Manuál v angličtině

Manuál v angličtině
Finnigan™
LCQ™ DUO
Getting Started
97033-97032 Revision B
ZoomScan™, WideBand™, LCQ™DUO, and Finnigan™ are trademarks of and Xcalibur® is a registered trademark of Thermo Electron Corporation.
Microsoft® and Windows® are registered trademarks of Microsoft Corporation. PEEK™ is a trademark of Victrex, plc. Kel-F® is a registered
trademark of 3M Company, Inc. Teflon® and Tefzel® are registered trademarks of E.I. du Pont de Nemours & Company. Unimetrics® is a registered
trademark of Unimetrics Company.
Technical information contained in this publication is for reference purposes only and is subject to change
without notice. Every effort has been made to supply complete and accurate information; however,
Thermo Electron Corporation assumes no responsibility and will not be liable for any errors, omissions,
damage, or loss that might result from any use of this manual or the information contained therein (even if this
information is properly followed and problems still arise).
This publication is not part of the Agreement of Sale between Thermo Electron Corporation and the purchaser
of an LC/MS system. In the event of any conflict between the provisions of this document and those contained
in Thermo Electron Corporation’s Terms and Conditions, the provisions of the Terms and Conditions
shall govern.
System Configurations and Specifications supersede all previous information and are subject to change
without notice.
Printing History: Revision A printed in October 2003. Revision B printed in July 2005.
Software Version: Xcalibur 1.4, LCQDUO 1.4
The products of Thermo Electron San Jose are produced under ISO 9001 accredited quality management systems.
Australia: P.O. Box 239 Rydalmere • Unit 14, 38 – 46 South Street • Rydalmere, N.S.W. 2116 • [61] (02) 9898-9000
Austria: Wehlistrasse 27b • A-1200 Wien • [43] (01) 333 50 34-0
Belgium: Technologiestraat 47 • B-1082 Brussels • [32] (02) 482 30 30
Canada: 5716 Coopers Avenue, Unit 1 • Mississauga, Ontario • L4Z 2E8 • [1] (905) 712-2258
France: 16 Avenue du Québec • Silic 765 • Z.A. de Courtaboeuf • F-91963 Les Ulis Cédex • [33] (01) 60 92 48 00
Germany: Im Steingrund 4-6 • D-63303 Dreieich • [49] (06103) 408 0
Italy: Strada Rivoltana • I-20090 Rodano (Milano) • [39] (02) 95059 226
Japan: C-2F • 3-9, Moriya-cho, Kanagawa-ku • Yokohama, Kanagawa • 221-0022 • [81] (45) 453 9100
Japan: Esaka Grand Building • 2-3-1 Esaka-cho, Suita City • Osaka 564-0063 • [81] (06) 6387-6681
Netherlands: Takkebijsters 1 • NL-4817 BL Breda • [31] (076) 5878 722
P.R. China: Room 901, Ping-an Mansion • No. 23, Jin Rong Street • Xi Cheng District • Beijing 100032 • [86] (010) 6621 0839
Spain: Sepulveda 7 A • ES-28108 Alcobendas (Madrid) • [34] (091) 657 4930
Spain: Acer 30 – 32 • Edificio Sertram – Planta 2, Modulo 3 • ES-08038 Barcelona • [34] (093) 223 0918
Sweden: Pyramidbacken 3 • S-141 75 Kungens Kurva (Stockholm) • [46] (08) 556 468 00
United Kingdom: Stafford House • 1 Boundary Park • Boundary Way • Hemel Hempstead • Hertfordshire HP2 7GE • [44] (01442) 233 555
U.S.A.: 355 River Oaks Parkway • San Jose, CA 95134-1991 • [1] (408) 965-6000
Notes: The country code is enclosed in square brackets [ ]. The city code or area code is enclosed in parenthesis ( ). For countries other than the U.S.A.,
when you are dialing from within the specified country, dial the 0 of the city code. For countries other than Italy, when you are dialing from outside the country,
do not dial the 0 of the city code.
Published by Technical Publications, Thermo Electron Corporation, San Jose, California.
Copyright© 2003 Thermo Electron Corporation. All rights reserved. Printed in the United States of America.
For your convenience, you can fill in this form electronically at the website: www.thermo.com/lcms-techpubs.
Customer Registration... Register now and receive all the privileges associated with being a Thermo Electron,
Finnigan product user, including application reports and technical reports.
Name _____________________________________________________________________________________________
Organization _______________________________________________________________________________________
Division/Dept. ______________________________________________________________________________________
Address ___________________________________________________________________________________________
City ____________________________________________ State ________________ Zip Code_____________________
Country ___________________________________________________________________________________________
Telephone _____________________________________________________________ Ext._________________________
Email ________________________________________________________________
System Type___________________________________________________________ Serial # _____________________
Tell us more... Let us know more about how you use this product:
My Organization Is: (Check one only)
‰ Agricultural lab
‰ Chemical Manufacturing lab
‰ Commercial (for profit) lab
‰ Electronic Manufacturing / Test lab
‰ Environmental Testing lab
‰ Government lab
‰ Medical / Diagnostic lab
‰ Pharmaceutical Manufacturing lab
‰ Research and Development lab
‰ Research Institute
‰ University / College lab
‰ Veterinary lab
‰ Other______________________
Job Role: (Check one only)
‰ Administrator
‰ Consultant
‰ Educator / Professor
‰ Executive
‰ Graduate Student / Post Doc
‰ Group Manager
‰ Laboratory Manager
‰ Purchasing Agent
‰ Project Manager
‰ Scientist / Technical Expert
‰ User / Technician
‰ Other __________________
‰
Primary Application: (Check one only)
‰ ADME / TOX
‰ Analytical
‰ Biomarkers
‰ Biomedical
‰ Clinical / Toxicology
‰ Drug Metabolism and Pharmacokinetics
‰ Drug Discovery
‰ Environmental
‰ Pharmaceutical
‰ Proteomics
‰ Research
‰ Structural Elucidation
‰ Other ___________________________
Reader Survey... Help us to improve the quality of our documentation by answering a few questions:
Finnigan LCQ Duo
Getting Started
The manual is well organized.
The manual is clearly written.
The manual contains all of the information I need.
The instructions are easy to follow.
The instructions are complete.
The technical information is easy to understand.
The figures are helpful.
The index and table of contents helped me find information quickly.
Revision B
97033-97032
Strongly
Agree
Agree
Disagree
Strongly
Disagree
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
Additional Comments: (Attach additional sheets if necessary.)
___________________________________________________________________________________
___________________________________________________________________________________
___________________________________________________________________________________
___________________________________________________________________________________
Tear this sheet from the manual, fold it closed, stamp it, and drop it in the mail.
fold
From _______________________________
____________________________________
Place
Stamp
Here
____________________________________
EDITOR, TECHNICAL PUBLICATIONS
THERMO ELECTRON SAN JOSE
355 RIVER OAKS PARKWAY
SAN JOSE, CA 95134-1991
UNITED STATES OF AMERICA
fold
Regulatory Compliance
Thermo Electron San Jose performs complete testing and evaluation of its products to ensure full
compliance with applicable domestic and international regulations. When your system is
delivered to you, it meets all pertinent electromagnetic compatibility (EMC) and safety standards
as follows:
EMC Certification
EN 55011
(1991)
EN 50082-1
(1992)
EN 61000-4-2
(1995)
EN 61000-4-3
(1996)
ENV 50204
(1995)
EN 61000-4-4
(1995)
EN 61000-4-5
(1995)
FCC Class A
EMC issues have been evaluated by EMC TECHNOLOGY SERVICES, A Subsidiary of
UNDERWRITERS LABORATORY, INC (UL)
Safety Compliance
Low Voltage Directive EN 61010-1 1993/A2
Please be aware that any changes that you make to your system may void compliance with one or
more of these EMC and/or safety standards.
Making changes to your system includes replacing a part. Thus, to ensure continued compliance
with EMC and safety standards, replacement parts should be ordered from Thermo Electron or
one of its authorized representatives.
FCC Compliance Statement
Note: This equipment has been tested and found to comply with the limits for a Class A
digital device, pursuant to part 15 of the FCC rules. These limits are designed to provide
reasonable protection against harmful interference when the equipment is operated in a
commercial environment. This equipment generates, uses, and can radiate radio frequency
energy. If it is not installed and used in accordance with the instruction manual, it may cause
harmful interference to radio communications. Operation of this equipment in a residential
area is likely to cause harmful interference. In this case, the user will be required to correct the
interference at his/her own expense.
Notice on Lifting and Handling of
Thermo Electron San Jose Instruments
For your safety, and in compliance with international regulations, the physical handling of this
Thermo Electron San Jose instrument requires a team effort for lifting and/or moving the
instrument. This instrument is too heavy and/or bulky for one person alone to handle safely.
Notice on the Proper Use of
Thermo Electron San Jose Instruments
In compliance with international regulations: If this instrument is used in a manner not
specified by Thermo Electron San Jose, the protection provided by the instrument could be
impaired.
97033/97133/97044/97144/70111
TE SJ - 31 July 2003
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 Electron has contracted with one or more recycling/disposal companies in each
EU Member State, and this product should be disposed of or recycled through them.
Further information on Thermo Electron’s compliance with these Directives, the
recyclers in your country, and information on Thermo Electron products which may
assist the detection of substances subject to the RoHS Directive are available at
www.thermo.com/WEEERoHS.
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 Electron hat Vereinbarungen getroffen mit Verwertungs-/Entsorgungsanlagen in
allen EU-Mitgliederstaaten und dieses Produkt muss durch diese Firmen
wiederverwertet oder entsorgt werden. Mehr Informationen über die Einhaltung dieser
Anweisungen durch Thermo Electron, die Verwerter und Hinweise die Ihnen nützlich
sein können, die Thermo Electron 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 Electron 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 Electron à ces
directives, les recycleurs dans votre pays et les informations sur les produits Thermo
Electron qui peuvent aider la détection des substances sujettes à la directive RoHS sont
disponibles sur www.thermo.com/WEEERoHS.
'JOOJHBO-$2 %60
Contents
________________________________________________________________________
Contents
Read This First...............................................................................................................................v
Changes to the Manual and Online Help ................................................................................................ vi
Abbreviations ......................................................................................................................................... vii
Typographical Conventions .................................................................................................................... xi
Data Input .............................................................................................................................. xi
Notes, Cautions, and CAUTIONS........................................................................................ xii
Topic Headings.................................................................................................................... xiii
Reply Cards ........................................................................................................................................... xiv
Introduction................................................................................................................................ 1-1
1.1
Why Use the LCQDUO MS Detector? ........................................................................................ 1-2
1.2
Which MS Detector Technique—ESI or APCI—Is Better for Analyzing My Samples? ......... 1-3
Using ESI/MS...................................................................................................................... 1-3
Using APCI/MS................................................................................................................... 1-4
1.3
How Can I Introduce My Samples into the MS Detector? ........................................................ 1-6
1.4
What Types of Buffers Should I Use? What Types Should I Avoid? ....................................... 1-8
1.5
How Should I Set Up the MS Detector for Various LC Flow Rates?........................................ 1-9
1.6
What Is Tuning and Calibration of the MS Detector All About? ............................................ 1-11
1.7
What Types of Experiments Can I Perform With the LCQDUO MS Detector?....................... 1-13
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode ........................ 2-1
2.1
Removing the APCI Probe Assembly........................................................................................ 2-2
2.2
Connecting the PEEK Safety Sleeve and Sample Tube to the ESI Probe ................................. 2-4
2.3
Installing the ESI Probe Assembly ............................................................................................ 2-6
2.4
Setting Up the Syringe Pump for Tuning and Calibrating ......................................................... 2-8
2.5
Setting Up the MS Detector for Tuning and Calibrating ......................................................... 2-11
Tuning and Calibrating Automatically in the ESI/MS Mode ................................................ 3-1
3.1
Testing the Operation of the MS Detector in the ESI/MS Mode ............................................... 3-3
5IFSNP
&-&$530/$03103"5*0/
______________________ Finnigan LCQDUO Getting Started _________________________
i
Contents
@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@ 'JOOJHBO-$2 %60
3.2
Tuning the MS Detector Automatically in the ESI/MS Mode .................................................. 3-6
3.3
Saving Your ESI/MS Tune Method......................................................................................... 3-10
3.4
Calibrating the MS Detector Automatically............................................................................ 3-12
3.5
Cleaning the MS Detector after Tuning and Calibrating......................................................... 3-16
Tuning with Your Analyte in LC/ESI/MS Mode..................................................................... 4-1
4.1
Setting Up to Introduce Sample by Syringe Pump into Solvent Flow from an LC ................. 4-3
4.2
Setting Up to Tune the MS Detector with Your Analyte........................................................... 4-7
4.3
Optimizing the MS Detector Tune Automatically with Your Analyte .................................... 4-10
4.4
Saving the ESI/MS Tune Method............................................................................................ 4-12
Acquiring ESI Sample Data Using the Tune Plus Window ................................................... 5-1
5.1
Setting Up to Introduce Sample by Loop Injection into Solvent Flow from an LC................. 5-2
5.2
Acquiring MS Data in the SIM Scan Type................................................................................ 5-5
5.3
Setting Up to Acquire MS/MS Data in the Full Scan Type..................................................... 5-10
Setting Up to Optimize Collision Energy ......................................................................... 5-10
Optimizing the Collision Energy Automatically for an MS/MS Experiment................... 5-13
Setting Up to Acquire Data in the APCI/MS Mode................................................................ 6-1
6.1
Removing the ESI Probe Assembly .......................................................................................... 6-2
6.2
Installing the APCI Probe Assembly......................................................................................... 6-3
6.3
Setting Up the Inlet for Tuning Using High-Flow Infusion ...................................................... 6-5
6.4
Setting Up the MS Detector for APCI/MS Operation............................................................... 6-8
Optimizing the MS Detector with Your Analyte in APCI/MS Mode.................................... 7-1
7.1
Optimizing the Tune of the MS Detector Automatically in APCI/MS Mode........................... 7-2
7.2
Saving the APCI/MS Tune Method........................................................................................... 7-4
7.3
Cleaning the MS Detector after Tuning in APCI Mode ............................................................ 7-6
Acquiring APCI Sample Data Using the Tune Plus Window ................................................ 8-1
8.1
Setting Up to Introduce Sample by Loop Injection into Solvent Flow from an LC................. 8-2
5IFSNP
ii __________________________Finnigan LCQDUO Getting Started_______________________
&-&$530/$03103"5*0/
'JOOJHBO-$2 %60
8.2
Contents
________________________________________________________________________
Acquiring APCI Data in the SIM Scan Mode............................................................................ 8-4
Instrument Configuration ........................................................................................................ A-1
A.1
Configuring Xcalibur Using the Instrument Configuration Dialog Box .................................. A-2
A.2
Using the LCQ Configuration Dialog Box ............................................................................... A-4
Sample Formulations.................................................................................................................B-1
B.1
Caffeine, MRFA, and Ultramark 1621 Stock Solutions............................................................ B-2
Stock Solution: Caffeine.................................................................................................... B-3
Stock Solution: MRFA ...................................................................................................... B-3
Stock Solution: Ultramark 1621 ........................................................................................ B-3
B.2
ESI Calibration Solution: Caffeine, MRFA, Ultramark 1621.................................................. B-4
B.3
Reserpine................................................................................................................................... B-5
Stock Solution: Reserpine ................................................................................................. B-5
ESI / APCI Sample Solution: Reserpine ........................................................................... B-5
5IFSNP
&-&$530/$03103"5*0/
______________________ Finnigan LCQDUO Getting Started ________________________
iii
Read This First
Welcome to the Thermo Electron, Finnigan™ LCQ™DUO MS detector! The
LCQDUO MS detector is a member of the Finnigan family of LC/MS
instruments.
Finnigan LCQDUO Getting Started includes the following chapters:
Chapter 1: Introduction answers typical questions about the LCQDUO MS
detector and lists LC/MS instrument parameters for typical analyses.
Chapter 2: Setting Up for Tuning and Calibrating the MS Detector in
ESI/MS Mode gives instructions for setting up the ESI probe assembly.
Chapter 3: Tuning and Calibrating Automatically in the ESI/MS Mode
provides procedures to tune and calibrate your LCQDUO MS detector using
calibration solution.
Chapter 4: Tuning with Your Analyte in LC/ESI/MS Mode describes how
to tune the LCQDUO MS detector in ESI mode using your compound of
interest.
Chapter 5: Acquiring ESI Sample Data Using the Tune Plus Window
describes a simple procedure for acquiring ESI sample data on your LCQDUO
system.
Chapter 6: Setting Up to Acquire Data in the APCI/MS Mode gives
instructions for setting up the APCI probe assembly.
Chapter 7: Optimizing the MS Detector with Your Analyte in APCI/MS
Mode describes how to optimize the tune in APCI mode using your
compound of interest.
Chapter 8: Acquiring APCI Sample Data Using the Tune Plus Window
describes a simple procedure for acquiring APCI sample data on your
LCQDUO system.
Appendix A: Instrument Configuration describes how to configure your
LCQDUO MS detector with Xcalibur® software.
Appendix B: Sample Formulations gives instructions about preparing
solutions you can use to acquire data with your LCQDUO.
If you want to perform analyses in ESI mode, read Chapters 2, 3, 4, and 5. If
you want to perform analyses in APCI mode, go to Chapters 2, 3, 6, 7, and 8.
To review the data you acquire using the procedures described in this manual,
refer to the manual: Finnigan Xcalibur Getting Productive: Qualitative
Analysis.
5IFSNP
&-&$530/$03103"5*0/
________________________ Finnigan LCQDUO Getting Started ______________________
v
Read This First
Changes to the Manual and Online Help
@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@ 'JOOJHBO-$2 %60
Changes to the Manual and Online Help
To suggest changes to this manual or the online Help, please send your
comments to:
Editor, Technical Publications
Thermo Electron San Jose
355 River Oaks Parkway
San Jose, CA 95134-1991
U.S.A.
You are encouraged to report errors or omissions in the text or index.
Thank you.
5IFSNP
vi _________________________Finnigan LCQDUO Getting Started_______________________
&-&$530/$03103"5*0/
'JOOJHBO-$2 %60
Read This First
_____________________________________________________________ Abbreviations
Abbreviations
The following abbreviations are used in this and other manuals and in the
online Help.
5IFSNP
&-&$530/$03103"5*0/
A
ampere
ac
alternating current
ADC
analog-to-digital converter
AP
acquisition processor
APCI
atmospheric pressure chemical ionization
API
atmospheric pressure ionization
ASCII
American Standard Code for Information
Interchange
b
bit
B
byte (8 b)
baud rate
data transmission speed in events per second
°C
degrees Celsius
CD
compact disc
CD-ROM
compact disc read-only memory
cfm
cubic feet per minute
CI
chemical ionization
CIP
carriage and insurance paid to
cm
centimeter
cm3
cubic centimeter
CPU
central processing unit (of a computer)
CRC
cyclic redundancy check
CRM
consecutive reaction monitoring
<Ctrl>
control key on the terminal keyboard
d
depth
Da
dalton
DAC
digital-to-analog converter
dc
direct current
DDS
direct digital synthesizer
DEP™
direct exposure probe
DS
data system
DSP
digital signal processor
________________________ Finnigan LCQDUO Getting Started _____________________
vii
Read This First
Abbreviations
@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@ 'JOOJHBO-$2 %60
EI
electron ionization
EMBL
European Molecular Biology Laboratory
<Enter>
enter key on the terminal keyboard
ESD
electrostatic discharge
ESI
electrospray ionization
eV
electron volt
f
femto (10-15)
°F
degrees Fahrenheit
.fasta file
extension of a SEQUEST search database file
FOB
free on board
ft
foot
FTP
file transfer protocol
g
gram
G
giga (109)
GC
gas chromatograph; gas chromatography
GC/MS
gas chromatograph / mass spectrometer
GND
electrical ground
GPIB
general-purpose interface bus
GUI
graphical user interface
h
hour
h
height
HPLC
high-performance liquid chromatograph
HV
high voltage
Hz
hertz (cycles per second)
ICIS™
Interactive Chemical Information System
ICL™
Instrument Control Language™
ID
inside diameter
IEC
International Electrotechnical Commission
IEEE
Institute of Electrical and Electronics Engineers
in.
inch
I/O
input/output
k
kilo (103, 1000)
K
kilo (210, 1024)
KEGG
Kyoto Encyclopedia of Genes and Genomes
kg
kilogram
5IFSNP
viii ________________________Finnigan LCQDUO Getting Started_______________________
&-&$530/$03103"5*0/
'JOOJHBO-$2 %60
5IFSNP
&-&$530/$03103"5*0/
Read This First
_____________________________________________________________ Abbreviations
l
length
L
liter
LAN
local area network
lb
pound
LC
liquid chromatograph; liquid chromatography
LC/MS
liquid chromatograph / mass spectrometer
LED
light-emitting diode
µ
micro (10-6)
m
meter
m
milli (10-3)
M
mega (106)
M+
molecular ion
MB
Megabyte (1048576 bytes)
MH+
protonated molecular ion
min
minute
mL
milliliter
mm
millimeter
MS
mass spectrometer; mass spectrometry
MS
MSn power: where n = 1
MS/MS
MSn power: where n = 2
MSn
MSn power: where n = 1 through 10
m/z
mass-to-charge ratio
n
nano (10-9)
NCBI
National Center for Biotechnology Information
(USA)
NIST
National Institute of Standards and Technology
(USA)
OD
outside diameter
Ω
ohm
p
pico (10-12)
Pa
pascal
PCB
printed circuit board
PID
proportional / integral / differential
P/N
part number
P/P
peak-to-peak voltage
________________________ Finnigan LCQDUO Getting Started _____________________
ix
Read This First
Abbreviations
@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@ 'JOOJHBO-$2 %60
ppm
parts per million
psig
pounds per square inch, gauge
RAM
random access memory
RF
radio frequency
RMS
root mean square
ROM
read-only memory
RS-232
industry standard for serial communications
s
second
SIM
selected ion monitoring
solids probe
direct insertion probe
SRM
selected reaction monitoring
SSQ®
single stage quadrupole
TCP/IP
transmission control protocol / Internet protocol
TIC
total ion current
Torr
torr
TSQ®
triple stage quadrupole
u
atomic mass unit
URL
uniform resource locator
V
volt
V ac
volts alternating current
V dc
volts direct current
vol
volume
w
width
W
watt
WWW
World Wide Web
Note. Exponents are written as superscripts. In the corresponding online
Help, exponents are sometimes written with a caret (^) or with e notation
because of design constraints in the online Help. For example:
MSn (in this manual) MS^n (in the online Help)
105 (in this manual)
10^5 (in the online Help)
5IFSNP
x __________________________Finnigan LCQDUO Getting Started_______________________
&-&$530/$03103"5*0/
'JOOJHBO-$2 %60
Read This First
___________________________________________________Typographical Conventions
Typographical Conventions
Typographical conventions have been established for Thermo Electron
San Jose manuals for the following:
•
Data input
•
Boxed information
•
Topic headings
Data Input
Throughout this manual, the following conventions indicate data input and
output via the computer:
5IFSNP
&-&$530/$03103"5*0/
•
Messages displayed on the screen are represented by capitalizing the
initial letter of each word and by italicizing each word.
•
Input that you enter by keyboard is represented in bold face letters.
(Titles of topics, chapters, and manuals also appear in bold face letters.)
•
For brevity, expressions such as “choose File > Directories” are used
rather than “pull down the File menu and choose Directories.”
•
Any command enclosed in angle brackets < > represents a single
keystroke. For example, “press <F1>” means press the key labeled F1.
•
Any command that requires pressing two or more keys simultaneously is
shown with a plus sign connecting the keys. For example, “press
<Shift> + <F1>” means press and hold the <Shift> key and then press the
<F1> key.
•
Any button that you click on the screen is represented in bold face letters
and a different font. For example, “click on Close”.
________________________ Finnigan LCQDUO Getting Started _____________________
xi
Read This First
Typographical Conventions
@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@ 'JOOJHBO-$2 %60
Boxed Information
Information that is important, but not part of the main flow of text, is
displayed in a box such as the one below.
Note. Boxes such as this are used to display information.
Boxed information can be of the following types:
•
Note – information that can affect the quality of your data. In addition,
notes often contain information that you might need if you are having
trouble.
•
Tip – helpful information that can make a task easier.
•
Important – critical information that can affect the quality of your data.
•
Caution – information necessary to protect your instrument from
damage.
•
CAUTION – hazards to human beings. Each CAUTION is accompanied
by a CAUTION symbol. Each hardware manual has a blue CAUTION
sheet that lists the CAUTION symbols and their meanings.
•
DANGER – laser-related hazards to human beings. It includes
information specific to the class of laser involved. Each DANGER is
accompanied by the international laser radiation symbol.
5IFSNP
xii _________________________Finnigan LCQDUO Getting Started_______________________
&-&$530/$03103"5*0/
'JOOJHBO-$2 %60
Read This First
___________________________________________________Typographical Conventions
Topic Headings
The following headings are used to show the organization of topics within a
chapter:
Chapter 1
Chapter Name
1.2 Second Level Topics
Third Level Topics
Fourth Level Topics
Fifth Level Topics
5IFSNP
&-&$530/$03103"5*0/
________________________ Finnigan LCQDUO Getting Started ____________________
xiii
Read This First
Reply Cards
@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@ 'JOOJHBO-$2 %60
Reply Cards
Thermo Electron San Jose manuals contain one or two reply cards. All
manuals contain a Customer Registration / Reader Survey card and some
contain a Change of Location card. These cards are located at the front of each
manual.
The Customer Registration / Reader Survey card has two functions. First,
when you return the card, you are placed on the Thermo Electron San Jose
mailing list. As a member of this list, you receive application reports and
technical reports in your area of interest, and you are notified of events of
interest, such as user meetings. Second, it allows you to tell us what you like
and do not like about the manual.
The Change of Location card allows us to track the whereabouts of the
instrument. Fill out and return the card if you move the instrument to another
site within your company or if you sell the instrument. Occasionally, we need
to notify owners of our products about safety or other issues.
5IFSNP
xiv ________________________Finnigan LCQDUO Getting Started_______________________
&-&$530/$03103"5*0/
Chapter 1
1. Introduction
Welcome to the Thermo Electron, Finnigan™ LCQ™DUO MS detector. The
LCQDUO MS detector is a member of the Finnigan family of LC/MS
instruments.
The LCQDUO MS detector is an advanced analytical instrument that includes
a syringe pump, an optional divert/inject valve, an atmospheric pressure
ionization (API) source, a mass spectrometer (MS) detector, and the
Xcalibur® data system. In a typical analysis, a sample can be introduced in
any of the following ways:
•
Using the syringe pump (direct infusion)
•
Using the inject valve fitted with a loop and an LC (flow injection
analysis)
•
Using a divert valve and HPLC fitted with a column (LC/MS)
In analysis by LC/MS, a sample is injected onto an LC column. The sample
is then separated into its various components. The components elute from
the LC column and pass into the MS detector where they are analyzed.
Analysis by direct infusion or flow injection provides no chromatographic
separation of components in the sample before it passes into the MS
detector. The data from the MS detector are then stored and processed by
the data system.
This introduction answers the following questions:
•
Why use the LCQDUO MS detector?
•
Which MS detector technique—ESI or APCI—is better for analyzing
my samples?
•
How can I introduce my samples into the MS detector?
•
What types of buffers should I use? What types should I avoid?
•
How should I set up the MS detector for various LC flow rates?
•
What is calibrating and tuning the MS detector all about?
•
What types of experiments can I perform with the LCQDUO MS
detector?
____________________ Finnigan LCQDUO Getting Started _______________________
1-1
Introduction
Why Use the LCQDUO MS Detector? _________________________________________
'JOOJHBO-$2 %60
1.1 Why Use the LCQDUO MS
Detector?
The attribute that sets the LCQDUO MS detector apart from other LC
detectors is the high level of analytical specificity that it provides. The
LCQDUO can provide five levels of analysis. Each level of analysis adds a
new dimension of specificity for positive compound identification. The five
levels of analysis are as follows:
•
•
•
•
•
Chromatographic separation and compound detection (retention time)
Mass analysis (molecular weight information)
Two-stage mass analysis (structural information)
WideBand™ Activation (structural information)
ZoomScan™ analysis (higher resolution scan)
Chromatographic separation and compound detection are obtained by all LC
detectors. Retention time alone, however, does not positively identify a
compound because many compounds can have the same retention time
under the same experimental conditions. In addition, even if a compound is
identified correctly by retention time, quantitation results can be in error
because other compounds in the sample might coelute with the compound of
interest.
Mass analysis allows for the identification of analytes of interest.
Atmospheric pressure ionization typically produces mass spectra that
provide molecular weight information, either directly for relatively nonpolar small molecules or after a mathematical manipulation for proteins or
peptides.
Two-stage mass analysis allows for even more positive compound
identification. MS/MS analysis monitors a reaction path: the production of
a specific product ion from a specific parent ion [called selective reaction
monitoring (SRM)]. Using SRM analysis, for example, you can easily
quantitate target analytes in complex matrices such as plant or animal tissue,
plasma, urine, groundwater, and soil. Because of the specificity of MS/MS
measurements and the ability to eliminate interferences by an initial mass
selection stage, quantitative target compound analysis is easily
accomplished using the LCQDUO MS detector.
The WideBand Activation option adds energy, within a range of 20 u,
during MS/MS fragmentation. This is useful when a loss of water or
ammonia is observed. Choose this feature for qualitative MS/MS when you
want enhanced structural information. Signal sensitivity is somewhat
reduced when you choose this option.
ZoomScan analysis provides information about the charge state of one or
more mass ions of interest. ZoomScan data is collected by using slower
scans in a narrow range at higher resolution. This can improve the
resolution of the 12C / 13C isotopes of the analyte ion, which allows for
unambiguous determination of charge state, which in turn allows for the
correct determination of molecular weight.
1-2
_______________________ Finnigan LCQDUO Getting Started ____________________
Introduction
'JOOJHBO-$2 %60 _____Which MS Detector Technique—ESI or APCI—Is Better for Analyzing My Samples?
1.2 Which MS Detector Technique—
ESI or APCI—Is Better for
Analyzing My Samples?
You can operate the MS detector in either of two atmospheric pressure
ionization modes:
•
Electrospray ionization (ESI)
•
Atmospheric pressure chemical ionization (APCI)
Typically, more polar compounds such as amines, peptides, and proteins are
best analyzed by ESI, and non-polar compounds such as steroids are best
analyzed by APCI.
Sample ions can carry a single charge or multiple charges. The number of
charges carried by the sample ions depends on the structure of the analyte of
interest and the mobile phase.
Using ESI/MS
The ESI mode typically produces mass spectra consisting of multiply
charged ions (for proteins and peptides) depending on the structure of the
analyte and the solvent. For example, the resulting mass spectrum of a
higher molecular weight protein or peptide typically consists of a
distribution of multiply charged analyte ions. The resulting mass spectrum
can be mathematically manipulated to determine the molecular weight of the
sample.
The ESI mode transfers ions in solution into the gas phase. Many samples
that previously were not suitable for mass analysis (for example, heat-labile
compounds or high molecular weight compounds) can be analyzed by ESI.
ESI can be used to analyze any polar compound that makes a preformed ion
in solution. The term preformed ion can include adduct ions. For example,
polyethylene glycols can be analyzed from a solution containing ammonium
acetate, because of adduct formation between the NH4+ ions in the solution
and oxygen atoms in the polymer. With ESI, the range of molecular weights
that can be analyzed by the LCQDUO MS detector is greater than 100,000 u,
due to multiple charging. ESI is especially useful for the mass analysis of
polar compounds, which include: biological polymers (for example,
proteins, peptides, glycoproteins, and nucleotides); pharmaceuticals and
their metabolites; and industrial polymers (for example, polyethylene
glycols).
You can use the ESI mode in either positive or negative ion polarity mode.
The ion polarity mode is determined by the polarity of the preformed ions in
solution: Acidic molecules form negative ions in high pH solution, and
basic molecules form positive ions in low pH solution. A positively charged
ESI needle is used to generate positive ions and a negatively charged needle
is used to generate negative ions.
____________________ Finnigan LCQDUO Getting Started _______________________
1-3
Introduction
Which MS Detector Technique—ESI or APCI—Is Better for Analyzing My Samples?_____
'JOOJHBO-$2 %60
You can vary the flow rate from the LC into the MS detector over a range
from 1 /PLQWR L/min. See Table 1-2. (In ESI, the buffer and the
buffer strength both have a noticeable effect on sensitivity. Therefore, it is
important to choose these variables correctly.) In the case of higher
molecular weight proteins or peptides, the resulting mass spectrum consists
typically of a series of peaks corresponding to a distribution of multiply
charged analyte ions.
The ESI process is affected by droplet size, surface charge, liquid surface
tension, solvent volatility, and ion solvation strength. Large droplets with
high surface tension, low volatility, strong ion solvation, low surface charge,
and high conductivity prevent good electrospray.
Organic solvents such as methanol, acetonitrile, and isopropyl alcohol are
superior to water for ESI. Volatile acids and bases are good, but salts above
10 mM and strong acids and bases are extremely detrimental.
The rules for a good electrospray are as follows:
•
Keep salts and non-volatile buffers out of the solvent system. For
example, avoid the use of salts containing sodium or potassium and
avoid the use of phosphates.
•
Use organic/aqueous solvent systems and volatile acids and bases.
•
Optimize the pH of the solvent system for your analyte of interest. For
example, if your analyte of interest is a compound containing carboxylic
acid, your mobile phase should be slightly alkaline (pH 8 or 9) to keep
the acid functional groups in solution as negative ions.
Using APCI/MS
Like ESI, APCI is a soft ionization technique. APCI provides molecular
weight information for compounds of medium polarity that have some
volatility.
APCI is a gas phase ionization technique. Therefore, the gas phase acidities
and basicities of the analyte and solvent vapor play an important role in the
APCI process.
APCI is typically used to analyze small molecules with molecular weight up
to about 2000 u. APCI is a very robust ionization technique. It is not
affected by minor changes in most variables such as changes in buffer or
buffer strength. The rate of solvent flowing from the LC into the MS
detector in APCI mode is typically high (between 0.2 and 2 mL/min). See
Table 1-3.
You can use APCI in positive or negative ion polarity mode. For most
molecules, the positive-ion mode produces a stronger ion current. This is
especially true for molecules with one or more basic nitrogen (or other
basic) atoms. Molecules which generally produce strong negative ions, with
acidic sites such as carboxylic acids and acid alcohols, are an exception to
this general rule.
1-4
_______________________ Finnigan LCQDUO Getting Started ____________________
Introduction
'JOOJHBO-$2 %60 _____Which MS Detector Technique—ESI or APCI—Is Better for Analyzing My Samples?
Although, in general, fewer negative ions are produced than positive ions,
negative ion polarity can be more specific. This is because the negative ion
polarity mode sometimes generates less chemical noise than does the
positive mode. Thus, selectivity might be better in the negative ion mode
than in the positive ion mode.
____________________ Finnigan LCQDUO Getting Started _______________________
1-5
Introduction
How Can I Introduce My Samples into the MS Detector? __________________________
'JOOJHBO-$2 %60
1.3 How Can I Introduce My Samples
into the MS Detector?
You can introduce your samples into the MS detector in a variety of ways.
Refer to Table 1-1.
The syringe pump is often used to introduce calibration solution for
automatic tuning and calibrating in ESI mode. You can also use this
technique to introduce a solution of pure analyte at a steady rate in ESI
mode, for example, for determining the structure of an unknown compound.
You can also use a Tee union to direct samples from the syringe pump into
an LC flow (without a column), which then enters the MS detector. This
technique is used to introduce sample at a steady rate at higher solvent flow
rates for tuning in ESI or APCI on an analyte of interest. You can also use
this technique to introduce a solution of pure analyte at a steady rate in ESI
or APCI.
You can introduce samples from a syringe into the loop of the injector
valve. You can then use the valve to introduce the sample into an LC flow,
which then enters the MS detector. This technique is used in ESI or APCI
to introduce pure analytes into the MS detector in a slug. It is useful when
you have a limited quantity of pure analyte.
You can also use an LC autosampler to introduce samples into an LC flow.
This technique is also used in ESI or APCI to introduce a solution of pure
analyte into the MS detector in a slug.
Finally, you can use an LC autosampler to introduce a mixture onto an LC
column. This technique is used with ESI or APCI to separate the analytes
before they are introduced sequentially into the MS detector.
You can refer to subsequent chapters in this manual and to Finnigan
LCQDUO Getting Connected for plumbing diagrams for methods of sample
introduction.
1-6
_______________________ Finnigan LCQDUO Getting Started ____________________
Introduction
'JOOJHBO-$2 %60 ___________________________How Can I Introduce My Samples into the MS Detector?
Table 1-1. Sample introduction techniques
Syringe Pump Flow
(no LC Flow)
Sample Introduction
Technique
Analytical Technique
Figure Reference
Syringe pump*
ESI automatic tuning
and calibrating
Finnigan LCQDUO
Getting Started
Figure 2-4
ESI analysis of a pure
analyte solution
LC Flow Without
Chromatographic
Separation (no
column)
LC Flow With
Chromatographic
Separation
Syringe pump into LC flow
(connected by Tee
union)*
ESI or APCI automatic
optimization of tuning on
analyte of interest
ESI or APCI analysis of
a pure analyte solution
Finnigan LCQDUO
Getting Started
Figure 4-1 (ESI)
Figure 6-1 (APCI)
Loop injection into LC flow
ESI or APCI analysis of
a pure analyte solution
Finnigan LCQDUO
Getting Started
Figure 5-1 (ESI)
Figure 8-1 (APCI)
Autosampler injection into
LC flow (one or multiple
injections)
ESI or APCI analysis of
a pure analyte solution
Autosampler injections
into LC column via LC
flow (one or multiple
injections)
ESI or APCI analysis of
mixtures
Finnigan LCQDUO
Getting
Connected
Figure 11-5 (ESI)
Figure 11-8 (APCI)
*Provides steady state introduction of sample (direct infusion)
____________________ Finnigan LCQDUO Getting Started _______________________
1-7
Introduction
What Types of Buffers Should I Use? What Types Should I Avoid? __________________
'JOOJHBO-$2 %60
1.4 What Types of Buffers Should I
Use? What Types Should I Avoid?
Many LC applications use nonvolatile buffers such as phosphate and borate
buffers. Use of these nonvolatile buffers with the MS detector, however,
causes the following problems:
•
Blocks the capillary in the probe
•
Causes salt buildup on the spray head and thus compromises the
integrity of the spray
•
Blocks the heated capillary
Therefore, you need to avoid the use of nonvolatile buffers.
Many volatile buffer solutions are available that can be used instead of
nonvolatile ones. Volatile buffer solutions include the following:
•
Trifluoroacetic acid
•
Acetic acid
•
Ammonium acetate
•
Ammonium formate
•
Ammonium hydroxide
Whenever possible, use volatile buffers when you use the MS detector.
1-8
_______________________ Finnigan LCQDUO Getting Started ____________________
Introduction
'JOOJHBO-$2 %60 ___________________How Should I Set Up the MS Detector for Various LC Flow Rates?
1.5 How Should I Set Up the MS
Detector for Various LC Flow
Rates?
The ESI probe can generate ions from liquid flows1 of 1 µL/min to
1.0 mL/min. This flow rate range allows you to use a wide range of
separation techniques, such as, CE, CEC, capillary LC, microbore LC, and
analytical LC.
The APCI probe can generate ions from liquid flows2 of 50 µL/min to
2.0 mL/min. This flow range allows you to use microbore LC, analytical
LC, and semipreparative LC.
As you change the rate of flow of solvents entering the MS detector, you
need to adjust several of the MS detector parameters. Specifically, for ESI
you need to adjust the temperature of the heated capillary and adjust the gas
flow rates for the sheath gas and auxiliary gas. And for APCI, you need to
adjust the heated capillary temperature and vaporizer temperature and adjust
the gas flow rates for the sheath gas and auxiliary gas.
In general, the higher the rate of liquid flowing into the MS detector, the
higher the temperature of the heated capillary (and vaporizer) and the higher
the gas flows.
Note. The syringe pump on the LCQDUO MS detector operates at a single
speed. The the volume of your syringe determines the flow rate of solution
into the MS detector. Specifically, the syringe pump delivers solution into
the MS detector at a rate equal to 1% of the total volume of a syringe per
minute. For example, if you use a 500-µL syringe, solution flows into the
ion source at 5 µL/min.
Table 1-2 provides guidelines for ESI operation for heated capillary
temperatures and gas flow rates for various LC solvent flow rates.
Table 1-3 provides guidelines for APCI operation for the heated capillary
temperature, vaporizer temperature, and gas flow rate for a range of LC
solvent flow rates.
The ESI probe can generate ions from liquid flows of as low as 1 µL/min. However, flows
below 5 µL/min require more care, especially with the position of the fused silica sample
tube within the ESI probe.
1
2
For the APCI probe, flows below 200 µL/min require more care to maintain a stable spray.
____________________ Finnigan LCQDUO Getting Started _______________________
1-9
Introduction
How Should I Set Up the MS Detector for Various LC Flow Rates?___________________
'JOOJHBO-$2 %60
Table 1-2. Guidelines for LC/ESI/MS Operation
LC Flow Rates
Infusion or LC at flow
rates of < 10 µL/min
LC at flow rates from
50 to 100 µL/min
Suggested
Column Size
Capillary
1 mm ID
Heated Capillary
Temperature
Typical setting:
150 to 200 °C
Sheath Gas
Auxiliary Gas
Required
Not required
Typical setting:
20 to 40 units
Typical setting:
0 units
Typical setting:
200 to 225 °C
Required
Not required, but
may help
depending on
conditions
Typical setting:
40 to 60 units
Typical setting:
0 units
LC at flow rates from
200 to 500 µL/min
2 to 3 mm ID
Typical setting:
225 to 250 °C
Required
Typical setting:
50 to 80 units
Usually helps to
reduce solvent
background ions
Typical setting:
20 units
LC at flow rates from
0.5 to 1 mL/min
4.6 mm ID
Typical setting:
250 to 290 °C
Required
Required
Typical setting:
80 to 100 units
Typical setting:
20-40 units
Vaporizer
Temperature
Sheath Gas
Auxiliary Gas
Typical setting:
400 to 500 °C
Required
Usually helps to
reduce solvent
background ions
Table 1-3. Guidelines for LC/APCI/MS Operation
LC Flow Rate
LC at flow rates
from 0.2 to
2 mL/min
Heated
Capillary
Temperature
Typical setting:
150 to 200 °C
Typical setting:
50 to 100 units
Typical setting:
0 to 20 units
1-10 ______________________ Finnigan LCQDUO Getting Started
____________________
Introduction
'JOOJHBO-$2 %60 ______________________ What Is Tuning and Calibration of the MS Detector All About?
1.6 What Is Tuning and Calibration of
the MS Detector All About?
You tune and then calibrate the MS detector in the ESI mode with
calibration solution to ensure its optimum performance.
You can then optimize the tune of the MS detector with your analyte of
interest in either the ESI or APCI mode, if you want to maximize the
detection of one or more particular ions. (It is sometimes possible to acquire
qualitative data without optimizing the parameters, but detection sensitivity
is compromised.)
Note. Before you calibrate, you need to tune to demonstrate that the
transmission of ions into the MS detector is optimum. To perform this
tune, you use the automatic tuning procedure in the Tune Plus window
while infusing a calibration solution into the MS detector at a steady rate of
5 /PLQIRUVHYHUDOPLQXWHV<RXFKRRVHRQHRIWKHLRQVLQWKHFDOLEUDWLRQ
solution that is closest to the mass-to-charge ratio of interest in your
analyte. You observe the Tune Plus window as Xcalibur tunes your
LCQDUO MS detector automatically. You then go on to perform a
calibration.
Calibration parameters are instrument parameters whose values do not vary
with the type of experiment. It is recommended that you calibrate the MS
detector about once every three months and that you check the calibration
about once a week. (To perform a calibration, in almost all cases, the
settings in the existing Calibration file are a sufficient starting place.)
Automatic and semi-automatic calibration (including checking the
calibration) require that you introduce calibration solution into the MS
detector at a steady rate while the procedure is running. You introduce the
solution directly from the syringe pump into the MS detector in the ESI/MS
mode.
Tune parameters are instrument parameters whose values can vary with the
type of experiment. For example, if your experiment requires quantitative
data on one or more particular ions, you need to tune the MS detector with
your analyte if you change any one of the parameters specific to the
experiment or analyte.
Note. The most important parameters that affect the signal quality during
ESI/MS operation are the heated capillary temperature, capillary voltage,
tube lens offset voltage, gases, and solution flow rate. If any one of these
parameters is changed, you need to reoptimize MS detector parameters.
____________________ Finnigan LCQDUO Getting Started ______________________
1-11
Introduction
What Is Tuning and Calibration of the MS Detector All About? ______________________
'JOOJHBO-$2 %60
Automatic and semi-automatic tuning procedures (including optimizing the
collision energy) require that you introduce calibration solution, or a tuning
solution of your analyte of interest, into the MS detector at a steady rate in
either of two ways:
•
Introduce the solution directly from the syringe pump. Refer to the
topic: Setting Up the Syringe Pump for Tuning and Calibrating in
Chapter 2.
•
Introduce the sample from the syringe pump into the effluent of the LC
by using a Tee union. Refer to the topic: Setting Up to Introduce
Sample by Syringe Pump into Solvent Flow from an LC in
Chapter 4.
The first method is good for tuning if you intend to use an experiment type
at a low flow rate involving the syringe pump. The second method is useful
if you intend to use an experiment type at a higher flow rate involving the
LC. However, the second method of introduction puts a comparatively large
amount of analyte into the MS detector. Therefore, before you can perform
an analytical run to analyze for the analyte, you might need to clean the API
spray shield.
Caution. Do not use calibrDWLRQVROXWLRQDWIORZUDWHVDERYH /PLQ
Ultramark 1621 can contaminate your system at high concentrations.
In most cases, you can use the tune you obtain from the automatic or semiautomatic tuning procedures for your analytical experiments. However, for
some applications, you might need to tune several MS detector parameters.
In that case, you would tune manually. With the manual tuning process, you
introduce a tuning solution at a steady rate.
It is best to tune with the MS detector in the same operational mode as that
for the analytical experiment.
Table 1-4 summarizes methods of sample introduction for each of the
calibration and tuning procedures.
Table 1-4. Summary of methods of sample introduction for calibration and tuning
Sample/
Sample Intro
Calibration solution/
Syringe pump
Check
9
Calibrating
Auto
Semiauto
9
Auto
9
Semiauto
Tuning
Manual
Collision
Energy
9
9
9
9
Your tune solution/
Syringe pump
9
9
9
9
Your tune solution/
Syringe pump into LC
flow by using Tee union
9
9
9
9
1-12 ______________________ Finnigan LCQDUO Getting Started
____________________
Introduction
'JOOJHBO-$2 %60 ____________What Types of Experiments Can I Perform With the LCQDUO MS Detector?
1.7 What Types of Experiments Can I
Perform With the LCQDUO MS
Detector?
The LCQDUO MS detector with the standard instrument license is capable of
performing MS and MS/MS experiments, and can analyze ions of m/z 50.00
to 2000.00 (Normal mass range). An optional Advanced Features License
enables the LCQDUO MS detector to perform MSn experiments, and to
analyze ions of m/z 15.00 to 200.00 (Low mass range). For more
information on Low mass range and MSn experiments, refer to the LCQDUO
online Help.
____________________ Finnigan LCQDUO Getting Started ______________________
1-13
Chapter 2
2. Setting Up for Tuning and Calibrating
the MS Detector in ESI/MS Mode
You tune and calibrate your LCQDUO MS detector in ESI mode before you
acquire data in either the ESI or the APCI mode.
This chapter contains the following sections:
•
Removing the APCI Probe Assembly
•
Connecting the PEEK™ Safety Sleeve and Sample Tube to the ESI
Probe
•
Installing the ESI Probe Assembly
•
Setting Up the Syringe Pump for Tuning and Calibrating
•
Setting Up the MS Detector for Tuning and Calibrating
Note. If you have not yet configured the Xcalibur data system for your
LCQDUO MS detector, do so by following the instructions in Appendix A:
Instrument Configuration.
If the ESI probe and the PEEK safety sleeve are installed on the API source,
go to the topic: Setting Up the Syringe Pump for Tuning and
Calibrating.
If no probe is installed, or if the ESI probe is installed without the PEEK
safety sleeve, go to the topic: Connecting the PEEK Safety Sleeve and
Sample Tube to the ESI Probe.
If the APCI probe is installed on the API source, go on to the next topic:
Removing the APCI Probe Assembly.
____________________ Finnigan LCQDUO Getting Started _______________________
2-1
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
Removing the APCI Probe Assembly__________________________________________
'JOOJHBO-$2 %60
2.1 Removing the APCI Probe
Assembly
To remove the APCI probe assembly, proceed as follows. See Figure 2-1
for the location of the components of the APCI probe assembly.
Note. The following procedures assume that you are familiar with your
LCQDUO instrument. If you need assistance, refer to LCQDUO online Help,
Finnigan LCQDUO Getting Connected and/or Finnigan LCQDUO
Hardware Manual.
ON
STANDBY
1. If the MS detector is On, put it in Standby mode, as follows:
Click on the On/Standby button. When the MS detector is in Standby,
the LCQDUO turns Off the sheath gas, auxiliary gas, and high voltage.
APCI
FLANGE
SPRAY
SHIELD
FLANGE
VAPORIZER
RETAINER BOLT
HEATER CABLE
(2 X)
PROBE SLIDE
ADAPTER
FASTENER
(UNDERSIDE)
AUXILIARY GAS
LINE FITTING
(GREEN)
CORONA NEEDLE
HIGH VOLTAGE
CONNECTOR
Figure 2-1. APCI probe assembly
2-2
_______________________ Finnigan LCQDUO Getting Started ____________________
SAMPLE
TRANSFER
LINE FITTING
(RED)
SHEATH GAS
LINE FITTING
(BLUE)
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
'JOOJHBO-$2 %60 __________________________________________Removing the APCI Probe Assembly
2. Loosen the two flange retainer bolts that secure the APCI probe
assembly to the API spray shield.
3. Pull back the APCI probe assembly from the spray shield.
4. Disconnect the corona needle high voltage cable from the corona needle
high voltage connector. To disconnect the cable, turn the locking ring
on the cable counterclockwise until you can pull the cable free.
5. Disconnect the vaporizer heater cable from the connector on the probe,
and then allow it to hang down.
6. Disconnect the sample transfer line from the APCI probe by turning the
sample transfer line fitting counterclockwise until you can pull the
transfer line and fitting free from the probe.
7. Disconnect the sheath gas line from the APCI probe by turning the
sheath gas line fitting counterclockwise until you can pull the sheath gas
line and fitting free from the probe.
8. Disconnect the auxiliary gas line from the APCI probe by turning the
auxiliary gas line fitting counterclockwise until you can pull the
auxiliary gas line and fitting free from the probe.
CAUTION. AVOID BURNS. The APCI vaporizer heater can reach
temperatures of 600 °C. Always allow the APCI probe to cool to
ambient temperature, for approximately 20 min, before handling or
removing the APCI probe from the APCI flange.
9. With one hand holding the APCI flange, loosen the knurled fastener that
secures the APCI flange to the probe slide adapter.
10. Remove the APCI probe assembly from the probe slide adapter by
sliding the probe off the slide adapter. Place the APCI probe assembly
on a lint-free tissue and allow it to cool to ambient temperature
(approximately 20 min).
11. Remove the corona discharge needle from the APCI probe assembly by
pulling it free from the corona discharge needle assembly. Store the
corona discharge needle by inserting it into one of the foam walls of the
APCI probe assembly storage container.
12. Store the APCI probe assembly in its foam storage container. (Make
sure that the APCI probe assembly is at ambient temperature before you
place it in its storage container.)
Go on to the next topic: Connecting the PEEK Safety Sleeve and Sample
Tube to the ESI Probe.
____________________ Finnigan LCQDUO Getting Started _______________________
2-3
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
Connecting the PEEK Safety Sleeve and Sample Tube to the ESI Probe ______________
'JOOJHBO-$2 %60
2.2 Connecting the PEEK Safety
Sleeve and Sample Tube to the ESI
Probe
Before you operate your LCQDUO MS detector, connect the PEEK safety
sleeve and sample tube to the ESI probe.
CAUTION. AVOID ELECTRICAL SHOCK. When you are operating
your instrument in the ESI mode, there are two situations in which you
could receive an electrical shock unless you install the safety kit discussed
below. When you are using the optional Metal Needle Kit
(P/N 70001-62217 or 70005-62013), you might receive an electrical shock
if you touch the fused-silica capillary tube. You could also receive an
electrical shock if the fused-silica capillary tube breaks during ESI
operation, with or without the metal needle installed. Therefore, for your
safety and in compliance with international safety standards, you must
cover the fused-silica capillary tube with the PEEK safety sleeve
(P/N 00301-22806) and associated PEEK ferrules (P/N 00101-18119)
provided in the Safety Sleeve Kit (P/N 70005-62015) before you operate
the instrument. Installation instructions (P/N 70005-97009) are included in
the kit. Operation of the instrument without the safety sleeve impairs the
safety protection provided by the instrument and, thus, could lead to serious
injury.
Connect the PEEK safety sleeve and sample tube to the ESI probe, as
follows. See Figure 2-2.
10-32 x 1/4-28 FITTING
(00101-18080)
FINGERTIGHT FITTING
(00101-18195)
PEEK SAFETY SLEEVE
(00301-22806)
GROUNDED FITTING
(00101-18182)
ESI PROBE
FERRULE
(00101-18119)
FINGERTIGHT FITTING
(00101-18081)
FERRULE
(00101-18119)
SAMPLE TUBE
FUSED-SILICA CAPILLARY
(00106-10499)
Figure 2-2. ESI/MS plumbing connections for the PEEK safety sleeve and sample tube
2-4
_______________________ Finnigan LCQDUO Getting Started ____________________
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
'JOOJHBO-$2 %60 ______________ Connecting the PEEK Safety Sleeve and Sample Tube to the ESI Probe
1. Use a fused-silica cutting tool to cut a 12-in. (30 cm) piece of sample
tube (P/N 00106-10499). Ensure that you squarely cut the ends of the
sample tube.
2. Insert the sample tube through the exit end of the ESI needle and into
the ESI probe.
3. Push the sample tube through the ESI probe until approximately 3.5 cm
(1.5-in.) is left protruding from the exit end of the ESI needle. The
remaining length of sample tube should exit the ESI probe sample inlet.
4. Slide the (clear) 10-32 [ 1/4-28 Kel-F fitting adapter over the sample
tube and tighten the fitting in the ESI probe sample inlet.
5. Slide the 25.4 cm (10.0-in.) precut (brown) PEEK safety sleeve over the
sample tube.
6. Slide a (brown) ferrule (P/N 00101-18119), narrow end first, over the
PEEK safety sleeve and up to the 10-32 [ 1/4-28 Kel-F fitting.
7. Slide a (red) Fingertight fitting onto the PEEK safety sleeve and push it
to the 10-32 [ 1/4-28 Kel-F® fitting. Tighten the Fingertight fitting
slightly, but not completely.
8. Push the PEEK safety sleeve over the sample tube and into the ESI
probe until it stops against the Teflon® needle seal inside the ESI probe.
9. Pull the sample tube (from the ESI needle end) until the sample tube is
flush with the precut square end of the PEEK safety sleeve.
10. Slide a (brown) Fingertight fitting and (brown) ferrule
(P/N 00101-18119), wide end first, over the free end of the PEEK safety
sleeve.
11. Connect the PEEK safety sleeve and ferrule to the grounded fitting by
tightening the Fingertight fitting. Ensure that the Fingertight fitting is
securely tightened around the PEEK safety sleeve, otherwise the sample
stream might enter between the sample tube and the PEEK safety
sleeve.
12. Ensure the sample tube is held tightly in the grounded fitting; check by
pulling the sample tube from the exit end of the ESI needle.
13. Use a fused-silica cutting tool to cut the sample tube at the ESI needle
so that only 2.5 cm (1-in.) remains protruding from the exit end of ESI
needle.
14. From the ESI sample inlet carefully pull the PEEK safety sleeve back
until the exit end of the sample tube is recessed inside the ESI needle
(approximately 1 mm).
15. Tighten the (red) Fingertight fitting to securely hold the PEEK safety
sleeve and sample tube in place in the ESI sample inlet.
Go on to the next topic: Installing the ESI Probe Assembly.
____________________ Finnigan LCQDUO Getting Started _______________________
2-5
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
Installing the ESI Probe Assembly ____________________________________________
'JOOJHBO-$2 %60
2.3 Installing the ESI Probe Assembly
To install the ESI probe assembly, proceed as follows. See Figure 2-3 for
the location of the components of the ESI probe assembly.
1. If your ESI probe assembly does not already contain a sample tube
(fused-silica capillary), you need to follow the procedure for installing a
sample tube that is outlined in the previous topic: Connecting the
PEEK Safety Sleeve and Sample Tube to the ESI Probe.
PROBE
RETAINER
BOLT
PROBE SLIDE
ADAPTER
FLANGE
RETAINER
BOLT (2 X)
AUXILIARY
GAS INLET
(GREEN)
ESI
PROBE
SHEATH
GAS INLET
(BLUE))
GROUNDED
FITTING
HOLDER
SAMPLE
INLET (RED)
Figure 2-3. ESI probe assembly
2-6
_______________________ Finnigan LCQDUO Getting Started ____________________
ESI NEEDLE
HIGH VOLTAGE
CONNECTOR
SHEATH
LIQUID
INLET
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
'JOOJHBO-$2 %60 ____________________________________________ Installing the ESI Probe Assembly
2. With one hand holding the ESI flange, align the ESI flange with the
probe slide adapter, and place it onto the guide rails.
3. Slide the ESI probe assembly onto the probe slide adapter. Secure the
ESI probe assembly to the probe slide adapter with the knurled fastener
that is located on the underside of the probe slide adapter.
4. Remove the Teflon coated septum from the entrance end of the heated
capillary.
5. With one hand, hold the ESI probe on the nozzle side of the probe to
keep it from moving in the flange. With the other hand, connect the
high voltage power cable to the connector labeled HV on the ESI probe.
Turn the locking-ring on the cable clockwise to secure the cable.
6. Push the ESI probe assembly against the spray shield.
7. Secure the ESI flange to the spray shield with the two (black) flange
retainer bolts.
8. Connect the sheath gas line and (blue) fitting to the inlet labeled Sheath
Gas on the ESI probe.
9. Connect the auxiliary gas line and (green) fitting to the inlet labeled
Aux Gas on the ESI probe.
10. Connect the sample transfer line (coming from the syringe) to the
grounded fitting.
11. If you are using sheath liquid, connect the sheath liquid line and fitting
to the inlet labeled Sheath Liquid on the ESI probe. If you are not using
sheath liquid, ensure that the 1/4-28 Tefzel® plug fitting
(P/N 00101-18075) is screwed into the sheath liquid inlet on the ESI
probe.
Go on to the next topic: Setting Up the Syringe Pump for Tuning and
Calibrating.
____________________ Finnigan LCQDUO Getting Started _______________________
2-7
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
Setting Up the Syringe Pump for Tuning and Calibrating ___________________________
'JOOJHBO-$2 %60
2.4 Setting Up the Syringe Pump for
Tuning and Calibrating
The inlet that you use for ESI tuning and calibrating is a syringe infusion
pump. A syringe pump allows you to infuse a sample solution into the ESI
source for extended periods of time.
The syringe pump and syringe are located on the front panel of your
LCQDUO MS detector. The plumbing connections for ESI/MS sample
introduction from the syringe pump are shown in Figure 2-4. To infuse
solution for tuning and calibrating, you install on the pump a 500- /
Unimetrics syringe containing the calibration solution.
Note. The syringe pump on the LCQDUO MS detector operates at a single
speed. The volume of your syringe determines the flow rate of solution
into the MS detector. Specifically, the syringe pump delivers solution into
the MS detector at a rate equal to 1% of the total volume of a syringe per
minute. For example, if you use a 500-µL syringe, solution flows into the
ion source at 5 µL/min.
GROUNDED
FITTING
LC UNION
SYRINGE PUMP
TO WASTE
Figure 2-4. ESI/MS plumbing connections for sample introduction
from the syringe pump
2-8
_______________________ Finnigan LCQDUO Getting Started ____________________
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
'JOOJHBO-$2 %60 ___________________________ Setting Up the Syringe Pump for Tuning and Calibrating
Use the following procedure to set up the syringe pump for infusion.
1. Connect a 4 cm (1.5 in.) segment of Teflon tube (00301-22803) with a
(brown) Fingertight fitting and a (brown) ferrule to the (black) LC
union. See Figure 2-5.
LC UNION
(00101-18202)
FINGERTIGHT FITTING
SYRINGE NEEDLE
(00101-18081)
SYRINGE NEEDLE
FERRULE
(00101-18196)
TEFLON TUBE
(00301-22803)
Figure 2-5. Plumbing connections for the syringe
2. Load a clean, 500- /8QLPHWULFVV\ULQJH31-19012) with
420 /RIWKHFDOLEUDWLRQVROXWLRQ5HIHUWRAppendix B: Sample
Formulations for a procedure for making the calibration solution.)
3. Insert the needle of a syringe into the segment of Teflon tube. Check
that the needle tip of the syringe fits readily into the opening in the free
end of the Teflon tubing. If necessary, you can enlarge the opening in
the end of the tubing slightly.
4. Place the syringe into the syringe holder of the syringe pump.
5. While squeezing the silver release button on the syringe pump handle,
push the handle forward until it just contacts the syringe plunger.
6. Connect a fused-silica infusion line from the LC union to the (stainless
steel) grounded fitting, as follows. See Figure 2-6.
a. Connect the infusion line (P/N 00106-10504) with a (brown)
Fingertight fitting and a (brown) ferrule to the free end of the LC
union.
b. Connect the other end of the infusion line with a (red) Fingertight
fitting and a (brown) ferrule to the grounded fitting.
____________________ Finnigan LCQDUO Getting Started _______________________
2-9
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
Setting Up the Syringe Pump for Tuning and Calibrating ___________________________
GROUNDED FITTING
(00101-18182)
FINGERTIGHT FITTING
(00101-18195)
FINGERTIGHT FITTING
(00101-18081)
INFUSION LINE
FUSED-SILICA CAPILLARY
(00106-10504)
'JOOJHBO-$2 %60
LC UNION
(00101-18202)
FERRULE
(00101-18120)
Figure 2-6. ESI/MS plumbing connections for the fused-silica
infusion line
Caution. Prevent solvent waste from backing up into the API ion source
and MS detector. Always ensure that the PVC drain hose is above the level
of liquid in the waste container.
7. Connect an appropriate length of (clear) 3/8 in. ID PVC hose to the ESI
probe outlet drain. Insert the end of the hose into a suitable container
(such as a gallon jug). Ideally, the container should be vented to a fume
exhaust system.
Go on to the next topic: Setting Up the MS Detector for Tuning and
Calibrating.
2-10 ______________________ Finnigan LCQDUO Getting Started
____________________
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
'JOOJHBO-$2 %60 ____________________________ Setting Up the MS Detector for Tuning and Calibrating
2.5 Setting Up the MS Detector for
Tuning and Calibrating
Use the following procedure to set up the MS detector for tuning and
calibrating in the ESI/MS mode.
You tune first with calibration solution to ensure detection of a sufficient
quantity of ions on which to calibrate. You then calibrate automatically
some of the LCQDUO instrument parameters with calibration solution. The
parameters that are optimized during calibration need to be updated only
occasionally, unless your experiments involve high resolution mass
assignment. Then, you might want to calibrate frequently.
Note. The following procedures assume that you are familiar with your
LCQDUO instrument and the Tune Plus window. If you need assistance,
refer to: LCQDUO online Help, Finnigan LCQDUO Getting Connected,
and/or Finnigan LCQDUO Hardware Manual.
CAUTION. Before you begin normal operation each day, ensure that you
have sufficient nitrogen for your API source. If you run out of nitrogen, the
LCQDUO system automatically turns the MS detector Off to prevent the
possibility of atmospheric oxygen from entering the ion source. The
presence of oxygen in the ion source when the MS detector is ON could be
unsafe. (In addition, if the LCQDUO system turns Off the MS detector
during an analytical run, you could lose data.)
1. Open the Tune Plus window from the Start button on your
Windows XP® Desktop, as follows:
a. Choose Start > All Programs > Xcalibur > Xcalibur. The
Xcalibur Roadmap – Home Page appears.
b. Click on the Instrument Setup button. The Instrument Setup
window appears.
c. Click on the LCQ Duo MS button. The New Method page is
displayed.
d. Click on the Tune Plus button. The Tune Plus window is displayed.
See Figure 2-7.
____________________ Finnigan LCQDUO Getting Started ______________________
2-11
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
Setting Up the MS Detector for Tuning and Calibrating ____________________________
'JOOJHBO-$2 %60
Figure 2-7. Tune Plus window, showing the MS detector in the Standby mode. (This Tune Plus
window shows abbreviated toolbars. Refer to the Note on page 3-4 for more
information.)
STANDBY
ON
2. In Tune Plus, take the MS detector out of Standby mode and turn it On,
as follows:
Click on the On/Standby button on the Control/Scan Mode toolbar. The
MS detector begins scanning, the LCQDUO applies high voltage to the
ESI probe, and the LCQDUO shows a real-time display in the Spectrum
view.
Note. The LCQDUO data system contains customized tune files for
different applications in the directory C:\Xcalibur\methods, including an
ESI low flow LC/MS experiment.
2-12 ______________________ Finnigan LCQDUO Getting Started
____________________
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
'JOOJHBO-$2 %60 ____________________________ Setting Up the MS Detector for Tuning and Calibrating
3. Open the Tune Method file that stores the factory default tune settings
for low-flow ESI operation, as follows:
a. Display the Open dialog box:
Choose File > Open.
b. Select the file AutoTune.LCQTune in the directory
C:\Xcalibur\methods:
Scroll down in the File Name combo box until you see
AutoTune.LCQTune. Then, click on the file name.
c. Open the file, and close the dialog box:
Click on Open. The LCQDUO downloads the Tune Method
parameters to the MS detector.
4. Examine the pre-tune ESI source settings, as follows:
a. If necessary, specify the ESI source:
Choose Setup > Change API Source Type. Then, in the
Change Source Type dialog box, which appears, select the ESI
option button.
b. Return to the Tune Plus window:
Click on OK.
c. Open the ESI Source dialog box from the Instrument Setup toolbar:
Click on the API Source button. See Figure 2-8, and verify that the
settings in your dialog box are the same as those shown in the
figure.
d. Return to the Tune Plus window:
Click on OK.
Figure 2-8. ESI Source dialog box, showing the settings to start a
typical low flow experiment
____________________ Finnigan LCQDUO Getting Started ______________________
2-13
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
Setting Up the MS Detector for Tuning and Calibrating ____________________________
5.
'JOOJHBO-$2 %60
Set the scan parameters for tuning and calibration, as follows:
a. Open the Define Scan dialog box:
Click on the Define Scan button on the Control/Scan Mode toolbar.
See Figure 2-9.
b. Select the MS scan mode in the Scan Description group box:
Click on the Scan Mode: MS option button.
c. Select the Full scan type:
Click on the Scan Type: Full option button.
d. Set the total number of microscans to 3 in the Scan Time group box:
Double-click in the Number of Microscans spin box, then type 3.
e. Set the maximum injection time to 200.00 ms:
Double click in the Maximum Inject Time spin box, then type 200.
f.
Specify that the ion source fragmentation option is turned off:
Confirm that the Turn On check box is not selected ( ).
g. Set the first mass for the scan range to m/z 150 in the Scan Ranges
group box:
Double-click in the First Mass text box, then type 150.
h. Set the last mass for the scan range to m/z 2000:
Double-click in the Last Mass text box, then type 2000.
i.
Ensure that the settings in your Define Scan dialog box are the same
as those shown in Figure 2-9.
j.
Save the MS detector scan parameters, and return to the Tune Plus
window:
Click on OK.
6. Select the profile data type, as follows:
Click on the Centroid/Profile button in the Control/Scan Mode toolbar
to toggle the data type to profile. (The picture on the button should be
the same as that shown here.)
7. Select the positive ion polarity mode, as follows:
Click on the Positive/Negative button in the Control/Scan Mode toolbar
to toggle the ion polarity mode to positive. (The picture on the button
should be the same as that shown here).
2-14 ______________________ Finnigan LCQDUO Getting Started
____________________
Setting Up for Tuning and Calibrating the MS Detector in ESI/MS Mode
'JOOJHBO-$2 %60 ____________________________ Setting Up the MS Detector for Tuning and Calibrating
Figure 2-9. Define Scan dialog box, showing the typical settings for ESI/MS operation
Go on to the chapter: Tuning and Calibrating Automatically in the
ESI/MS Mode.
____________________ Finnigan LCQDUO Getting Started ______________________
2-15
Chapter 3
3. Tuning and Calibrating Automatically
in the ESI/MS Mode
This chapter provides information on how to tune and calibrate the LCQDUO
MS detector in the ESI/MS mode. For most applications, you tune and
calibrate in the ESI mode of operation through automatic procedures.
The automatic tuning and calibrating procedures properly tune and calibrate
the MS detector for ESI operation for the entire mass range of the MS
detector. The procedures use a calibration solution introduced into the MS
detector in low flow mode. (See Table 1-2.)
In general, once you have tuned and calibrated the MS detector, you do not
need to repeat these procedures very often. However, if your particular
analyses require exact molecular weight determination, you will need to
check the MS detector calibration periodically.
To tune and calibrate your MS detector automatically in the ESI/MS mode,
you do the following:
•
Infuse a low concentration calibration solution containing caffeine,
MRFA, and Ultramark 1621 into the ESI source by using the syringe
pump. (Refer to the topic: Setting Up the Syringe Pump for Tuning
and Calibrating.)
•
Test the operation of the ESI source by observing the singly-charged,
positive ions for caffeine, MRFA, and Ultramark 1621 in the Tune Plus
window.
•
Tune the MS detector from the Tune Plus window to ensure efficient
transmission of ions.
•
Calibrate the MS detector.
Note. Before you begin any of the procedures in this chapter, ensure that
you have completed the procedures in the chapter: Setting Up for Tuning
and Calibrating the MS Detector in ESI/MS Mode.
____________________ Finnigan LCQDUO Getting Started _______________________
3-1
Tuning and Calibrating Automatically in the ESI/MS Mode
_______________________________________________________________________
'JOOJHBO-$2 %60
This chapter contains the following sections:
3-2
•
Testing the Operation of the MS Detector in the ESI/MS Mode
•
Tuning the MS Detector Automatically in the ESI/MS Mode
•
Saving Your ESI/MS Tune Method
•
Calibrating the MS Detector Automatically
•
Cleaning the MS Detector after Tuning and Calibrating
_______________________ Finnigan LCQDUO Getting Started ____________________
Tuning and Calibrating Automatically in the ESI/MS Mode
'JOOJHBO-$2 %60 ______________________ Testing the Operation of the MS Detector in the ESI/MS Mode
3.1 Testing the Operation of the MS
Detector in the ESI/MS Mode
You are now ready to test if your MS detector is operating properly. To test
for proper operation, you infuse the calibration solution into the ESI source,
and then you monitor the real-time display of the mass spectrum of
calibration solution.
1. Display the Syringe Pump dialog box, as follows:
Click on the Syringe Pump button. See Figure 3-1.
Figure 3-1. Syringe Pump dialog box
2. Start the syringe pump from the Syringe Pump dialog box, as follows:
a. Select the On option button in the Flow Control group box to turn
on the syringe pump. The Purge checkbox becomes active.
b. Select
the Purge checkbox to deliver solution into the API
source at a rate five times faster than when the checkbox is not
selected. (Use the Purge option to expedite the flow of solution into
the ion source, for example, when you start infusion.)
3. Ensure that the Spectrum view is displayed:
Click on the Display Spectrum View button on the File/Display toolbar.
4. Observe the singly-charged ions of calibration solution in the Spectrum
view of the Tune Plus window. The ions are as follows. See
Figure 3-2.
•
Caffeine: m/z 195
•
MRFA: m/z 524
•
Ultramark 1621: m/z 1022, 1122, 1222, 1322, 1422, 1522, 1622,
1722, 1822
____________________ Finnigan LCQDUO Getting Started _______________________
3-3
Tuning and Calibrating Automatically in the ESI/MS Mode
Testing the Operation of the MS Detector in the ESI/MS Mode ______________________
'JOOJHBO-$2 %60
5. While you observe the Spectrum view, ask yourself the following
questions:
• Is the ion current signal present?
• Is the signal stable, varying by less than about 15% from scan to scan?
6. Deselect
the Purge checkbox when the signal becomes stabilized.
The calibration solution enters the ESI source at 5 µL/min.
7. Click on Close in the Syringe Pump dialog box.
If you answered “yes” to the questions in step 5, your MS detector is
operating properly.
If you answered “no” to either of these questions try the following
troubleshooting measures:
•
Ensure the fused-silica sample tube does not extend beyond the tip of
the ESI needle.
•
Ensure the entrance to the heated capillary is clean, and isn’t covered
with a piece of septum.
•
Ensure that the automatic gain control feature is On, and the Full MS
Target is set to 5e+007 (5 × 107). Verify the setting by choosing
Setup > Injection Control from the Tune Plus window.
Note. You can select to display one of two sets of icons in the following
toolbars of the Tune Plus window: the Instrument Setup toolbar and the
Control/Scan Mode toolbar. You can specify to display either a full or a
partial user interface for the Tune Plus window from the Instrument
Configuration dialog box. Refer to Appendix A: Instrument
Configuration for more information.
3-4
_______________________ Finnigan LCQDUO Getting Started ____________________
Tuning and Calibrating Automatically in the ESI/MS Mode
'JOOJHBO-$2 %60 ______________________ Testing the Operation of the MS Detector in the ESI/MS Mode
Figure 3-2. Spectrum view of the Tune Plus window, showing a real-time display of calibration
solution. (Note that the toolbars are not abbreviated.)
Congratulations! You have demonstrated that your MS detector is operating
properly in the ESI mode. You are now ready to tune and calibrate the MS
detector. Leave your LCQDUO MS detector as it is, and go to the next topic:
Tuning the MS Detector Automatically in the ESI/MS Mode.
____________________ Finnigan LCQDUO Getting Started _______________________
3-5
Tuning and Calibrating Automatically in the ESI/MS Mode
Tuning the MS Detector Automatically in the ESI/MS Mode ________________________
'JOOJHBO-$2 %60
3.2 Tuning the MS Detector
Automatically in the ESI/MS Mode
You tune the MS detector automatically in the ESI/MS mode to optimize
important parameters, including capillary voltage and tube lens offset
voltage.
Use the following procedure to tune the MS detector automatically:
1. Display the automatic tuning page in the Tune dialog box as follows:
Click on the Tune button on the Control/Scan Mode toolbar. Then,
click on the Automatic tab. See Figure 3-3.
Figure 3-3. Tune dialog box, showing the automatic tuning page
3-6
_______________________ Finnigan LCQDUO Getting Started ____________________
Tuning and Calibrating Automatically in the ESI/MS Mode
'JOOJHBO-$2 %60 _________________________Tuning the MS Detector Automatically in the ESI/MS Mode
2. Optimize your tune on the peak at m/z 524.3, as follows:
Double-click on the Mass spin box, then type 524.3.
Note. In this example, you use the mass peak at m/z 524.3 to optimize the
tune. However, you can optimize the tune on any of the mass peaks of the
calibration solution. Choose the mass peak of the calibration solution that
is nearest the mass peak of interest in your analytical sample.
3. Start the automatic tuning procedure, as follows:
a. Click on Start. A message box displays the following message:
Please ensure that the 500 microliter syringe is full.
Ensure the syringe contains at least 420 µL calibration solution.
b. Close the message box, and return to the Tune dialog box:
Click on OK.
4.
Display the Graph view, as follows:
Click on the Graph View button in the File/Display toolbar. See
Figure 3-4.
5.
Observe the Tune Plus window and the Tune dialog box. While
automatic tuning is in progress, the LCQDUO displays various tests in
the Spectrum and Graph views in Tune Plus and displays various
messages in the Status group box in the Tune dialog box. Your Tune
Plus window should now look like the one shown in Figure 3-4.
Note. If your MS detector had failed the automatic calibration procedure,
you can now return to the topic: Calibrating the MS Detector
Automatically, and repeat the procedure. Now that you have tuned the MS
detector, the calibration procedure will probably run successfully.
6.
Examine the ESI source parameters after tuning, as follows:
Click on the Advanced button in the ESI Source dialog box, and
compare the settings shown in Figure 3-5 with the pre-tune settings
shown in Figure 2-8.
____________________ Finnigan LCQDUO Getting Started _______________________
3-7
Tuning and Calibrating Automatically in the ESI/MS Mode
Tuning the MS Detector Automatically in the ESI/MS Mode ________________________
'JOOJHBO-$2 %60
Figure 3-4. Tune Plus window, showing results of a typical automatic tune procedure
3-8
_______________________ Finnigan LCQDUO Getting Started ____________________
Tuning and Calibrating Automatically in the ESI/MS Mode
'JOOJHBO-$2 %60 _________________________Tuning the MS Detector Automatically in the ESI/MS Mode
Figure 3-5.
ESI Source dialog box, showing parameters after
automatic tuning
You have now successfully tuned the MS detector in ESI/MS mode using
the calibration solution. Go on to the next topic: Saving Your ESI/MS
Tune Method.
____________________ Finnigan LCQDUO Getting Started _______________________
3-9
Tuning and Calibrating Automatically in the ESI/MS Mode
Saving Your ESI/MS Tune Method____________________________________________
'JOOJHBO-$2 %60
3.3 Saving Your ESI/MS Tune Method
You can save the parameters you just set in a tune method specific to your
particular analyte and solvent flow rate. (In this case, you save settings
obtained using calibration solution.) You can recall the tune method and use
it as a starting point for optimizing the MS detector on a different analyte of
interest or at a different flow rate.
Note. You must save the Tune Method while the MS detector is On.
Save your ESI/MS tune method (for low-flow operation) when automatic
tuning is complete, as follows:
1. Display the Save As dialog box, as follows:
Choose File > Save As. See Figure 3-6.
Figure 3-6.
Save As dialog box, showing files in the directory
C:\Xcalibur\methods
3-10 ______________________ Finnigan LCQDUO Getting Started
____________________
Tuning and Calibrating Automatically in the ESI/MS Mode
'JOOJHBO-$2 %60 ____________________________________________Saving Your ESI/MS Tune Method
2. Select the C:\Xcalibur\methods directory.
3. Name the Tune Method, as follows:
Click on the File Name text box, and type ESImyTune.
4. Save the Tune Method, close the dialog box, and return to the Tune Plus
window, as follows:
Click on Save. Note that the Tune Method is named
ESImyTune.LCQTune.
After you have tuned the MS detector, you are ready to calibrate. Go on to
the next topic: Calibrating the MS Detector Automatically.
____________________ Finnigan LCQDUO Getting Started ______________________
3-11
Tuning and Calibrating Automatically in the ESI/MS Mode
Calibrating the MS Detector Automatically ______________________________________
'JOOJHBO-$2 %60
3.4 Calibrating the MS Detector
Automatically
Use the following procedure to calibrate the MS detector automatically:
1. Display the automatic calibration page from the Tune Plus menu bar, as
follows:
Choose Control > Calibrate. See Figure 3-7.
Figure 3-7. Calibrate dialog box, showing the Automatic page
3-12 ______________________ Finnigan LCQDUO Getting Started
____________________
Tuning and Calibrating Automatically in the ESI/MS Mode
'JOOJHBO-$2 %60 ______________________________________ Calibrating the MS Detector Automatically
2. Start the automatic calibration procedure, as follows:
a. Click on Start. A message box displays the following message:
Please ensure that the 500 microliter syringe is full.
Ensure the syringe contains at least 420 µL calibration solution.
b. Close the message box, and return to the Calibrate dialog box:
Click on OK.
3. Observe the Tune Plus window and the Calibrate dialog box. While the
automatic calibration is in progress, the LCQDUO displays a variety of
tests in the Spectrum and Graph views and displays a variety of
messages in the Status box of the Calibrate dialog box. See Figure 3-8
for a typical display.
Figure 3-8. Tune Plus window, showing a typical automatic calibration procedure in progress
____________________ Finnigan LCQDUO Getting Started ______________________
3-13
Tuning and Calibrating Automatically in the ESI/MS Mode
Calibrating the MS Detector Automatically ______________________________________
'JOOJHBO-$2 %60
The automatic calibration procedure typically takes about 40 min. See
Figure 3-9.
When the calibration procedure is complete, the LCQDUO restores the full
scan ESI mass spectrum in the Spectrum view, and the Instrument Messages
dialog box is displayed, which indicates whether or not the calibration
procedure was successful.
•
If the calibration procedure was successful, the LCQDUO saves the new
calibration parameters automatically to the hard disk.
•
If the calibration procedure failed, you can try it again after you have
performed the automatic tuning procedure in the topic: Tuning the MS
Detector Automatically in the ESI/MS Mode. In the tuning
procedure, the LCQDUO optimizes several MS detector parameters to
increase the intensity of the signal and improve the overall quality of the
mass spectrum. You can then select the parameters that failed
calibration, and use the semi-automatic calibration procedure for those
specific parameters.
When the calibration procedure is successful, your MS detector is properly
tuned and calibrated for low-flow experiments. In many cases, fine tuning
on your particular analyte is not necessary. You are ready to analyze
samples if the following two conditions are true:
•
You do not need to maximize the intensity of the ion signals for your
analyte.
•
You do not need to change any of the following parameters that affect
signal quality: heated capillary temperature, capillary voltage, tube lens
offset voltage, gases, and solution flow rate.
In Chapter 4: Tuning with Your Analyte in LC/ESI/MS Mode, you
change the solution flow rate and optimize the MS detector parameters on
reserpine, or on your particular analyte.
Before you tune with your analyte, go to the next topic: Cleaning the MS
Detector after Tuning and Calibrating.
3-14 ______________________ Finnigan LCQDUO Getting Started
____________________
Tuning and Calibrating Automatically in the ESI/MS Mode
'JOOJHBO-$2 %60 ______________________________________ Calibrating the MS Detector Automatically
Figure 3-9. Tune Plus window, showing results of a typical automatic calibration procedure
____________________ Finnigan LCQDUO Getting Started ______________________
3-15
Tuning and Calibrating Automatically in the ESI/MS Mode
Cleaning the MS Detector after Tuning and Calibrating ____________________________
'JOOJHBO-$2 %60
3.5 Cleaning the MS Detector after
Tuning and Calibrating
Use the following procedure to clean the MS detector after calibrating. This
procedure describes how to clean your MS detector after using the
calibration solution, in preparation for acquiring data on your analyte of
interest.
ON
1. Put the MS detector in Standby, as follows:
Click on the On/Standby button. When the MS detector is in Standby,
the LCQDUO turns Off the sheath gas, auxiliary gas, ESI high voltage,
and syringe pump. The MS detector stops scanning, and the LCQDUO
freezes the displays for the Spectrum and Graph views.
STANDBY
CAUTION. Always place the MS detector in Standby (or Off) before you
open the API source to atmospheric oxygen. The presence of oxygen in the
ion source when the MS detector is ON could be unsafe. (LCQDUO
automatically turns the MS detector Off when you open the API source,
however, it is best to take this added precaution.)
2. Remove the syringe from the syringe pump holder, as follows:
a. Press the silver button, and pull back on the silver syringe pump
handle to free the syringe.
b. Remove the syringe from the holder.
c. Disconnect the tip of the syringe needle from the Teflon tubing.
3. Clean the syringe thoroughly, as follows:
First, clean the syringe with a solution of 5% acetic acid in water. Next,
rinse the syringe with acetone. Then, rinse the syringe with a solution
of 50:50 methanol:water.
4. Loosen completely the two ESI flange retainer bolts.
CAUTION. AVOID BURNS. At operating temperatures, the heated
capillary can severely burn you! The heated capillary typically operates
between 200 and 300 °C. Always allow the heated capillary to cool to
room temperature, for approximately 20 min, before you touch or
remove this component. Always be careful not to touch the entrance end
of the heated capillary when it is exposed.
3-16 ______________________ Finnigan LCQDUO Getting Started
____________________
Tuning and Calibrating Automatically in the ESI/MS Mode
'JOOJHBO-$2 %60 ____________________________ Cleaning the MS Detector after Tuning and Calibrating
5. Slide the ESI flange out, away from the MS detector chassis, on the
slide rails.
6. Being careful not to touch the heated capillary with your hand, place a
small Teflon coated septum over the entrance end of the heated capillary
to seal the vacuum chamber of the MS detector.
7. Flush the sample transfer line, sample tube, and ESI probe thoroughly
with a solution of 5% acetic acid in water (or with another appropriate
solvent), as follows:
Note. The solvent that you use to flush the sample transfer line, sample
tube, and ESI probe assembly depends on the solvent system you use to
dissolve your samples. For example, if you are using a buffered solution of
a high concentration, an acidic solution is appropriate.
a. Fill a clean, 250 µL Unimetrics syringe with a solution an
appropriate solvent.
b. While holding the plunger of the syringe in place, carefully insert
the needle of the syringe into the free end of the Teflon tube.
c. Flush the sample transfer line, sample tube, and ESI probe with the
solution by slowly depressing the syringe plunger. Visually check
that the solution is exiting the tip of the ESI probe on the inside of
the probe assembly. Use a lint-free tissue to gently remove the
excess solution as it exits the probe.
d. Remove the needle of the syringe from the Teflon tube.
8. Repeat step 7 with acetone.
9. Repeat step 7 with a solution of 50:50 methanol:water.
10. Being careful not to touch the heated capillary with your hand, remove
the septum from the entrance end of the heated capillary.
11. Slide the ESI flange forward along the slide rails and into the ion source
housing.
12. Secure the ESI flange with the two flange retainer bolts.
If you plan to run analytical samples in high-flow ESI mode (using flow
UDWHVEHWZHHQDQG /PLQRSWLPLze the tune further by following
the procedures in the next chapter: Tuning with Your Analyte in
LC/ESI/MS Mode.
____________________ Finnigan LCQDUO Getting Started ______________________
3-17
Chapter 4
4. Tuning with Your Analyte in
LC/ESI/MS Mode
This chapter provides information on tuning the MS detector in the
LC/ESI/MS mode using your analyte. You optimize the sensitivity of your
analyte in the MS detector through an automatic procedure.
The customized Tune Methods contained in your LCQDUO data system are
optimized for a wide range of applications, and they can often be used
without further tuning of your MS detector. However, for certain
applications you might need to tune and optimize several MS detector
parameters.
For instance, the most important parameters that interact with the ESI
interface and signal quality are as follows:
•
Heated capillary temperature
•
Tube lens offset voltage
•
Capillary voltage
•
Sheath gas flow rate
The settings for these parameters depend on the solvent flow rate and target
analyte composition. In general, you should fine tune your MS detector
whenever you change the solvent flow rate conditions of your particular
application. In this procedure, you use the ESI low-flow Tune Method
ESImyTune.LCQTune as a starting point, and then further optimize the MS
detector parameters using an automatic procedure. The automatic procedure
adjusts the tube lens offset voltage, capillary voltage, and voltages applied to
the ion optics until the ion transmission of your analyte is maximized.
The capillary is heated to maximize the ion transmission to the MS detector.
For ESI only, you set the heated capillary temperature proportional to the
infusion flow rate of your solution. See Table 1-2. For this procedure, the
heated capillary temperature is set to 200 °C, and sheath gas is set to 60.
____________________ Finnigan LCQDUO Getting Started _______________________
4-1
Tuning with Your Analyte in LC/ESI/MS Mode
_______________________________________________________________________
'JOOJHBO-$2 %60
Note. If your experiment is performed at a flow rate bHORZ /PLQDQG
the results you want can be obtained without optimizing the MS detector on
your particular analyte, you can go to Chapter 5: Acquiring ESI Sample
Data Using the Tune Plus Window to acquire sample data.
Note. Before you begin any of the procedures in this chapter, ensure that
you have completed the procedures in Chapter 3: Tuning and
Calibrating Automatically in the ESI/MS Mode.
To tune the MS detector in the ESI/MS (high-flow) mode using your
analyte, you do the following:
•
Set up the MS detector for your specific analyte from the Tune Plus
window.
•
Infuse your analyte into the MS detector using a syringe pump
connected to the LC with a Tee union.
•
Optimize the MS detector parameters for your analyte while the solution
flows into the MS detector.
This chapter contains the following sections:
4-2
•
Setting Up to Introduce Sample by Syringe Pump into Solvent Flow
from an LC
•
Setting Up to Tune the MS Detector with Your Analyte
•
Optimizing the MS Detector Tune Automatically with Your Analyte
•
Saving the ESI/MS Tune Method
_______________________ Finnigan LCQDUO Getting Started ____________________
Tuning with Your Analyte in LC/ESI/MS Mode
'JOOJHBO-$2 %60 _________Setting Up to Introduce Sample by Syringe Pump into Solvent Flow from an LC
4.1 Setting Up to Introduce Sample by
Syringe Pump into Solvent Flow
from an LC
Set up to introduce your analyte by syringe pump into solvent flow from an
LC. The plumbing connections for ESI/MS sample introduction from the
syringe pump into solvent flow from an LC are shown in Figure 4-1.
To make the plumbing connections for ESI/MS sample introduction from
the syringe pump into solvent flow from an LC, do the following.
1. Connect a 4 cm (1.5 in.) segment of Teflon tubing with a (brown)
Fingertight fitting and a (brown) ferrule to the (black) LC union. (See
Figure 2-5.)
2. Load a clean, 500- /8QLPHWULFVV\ULQJH31 00301-19012) with
420 /RIWKHQJ /VROXWLRQRIUHVHUSLQHRU\RXUDQDO\WHRILQWHUHVW
(Refer to Appendix B: Sample Formulations for a procedure for
making the reserpine tuning solution.)
LC TEE UNION
FROM LC
LC UNION
SYRINGE PUMP
TO WASTE
Figure 4-1. ESI/MS plumbing connections for sample introduction
from the syringe pump into solvent flow from an LC
____________________ Finnigan LCQDUO Getting Started _______________________
4-3
Tuning with Your Analyte in LC/ESI/MS Mode
Setting Up to Introduce Sample by Syringe Pump into Solvent Flow from an LC ________
'JOOJHBO-$2 %60
3. Insert the needle of a syringe into the segment of Teflon tube. Check
that the needle tip of the syringe fits readily into the opening in the free
end of the Teflon tubing. If necessary, you can enlarge the opening in
the end of the tubing slightly.
4. Place the syringe into the syringe holder of the syringe pump.
5. While pressing the silver release button on the handle, push the handle
forward until it just contacts the syringe plunger.
6. Connect the fused-silica infusion line from the (black) LC union to the
(black) LC Tee union, as follows. See Figure 4-2.
a. Connect the infusion line (P/N 00106-10504) with a (brown)
Fingertight fitting and a (brown) ferrule to the free end of the LC
union.
b. Connect the other end of the infusion line with a (red) Fingertight
fitting and a (brown) ferrule to the side arm of the LC Tee union.
FINGERTIGHT FITTING
(00101-18081)
FERRULE
(00101-18196)
PEEK TUBING
(00301-22912)
TO GROUNDED FITTING
LC TEE UNION
(00101-18204)
PEEK TUBING
FROM DIVER/INJECT VALVE
(00301-22912)
FERRULE
(00101-18120)
FINGERTIGHT FITTING
(00101-18195)
INFUSION LINE
FUSED-SILICA CAPILLARY
(00106-10504)
FROM LC UNION
AND SYRINGE PUMP
Figure 4-2. ESI/MS plumbing connections for the LC Tee union
7. Connect an appropriate length of (red) PEEK tubing from the (stainless
steel) grounded fitting to the (black) LC Tee union, as follows. (See
Figure 4-2.)
a. Use a PEEK tubing cutter to cut a 4 cm (1.5 in.) length of the PEEK
tubing.
b. Connect the PEEK tubing with a (brown) Fingertight fitting and a
(brown) ferrule to the grounded fitting.
c. Connect the other end of the PEEK tubing with a (brown)
Fingertight fitting and a (brown) ferrule to the LC Tee union.
4-4
_______________________ Finnigan LCQDUO Getting Started ____________________
Tuning with Your Analyte in LC/ESI/MS Mode
'JOOJHBO-$2 %60 _________Setting Up to Introduce Sample by Syringe Pump into Solvent Flow from an LC
8. If you have not already done so, connect the PEEK safety sleeve and
fused-silica sample tube from the grounded fitting to the ESI probe
sample inlet following the procedure in the topic: Connecting the
PEEK Safety Sleeve and Fused-Silica Sample Tube to the ESI
Probe.
If you have installed the stainless steel needle in the ESI probe, connect
the PEEK safety sleeve and fused-silica capillary tube from the
grounded fitting to the ESI probe sample inlet following the procedure
in the topic: Installing the Stainless Steel Needle and Connecting the
PEEK Safety Sleeve and Fused-Silica Capillary Tube to the ESI
Probe in LCQDUO Getting Connected.
When the connections between the grounded fitting and the LC Tee
union are complete, they should appear as in Figure 4-3.
PEEK SAFETY SLEEVE
AND SAMPLE TUBE
TO ESI PROBE
SAMPLE INLET
FINGERTIGHT FITTING
(00101-18081)
GROUNDED FITTING
(00101-18182)
LC TEE UNION
(00101-18204)
PEEK TUBING
(00301-22912)
PEEK TUBING
(00301-22912)
FROM DIVERT/INJECT VALVE
FINGERTIGHT FITTING
(00101-18081)
INFUSION LINE
FUSED-SILICA CAPILLARY
(00106-10504)
FROM LC UNION
AND SYRINGE PUMP
Figure 4-3. ESI/MS plumbing connections for the LC Tee union (shown
fully connected)
9. Connect an appropriate length of PEEK tubing (transfer line from the
divert/inject valve) from the divert/inject valve to the free end of the
(black) LC Tee union, as follows. See Figure 4-4.
a. Connect a length of PEEK tubing with a (stainless steel) nut and a
(stainless steel) ferrule to port 3 of the divert/inject valve.
b. Connect the other end of the PEEK tubing with a (brown)
Fingertight fitting and a (brown) ferrule to the free end of the LC
Tee union.
____________________ Finnigan LCQDUO Getting Started _______________________
4-5
Tuning with Your Analyte in LC/ESI/MS Mode
Setting Up to Introduce Sample by Syringe Pump into Solvent Flow from an LC ________
FINGERTIGHT FITTING
(00101-18081)
'JOOJHBO-$2 %60
VALCO NUT
(00101-16008)
TO LC TEE UNION
TO DIVERT/INJECT VALVE
FERRULE
(00101-18196)
PEEK TUBING
(00301-22912)
FERRULE
(00101-18122)
Figure 4-4. Plumbing connections for the the divert/inject valve and
LC Tee union
10. Connect an appropriate length of PEEK tubing (transfer line from the
LC) from the divert/inject valve to the LC, as follows:
a. Connect a length of PEEK tubing with a (stainless steel) nut and a
(stainless steel) ferrule to port 2 of the divert/inject valve.
b. Connect the other end of the PEEK tubing with a proper fitting and
a ferrule to the outlet of the LC.
11. Connect an appropriate length of PEEK tubing (waste line) from the
divert/inject valve to a waste container, as follows:
a. Connect a length of PEEK tubing with a (stainless steel) nut and a
(stainless steel) ferrule to port 1 of the divert/inject valve.
b. Insert the other end of the PEEK tubing in a suitable waste
container.
Caution. Prevent solvent waste from backing up into the API ion source
and MS detector. Always ensure that the PVC drain hose is above the level
of liquid in the waste container.
12. Connect an appropriate length of (clear) 3/8 in. ID PVC hose to the ESI
probe outlet drain. Insert the other end of the hose into a suitable
container (such as a gallon jug). Ideally, the waste container should be
vented to a fume exhaust system.
Go on to the next topic: Setting Up to Tune the MS Detector with Your
Analyte.
4-6
_______________________ Finnigan LCQDUO Getting Started ____________________
Tuning with Your Analyte in LC/ESI/MS Mode
'JOOJHBO-$2 %60 ____________________________ Setting Up to Tune the MS Detector with Your Analyte
4.2 Setting Up to Tune the MS Detector
with Your Analyte
Use the following procedure to set up the MS detector to tune automatically
on your analyte in ESI/MS mode. (In this example, you can use the
reserpine solution described in Appendix B: Sample Formulations, or you
can use a solution of your analyte of interest.)
Caution'RQRWXVHFDOLEUDWLRQVROXWLRQDWIORZUDWHVDERYH /PLQ
Ultramark 1621 can contaminate your system at high concentrations.
Note. The following procedures assume that you are familiar with your
LCQDUO instrument and the Tune Plus window. If you need assistance,
refer to: LCQDUO online Help and/or Finnigan LCQDUO Hardware
Manual.
1. If you have not already done so, open the Tune Plus window from the
Start button on your Windows XP Desktop, as follows:
a. Choose Start > All Programs > Xcalibur > Xcalibur. The
Xcalibur Roadmap – Home Page appears.
b. Click on the Instrument Setup button. The Instrument Setup
window appears.
c. Click on the LCQ Duo MS button. The New Method page is
displayed.
d. Click on the Tune Plus button. The Tune Plus window appears.
STANDBY
ON
2. In Tune Plus, take the MS detector out of Standby mode and turn it On,
as follows:
Click on the On/Standby button in the Control/Scan Mode toolbar. The
MS detector begins scanning, the LCQDUO applies high voltage to the
ESI probe, and the LCQDUO shows a real-time display in the Spectrum
view.
____________________ Finnigan LCQDUO Getting Started _______________________
4-7
Tuning with Your Analyte in LC/ESI/MS Mode
Setting Up to Tune the MS Detector with Your Analyte ____________________________
'JOOJHBO-$2 %60
3. Open the ESImyTune.LCQTune Tune Method, the Tune Method you
saved in Chapter 3, as follows:
a. Display the Open dialog box from the File/Display toolbar:
Click on the Open File icon.
b. Select the file ESImyTune.LCQTune in the directory
C:\Xcalibur\methods:
Scroll down in the File Name combo box until you see
ESImyTune.LCQTune. Then, click on the file name.
c. Open the file, and close the dialog box:
Click on Open. The LCQDUO downloads the Tune Method
parameters to the MS detector, and the title bar in the Tune Plus
window should read as follows:
C:\Xcalibur\methods\ESImyTune.LCQTune – Tune Plus
4. Define the scan parameters for tuning with your analyte in the ESI/MS
mode, as follows:
a. Open the Define Scan dialog box:
Click on the Define Scan button in the Instrument Control toolbar.
See Figure 4-5.
b. Select the MS scan mode in the Scan Description group box:
Click on the Scan Mode: MS option button.
c. Select the SIM scan type:
Click on the Scan Type: SIM option button. Note that the LCQDUO
sets the Number of Scan Ranges to 1.
d. Set the total number of microscans to 3 in the Scan Time group box:
Double-click in the Number of Microscans spin box, then type 3.
e. For this example, leave the maximum injection time set to its
default value of 200.00 ms.
f.
Specify that the ion source fragmentation option is turned off:
Confirm that the Turn On check box is not selected .
g. Set the center mass for the scan range to m/z 609.2 (for reserpine) in
the Scan Ranges group box:
Double-click in the Center Mass text box, then type 609.2.
h. Set the width of the scan range to m/z 2:
Double-click in the Width text box, then type 2.
i.
4-8
Ensure that the settings in your Define Scan dialog box are the same
as those shown in Figure 4-5.
_______________________ Finnigan LCQDUO Getting Started ____________________
Tuning with Your Analyte in LC/ESI/MS Mode
'JOOJHBO-$2 %60 ____________________________ Setting Up to Tune the MS Detector with Your Analyte
Figure 4-5. Define Scan dialog box, showing typical settings for acquiring reserpine data of the
SIM type
j.
Save the MS detector scan parameters, and close the Define Scan
dialog box:
Click on OK.
5. Select the profile data type from the Control/Scan Mode toolbar, as
follows:
Click on the Centroid/Profile button to toggle the data type to profile.
(The picture on the button should be the same as that shown here).
6. Select the positive ion polarity mode, as follows:
Click on the Positive/Negative button in the to toggle the ion polarity
mode to positive. (The picture on the button should be the same as that
shown here).
You have completed setting up to tune your MS detector with your analyte
in ESI/MS mode. Go to the next topic: Optimizing the MS Detector Tune
Automatically with Your Analyte.
____________________ Finnigan LCQDUO Getting Started _______________________
4-9
Tuning with Your Analyte in LC/ESI/MS Mode
Optimizing the MS Detector Tune Automatically with Your Analyte ___________________
'JOOJHBO-$2 %60
4.3 Optimizing the MS Detector Tune
Automatically with Your Analyte
Optimize the MS detector tune automatically to maximize the ion
transmission of reserpine, or your analyte of interest, for a high-flow
experiment. It is recommended that you begin optimizing after you have
successfully passed an automatic tuning procedure and an automatic
calibration procedure with the calibration solution infused at 5 µL/min.
Use the following procedure to tune the MS detector automatically on the
reserpine m/z 609.2 at your particular flow rate, for example, 1 mL/min.
(See Table 1-2 for guidelines about flow rates and temperatures.)
1. Display the automatic tuning page in the Tune dialog box, as follows:
Click on the Tune button on the Control/Scan Mode toolbar. Then,
click on the Automatic tab.
2. Optimize your tune on the peak at m/z 609.2, as follows:
Double-click on the Mass spin box, then type 609.2.
3. Ensure that the Divert/Inject valve is in the Detector position, as
follows:
a. Click on the Divert/Inject button to open the Divert/Inject Valve
dialog box. See Figure 4-6.
b. Select the Detector option button.
c. Click on Close.
Figure 4-6. Divert/Inject Valve dialog box
4. Start the automatic tuning procedure from the Tune dialog box, as
follows:
a. Click on Start. A message box displays the following message:
Please ensure that the 500 microliter syringe is full.
Ensure the syringe pump contains at least 420 µL of the 1 ng/µL
reserpine tuning solution.
b. Close the message box, and return to the Tune Plus window:
Click on OK.
4-10 ______________________ Finnigan LCQDUO Getting Started
____________________
Tuning with Your Analyte in LC/ESI/MS Mode
'JOOJHBO-$2 %60 ___________________ Optimizing the MS Detector Tune Automatically with Your Analyte
5. Display the Graph view, as follows:
Click on the Graph View button in the File/Display toolbar.
6. Observe the Tune Plus window and the Tune dialog box. While
automatic tuning is in progress, the LCQDUO displays various tests in
the Spectrum and Graph views in the Tune Plus window and displays
various messages in the Status group box in the Tune dialog box. Your
Tune Plus window should now look like the one shown in Figure 4-7.
Note. The most important parameters that affect the signal quality during
ESI/MS operation are the heated capillary temperature, capillary voltage,
tube lens offset voltage, gases, and solution flow rate. If any one of these
parameters is changed, you need to reoptimize MS detector parameters.
You can use the Semi-Automatic tune procedure to tune the MS detector on
individual parameters.
You have now successfully tuned the MS detector in ESI/MS mode for the
compound reserpine (or your analyte of interest). Go on to the next topic:
Saving the ESI/MS Tune Method.
Figure 4-7. Tune Plus window with the Tune dialog box, showing the Automatic page
____________________ Finnigan LCQDUO Getting Started ______________________
4-11
Tuning with Your Analyte in LC/ESI/MS Mode
Saving the ESI/MS Tune Method _____________________________________________
'JOOJHBO-$2 %60
4.4 Saving the ESI/MS Tune Method
Save your ESI/MS tune method (for a high-flow experiment using your
analyte) when automatic tuning is complete, as follows:
Note. Save the Tune Method while the MS detector is On, if any of the ion
source parameters are different than those with which you started.
1. Display the Save As dialog box, as follows:
Choose File > Save As. See Figure 4-8.
Figure 4-8.
Save As dialog box, showing files in the directory
C:\Xcalibur\methods
4-12 ______________________ Finnigan LCQDUO Getting Started
____________________
Tuning with Your Analyte in LC/ESI/MS Mode
'JOOJHBO-$2 %60 _____________________________________________ Saving the ESI/MS Tune Method
2. Select the C:\Xcalibur\methods directory.
3. Name the Tune Method for the analyte of interest, as follows:
Click on the File Name text box, and type reserpine.
4. Save the Tune Method, close the dialog box, and return to the Tune Plus
window, as follows:
Click on Save. Note that the Tune Method is named
reserpine.LCQTune.
After you have optimized the tune of the MS detector, you are ready to
perform your ESI/MS experiment in MS scan modes. Go on to the next
chapter: Acquiring ESI Sample Data using the Tune Plus Window.
____________________ Finnigan LCQDUO Getting Started ______________________
4-13
Chapter 5
5. Acquiring ESI Sample Data Using the
Tune Plus Window
This chapter provides information on acquiring LC/ESI/MS sample data
using the Tune Plus window. This experiment uses reserpine but you can
follow the same procedure with your analyte of interest.
Note. The following procedures assume that you are familiar with your
LCQDUO instrument and the Tune Plus window. If you need information,
refer to the LCQDUO online Help, Finnigan LCQDUO Getting Connected,
and/or Finnigan LCQDUO Hardware Manual.
Ensure that you have completed the procedures in the Chapter 3: Tuning
and Calibrating Automatically in the ESI/MS Mode and Chapter 4:
Tuning with Your Analyte in LC/ESI/MS Mode.
This chapter contains the following sections:
•
Setting Up to Introduce Sample by Loop Injection into Solvent Flow
from an LC
•
Acquiring MS Data in the SIM Scan Type
•
Setting Up to Acquire MS/MS Data in the Full Scan Type
____________________ Finnigan LCQDUO Getting Started _______________________
5-1
Acquiring ESI Sample Data Using the Tune Plus Window
Setting Up to Introduce Sample by Loop Injection into Solvent Flow from an LC_________
'JOOJHBO-$2 %60
5.1 Setting Up to Introduce Sample by
Loop Injection into Solvent Flow
from an LC
Set up to introduce sample by loop injection into solvent flow from an LC.
The plumbing connections for ESI/MS sample introduction by loop
injection are shown in Figure 5-1.
To make the plumbing connections for ESI/MS sample introduction by loop
injection into solvent flow from an LC, do the following:
1. If you have not already done so, connect the PEEK safety sleeve and
fused-silica sample tube from the grounded fitting to the ESI probe
sample inlet following the procedure in the topic: Connecting the
PEEK Safety Sleeve and Fused-Silica Sample Tube to the ESI
Probe.
FROM LC
INJECTION PORT
(CENTER, POSITION 5)
TO WASTE
Figure 5-1. ESI/MS plumbing connections for sample introduction by
loop injection into solvent flow from an LC
5-2 _________________
Finnigan LCQDUO Getting Started _______________
Acquiring ESI Sample Data Using the Tune Plus Window
'JOOJHBO-$2 %60 _________Setting Up to Introduce Sample by Loop Injection into Solvent Flow from an LC
If you have installed the stainless steel needle in the ESI probe, connect
the PEEK safety sleeve and fused-silica capillary tube from the
grounded fitting to the ESI probe sample inlet following the procedure
in the topic: Installing the Stainless Steel Needle and Connecting the
PEEK Safety Sleeve and Fused-Silica Capillary Tube to the ESI
Probe in Finnigan LCQDUO Getting Connected.
2. Connect an appropriate length of (red) PEEK tubing (transfer line from
the divert/inject valve) from the divert/inject valve to the (stainless steel)
grounded fitting, as follows. See Figure 5-2.
a. Connect a length of PEEK tubing fitted with a (stainless steel) nut
and a (stainless steel) ferrule to port 3 of the divert/inject valve.
b. Connect the other end of the PEEK tubing with a (brown)
Fingertight fitting and a (brown) ferrule to the free end of the
grounded fitting.
FINGERTIGHT FITTING
(00101-18081)
VALCO NUT
(00101-16008)
TO GROUNDED FITTING
TO DIVERT/INJECT VALVE
FERRULE
(00101-18196)
PEEK TUBING
(00301-22912)
FERRULE
(00101-18122)
Figure 5-2. ESI/MS plumbing connections for the divert/inject valve
and grounded fitting
3. Connect a 5 µL sample loop (P/N 00110-16010) with (stainless steel)
nuts and (stainless steel) ferrules to ports 1 and 4 of the divert/inject
valve.
4. Connect an appropriate length of PEEK tubing (transfer line from the
LC) from the divert/inject valve to the LC, as follows:
a. Connect a length of the PEEK tubing with a (stainless steel) nut and
(stainless steel) ferrule to port 2 of the divert/inject valve.
b. Connect the other end of the PEEK tubing with a proper fitting and
a ferrule to the outlet of the LC.
____________________ Finnigan LCQDUO Getting Started _______________________
5-3
Acquiring ESI Sample Data Using the Tune Plus Window
Setting Up to Introduce Sample by Loop Injection into Solvent Flow from an LC_________
'JOOJHBO-$2 %60
5. Connect an appropriate length of PEEK tubing (waste line) from the
divert/inject valve to a waste container, as follows:
a. Connect a length of PEEK tubing with a (stainless steel) nut and
(stainless steel) ferrule to port 6 of the divert/inject valve.
b. Insert the other end of the PEEK tubing into a suitable waste
container.
Caution. Prevent solvent waste from backing up into the API ion source
and MS detector. Always ensure that the PVC drain hose is above the level
of liquid in the waste container.
6. Connect an appropriate length of (clear) 3/8 in. ID PVC hose to the ESI
probe outlet drain. Insert the end of the hose into a suitable waste
container (such as a gallon jug). Ideally, the container should be vented
to a fume exhaust system.
Go on to the next topic: Acquiring MS Data in the SIM Scan Type.
5-4 _________________
Finnigan LCQDUO Getting Started _______________
Acquiring ESI Sample Data Using the Tune Plus Window
'JOOJHBO-$2 %60 ______________________________________
Acquiring MS Data in the SIM Scan Type
5.2 Acquiring MS Data in the SIM Scan
Type
Use the following procedure to acquire a file of reserpine data in the single
ion monitoring (SIM) type. LCQDUO automatically saves the data you
acquire on your hard disk.
STANDBY
1. If you have not already done so, take the MS detector out of Standby
mode and turn it On from the Tune Plus window, as follows:
Click on the On/Standby button in the Control/Scan Mode toolbar. The
MS detector begins scanning, LCQDUO applies high voltage to the ESI
probe, and a real-time display shows in the Spectrum view.
ON
2. Ensure that the Centroid data type is selected. (The picture on the
button should be the same as that shown here.)
3. Ensure that the scan parameters are defined to acquire SIM data for
reserpine (or your analyte of interest), as follows:
a. Open the Define Scan dialog box:
Click on the Define Scan button. See Figure 5-3.
b. Compare the values in your dialog box to those in Figure 5-3. Then,
click on OK.
Figure 5-3. Define Scan dialog box, showing typical settings for acquiring reserpine data in the
SIM scan type
____________________ Finnigan LCQDUO Getting Started _______________________
5-5
Acquiring ESI Sample Data Using the Tune Plus Window
Acquiring MS Data in the SIM Scan Type ______________________________________
'JOOJHBO-$2 %60
4. Turn on the LC pump, and specify your flow rate of 1 mL/min, for
example. Ensure that your system is free of leaks.
5. Open the Acquire Data dialog box, as follows:
Click on the Acquire Data button on the Control/Scan Mode toolbar.
See Figure 5-4.
Figure 5-4. Acquire Data dialog box, showing the acquisition status of the raw data file
6. Specify the acquisition parameters, as follows:
a. Specify a filename, as follows:
Double-click in the Filename text box, then type
C:\Xcalibur\Data\reserpine1.raw.
b. Specify the sample identity, as follows:
Double-click in the Sample Name text box, then type reserpine. If
you are not using reserpine, type the name of your particular
analyte.
c. Specify a comment about your experiment, as follows. (For
example, describe the scan mode, scan type, ionization mode,
sample amount, or method of sample introduction.)
Double-click in the Comment text box, then type SIM, ESI, 50 pg,
loop. Xcalibur includes the comment on hard copies of your data.
d. Select to acquire data continuously (until you stop the acquisition)
in the Acquire Time group box, as follows:
Select the Continuously option button.
7. Leave the Acquire Data dialog box open during data acquisition, but
move it to a corner of the Tune Plus window.
8. Click on Start in the Acquire Data dialog box to begin acquiring data to
the file reserpine1.raw. The Acquisition Status group box displays the
following message.
State: Acquiring
Time (min):
5-6 _________________
Finnigan LCQDUO Getting Started _______________
Acquiring ESI Sample Data Using the Tune Plus Window
'JOOJHBO-$2 %60 ______________________________________
Acquiring MS Data in the SIM Scan Type
9. Click on the Divert/Inject Valve button to open the Divert/Inject Valve
dialog box. See Figure 5-5.
10. Select the Load option button, and overfill the 5-µL injector loop with
the 10 pg/µL solution of reserpine, or a solution of your analyte of
interest.
Figure 5-5. Divert/Inject Valve dialog box
11. Inject the reserpine solution into the ESI source from the Instrument
Setup toolbar, as follows:
Select the Inject option button. Leave the Divert/Inject Valve dialog
box open.
12. Observe the reserpine peak (m/z 609.2), or that of your analyte of
interest, in the Spectrum view. See Figure 5-6. Wait about 1 min
before you inject again (step 13b, below).
____________________ Finnigan LCQDUO Getting Started _______________________
5-7
Acquiring ESI Sample Data Using the Tune Plus Window
Acquiring MS Data in the SIM Scan Type ______________________________________
'JOOJHBO-$2 %60
Figure 5-6. Tune Plus window, showing the SIM spectrum of reserpine during analysis by loop
injection
13. Perform the following repetitive sequence to obtain a total of four
consecutive loop injections of reserpine.
a. Select the Load option button, and overfill the injector loop with the
10 pg/µL solution of reserpine.
b. Select the Inject option button to inject the reserpine solution into
the ESI source, then observe the Spectrum view.
c. With the Load option selected, overfill the injector loop with the
10 pg/µL solution of reserpine.
d. Again, select the Inject option button, and observe the Spectrum
view.
e. For the fourth time, select the Load option button, then overfill the
injector loop with the 10 pg/µL solution of reserpine.
f.
5-8 _________________
Finally, select the Inject option button to inject the reserpine
solution.
Finnigan LCQDUO Getting Started _______________
Acquiring ESI Sample Data Using the Tune Plus Window
'JOOJHBO-$2 %60 ______________________________________
Acquiring MS Data in the SIM Scan Type
14. Click on the Close button in the Divert/Inject Valve dialog box.
15. Click on Stop in the Acquire Data dialog box to end the data
acquisition. Then, click on Cancel to close the Acquire Data dialog
box.
Review the mass spectrum and chromatogram in the Xcalibur Qual Browser
window. See Figure 5-7.
For more information about reviewing the data you acquired using LCQDUO
with Xcalibur, refer to the manual: Finnigan Xcalibur Getting
Productive: Qualitative Analysis.
Go on to the next topic: Setting Up to Acquire MS/MS Data in the Full
Scan Type.
Figure 5-7. Qual Browser window, showing loop injections of reserpine in the Chromatogram
view (left) and m/z 609 in the Spectrum view.
____________________ Finnigan LCQDUO Getting Started _______________________
5-9
Acquiring ESI Sample Data Using the Tune Plus Window
Setting Up to Acquire MS/MS Data in the Full Scan Type __________________________
'JOOJHBO-$2 %60
5.3 Setting Up to Acquire MS/MS Data
in the Full Scan Type
Prepare to acquire MS/MS data in the Full scan type on reserpine, or on
your analyte of interest. You need to optimize the relative collision energy
before you acquire MS/MS data. The relative collision energy for a
particular analysis depends on the type of sample you are analyzing.
The information in this topic applies to operation of the LCQDUO MS
detector in either the ESI or the APCI mode.
This topic contains the following topics:
•
Setting Up to Optimize Collision Energy
•
Optimizing the Collision Energy Automatically for an MS/MS
Experiment
Setting Up to Optimize Collision Energy
Set up to optimize the collision energy, as follows:
1. Configure the inlet with a Tee union, as described in the topic: Setting
Up to Introduce Sample by Syringe Pump into Solvent Flow from
an LC in Chapter 4.
STANDBY
2. If you have not already done so, take the MS detector out of Standby
mode and turn it On from the Tune Plus window, as follows:
Click on the On/Standby button in the Control/Scan Mode toolbar. The
MS detector begins scanning, the LCQDUO applies high voltage to the
ESI probe, and a real-time display shows in the Spectrum view.
ON
3. Ensure that the Centroid data type is selected. (The picture on the
button should be the same as that shown here.)
4. Ensure that the scan parameters are defined to acquire MS/MS Full scan
data for reserpine (or your analyte of interest), as follows:
a. Open the Define Scan dialog box:
Click on the Define Scan button. See Figure 5-8.
b. Ensure that the values in your dialog box are the same as those
shown in Figure 5-8. Then, click on OK.
5-10 ________________
Finnigan LCQDUO Getting Started _______________
Acquiring ESI Sample Data Using the Tune Plus Window
'JOOJHBO-$2 %60 __________________________ Setting Up to Acquire MS/MS Data in the Full Scan Type
Figure 5-8. Define Scan dialog box, showing typical settings for acquiring MS/MS data in the Full
scan type on reserpine. Note that the WideBand Activation checkbox is active when
the MS/MS option is selected.
5. Turn on your LC pump, and specify a flow rate of 1 mL/min, for
example. Ensure that your system is free of leaks.
6. 7XUQRQWKHV\ULQJHSXPSWRLQIXVHWKHQJ /UHVHUSLQHVROXWLRQDV
follows:
Click on the Syringe Pump icon. Then, select the On option button.
Move the Syringe Pump dialog box to the top of the monitor screen.
7. Display the Tune dialog box – Collision Energy page, as follows:
First, click on the Tune button. Then, click on the Collision Energy tab.
See Figure 5-9.
____________________ Finnigan LCQDUO Getting Started ______________________
5-11
Acquiring ESI Sample Data Using the Tune Plus Window
Setting Up to Acquire MS/MS Data in the Full Scan Type __________________________
'JOOJHBO-$2 %60
Figure 5-9. Tune dialog box, showing the Collision Energy page
8. Specify the product ion for the optimization procedure as the peak at
m/z 397.2, as follows:
Click on the Product Ion Mass option button. Then, double-click on the
spin box, and type 397.2.
Go on to the next topic: Optimizing the Collision Energy Automatically
for an MS/MS Experiment.
5-12 ________________
Finnigan LCQDUO Getting Started _______________
Acquiring ESI Sample Data Using the Tune Plus Window
'JOOJHBO-$2 %60 __________________________ Setting Up to Acquire MS/MS Data in the Full Scan Type
Optimizing the Collision Energy
Automatically for an MS/MS Experiment
The optimum relative collision energy is the one that produces the
maximum product ion intensity. Optimize the relative collision energy
automatically for the ESI/MS/MS analysis of reserpine, as follows:
1. Click on Start in the Tune dialog box to start the optimization
procedure. A message box displays the following message:
Please ensure that the 500 microliter syringe is full.
Ensure the syringe contains at least 420 µL of the 1 ng/µL reserpine
solution.
2. Close the message box, and return to the Tune dialog box:
Click on OK.
3. Your Tune Plus window should now look like the one shown in
Figure 5-10.
Figure 5-10. Tune Plus window, showing MS/MS product ions of reserpine in the Spectrum view
____________________ Finnigan LCQDUO Getting Started ______________________
5-13
Acquiring ESI Sample Data Using the Tune Plus Window
Setting Up to Acquire MS/MS Data in the Full Scan Type __________________________
'JOOJHBO-$2 %60
4. Leave the Tune dialog box open. Observe the MS/MS Full scan
spectrum of reserpine, or that of your analyte of interest, in the
Spectrum view.
5. When the collision energy is optimized, the Accept Optimized Value
dialog box appears. See Figure 5-11.
Figure 5-11. Accept Optimized Value dialog box
6. Click on the Accept button to accept the new collision energy value, and
close the dialog box.
7. Turn off the syringe pump, and close the Syringe Pump dialog box, as
follows:
Select the Off option button in the Syringe Pump dialog box. Then click
on Close.
8. Click on Cancel to close the Tune dialog box.
After you optimize the relative collision energy, you can acquire MS/MS
data on your analyte of interest.
Go on to the next chapter: Setting Up to Acquire Data in the APCI/MS
Mode.
5-14 ________________
Finnigan LCQDUO Getting Started _______________
Chapter 6
6. Setting Up to Acquire Data in the
APCI/MS Mode
To set up the LCQDUO MS detector to acquire data in the APCI/MS mode,
you do the following:
•
Install the APCI probe assembly
•
Set up to infuse your analyte into the MS detector using a syringe pump
connected to the LC with Tee union
•
Set up the MS detector for APCI/MS operation from the Tune Plus
window
This chapter contains the following sections:
•
Removing the ESI probe assembly
•
Installing the APCI probe assembly
•
Setting Up the Inlet for High-Flow Infusion
•
Setting Up the MS Detector for APCI/MS Operation
If the APCI probe is already installed on the API source, go to the topic:
Setting Up the Inlet for High-Flow Infusion.
If the ESI probe is installed on the API source, go on to the next topic
Removing the ESI Probe Assembly.
____________________ Finnigan LCQDUO Getting Started _______________________
6-1
Setting Up to Acquire Data in the APCI/MS Mode
Removing the ESI Probe Assembly ___________________________________________
'JOOJHBO-$2 %60
6.1 Removing the ESI Probe Assembly
To remove the ESI probe assembly, proceed as follows. See Figure 2-3 for
the location of the components of the ESI probe assembly.
1. Stop the flow of sample solution (from the LC or syringe pump) into the
ESI probe.
2. Disconnect the high voltage cable from the connector labeled HV on the
ESI probe. To disconnect the cable, turn the locking ring on the cable
counterclockwise until you can pull the cable free.
3. Disconnect the sample transfer line from the grounded transfer line
fitting on the ESI flange. (The sample transfer line is the line that
comes from the LC, divert/injector valve, or syringe pump. It is not the
fused silica capillary that enters the ESI probe.)
4. Disconnect the sheath gas line and fitting from the inlet labeled Sheath
Gas on the ESI probe by turning the sheath gas line fitting (blue)
counterclockwise until you can pull the sheath gas line and fitting free
from the probe.
5. Disconnect the auxiliary gas line and fitting from the inlet labeled Aux
Gas on the ESI probe by turning the auxiliary gas line fitting (green)
counterclockwise until you can pull the auxiliary gas line and fitting free
from the probe.
6. If the sheath liquid line is attached to the ESI probe, disconnect the
sheath liquid line and fitting from the inlet labeled Sheath Liquid on the
ESI probe by turning the sheath liquid line fitting counterclockwise until
you can pull the sheath liquid line and fitting free from the probe.
7. Loosen the two flange retainer bolts that secure the ESI probe assembly
to the spray shield.
8. Pull back the ESI probe assembly from the spray shield.
9. With one hand holding the ESI flange, loosen the knurled fastener that
secures the ESI flange to the probe slide adapter.
10. Slide the ESI probe assembly off the probe slide adapter. Store the ESI
probe assembly in its foam storage container.
Go on to the next topic: Installing the APCI Probe Assembly.
6-2
_______________________ Finnigan LCQDUO Getting Started ____________________
Setting Up to Acquire Data in the APCI/MS Mode
'JOOJHBO-$2 %60 ___________________________________________Installing the APCI Probe Assembly
6.2 Installing the APCI Probe
Assembly
To install the APCI probe assembly, remove the ESI probe assembly using
the procedure in the previous section; then proceed as follows. See
Figure 2-1 for the location of the components of the APCI probe assembly.
1. Remove the APCI probe assembly and corona discharge needle from the
storage container.
2. Insert the corona discharge needle into its socket in the corona discharge
needle assembly.
3. If your APCI probe assembly does not already contain a sample tube
(fused-silica capillary), follow the procedure for installing a sample tube
in the topic: Maintaining the APCI Probe in the MS Detector
Maintenance chapter of the Finnigan LCQDUO Hardware Manual.
Ensure that the probe retainer bolt is tight and the APCI probe is secured
to the APCI flange.
Caution. Inspect the APCI probe. Make sure that the green ground wire is
not touching the vaporizer casing. Reposition the ground wire if necessary.
4. Slide the APCI probe assembly onto the probe slide adapter. Secure the
APCI probe assembly to the probe slide adapter with the knurled
fastener.
5. Connect the sample transfer line and fitting to the inlet labeled LC on
the APCI probe.
6. Connect the auxiliary gas line and (green) fitting to the inlet labeled A
on the APCI probe.
7. Connect the sheath gas line and (blue) fitting to the inlet labeled S on the
APCI probe.
8. Connect the high voltage cable to the connector on the APCI probe
assembly. Turn the locking-ring on the cable clockwise to secure the
cable.
9. Connect the vaporizer heater cable to the connector on the front panel of
the MS detector (beneath the APCI probe assembly). Make sure that the
red dot on the cable is aligned with the red mark on the connector.
10. Push the APCI probe assembly against the spray shield.
11. Secure the APCI flange to the spray shield with the two flange retainer
bolts.
12. In the Tune Plus window, choose Setup > Change to APCI to
configure the software for APCI operation.
____________________ Finnigan LCQDUO Getting Started _______________________
6-3
Setting Up to Acquire Data in the APCI/MS Mode
Installing the APCI Probe Assembly___________________________________________
'JOOJHBO-$2 %60
Go on to the next topic: Setting Up the Inlet for Tuning Using High-Flow
Infusion.
6-4
_______________________ Finnigan LCQDUO Getting Started ____________________
Setting Up to Acquire Data in the APCI/MS Mode
'JOOJHBO-$2 %60 ___________________________ Setting Up the Inlet for Tuning Using High-Flow Infusion
6.3 Setting Up the Inlet for Tuning
Using High-Flow Infusion
The plumbing connections for APCI/MS sample introduction from the
syringe pump are shown in Figure 6-1.
To make the plumbing connections for APCI/MS sample introduction from
the syringe pump into solvent flow from an LC, proceed as follows:
1. Connect a 4 cm (1.5 in) segment of Teflon tubing with a (brown)
Fingertight fitting and a (brown) ferrule to the (black) LC union. (See
Figure 6-2.)
2. Load a clean, 500- /8QLPHWULFVV\ULQJH31301-19012) with
420 /RIDQJ /VROXWLRQRIUHVHUSLQHRU\RXUDQDO\WHRILQWHUHVW
(Refer to Appendix B: Sample Formulations for a procedure for
making the reserpine tuning solution.)
3. Insert the needle of a syringe into the segment of Teflon tubing, and
place the syringe in the syringe holder of the syringe pump.
LC TEE UNION
FROM LC
LC UNION
SYRINGE PUMP
TO WASTE
Figure 6-1. APCI/MS plumbing connections for sample introduction
from the syringe pump into solvent flow from an LC
____________________ Finnigan LCQDUO Getting Started _______________________
6-5
Setting Up to Acquire Data in the APCI/MS Mode
Setting Up the Inlet for Tuning Using High-Flow Infusion ___________________________
'JOOJHBO-$2 %60
4. Connect a fused-silica infusion line from the (black) LC union to the
(black) LC Tee union, as follows. See Figure 6-2.
a. Connect the infusion line (P/N 00106-10504) with a (brown)
Fingertight fitting and a (brown) ferrule to the free end of the LC
union.
b. Connect the other end of the infusion line with a (red) Fingertight
fitting and a (brown) ferrule to the side arm of the LC Tee union.
FINGERTIGHT FITTING
(00101-18081)
FERRULE
(00101-18196)
PEEK TUBING
(00301-22912)
TO APCI PROBE
LC TEE UNION
(00101-18204)
PEEK TUBING
FROM DIVER/INJECT VALVE
(00301-22912)
FERRULE
(00101-18120)
FINGERTIGHT FITTING
(00101-18195)
INFUSION LINE
FUSED-SILICA CAPILLARY
(00106-10504)
FROM LC UNION
AND SYRINGE PUMP
Figure 6-2.
APCI/MS plumbing connections for the LC Tee union and
syringe pump
Note. To cut the PEEK tubing used to connect your LC to the divert/inject
valve and the divert/inject valve to the APCI source, use a PEEK tubing
cutter. This ensures that the tubing is cut straight. In addition, make sure
your LC fittings, ferrules, and PEEK tubing are installed properly. By
using these precautions, you prevent void (dead) volumes. The exclusion
of void volumes is critical to microbore LC. Also, void volumes affect the
quality of the MS detector signal.
5. Connect a segment of PEEK tubing from the (black) LC Tee union to
the APCI LC inlet, as follows. (See Figure 6-2.)
a. Use a PEEK tubing cutter to cut a 4 cm (1.5 in.) length of the PEEK
tubing.
b. Connect the PEEK tubing with a (brown) Fingertight fitting and a
(brown) ferrule to a free end of the (black) LC Tee union.
c. Connect the other end of the PEEK tubing with a (red) Fingertight
fitting and a (brown) ferrule to the LC inlet located on the APCI
probe.
6-6
_______________________ Finnigan LCQDUO Getting Started ____________________
Setting Up to Acquire Data in the APCI/MS Mode
'JOOJHBO-$2 %60 ___________________________ Setting Up the Inlet for Tuning Using High-Flow Infusion
6. Connect an appropriate length of PEEK tubing (transfer line from the
divert/inject valve) from the divert/inject valve to the LC Tee union, as
follows. (See Figure 6-2.)
a. Connect a length of PEEK tubing with a (stainless steel) nut and a
(stainless steel) ferrule to port 3 of the divert/inject valve.
b. Connect the other end of the PEEK tubing with a (brown)
Fingertight fitting and a (brown) ferrule to the free end of the LC
Tee union.
7. Connect an appropriate length of PEEK tubing (transfer line from the
LC) from the divert/inject valve to the LC, as follows:
a. Connect a length of PEEK tubing with a (stainless steel) nut and a
(stainless steel) ferrule to port 2 of the divert/inject valve.
b. Connect the other end of the PEEK tubing with a proper fitting and
a ferrule to the outlet of the LC.
8. Connect an appropriate length of PEEK tubing (waste line) from the
divert/inject valve to a waste container, as follows:
a. Connect a length of PEEK tubing with a (stainless steel) nut and a
(stainless steel) ferrule to port 1 of the divert/inject valve.
b. Insert the other end of the PEEK tubing in a suitable waste
container.
Caution. Prevent solvent waste from backing up into the API ion source
and MS detector. Always ensure that the PVC drain hose is above the level
of liquid in the waste container.
9. Connect an appropriate length of (clear) 3/8 in. ID PVC hose to the ESI
probe outlet drain. Insert the other end of the hose into a suitable waste
container (such as a gallon jug). Ideally, the waste container should be
vented to a fume exhaust system.
Go to the next topic: Setting Up the MS Detector for APCI/MS
Operation.
____________________ Finnigan LCQDUO Getting Started _______________________
6-7
Setting Up to Acquire Data in the APCI/MS Mode
Setting Up the MS Detector for APCI/MS Operation ______________________________
'JOOJHBO-$2 %60
6.4 Setting Up the MS Detector for
APCI/MS Operation
Use the following procedure to set up the MS detector for APCI/MS
operation.
STANDBY
ON
1. In Tune Plus, take the MS detector out of Standby mode and turn it On,
as follows:
Click on the On/Standby button in the Control/Scan Mode toolbar. The
MS detector begins scanning, and the LCQDUO applies high voltage to
the corona needle and shows a real-time display in the Spectrum view.
2. Open the APCITune.LCQTune Tune Method, the Tune Method for
high-flow APCI operation, as follows:
a. Display the Open dialog box, as follows:
Choose File > Open.
b. Select the file APCITune.LCQTune in the directory
C:\Xcalibur\Methods, as follows:
Scroll down in the File Name combo box until you see
APCITune.LCQTune. Then, click on the file name.
c. Open the file, and close the dialog box, as follows:
Click on OK. LCQDUO downloads the Tune Method parameters to
the MS detector.
3. Verify that LCQDUO opened the Tune Method, as follows:
a. Open the APCI Source dialog box, as follows:
Click on the APCI Source button in the Instrument Setup toolbar.
See Figure 6-3.
b. Verify that the settings in your dialog box are similar to those
shown in Figure 6-3.
Figure 6-3.
6-8
APCI Source dialog box, showing the proper settings for
a typical high flow experiment
_______________________ Finnigan LCQDUO Getting Started ____________________
'JOOJHBO-$2 %60 ______________________________
Setting Up to Acquire Data in the APCI/MS Mode
Setting Up the MS Detector for APCI/MS Operation
c. Close the dialog box, as follows:
Click on OK.
4. Define the scan parameters for tuning the MS detector in the APCI/MS
mode, as follows:
a. Open the Define Scan dialog box, as follows:
Click on the Define Scan button in the Control/Scan Mode toolbar.
See Figure 6-4.
b. Select the MS scan mode in the Scan Description group box, as
follows:
Click on the Scan Mode: MS option button.
c. Select the SIM data type, as follows:
Click on the Scan Type: SIM option button. Note that LCQDUO sets
the Number of Scan Ranges to 1.
d. Set the total number of microscans to 1 in the Scan Time group box,
as follows:
Double-click in the Number of Microscans spin box, then type 1.
e. For this example, leave the maximum injection time set to its
default value of 200.00 ms.
f.
Specify that the ion source fragmentation option is turned off in the
Source Fragmentation group box:
Confirm that the Turn On check box is not selected (
).
g. Set the center mass for the scan range to m/z 609.2 in the Scan
Ranges group box, as follows:
Double-click in the Center Mass text box, then type 609.2.
h. Set the width of the scan range to m/z 2, as follows:
Double-click in the Width text box, then type 2.
i.
Ensure that the settings in your Define Scan dialog box are the same
as those shown in Figure 6-4.
j.
Save the MS detector scan parameters, and close the Define Scan
dialog box, as follows:
Click on OK.
____________________ Finnigan LCQDUO Getting Started _______________________
6-9
Setting Up to Acquire Data in the APCI/MS Mode
Setting Up the MS Detector for APCI/MS Operation ______________________________
'JOOJHBO-$2 %60
Figure 6-4. Define Scan dialog box, showing typical settings for APCI/MS operation
5. Select the centroid data type, as follows:
Click on the Centroid/Profile button in the Control/Scan Mode toolbar
to toggle the data type to centroid. (The picture on the button should be
the same as that shown here).
6. Select the positive ion polarity mode, as follows:
Click on the Positive/Negative button in the Control/Scan Mode toolbar
to toggle the ion polarity mode to positive. (The picture on the button
should be the same as that shown here).
You have now completed setting up your MS detector for APCI/MS
operation. Go to the next chapter: Optimizing the MS Detector with
Your Analyte in APCI/MS Mode.
6-10 ______________________ Finnigan LCQDUO Getting Started
____________________
Chapter 7
7. Optimizing the MS Detector with Your
Analyte in APCI/MS Mode
This chapter provides information on optimizing the tune of your MS
detector in the APCI/MS high flow mode. It is not necessary to recalibrate
the MS detector when you switch to APCI/MS operation. You can use the
calibration settings you obtained from the successful automatic calibration
procedure you performed in the ESI/MS mode.
For APCI/MS operation you simply open a default tune method located in
your C:\Xcalibur\methods directory, in this case APCIhighflow.LCQTune.
From this starting point, you optimize automatically the tube lens offset
voltage, capillary voltage, and heated capillary temperature for your
particular analyte.
Note. The following procedures assume that you are familiar with your
LCQDUO instrument and the Tune Plus window. If you need information,
refer to the LCQDUO online Help, Finnigan LCQDUO Getting Connected,
and/or Finnigan LCQDUO Hardware Manual.
Ensure that you have completed the procedures in the topics Tuning and
Calibrating Automatically in the ESI/MS Mode and Setting Up to
Acquire Data in the APCI/MS Mode.
This chapter includes the following sections:
•
Optimizing the Tune of the MS Detector Automatically in APCI/MS
Mode
•
Saving the APCI/MS Tune Method
•
Cleaning the Spray Shield after Optimization
____________________ Finnigan LCQDUO Getting Started _______________________
7-1
Optimizing the MS Detector with Your Analyte in APCI/MS Mode
Optimizing the Tune of the MS Detector Automatically in APCI/MS Mode ______________
'JOOJHBO-$2 %60
7.1 Optimizing the Tune of the MS
Detector Automatically in APCI/MS
Mode
You can optimize the tune of the MS detector automatically for APCI
operation.
The most important parameters that affect the signal quality during
APCI/MS operation are the vaporizer temperature, heated capillary
temperature, capillary voltage, tube lens offset voltage, gases, and solution
flow rate. If any one of these parameters is changed, you need to reoptimize
MS detector parameters. (You can use the Semi-Automatic tune procedure
to tune the MS detector on individual parameters.)
Use the following procedure to optimize the MS detector automatically on
the reserpine peak at m/z 609.2 at your particular flow rate, for example,
1 mL/min. (Refer to Table 1-2 for guidelines about flow rates and
temperatures.)
1. Display the automatic tuning page in the Tune dialog box, as follows:
Click on the Tune button on the Control/Scan Mode toolbar. Then,
click on the Automatic tab.
2. Specify that you want to tune on the peak at m/z 609.2, as follows:
Double-click on the Mass spin box, then type 609.2.
3. Ensure that the Divert/Inject valve is in the Detector position, as
follows:
a. Click on the Divert/Inject button to open the Divert/Inject Valve
dialog box.
b. Select the Detector option button.
c. Click on Close.
4. Start the automatic tuning procedure from the Tune dialog box, as
follows:
a. Click on Start. A message box displays the following message:
Please ensure that the 500 microliter syringe is full.
Ensure the syringe pump contains at least 420 µL of the 1 ng/µL
reserpine tuning solution.
b. Close the message box, and return to the Tune Plus window:
Click on OK.
5. Display the Graph view, as follows:
Click on the Graph View button in the File/Display toolbar.
7-2
_______________________ Finnigan LCQDUO Getting Started ____________________
Optimizing the MS Detector with Your Analyte in APCI/MS Mode
'JOOJHBO-$2 %60 ______________ Optimizing the Tune of the MS Detector Automatically in APCI/MS Mode
6. Observe the Tune Plus window and the Tune dialog box. While
automatic tuning is in progress, LCQDUO displays various tests in the
Spectrum and Graph views in the Tune Plus window and displays
various messages in the Status group box in the Tune dialog box. Your
Tune Plus window should now look similar to the one shown in
Figure 7-1.
You have now successfully tuned the MS detector in APCI/MS mode for the
compound reserpine (or your analyte of interest). Leave the LC pumps on
(with a flow rate of approximately 1 mL/min), and leave the Tune Plus
window open with APCIhighflow.LCQTune to complete the next topic:
Saving the APCI/MS Tune Method.
Figure 7-1. Tune Plus window with the Tune dialog box, showing the Automatic page
____________________ Finnigan LCQDUO Getting Started _______________________
7-3
Optimizing the MS Detector with Your Analyte in APCI/MS Mode
Saving the APCI/MS Tune Method____________________________________________
'JOOJHBO-$2 %60
7.2 Saving the APCI/MS Tune Method
You can save the settings you just obtained in a tune method specific to your
particular analyte and solvent flow rate. (In this case, you save settings
obtained using reserpine.) You can recall the tune method and use it as a
starting point for optimizing the MS detector on reserpine at a different flow
rate.
Note. Save the Tune Method while the MS detector is On.
Save your APCI/MS tune method, as follows:
1. Display the Save As dialog box, as follows:
Choose File > Save As. See Figure 7-2.
Figure 7-2.
7-4
Save As dialog box, showing files in the directory
C:\Xcalibur\methods
_______________________ Finnigan LCQDUO Getting Started ____________________
Optimizing the MS Detector with Your Analyte in APCI/MS Mode
'JOOJHBO-$2 %60 ____________________________________________Saving the APCI/MS Tune Method
2. Select the C:\Xcalibur\methods directory.
3. Name the Tune Method, as follows:
Click on the File Name text box, and type APCImyTune.
4. Save the Tune Method, close the dialog box, and return to the Tune Plus
window, as follows:
Click on Save. Note that the Tune Method is named
APCImyTune.LCQTune.
Before you acquire data, go to the next topic: Cleaning the MS Detector
after Tuning in APCI Mode.
____________________ Finnigan LCQDUO Getting Started _______________________
7-5
Optimizing the MS Detector with Your Analyte in APCI/MS Mode
Cleaning the MS Detector after Tuning in APCI Mode _____________________________
'JOOJHBO-$2 %60
7.3 Cleaning the MS Detector after
Tuning in APCI Mode
Use the following procedure to clean the MS detector after tuning on your
analyte of interest.
ON
1. Put the MS detector in Standby, as follows:
Click on the On/Standby button. When the MS detector is in Standby,
the LCQDUO turns Off the vaporizer heater and the corona discharge
voltage. The MS detector stops scanning, and the LCQDUO freezes the
displays for the Spectrum and Graph views.
STANDBY
CAUTION. Always place the MS detector in Standby (or Off) before you
open the API source to atmospheric oxygen. The presence of oxygen in the
ion source when the MS detector is ON could be unsafe. (The LCQDUO
automatically turns the MS detector Off when you open the API source,
however, it is best to take this added precaution.)
2. Turn Off the syringe pump to stop the flow of solution into the ion
source, as follows:
a. Display the Syringe Pump dialog box, as follows:
Click on the Syringe Pump button.
b. Select the Off option button in the Flow Control group box to turn
off the syringe pump.
c. Click on Close.
3. Remove the syringe from the syringe holder, as follows:
a. Pull back on the silver syringe handle to free the syringe.
b. Remove the syringe from the holder.
c. Disconnect the tip of the syringe needle from the Teflon tubing.
4. Clean the syringe thoroughly, as follows:
First, clean the syringe with a solution of 5% acetic acid in water. Then,
rinse the syringe with a solution of 50:50 methanol:water. Finally, rinse
the syringe with acetone.
5. Loosen completely the two APCI flange retainer bolts.
6. Slide the APCI flange out, away from the MS detector chassis, on the
slide rails.
7-6
_______________________ Finnigan LCQDUO Getting Started ____________________
Optimizing the MS Detector with Your Analyte in APCI/MS Mode
'JOOJHBO-$2 %60 _____________________________ Cleaning the MS Detector after Tuning in APCI Mode
7. Flush the sample transfer line, sample tube, and APCI probe thoroughly
with a solution of 5% acetic acid in water (or with another appropriate
solvent), as follows:
Note. The solvent that you use to flush the sample transfer line, sample
tube, and APCI probe assembly depends on the solvent system you use to
dissolve your samples. For example, if you are using a buffered solution of
a high concentration, an acidic solution is appropriate.
a. Fill a clean, 250 µL Unimetrics syringe with a solution an
appropriate solvent.
b. While holding the plunger of the syringe in place, carefully insert
the needle of the syringe into the free end of the Teflon tube.
c. Flush the sample transfer line, sample tube, and APCI probe with
the solution by slowly depressing the syringe plunger. Visually
check that the solution is exiting the tip of the APCI probe on the
inside of the probe assembly. Use a lint-free tissue to gently remove
the excess solution as it exits the probe.
d. Remove the needle of the syringe from the Teflon tube.
8. Repeat step 7 with a solution of 50:50 methanol:water.
9. Slide the APCI flange forward along the slide rails and into the ion
source housing.
10. Secure the APCI flange with the two flange retainer bolts.
If you plan to run analytical samples in APCI mode, go to the next chapter:
Acquiring APCI Sample Data Using the Tune Plus Window.
____________________ Finnigan LCQDUO Getting Started _______________________
7-7
Chapter 8
8. Acquiring APCI Sample Data Using
the Tune Plus Window
This chapter provides information on acquiring LC/APCI/MS sample data
using the Tune Plus window. This experiment uses reserpine but you can
follow the same procedure with your analyte of interest.
Note. The following procedures assume that you are familiar with your
LCQDUO instrument and the Tune Plus window. If you need information,
refer to the LCQDUO online Help, Finnigan LCQDUO Getting Connected,
and/or Finnigan LCQDUO Hardware Manual.
Ensure that you have completed the procedures in the topics Tuning and
Calibrating Automatically in the ESI/MS Mode and Optimizing the MS
Detector Automatically in APCI/MS Mode.
This chapter contains the following sections:
•
Setting Up to Introduce Sample by Loop Injection into Solvent Flow
from an LC
•
Acquiring APCI Data in the SIM Scan Mode
____________________ Finnigan LCQDUO Getting Started _______________________
8-1
Acquiring APCI Sample Data Using the Tune Plus Window
Setting Up to Introduce Sample by Loop Injection into Solvent Flow from an LC_________
'JOOJHBO-$2 %60
8.1 Setting Up to Introduce Sample by
Loop Injection into Solvent Flow
from an LC
This topic provides information on how to introduce sample by loop
injection into solvent flow from an LC. The plumbing connections for
APCI/MS sample introduction by loop injection are shown in Figure 8-1.
To make the plumbing connections, do the following:
1. Connect an appropriate length of (red) PEEK tubing (transfer line from
the divert/inject valve) from port 3 of the divert/inject valve to the
(stainless steel) fitting at the center of the APCI probe. See Figure 8-1.
2. Connect a 5 µL sample loop (P/N 00110-16010) with (stainless steel)
set nuts and (stainless steel) ferrules to ports 1 and 4 of the divert/inject
valve.
FROM LC
INJECTION PORT
(CENTER, POSITION 5)
TO WASTE
Figure 8-1. APCI/MS plumbing connections for sample introduction by
loop injection into solvent flow from an LC
8-2 _________________
Finnigan LCQDUO Getting Started _______________
Acquiring APCI Sample Data Using the Tune Plus Window
'JOOJHBO-$2 %60 _________Setting Up to Introduce Sample by Loop Injection into Solvent Flow from an LC
3. Connect an appropriate length of PEEK tubing (transfer line from the
LC) from the divert/inject valve to the LC, as follows:
a. Connect a length of the PEEK tubing with a (stainless steel) nut and
(stainless steel) ferrule to port 2 of the divert/inject valve.
b. Connect the other end of the PEEK tubing with a proper fitting and
a ferrule to the outlet of the LC.
4. Connect an appropriate length of PEEK tubing (waste line) from the
divert/inject valve to a waste container, as follows:
a. Connect a length of PEEK tubing with a (stainless steel) nut and
(stainless steel) ferrule to port 6 of the divert/inject valve.
b. Insert the other end of the PEEK tubing into a suitable waste
container.
Caution. Prevent solvent waste from backing up into the API ion source
and MS detector. Always ensure that the PVC drain hose is above the level
of liquid in the waste container.
5. Connect an appropriate length of (clear) 3/8 in. ID PVC hose to the
APCI probe outlet drain. Insert the end of the hose into a suitable waste
container (such as a gallon jug). Ideally, the container should be vented
to a fume exhaust system.
TO ION
SOURCE
Figure 8-2.
TO ION
SOURCE
Divert/inject (Valco injector) valve, showing the correct
set up for analysis by loop injection and showing the flow
of liquid through the valve in the LOAD and INJECT
positions
Go on to the next topic: Acquiring APCI Data in the SIM Scan Mode.
____________________ Finnigan LCQDUO Getting Started _______________________
8-3
Acquiring APCI Sample Data Using the Tune Plus Window
Acquiring APCI Data in the SIM Scan Mode ____________________________________
'JOOJHBO-$2 %60
8.2 Acquiring APCI Data in the SIM
Scan Mode
Use the following procedure to acquire a file of reserpine data in the single
ion monitoring (SIM) mode. The LCQDUO automatically saves the data you
acquire on your hard disk.
STANDBY
ON
1. If you have not already done so, take the MS detector out of Standby
mode and turn it On from the Tune Plus window, as follows:
Click on the On/Standby button in the Control/Scan Mode toolbar. The
MS detector begins scanning, and a real-time display shows in the
Spectrum view.
2. Ensure that the Centroid data type is selected. (The picture on the
button should be the same as that shown here.)
3. Ensure that the scan parameters are defined to acquire SIM data for
reserpine (or your analyte of interest), as follows:
a. Open the Define Scan dialog box:
Click on the Define Scan button. See Figure 8-3.
b. Ensure that the values in your dialog box to those in Figure 8-3.
Then, click on OK.
Figure 8-3. Define Scan dialog box, showing typical settings for acquiring data in the SIM scan
mode
4. Turn on your LC pump, and specify a flow rate of 1 mL/min, for
example. Ensure that your system is free of leaks.
8-4 _________________
Finnigan LCQDUO Getting Started _______________
Acquiring APCI Sample Data Using the Tune Plus Window
'JOOJHBO-$2 %60 ____________________________________
Acquiring APCI Data in the SIM Scan Mode
5. Open the Acquire Data dialog box, as follows:
Click on the Acquire Data button on the Control/Scan Mode toolbar.
See Figure 8-4.
Figure 8-4. Acquire Data dialog box, showing the acquisition status of the raw data file
6. Specify the acquisition parameters, as follows:
a. Specify a filename, as follows:
Double-click in the File Name text box, then type, for example,
C:\Xcalibur\Data\reserpine3.raw.
b. Specify the sample identity, as follows:
Double-click in the Sample Name text box, then type reserpine. If
you are not using reserpine, type the name of your particular
analyte.
c. Specify a comment about your experiment, as follows. (For
example, type the scan mode, ionization mode, sample amount,
and/or method of sample introduction.)
Double-click in the Comment text box, then type SIM, APCI,
50 pg, loop. Xcalibur includes the comment on hard copies of your
data.
d. Select to acquire data continuously (until you stop the acquisition)
in the Acquire Time group box, as follows:
Select the Continuously option button.
7. Leave the Acquire Data dialog box open during data acquisition, but
move it to a corner of the Tune Plus window.
8. Click on Start in the Acquire Data dialog box to begin acquiring data to
the file reserpine3.raw. See Figure 8-5. The Acquisition Status group
box displays the following message.
State: Acquiring
Time (min):
____________________ Finnigan LCQDUO Getting Started _______________________
8-5
Acquiring APCI Sample Data Using the Tune Plus Window
Acquiring APCI Data in the SIM Scan Mode ____________________________________
'JOOJHBO-$2 %60
Figure 8-5. Tune Plus window, showing the SIM spectrum of reserpine during analysis by loop
injection
9. Inject the reserpine solution into the APCI source from the Instrument
Setup toolbar, as follows:
a. Click on the Divert/Inject button. The Divert/Inject Valve dialog
box appears. See Figure 8-6.
Figure 8-6. Divert/Inject Valve dialog box
8-6 _________________
Finnigan LCQDUO Getting Started _______________
Acquiring APCI Sample Data Using the Tune Plus Window
'JOOJHBO-$2 %60 ____________________________________
Acquiring APCI Data in the SIM Scan Mode
b. Select the Load option button, and overfill the 5-µL injector loop
with the 10 pg/µL solution of reserpine, or a solution of your analyte
of interest.
c. Select the Inject option button.
10. Observe the reserpine peak (m/z 609.2), or that of your analyte of
interest, in the Spectrum view.
11. Perform the following repetitive sequence to obtain a total of four
consecutive loop injections of reserpine in the SIM scan mode. Wait
about 1 min between injections.
a. Select the Load option button to put the divert/inject valve in the
Load position. Overfill the injector loop with the 10 pg/µL solution
of reserpine.
b. Select the Inject option button to inject the reserpine solution into
the APCI source. Then, observe the Spectrum view.
c. Again, select the Load option. Again, overfill the injector loop with
the 10 pg/µL solution of reserpine.
d. Again, select the Inject option button, and observe the Spectrum
view.
e. For the fourth time, select the Load option button, then overfill the
injector loop with the 10 pg/µL solution of reserpine.
f.
Finally, select the Inject option button to inject the reserpine
solution.
12. Click on Stop in the Acquire Data dialog box to end the data
acquisition.
Review the mass spectrum and chromatogram in the raw file you just
acquired using the Xcalibur Qual Browser window. See Figure 8-7.
For more information about reviewing the data you acquire using the
LCQDUO MS detector with Xcalibur, refer to the manual: Finnigan
Xcalibur Getting Productive: Qualitative Analysis.
Note. If you want to acquire MS/MS Full scan data in APCI mode, refer to
the section Setting Up to Acquire MS/MS Data in the Full Scan Type.
____________________ Finnigan LCQDUO Getting Started _______________________
8-7
Acquiring APCI Sample Data Using the Tune Plus Window
Acquiring APCI Data in the SIM Scan Mode ____________________________________
'JOOJHBO-$2 %60
Figure 8-7. Qual Browser window, showing loop injections of reserpine in the Chromatogram
view (left) and m/z 609 in the Spectrum view
8-8 _________________
Finnigan LCQDUO Getting Started _______________
Appendix A
A. Instrument Configuration
This appendix provides instructions for configuring the Xcalibur data
system for your LCQDUO MS detector.
This appendix contains the following sections:
•
Configuring the LCQDUO MS detector using the Instrument
Configuration dialog box
•
Using the LCQDUO Configuration dialog box
Note. If you need more information, refer to the LCQDUO online Help,
Finnigan LCQDUO Getting Connected, and/or Finnigan LCQDUO
Hardware Manual.
____________________ Finnigan LCQDUO Getting Started _______________________
A-1
Instrument Configuration
Configuring Xcalibur Using the Instrument Configuration Dialog Box__________________
'JOOJHBO-$2 %60
A.1 Configuring Xcalibur Using the
Instrument Configuration Dialog
Box
Configure the Xcalibur data system for your LCQDUO MS detector by using
the Instrument Configuration dialog box, as follows:
1. Open the Instrument Configuration dialog box from the Windows XP
Desktop, as follows:
Double-click on the Instrument Configuration icon. See Figure A-1.
Figure A-1. Instrument Configuration dialog box, showing LCQDUO in the Configured Devices list
2. Click on the LCQ Duo MS button in the Available Devices group box.
3. Click on the Add button. Xcalibur moves the LCQ Duo MS button to
the Configured Devices group box.
A-2 _______________________ Finnigan LCQDUO Getting Started
____________________
Instrument Configuration
'JOOJHBO-$2 %60 __________________Configuring Xcalibur Using the Instrument Configuration Dialog Box
You need to repeat this procedure for each hardware device in your LC/MS
system: LC, autosampler, UV detector, and so on. See Figure A-2.
Go on to the next topic: Using the LCQ Configuration Dialog Box.
Figure A-2. Instrument Configuration dialog box, showing configured devices
____________________ Finnigan LCQDUO Getting Started _______________________
A-3
Instrument Configuration
Using the LCQ Configuration Dialog Box _______________________________________
'JOOJHBO-$2 %60
A.2 Using the LCQ Configuration
Dialog Box
Use the LCQ Configuration dialog box to specify, for example, the
following parameters:
•
Full Tune Plus user interface
•
[Ion] Source Type
•
Divert Valve Plumbing
Specify parameters after you display a page in the dialog box, as follows:
1. Open the LCQ Configuration dialog box from the Instrument
Configuration dialog box, as follows:
Double-click on the LCQ Duo MS button in the Configured Devices
group box. The LCQ Configuration dialog box is displayed. See
Figure A-3.
Figure A-3. LCQ Configuration dialog box, showing the MS detector
page
2. Select each page in turn to specify the parameters as you want them.
For example, specify to display all the available icons in the Tune Plus
window toolbars when you open the Tune Plus window, as follows:
a. Select the Tune Plus tab to display the Tune Plus page. See
Figure A-4.
A-4 _______________________ Finnigan LCQDUO Getting Started
____________________
Instrument Configuration
'JOOJHBO-$2 %60 _______________________________________ Using the LCQ Configuration Dialog Box
Figure A-4. LCQ Configuration dialog box, showing the Tune Plus
page
b. Select
the Full Tune Plus User Interface checkbox.
c. Click on OK to close the dialog box.
3. Click on the Done button in the Instrument Configuration dialog box to
return to the Windows XP Desktop.
____________________ Finnigan LCQDUO Getting Started _______________________
A-5
Appendix B
B. Sample Formulations
This appendix provides instructions for the preparation of several stock
solutions. These solutions are used for tuning, calibrating, and
demonstrating applications of the APCI / ESI system. Formulations for
sample solutions in this appendix are as follows:
•
Caffeine, MRFA, and Ultramark 1621 stock solutions
•
ESI Calibration Solution: Caffeine, MRFA, Ultramark 1621
•
Reserpine
Always take safety precautions when you handle chemicals and unknown
samples. ENSURE THAT YOU READ AND UNDERSTAND THE
HAZARDS OF THE CHEMICALS USED IN THE FOLLOWING
PREPARATIONS. Dispose of all laboratory reagents by the appropriate
method for a specific reagent or solvent.
Material Safety Data Sheets (MSDS) provide summarized information on
the hazard and toxicity of specific chemical compounds. MSDSs also
provides information on the proper handling of compounds, first aid for
accidental exposure, and procedures for the remedy of spills or leaks.
Producers and suppliers of chemical compounds are required by law to
provide their customers with the most current health and safety information
in the form of an MSDS. Read the material safety data sheets for each
chemical you use. Examples of potentially hazardous chemicals used in
procedures throughout this manual are as follows:
•
Acetic acid
•
Acetonitrile
•
Methyl alcohol (methanol)
•
Reserpine
____________________ Finnigan LCQDUO Getting Started _______________________
B-1
Sample Formulations
Caffeine, MRFA, and Ultramark 1621 Stock Solutions_____________________________
'JOOJHBO-$2 %60
B.1 Caffeine, MRFA, and
Ultramark 1621 Stock Solutions
For tuning and calibrating the ESI system, you use a solution of caffeine,
MRFA, and Ultramark 1621 in an acetonitrile:methanol:water solution
containing 1% acetic acid. You prepare the calibration solution from each
of the following:
•
Caffeine stock solution
•
MRFA stock solution
•
Ultramark 1621 stock solution
Note. Vials of caffeine, MRFA, and Ultramark 1621 are included in the
API accessory kit. To order more of these compounds, write or call:
Sigma Chemical Company
P. O. Box 14508
St. Louis, Missouri, USA 63178-9916
(800) 325-3010 (in the USA or Canada)
(314) 771-3750 (outside the USA or Canada)
CAUTION. AVOID EXPOSURE TO POTENTIALLY HARMFUL
MATERIALS. Always wear protective gloves and safety glasses when
you use solvents or corrosives. Also, contain waste streams and use proper
ventilation. Refer to your supplier’s Material Safety Data Sheets (MSDS)
for procedures that describe how to handle a particular solvent and/or
corrosive substance.
B-2 _______________________ Finnigan LCQDUO Getting Started
____________________
Sample Formulations
'JOOJHBO-$2 %60 _____________________________Caffeine, MRFA, and Ultramark 1621 Stock Solutions
Stock Solution: Caffeine
A 1 mg/mL stock solution of caffeine in 100% methanol is provided with
your LCQ system.
Stock Solution: MRFA
Prepare a 1 mL stock solution of 5 nmol/µL MRFA in 50:50 methanol:water
as follows:
1. Obtain the vial of L-methionyl-arginyl-phenylalanyl-alanine
acetate⋅H2O (MRFA) in your accessory kit. In this form, the MRFA
sample has an average molecular weight of 523.6 u. Carefully weigh
3.0 mg of the MRFA sample.
2. Dissolve the MRFA sample in a total volume of 1 mL of 50:50
methanol:water. Mix the solution thoroughly.
3. Label the vial MRFA stock solution.
Stock Solution: Ultramark 1621
Prepare a 10 mL stock solution of 0.1% Ultramark 1621 in acetonitrile as
follows:
1. Obtain the vial of Ultramark 1621 in your accessory kit.
2. Using a syringe, measure out 10 µL of Ultramark 1621, and dissolve it
in 10 mL of acetonitrile. Mix the solution thoroughly.
3. Label the vial Ultramark 1621 stock solution.
Go to the next topic: ESI Calibration Solution: Caffeine, MRFA,
Ultramark 1621.
____________________ Finnigan LCQDUO Getting Started _______________________
B-3
Sample Formulations
ESI Calibration Solution: Caffeine, MRFA, Ultramark 1621_________________________
'JOOJHBO-$2 %60
B.2 ESI Calibration Solution: Caffeine,
MRFA, Ultramark 1621
Prepare 5 mL of the calibration solution, as follows:
1. Pipet 100 µL of the stock solution of caffeine into a clean, dry vial.
2. Pipet 5 µL of the stock solution of MRFA into the vial.
3. Pipet 2.5 mL of the stock solution of Ultramark 1621 into the vial.
Note. Use only glass pipets or stainless steel syringes when measuring
glacial acetic acid. Using plastic pipet tips causes contamination of acid
stock solutions, which can introduce contaminants in the calibration
solution.
4. Pipet 50 µL of glacial acetic acid into the vial.
5. Pipet 2.34 mL of 50:50 methanol:water into the vial, and then mix the
solution thoroughly.
6. Label the vial and store it in a refrigerator until it is needed.
B-4 _______________________ Finnigan LCQDUO Getting Started
____________________
Sample Formulations
'JOOJHBO-$2 %60 _______________________________________________________________ Reserpine
B.3 Reserpine
Follow the directions given below to prepare a stock solution of reserpine.
Then use serial dilutions of the stock solution to make either an APCI or ESI
sample solution as described in: ESI / APCI Sample Solution: Reserpine.
Stock Solution: Reserpine
Prepare a stock solution of 1 µg/µL reserpine in 1% acetic acid in 50:50
methanol:water:
1. Obtain the 1 gram vial of reserpine in your accessory kit. (The average
molecular weight of resperine is 608.7 u). Weigh out 1 mg of reserpine
and transfer the sample to a polypropylene microcentrifuge tube.
2. Dissolve the reserpine sample in a total volume of 1 mL of 1% acetic
acid in 50:50 methanol:water.
3. Ensure the sample is thoroughly dissolved in solution.
ESI / APCI Sample Solution: Reserpine
Prepare 1 mL of the sample solution of 10 pg/µL (82 fmol/µL) in 1% acetic
acid in 50:50 methanol:water, as follows:
1. Pipet 100 µL of the stock solution (1µg/µL) of reserpine into a clean
polypropylene microcentrifuge tube.
2. Add 900 µL of 1% acetic acid in 50:50 methanol:water to the tube.
3. Mix this solution (100 ng/µL) thoroughly.
4. Transfer 10 µL of the 100 ng/µL solution into a clean polypropylene
tube.
5. Add 990 µL of 1% acetic acid in 50:50 methanol:water to the tube.
6. Mix this solution (1 ng/µL) thoroughly.
7. Transfer 10 µL of the 1 ng/µL solution into a clean polypropylene tube.
8. Add 990 µL of 1% acetic acid in 50:50 methanol:water to the tube.
9. Mix this solution (10 pg/µL) thoroughly.
10. Store the ESI / APCI sample solution (10 pg/µL) in a refrigerator until it
is needed.
____________________ Finnigan LCQDUO Getting Started _______________________
B-5
Index
'JOOJHBO-$2 %60 _______________________________________________________________________
Index
Note. Dialog box is abbreviated d.b.
A
Accept Optimized Value d.b.
figure, 5-14
Acquire Data d.b.
figure, 5-6, 8-5
acquiring data
prerequisite for MS/MS mode, 5-10
SIM scan type, 5-5, 8-4
using Tune Plus, 5-1, 8-1
APCI
plumbing
LC Tee union (figure), 6-6
plumbing for sample input
loop injection (figure), 8-2
Tee to LC (figure), 6-5
APCI probe assembly
figure, 2-2
installing, 6-3
removing, 2-2
APCI Source d.b.
figure, 6-8
APCI/MS
operational guidelines
table, 1-10
setting up MS detector for, 6-8
using, discussion, 1-4
API source
discussion, 1-3
sufficient nitrogen flow (CAUTION), 2-11
AutoTune file, 2-13
B
buffers, LC
discussion, 1-8
C
caffeine
stock solution, preparing, B-3
vial of sample formulation (note), B-2
Calibrate d.b.
(figure), 3-12
calibrating
discussion, 1-11
MS detector, 3-12
MS detector setup, 2-11
MS detector, in ESI/MS mode
automatically (note), 3-7
results (figure), 3-15
solutions
Caution, 4-7
ions, 3-3
preparing, B-4
real-time plot of, in Spectrum view (figure), 3-5
table, 1-12
syringe pump setup for, 2-8
capillary temperature
setting (figure), 2-13
capillary voltage
setting (figure), 2-13
capillary, heated
CAUTION, 3-16
temperature (note), 4-11
voltage (note), 4-11
Cautions
calibration solution
flow rate limit, 4-7
solvent waste backing into API2 source housing,
2-10, 4-6, 5-4, 8-3
solvent waste backing into API2 source housing,
6-7
CAUTIONS
avoid burns from APCI vaporizer heater, 2-3
avoid burns from heated capillary, 3-16
avoid electrical shock if fused silica transfer line
breaks, 2-4
avoid exposure to potentially harmful materials, B2
exposing ion source to oxygen, 3-16, 7-6
sufficient nitrogen flow for API source, 2-11
cleaning MS detector, 3-16, 7-6
collision energy
optimizing, procedure, 5-13
computer terms
directories, 3-11, 4-13, 7-5
files, 3-11, 4-13, 7-5
configuring
LCQDUO with Xcalibur, A-2
D
data
acquiring, in SIM scan type, 5-5, 8-4
reviewing, in Qual Browser window (figure), 5-9, 88
Define Scan d.b.
APCI settings
figure, 6-10
figure, 2-15, 4-9, 5-5, 8-4
MS/MS Full scan settings (figure), 5-11
dialog boxes
Accept Optimized Value (figure), 5-14
____________________ Finnigan LCQDUO Getting Started __________________________
I
Index
_______________________________________________________________________
Acquire Data, 5-6, 8-5
APCI Source (figure), 6-8
Calibrate (figure), 3-12
Define Scan, 2-15, 4-9, 5-5, 8-4
APCI settings (figure), 6-10
MS/MS Full scan settings, 5-11
Divert/Inject Valve, 4-10, 5-7, 8-6
ESI Source, 2-13, 3-9
Instrument Configuration, A-2, A-3
LCQ Configuration, A-4, A-5
Save As, 3-10, 4-12, 7-4
Syringe Pump
figure, 3-3
Tune, 3-6
figure, 4-11, 7-3
Tune, Collision Energy page (figure), 5-12
Divert/Inject valve
loop injection (figure), 8-3
Divert/Inject Valve d.b., 4-10, 5-7, 8-6
E
ESI
plumbing for sample input
loop injection (figure), 5-2
syringe pump infusion (figure), 2-8
Tee to LC (figure), 4-3
ESI probe assembly
figure, 2-6
installing, 2-6
removing, 6-2
ESI Source d.b.
figure, 2-13, 3-9
ESI/MS
defining scan parameters, 2-15
operational guidelines
table, 1-10
optimizing MS detector, 4-1
plumbing. See plumbing
preparing calibration solution for, B-4
setting up MS detector for, 4-7
testing MS detector, 3-3
using, discussion, 1-3
'JOOJHBO-$2 %60
ESI
syringe pump infusion, 2-8
ESI probe assembly, 2-6
ESI Source d.b., 2-13, 3-9
Instrument Configuration d.b., A-2, A-3
LCQ Configuration d.b., A-4, A-5
plumbing
loop injection, 5-3
loop injection, APCI, 8-2
loop injection, ESI, 5-2
syringe pump, 2-9, 2-10
Tee union, 4-4, 4-5, 4-6
Tee union, APCI, 6-5
Tee union, ESI, 4-3
Qual Browser window, 5-9, 8-8
results of loop injection, 5-8, 5-13, 8-6
Save As d.b., 3-10, 4-12, 7-4
Syringe Pump d.b., 3-3
Tune d.b., Collision Energy page, 5-12
Tune d.b., settings for automatic tuning, 3-6
Tune Plus window, 2-12
automatic calibration, 3-13
automatic calibration results, 3-15
Automatic page, Tune d.b., 4-11, 7-3
real-time plot of calibration solution in Spectrum
view, 3-5
results of automatic tune, 3-8
WideBand Activation checkbox, 5-11
flow rates
discussion, 1-9
note, 4-11
using LCQDUO syringe pump (note), 2-8
G
Graph view
displaying, 3-7
guidelines
LCQDUO operation (table), 1-10
H
Home Page, Xcalibur
displaying, 2-11
F
Figures
Accept Optimized Value d.b., 5-14
Acquire Data d.b., 5-6, 8-5
APCI probe assembly, 2-2
APCI Source d.b., 6-8
Calibrate d.b., 3-12
connecting PEEK safety sleeve, 2-4
Define Scan d.b., 2-15, 4-9, 5-5, 8-4
MS/MS Full scan settings, 5-11
Define Scan d.b., APCI settings, 6-10
Divert/Inject valve
positions for loop injection, 8-3
Divert/Inject Valve d.b., 4-10, 5-7, 8-6
I
Instrument Configuration d.b.
figure, A-2, A-3
Instrument Setup window
displaying, 2-11
introducing samples into LCQDUO
discussion, 1-6
ion source
exposing to atmosphere
CAUTION, 3-16, 7-6
II__________________________ Finnigan LCQDUO Getting Started
____________________
Index
'JOOJHBO-$2 %60 _______________________________________________________________________
refer to LCQDUO online Help, 2-2, 2-11, 4-7, 5-1
saving Tune Method, 3-10, 4-12, 7-4
solution flow rate, 4-11
solvents, 3-17, 7-7
syringe pump speed, 2-8
tube lens voltage, 4-11
use glass pipettes or stainless steel syringes with
glacial acetic acid, B-4
vials of sample formulations, B-2
L
LC
using buffers, discussion, 1-8
LCQ Configuration d.b
using, A-4
LCQ Configuration d.b.
figure, A-4, A-5
LCQDUO
flow rates, discussion, 1-9
operational guidelines
table, 1-10
sample introduction
discussion, 1-6
table, 1-7
using, discussion, 1-2
LCQDUO
configuring with Xcalibur, A-2
loop injection
reserpine results (figure), 5-8, 5-13, 8-6
setup for, 5-2, 8-2
M
methods
Tune, opening predefined, 2-13, 4-8, 6-8
Tune, saving (note), 3-10, 4-12, 7-4
MRFA
preparing stock solution, B-3
vial of sample formulation (note), B-2
MS detector
automatic calibration procedure (note), 3-7
calibrating, 3-12
cleaning for normal operation, 3-16, 7-6
ESI/MS mode, testing, 3-3
optimizing tune, LC/ESI/MS operation, 4-1
setting up for APCI/MS operation, 6-8
setting up for ESI/MS operation, 4-7
MS/MS
acquiring, data, 5-10
using, discussion, 1-2
N
nitrogen
sufficient flow for API source (CAUTION), 2-11
Notes
acquire data without optimizing, 4-2
configuring Xcalibur for LCQDUO, 2-1
cutting PEEK tubing, 6-6
displaying toolbars, Tune Plus window, 3-4
ensuring that MS detector is set up, tuned, and
calibrated, 4-2
ESI tune files, 2-12
heated capillary temperature, 4-11
MS detector, automatic calibration procedure, 3-7
optimizing MS detector tune, 3-7
refer to LCQDUO Getting Connected, 2-2, 2-11
refer to LCQDUO Hardware Manual, 2-2, 2-11, 4-7
O
optimizing
heated capillary and tube lens
automatically, 4-1
MS detector for LC/ESI/MS operation, 4-1
tune, automatically (figure), 3-8
optimizing collision energy
procedure, 5-13
P
PEEK safety sleeve
connecting, 2-4
figure, 2-4
connecting to ESI probe, 2-4
PEEK tubing
cutting (note), 6-6
pipettes
use glass, with glacial acetic acid (note), B-4
plumbing
divert/inject valve (figure), 4-6, 5-3
ESI, for syringe pump infusion (figure), 2-8
loop injection (figure), 5-3
loop injection, APCI (figure), 8-2
loop injection, ESI (figure), 5-2
syringe pump (figure), 2-9, 2-10
syringe pump, Tee to LC (figure), 4-4, 4-5
syringe pump, Tee to LC, APCI (figure), 6-5
syringe pump, Tee to LC, ESI (figure), 4-3
procedures
acquiring data using Tune Plus, 5-1, 8-1
acquiring data, SIM scan type, 5-5, 8-4
calibrating MS detector automatically, 3-12
cleaning MS detector after tuning, calibrating, 316, 7-6
configuring Xcalibur for LCQDUO, A-2
connecting PEEK safety sleeve to ESI probe, 2-4
installing ESI probe, 2-6
optimizing collision energy, 5-13
optimizing MS detector automatically, 4-10, 7-2
preparing a stock solution of reserpine, B-5
preparing an ESI sample solution of reserpine, B-5
preparing ESI calibration solution, B-4
preparing stock solutions, B-2
removing APCI probe, 2-2
saving Tune Method, 3-10, 4-12, 7-4
setting up MS detector for APCI/MS operation, 6-8
setup MS detector for tuning, calibrating, 2-1
____________________ Finnigan LCQDUO Getting Started _________________________
III
Index
_______________________________________________________________________
setup syringe pump for tuning, calibrating, 2-8
setup, loop injection, 5-2, 8-2
setup, tune MS detector with your analyte, 4-7
syringe pump, LC setup, 4-3
testing MS detector in ESI/MS mode, 3-3
tuning and calibrating
MS detector setup, 2-11
tuning and calibrating automatically, ESI mode, 3-1
tuning MS detector automatically with your analyte,
4-1
tuning MS detector automatically, ESI/MS mode,
3-6
Purge checkbox, 3-3
Q
Qual Browser window
figure, 5-9, 8-8
R
reserpine
acquiring, data, loop injection, 5-1, 8-1
preparing ESI sample solution, B-5
preparing stock solution, B-5
S
safety sleeve, PEEK
connecting, 2-4
figure, 2-4
sample formulations
preparing, B-2
sample solutions
ESI, reserpine, B-5
sample tube, 2-5
Save As d.b.
figure, 3-10, 4-12, 7-4
saving
files (figure), 3-10, 4-12, 7-4
saving Tune Method
note, 3-10, 4-12, 7-4
scan parameters
defining (figure), 2-15
sheath gas flow rate
figure, 2-13
SIM scan type
acquiring data in, 5-5, 8-4
solutions, preparing
caffeine, stock, B-3
calibrating, ESI, B-4
ESI sample, reserpine, B-5
MRFA, stock, B-3
reserpine, stock, B-5
stock, B-2
Ultramark 1621, stock, B-3
solvents
(note), 3-17, 7-7
'JOOJHBO-$2 %60
syringe pump
setup for tuning, calibrating, 2-8
speed (note), 2-8
Syringe Pump d.b.
figure, 3-3
syringes
use stainless steel, with glacial acetic acid (note),
B-4
T
Tables
APCI operational guidelines, 1-10
ESI operational guidelines, 1-10
introducing sample for calibrating, tuning, 1-12
sample introduction techniques, 1-7
toolbars
Tune Plus window (note), 3-4
tube lens
setting (figure), 2-13
voltage (note), 4-11
Tune d.b.
Automatic page, 4-11
Collision Energy page (figure), 5-12
figure, 3-6
showing settings for automatic tuning (figure), 3-6
Tune Methods
opening predefined, 2-13, 4-8, 6-8
saving (note), 3-10, 4-12, 7-4
Tune Plus window
figure, 2-12, 3-15
loop injection (figure), 5-8, 5-13, 8-6
opening, 2-11, 4-7
results of automatic calibration (figure), 3-13
showing automatic tune optimization (figure), 3-8
Spectrum view, 3-5
toolbars (note), 3-4
toolbars, extending, A-4
tuning (figure), 4-11, 7-3
tuning results (figure), 3-8
using to acquire data, 5-1, 8-1
tuning
automatically, 3-6
discussion, 1-11
ESI tune files (note), 2-12
MS detector
optimizing (note), 3-7
MS detector for LC/ESI/MS operation, 4-1
MS detector setup for, 2-11
syringe pump setup for, 2-8
voltages, 3-6
U
Ultramark 1621
preparing stock solution, B-3
vial of sample formulation (note), B-2
IV _________________________ Finnigan LCQDUO Getting Started
____________________
Index
'JOOJHBO-$2 %60 _______________________________________________________________________
W
Z
WideBand Activation
figure, 5-11
using, discussion, 1-2
ZoomScan
using, discussion, 1-2
X
Xcalibur
configuring, for LCQDUO, A-2
displaying Home Page, 2-11
____________________ Finnigan LCQDUO Getting Started __________________________ V
Was this manual useful for you? yes no
Thank you for your participation!

* Your assessment is very important for improving the work of artificial intelligence, which forms the content of this project

Download PDF

advertisement