Waters TQ Detector Quick Start Manual
Waters TQ Detector
Quick Start Guide
71500126803/Revision A
Copyright © Waters Corporation 2007.
All rights reserved.
Copyright notice
© 2007 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF
AMERICA AND IRELAND. ALL RIGHTS RESERVED. THIS DOCUMENT
OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM
WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER.
The information in this document is subject to change without notice and
should not be construed as a commitment by Waters Corporation. Waters
Corporation assumes no responsibility for any errors that may appear in this
document. This document is believed to be complete and accurate at the time
of publication. In no event shall Waters Corporation be liable for incidental or
consequential damages in connection with, or arising from, its use.
Waters Corporation
34 Maple Street
Milford, MA 01757
USA
Trademarks
Connections Insight, ESCi, and Waters are registered trademarks of Waters
Corporation. ACQUITY UPLC, IntelliStart, IonSABRE, MassLynx, T-Wave,
UPLC, and ZSpray are trademarks of Waters Corporation.
Other trademarks or registered trademarks are the sole property of their
respective owners.
Customer comments
Waters’ Technical Communications departme nt invites you to tell us of any
errors you encounter in this document or to suggest ide as for otherwise
improving it. Please help us better understand what you expect from our
documentation so that we can continuously improve its accuracy and
usability.
We seriously consider every customer comment we receive. You can reach us
at tech_comm@waters.com.
Waters Corporation
34 Maple Street
Milford, MA 01757
USA
iii
Safety considerations
Some reagents and samples used with Waters® instruments ca n po se
chemical, biological, and radiological hazard s. Be sure you are aware of the
potentially hazardous effects of all substances you work with. Always observe
Good Laboratory Practice (GLP) guidelines, published by the U.S. Food and
Drug Administration, and consult your organization’s safety representative
for guidance.
When you develop methods, follow the “Pro tocol for the Ado ption of Analyt ical
Methods in the Clinical Chemistry Laboratory,” Ame rican Journal of Medical
Technology, 44, 1, pages 30–37 (1978). This protocol addresses good operating
procedures and the techniques necessary to validate system and method
performance.
Instrument-specific safety consideratio ns
Solvent leakage hazard
The source exhaust system is designed to be robust and leak-tight. Waters
recommends you perform a hazard analysis, assuming a maximum leak into
the laboratory atmosphere of 10% HPLC eluate.
Warning:
• To confirm the integrity of the source exhaust system, the
source seals must be renewed at intervals not exceeding one
year.
• The source seals can withs tand exposure only to specific
solvents, see Appendix C in the Wat ers TQ Det ector Oper ator’ s
Guide. If you intend to use solvents other than those listed,
you must first determine whether those solvents are
compatible with the composition of the seals.
iv
Flammable solvents hazard
Warning: Where signif icant quantities of flammable solvents are
involved, the source must receive a continuous flow of nitrogen to
prevent possible ignition within that enclos ed space.
Never let the nitrogen supply pressure fall below 400 kPa (4 bar, 58 psi)
during analyses that require flammable solvents. Connect to the LC output
with a gas-f a il co n n e ctor to stop the LC solvent if th e ni t rogen suppl y fa i l s.
High temperature hazard
Warning: The source enclosure can be hot. To avoid burn injuries,
avoid touching the source enclosure when operating or servicing the
TQ Detector.
Waters TQ Detector high temperature hazard:
Source enclosure ass em bly
v
High voltage hazard
Warning:
• To avoid electric shock, do not remove the TQ Detector’s protective
panels. The components they cover are not user-serviceable.
• To avoid non-lethal electric shock, any equipment connected to the
ESI and IonSABRE™ APCI probes must be grounded.
• When the TQ Detector is in Operate mode, certain ext ernal surfaces
can conduct high voltages. To avoid non-lethal electric shock, make
sure the instrument is in Standby mode before touching areas
marked with this high voltage warning symbol.
Safety advisories
Consult Appendix A for a comprehensive list of warning and caution
advisories.
vi
Operating this device
When operating this device, follow standard quality cont rol procedures and
the guidelines presented in this section.
Symbols
Symbol Definition
Intended use
The Waters Tandem Quadrupole (TQ) Detector is intended to be used as a
research tool to deliver authentica ted mass measurement in both MS and
MS/MS modes.
The Waters TQ Detector can be used for general In Vitro Diagnostic
applications, only by professionally trained and qualified laboratory
personnel.
E C RE P
Authorized Representative of the
European Community
The CE symbol serves as
confirmation of the conformity of a
product with all European
Community directives applicable to
that product.
For in vitro diagnostic use.
IV D
The Waters TQ Detector is CE-marked according to the
European Union In Vitro Diagnostic Device Directive 98/79/EC.
vii
Calibration
To calibrate LC systems, follow acceptable calibration method s using at least
five standards to generate a standard curve . The concentration range for
standards should cover the entire range of quali ty-control samples, typical
specimens, and atypical specimens.
To calibrate mass spectrometers, consult the calibration section of the
operator’s guide of the instrument you are calibrating.
Quality control
Routinely run three quality-control samples that represent subnormal,
normal, and above-normal levels of a compound. Ensure that quality-control
sample results fall within an acceptable range, and evaluate precision from
day to day and run to run. Data col lected when qualit y control samples are ou t
of range might not be valid. Do not r eport the se dat a until you ar e cert ain that
the instrument performs satisfactorily.
When analyzing samples from a co m plex matrix such as soil, tissue,
serum/plasma, whole blood, etc., note tha t the mat rix components can
adversely affect LC/MS results, enhancing or suppres sing ionization. To
minimize these matrix effects, Wate rs recommends you adopt the following
measures:
viii
• Prior to the instrumental analysis, use appropria te sample
pretreatment such as protein precipitation, liquid/liquid extraction
(LLE), or solid phase extraction (SPE) to remove matrix interferences.
• Whenever possible, verify method accuracy and precision using
matrix-matched calibrators and QC samples.
• Use one or more internal standard compounds, preferably
isotopically-labeled analytes.
IVD authorized representative information
IVD authorized representative
Waters Corporation (Micromass UK Limited) is
registered in the United Kingdom with the
Medicines and Healthcare P roducts Regulatory
Agency (MHRA) at market Towers, 1 Nine Elms
Lane, London, SW8 5NQ. The referenc e number
is IVD000167.
Waters Corp o ra tion (Microm a ss UK Ltd.)
Floats Road
Wythenshawe
Manchester M23 9LZ
United Kingdom
Telephone: +44-161-946-2400
Fax: +44-161-946-2480
Contact: Quality manager
ix
x
Table of Contents
Safety considerations .......................................................................................... iv
Instrument-s pe c if i c sa f et y co n si d erations............. ... ............. .. .......................... iv
Flammable solvents hazard ................................................................................ v
High temperature hazard.................................................................................... v
High voltage haza r d......... .. .......................... ... ............. .. .. ............. ... ................... vi
Safety advisories................................................................................................. vi
Operating this device ......................................................................................... vii
Symbols ............................................................................................................. vii
Intended use...................................................................................................... vii
Calibration ....................................................................................................... viii
Quality control ................................................................................................. viii
IVD authorized representative information ................................................. ix
IVD authorized representative .......................................................................... ix
1 Waters TQ Detector Overview ............................................................ 1-1
Overview ............................................................................................................. 1-2
Waters TQ Dete c to r.......................... ... ............. .. ............. .. .............. .. ............. . 1-2
ACQUITY TQD UPLC/MS system .......... .. .. ................................................... 1-4
MassLynx mass spectrometry software and data system ............................. 1-5
ACQUITY UPLC Console......................................................................... ....... 1-6
Ionization techniques and source probes ................................................... 1-7
Electrospray i on i za tion (ESI)..... .. ............. .. .............. .. ............. .. .............. .. ..... 1-7
Combined electrospray ionization and atmospheric p ressure chemical ionization
(ESCi) ......................................................................................................... 1-7
Atmospheric pressure chemical ionization..................................................... 1-8
Atmospheric pressure photoionization ........................................................... 1-8
Table of Contents xi
Ion optics ............................................................................................................. 1-9
MS operating modes ....................................................................................... 1-10
MS/MS operating modes ................................................................................ 1-11
Product (daug h t er) ion mode. .............. .. ............. .. ............. ... ............. .. .......... 1-11
Precursor (pare n t) ion mode...... ............. .. ............. ... ............. .. .. .............. .. ... 1-1 2
Multiple reaction monitoring mode .............................................................. 1-12
Constant neutral loss mode........................................................................... 1-13
Sample inlet ...................................................................................................... 1-14
Vacuum system ................................................................................................ 1-14
Rear panel .. ... ............. .. .. .. ... .. .. ............. ... .. .. .. .. .............. .. .. .. .. ... ............. .. .. ... .. .. . 1-15
IntelliStart fluidics system overview ......................................................... 1-16
IntelliStart fluidics system operation ........................................................ 1-17
Operating the IntelliStart fluidics system from the ACQUITY UPLC
Console . .. ............. .. .............. .. ............. .. .............. .. ............. .. .............. .. .. ... 1-1 7
Operating the IntelliStart fluidics system from the Tune window............. 1-18
Programming the MS method to operate the IntelliStart fluidics system. 1-18
2 Preparing the Waters TQ Detector for Operation ......................... 2-1
Starting the TQ Detector ................................................................................. 2-2
Configuring IntelliStart..................... ............................................... .. ............. 2-6
Verifying the instrument’s state of readiness................................................ 2-6
Tuning and calibration information ............................................................... 2-6
Running the TQ Detector at high flow rates.................................................. 2-7
Monitoring the TQ Detector LEDs.................................................................. 2-8
Preparing the IntelliStart fluidics system .................................................. 2-9
Installing the solvent manifold drip tray........................... .. .......................... 2-9
Installing the reservoir bottles..................... .......................................... ....... 2-10
Diverter valve positions................................................................................. 2-11
Purging the infusion syringe......................................................................... 2-13
Rebooting the TQ Detector ........................................................................... 2-14
Rebooting the TQ Detector by pressing the reset button ............................ 2-14
xii Table of Contents
Shutting down the TQ Detector .................................................................. 2-15
Putting the TQ Detector in Standby mode for overnight shutdown........... 2-15
Complete TQ Detector shutdown.................................................................. 2-16
Emergency TQ Detector shutdown............................................................... 2-17
3 ESI and ESCi Modes of Operation ..................................................... 3-1
Introduction ....................................................................................................... 3-2
Installing the ESI probe .................................................................................. 3-2
Required materials .......................................................................................... 3-2
Installing the corona pin ................................................................................. 3-5
Required materials .......................................................................................... 3-5
Optimizing the ESI probe for ESCi operation ........................................... 3-7
Required materials .......................................................................................... 3-7
Removing the corona pin ................................................................................ 3-9
Required materials .......................................................................................... 3-9
Removing the ESI probe ............................................................................... 3-11
Required materials ........................................................................................ 3-11
4 Operating the Waters TQ Detector .................................................... 4-1
Setting-up the instrument .............................................................................. 4-2
Required materials .......................................................................................... 4-2
Performing a sample tune .............................................................................. 4-7
Required materials .......................................................................................... 4-7
Developing experiment methods .................................................................. 4-9
Required materials .......................................................................................... 4-9
Verifying the sys t em us in g Sys t em QC ........... ............. .. ... .. .. .. .. .............. .. . 4-11
A Safety Advisories ..................... ..... ..... ................................................... A-1
Warning symbols ............................................................................................... A-2
Task-specific hazard warnings........................................................................ A-2
Warnings that apply to particular instruments, instrument components, and
sample types............................................................................................... A-3
Table of Contents xiii
Caution symbol .................................................................................................. A-5
Warnings that apply to all Waters instruments ......................................... A-5
Electrical and handling symbo l s ............. .. ... .. .. .. ............. ... .. .. .. .. ... ............. . A-13
Electrical symbols.......................................................................................... A-13
Handling symbols .......................................................................................... A-14
xiv Table of Contents
1 Waters TQ Detector Overview
This chapter describes the instrument, including its controls and gas
and plumbing connections.
Contents:
Topic Page
Overview 1-2
Ionization techniques and source probes 1-7
Ion optics 1-9
MS operating modes 1-10
MS/MS operating modes 1-11
Sample inlet 1-14
Vacuum system 1-14
Rear panel 1-15
IntelliStart fluidics system overview 1-16
IntelliStart fluidics system operation 1-17
1-1
Overview
Waters TQ Detector
The Waters® TQ Detector is a tandem quadrupole, atmospheric pressure
ionization (API) mass spectrometer. Designed for routine UPLC™/MS/MS
analyses in quantitative and qualitative applications, it can operate at fast
acquisition speeds compatible with ultra performance LC.
Waters provides these ion sources with the instrument as standard
equipment:
• ZSpray™ (dual orthogonal sampling) interface.
• Multi-mode ESCi
chemical ionization (APCI) and electrospray ionization (ESI).
Optional ionization modes are IonSABRE™ APCI and APPI (atmospheric
pressure photoionization) .
For TQ Detector specifications, see the Waters TQ Detector Site Preparation
Guide.
Waters TQ Detector:
®
ionization switching for atmospheric pressure
1-2 Waters TQ Detector Overview
TP02592
Waters TQ Detector with doors open:
HV
PROBE
NEBULIZER
POWER OPERATE
DESOLVATION
V
H
E
B
O
R
P
IntelliStart technology
TP02627
IntelliStart™ technology monitors LC/MS/MS performance and reports when
the TQ Detect or is ready for use.
The IntelliStart software automatically tune s and mass calibrates the TQ
Detector and displays performance readbacks . In tegrated with MassLynx™
mass spectrometry software and ACQUITY UPLC™ Console software,
IntelliStart enables simplified set-up of the system for use in routine
analytical and open access applications.
The IntelliStart fluidics system is built into the TQ Detector. It delivers
sample directly to the MS probe from the LC column or from two integral
reservoirs. The integral reservoirs can also deliver sample through direct or
combined infusion so that you can optimize instrument performance at
analytical flow rates.
See also: The TQ Detector online Help for further details of IntelliStart.
Overview 1-3
ACQUITY TQD UPLC/MS system
Note: The Waters TQ Detector is designed for compatib ility with the
ACQUITY UPLC system; if you are not using an ACQUITY UPLC system,
refer to the documentation relevant to the LC syst em being used.
The ACQUITY TQD UPLC/MS system includes an ACQUITY UPLC system
and the Waters TQ Detector.
ACQUITY UPLC system
The ACQUITY UPLC system includes a binary solvent manager, sample
manager, column heater, optional sample organizer, op tional detectors, and a
specialized ACQUITY UPLC column. Waters MassLynx mass spectrometry
software controls the system.
See also:
• ACQUITY UPLC System Operator’s Guide
• Controlling Contamination in LC/MS System s (part number
715001307). You can find this document on http: //www.waters.com; click
Services and Support and then Support Center.
1-4 Waters TQ Detector Overview
Waters ACQUITY TQD:
Sample organizer (option al)
manager
Solvent tray
Column heater
TP02597
Sample managerBinary solvent
TQ Detector
MassLynx mass spectrometry software and data system
MassLynx is a high-performance mass spect rometry application that acquires,
analyzes, manages, and distributes ultra-violet (UV), evaporative light
scattering, analog, and mass spectrometry data.
MassLynx software permits these major operations:
• Configuring the instrument.
• Creating LC inlet and MS/MS methods that define operating
parameters for a run.
• Using IntelliStart softwar e to tune and m a ss calibrate the TQ Detector.
• Running samples.
• Monitoring the run.
• Acquiring data.
• Processing data.
Overview 1-5
•Reviewing data.
•Printing data.
See also: MassLynx 4.1 user documentation and online Help for more
information on installing and using MassLynx software.
ACQUITY UPLC Console
The ACQUITY UPLC Console is a software application with which you
configure settings, monitor perfor manc e, run diagnostic tests, and maintain
the system and its modules. The ACQUITY UPLC Console functions
independently of MassLynx and does not recognize or control the data system.
See also: ACQUITY UPLC System console online Help for details of the TQ
Detector.
TQ Detector ACQUITY UPLC Console page:
1-6 Waters TQ Detector Overview
Ionization techniques and source probes
Electrospray ionization (ESI)
In electrospray ionization (ESI), a strong electrical charge is given the eluent
as it emerges from a nebulizer. The droplets that compose the resultant
aerosol undergo a reduction in size (solvent evaporat ion). As solvent continues
to evaporate, the charge density incre ases until the dr oplet su rfac es eject io ns
(ion evaporation). The ions can be singly or multiply charged. The multiply
charged ions are of particular interest because the TQ Detector separates
them according to their mass-to-charge ratios (m/z), permitting the detection
of high-molecular-weight compounds.
The instrument can accommodate eluent flow rates of up to 1 mL/min.
Combined electrospray ionization and atmospheric pressure chemical ionization (ESCi)
Combined electrospray ionization and atmospheric pressure chemical
ionization (ESCi) is supplied as standard equipment on the TQ Detector. In
ESCi, the standard ESI probe is used in conjunction with a corona pi n to allow
alternating acquisition of ESI and APCI ionization data, facilitating high
throughput and wider compound coverage.
Ionization techniques and source probes 1-7
ESCi mode:
Corona pin
TP02695
Sample cone tip
See also: “Electrospray ionization (ESI)” on page 1-7.
Atmospheric pressure chemical ionization
A dedicated high performance atmospheric pressure chemical ionization
(APCI) probe is offered as an option.
See also: The Waters TQ Detector Operator’s Guide for full details.
Atmospheric pressure photoionization
Atmospheric pressure photoionization (APPI) is offered as an option. It uses
photons generated by a krypton-dischar ge ultra-violet (UV) lamp ( ∼10.2 eV) to
produce sample ions from vaporized LC eluent.
1-8 Waters TQ Detector Overview
Ion optics
The TQ Detector’s ion optics operate as follows:
• Samples from the LC or Intellistart fluidics system are introduced at
atmospheric pressure into the ionization source.
• The ions pass through the sample cone into the vacuum system.
• The ions pass through the transfer optics to the fir st quadrupole where
they are filtered according to their mass-to-charge ratio (m/z).
• The mass-separated ions pass into the T-Wave™ collision cell where
they either undergo collision-induced dissociation (CID) or pass to the
second quadrupole. Any fragment ions are then mass-analyzed by the
second quadrupole.
• The transmitted ions are detected by the photomultiplier detection
system.
• The signal is amplified, digitized, and sent to the MassLynx mass
spectrometry software.
Ion optics overview:
Sample cone
Sample inlet Transfer optics
T-W ave co llision
cell
Conversion dynode
Isolation valve
Z-Spray ion sourc e Quadrupole 1
(MS1)
Quadrupole 2
(MS2)
Detector
Ion optics 1-9
MS operating modes
The following table shows the MS operating modes.
MS operating modes :
Operating mode MS1 Collision cell MS2
MS1 Resolving
(scanning)
MS2 Pass all masses Resolving
SIR Resolving (static) Pass all masses
The MS1 mode, in which MS1 is used as the mass filter, is the most common
and most sensitive method of per formi ng MS analysis. It i s di rectly analogous
to using a single-quadrupole mass spectrometer.
The MS2 mode of operation is used, with collision gas p resent, when switching
rapidly between MS and MS/MS operat ion (for example, in survey scan mode).
It also provides a useful tool for instrument tuning and calibration before
MS/MS analysis and for fault diagnosis.
The selected ion recording (SIR) mode of operation is used as a quantitation
mode when no suitable fragment ion can be found to perform a more specific
multiple reaction monitoring (MRM) analysis.
Pass all masses
(scanning)
1-10 Waters TQ Detector Overview
MS/MS operating modes
The following table shows the MS/MS operating modes.
MS/MS operating modes :
Operating mode MS1 Collision cell MS2
Product
(daughter) ion
Static (at
precursor mass)
spectrum
Precursor
Scanning Static (at product
(parent) ion
spectrum
MRM Static (at
precursor mass)
Constant neutral
loss spectrum
Scanning
(synchronized
with MS2)
Product (daughter) ion mode
Product ion mode is the most commonly used MS/MS operating mode. An ion
of interest is selected for fragment ation in the collision cell, thus yielding
structural information.
Product ion mode:
Pass all masses Scanning
mass)
Static (at product
mass)
Scanning
(synchronized
with MS1)
MS1
Static (at precursor mass)
Collision cell
Pass all masses
Typical applications
• Structural elucidation (for example, peptide sequencing)
MS/MS operating modes 1-11
MS2
Scanning
• Method development for MRM screening studies:
– Identifying product ions for use in MRM transitions.
– Optimizing CID tuning conditions to maximize the yield of a specific
product ion to be used in MRM analysis.
Precursor (par ent) ion mode
Precursor ion mode:
MS1
Scanning
Typical application
You typically use the precursor ion mode for structural elucidation–that is, to
complement or confirm product scan data–by scanning for all the precursors of
a common product ion.
Multiple reaction monitoring mode
MRM mode is a highly selectiv e MS/MS equivalent of SIR. As both MS1 and
MS2 are static, greater dwell time on the ions of interest is allowed, and
therefore better sensitivity compared to scanning MS/MS. This is the most
commonly used acquisition mode for quantitative analysis, allowing the
compound of interest to be isolated from the chemical background noise.
Collision cell
Pass all masses
MS2
Static (at product mass)
1-12 Waters TQ Detector Overview
Multiple reaction monitoring mode:
MS1
Static (at precursor mass)
Typical application
You typically use MRM to quantify known analytes in complex samples:
• Drug metabolite and pharmacokinetic studies.
• Environmental, for example, pest icide and herbicide analysis.
• Forensic or toxicology, for example, screening for target drugs in sport.
• MRM does not produce a spectrum because only one transition is
monitore d at a time. As in SI R , a c h romatogram is produce d.
Constant neutral loss mode
Constant neutral loss mode detects the loss of a specific neutral fragment or
functional group from an unspecified precur sor or precursors.
The scans of MS1 and MS2 are synchronized. When MS1 transmits a specific
precursor ion, MS2 “lo oks” to s ee whether that p recu rsor lo ses a fra gment of a
certain mass. If it does, the loss registers at the detector.
In constant neutral loss mode, the spectrum show s the masses of all
precursors that actually lost a fragment of a certain mass .
Collision cell
Pass all masses
MS2
Static (at product mass)
MS/MS operating modes 1-13
Constant neutral lo ss mode:
Typical application
You typically use constant neutral loss mode to screen mixtures for a specific
class of compound that is character ized by a common f ragmentat ion pathway,
indicating the presence of compounds containing a common functional group.
Sample inlet
Either of two methods delivers solvent and sample to the installed probe:
• An LC system, which delivers the eluent from an LC analysis.
• IntelliStart fluidics system, which uses on-board solutions to automate
instrument optimization. You can de liver solutions by direct or
combined infusion.
Vacuum system
MS1
Scanning
(synchronized with MS2)
Collision cell
Pass all masses
MS2
Scanning
(synchronized with MS1)
An external roughing (rotary vane) pump and an internal split- flow
turbomolecular pump combine to create the source vacuum. The
turbomolecular pump evacuates the analyzer and ion transfer region.
Vacuum leaks and electrical or vacuum pump failures cause vacuum loss,
which protective interlocks guard against. The system monitors
turbomolecular pump speed and continuously measures vacuum pressure
with a built-in Pirani gauge. The gauge also serves as a switch, discontinuing
operation when it senses vacuum loss.
A vacuum isolation valve isolates the source from the mass analyzer, allowing
routine source maintenance without venting.
1-14 Waters TQ Detector Overview
Loading...