray temperature control, description 15
trays, loading 51–52
troubleshooting
band broadening 113
baseline drift 112
blank injections 71, 121
blockage in the injection system 79
carryover 114
contact closure 37, 116
dead volume configuration setting 122
Ethernet communication 116
general chromatography (table) 110
injection valve 117
injection volume, lower than expected 120
irregular peak shapes 113
irreproducible peak areas 115
leaks 118
needle, plugged 118
no peaks 114
overview 109
peak heights smaller than expected 113
precision and accuracy, lack of 114, 118
restricted XYZ arm movement 116
sensitivity changes 112
shifting retention times 112
startup 116
syringe 118
vial caps 117
tubing
behind column oven 15
high pressure 43
needle 7
transfer tube 11
wash bottle 41
148
Accela Autosampler Hardware Manual
U
unit ID
description 39
setting for firmware downloads 132
unpacking your autosampler 35
URLs
customer manuals Web site xxii
documentation survey for customer feedback xxii
microplate ordering information 59
V
Valco injection valve 139
Validation dialog box, Xcalibur data system 107
variable loop injection, description 18
vial sensor (note) 53
vials
oversized 53
recommended 57
volume
excess volume drawn for a partial loop injection 18
minimum injection volume for the no waste injection
mode 17
withdrawn for a full loop injection (table) 19
W
wash bottle
allowing to run dry 120
assembly 41
location 5
wash station, description 10
WEEE compliance v
X
Xcalibur data system
control 30
direct control commands 141
firmware downloads 133
LC device drivers 129
maintenance cycles 98
XYZ arm
description 5
homing 91
moving to the needle removal position 85
Thermo Scientific
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