AB SCIEX TripleTOF® 5600/5600+
Instruments
System User Guide
Document Number: D5033331 A
Release Date: May 2012
This document is provided to customers who have purchased
AB Sciex equipment to use in the operation of such AB Sciex
equipment. This document is copyright protected and any reproduction
of this document or any part of this document is strictly prohibited,
except as AB Sciex may authorize in writing.
Software that may be described in this document is furnished under a
license agreement. It is against the law to copy, modify, or distribute
the software on any medium, except as specifically allowed in the
license agreement. Furthermore, the license agreement may prohibit
the software from being disassembled, reverse engineered, or
decompiled for any purpose.
Portions of this document may make reference to other manufacturers
and/or their products, which may contain parts whose names are
registered as trademarks and/or function as trademarks of their
respective owners. Any such use is intended only to designate those
manufacturers' products as supplied by AB Sciex for incorporation into
its equipment and does not imply any right and/or license to use or
permit others to use such manufacturers' and/or their product names
as trademarks.
AB Sciex makes no warranties or representations as to the fitness of
this equipment for any particular purpose and assumes no
responsibility or contingent liability, including indirect or consequential
damages, for any use to which the purchaser may put the equipment
described herein, or for any adverse circumstances arising therefrom.
For research use only. Not for use in diagnostic procedures.
The trademarks mentioned herein are the property of
AB Sciex Pte. Ltd. or their respective owners.
AB SCIEX™ is being used under license.
AB SCIEX
71 Four Valley Dr., Concord, Ontario, Canada. L4K 4V8.
AB Sciex LP is ISO 9001 registered.
© 2012 AB SCIEX.
Printed in Canada.
Contents
Safety and Regulatory Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
General Safety Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7
Symbols and Conventions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7
Qualified Personnel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8
Equipment Use and Modification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8
Shipping Crate Labels and Indicators . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9
Regulatory Compliance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10
Australia and New Zealand . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10
Canada . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10
Europe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10
United States . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10
International . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11
Symbols and Labels on the Mass Spectrometer . . . . . . . . . . . . . . . . . . . . . . . . .11
Occupational Health and Safety Symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .13
Mains Supply . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14
Laboratory Ventilation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15
Environmental Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15
Instrument Disposal (Waste Electrical and Electronic Equipment) . . . . . . . . . . .16
Chapter 1 System Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Theory of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .17
Data Handling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .17
System Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .18
Dress Panel LEDs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .20
Start Up the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .20
Shut Down the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .21
Use the Integrated Syringe Pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .22
Edit the Hardware Profile for the Integrated Syringe Pump . . . . . . . . . . . . . . .22
Adjust the Integrated Syringe Pump Position . . . . . . . . . . . . . . . . . . . . . . . . .23
Configure the Integrated Syringe Pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . .25
Reset the Syringe Pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .26
Instrument Safe Fluids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .26
Chapter 2 Hardware Profiles and Projects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Sample Workflows . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .27
Hardware Profiles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .29
Create a Hardware Profile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .29
Add Peripheral Devices to Hardware Profiles . . . . . . . . . . . . . . . . . . . . . . . . .33
Troubleshoot Hardware Profile Activation . . . . . . . . . . . . . . . . . . . . . . . . . . . .34
Create Projects and Subprojects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .35
Create a New Subproject . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .36
Copy a Subproject . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .36
Switch Between Projects and Subprojects . . . . . . . . . . . . . . . . . . . . . . . . . . .37
Installed Project Folders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .37
Back up the API Instrument Project . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .38
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Chapter 3 Instrument Tuning and Calibrating. . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Required material . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .39
Prerequisites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .39
Optimize the Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .39
Chapter 4 Basic Acquisition Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
Create an Acquisition Method using the Method Wizard . . . . . . . . . . . . . . . . . .41
Create Acquisition Methods using the Acquisition Method Editor . . . . . . . . . .42
Add an Experiment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .43
Add a Period . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .43
Copy an Experiment into a Period . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .43
Copy an Experiment within a Period . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .43
Scan Techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .44
Single Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .44
Quadrupole-Based Single Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . .44
Time-of-Flight Single Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . .44
Tandem Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .44
Product Ion Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .45
Precursor Ion Mass Spectrometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .45
About Spectral Data Acquisition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .45
About Instrument Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .45
Acquisition Method Editor Icons . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .49
Chapter 5 Batches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
Create and Submit a Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .51
Set Queue Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .51
Add Sets and Samples to a Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .53
Set up Sample Calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .54
Submit a Sample or a Set of Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .55
Change Sample Order . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .55
Acquire Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .55
Set Sample Locations in the Batch Editor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .56
Select Vial Positions using the Locations Tab (Optional) . . . . . . . . . . . . . . . . . .57
Stop Sample Acquisition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .57
Batch and Acquisition Method Editor Tips . . . . . . . . . . . . . . . . . . . . . . . . . . . . .58
Batch Editor Right-Click Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .58
Queue States and Device Status . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .59
Queue States . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .59
View Instrument and Device Status Icons . . . . . . . . . . . . . . . . . . . . . . . . . . . .60
Queue Right-Click Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .62
Chapter 6 Analyzing and Processing Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
Open Data Files . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .65
Navigate Between Samples in a Data File . . . . . . . . . . . . . . . . . . . . . . . . . . .65
Show Experimental Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .66
Show Experimental Data in Tables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .67
Show ADC Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .67
Show Basic Quantitative Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .68
Chromatograms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .68
Show TICs from a Spectrum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .69
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Show a Spectrum from a TIC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .69
Generate XICs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .69
Generate BPCs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .72
Generate XWCs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .74
View DAD Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .74
Generate TWCs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .74
Adjust the Threshold . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .75
Data Processing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .77
Graphs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .77
Zoom in on the y-axis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .79
Zoom in on the x-axis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .79
Chapter 7 DuoSpray™ Ion Source User Reference . . . . . . . . . . . . . . . . . . . . . . 83
Introduction to the Ion Source . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .83
Probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .84
Gas and Electrical Connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .86
Ion Source Latch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .86
Source Exhaust System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .86
Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .87
Install the Probe in the Ion Source Housing . . . . . . . . . . . . . . . . . . . . . . . . . .87
Install the Ion Source . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .88
Connect the Sample Tubing and Cables for Sample Introduction with
the TurboIonSpray Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .88
Connect the Sample Tubing and Cables for Sample Introduction with
the APCI Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .89
Optimization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .89
Optimize the TurboIonSpray Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .90
Optimize the APCI Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .92
Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .95
Clean the Probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .95
Remove the Ion Source . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .96
Remove the Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .96
Clean the Electrode Tube . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .97
Adjust the Electrode Tip Extension . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .98
Remove the Corona Discharge Needle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .99
Replace the Sample Tubing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .100
Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .101
Consumables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .102
Chapter 8 Cleaning and Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103
Health and Safety Precautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .103
Clean the Surfaces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .104
Empty the Drain Bottle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .104
Front-End Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .105
Best Practices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .106
Prepare for Routine Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .107
Clean the Curtain Plate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .108
Clean the Front of the Orifice Plate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .109
Put the Instrument Back into Service . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .109
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Chapter 9 Basic System Troubleshooting. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
Appendix A Recommended Calibration Ions . . . . . . . . . . . . . . . . . . . . . . . . . . .113
Appendix B Exact Masses and Chemical Formulas . . . . . . . . . . . . . . . . . . . . .115
PPG . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .115
Reserpine (C33H40N2O9) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .116
Taurocholic Acid (C26H45NO7S) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .116
TOF Calibration Solution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .116
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Safety and Regulatory Information
This section contains general safety-related information, describes the symbols and conventions
used in the documentation, and provides regulatory compliance information. It also describes
potential hazards and associated warnings for the system, and the precautions that should be
taken to minimize the hazards. In addition to this section, refer to the Site Planning Guide. It
provides site requirements, including requirements for the mains supply, source exhaust,
ventilation, compressed air, nitrogen, and the roughing pump.
General Safety Information
Before operating any instrument, become familiar with its operation and with the potential
hazards. To prevent personal injury or instrument damage, read, understand, and obey all safety
precautions. Warnings in this document and labels on the mass spectrometer are shown with
international symbols. Failure to heed these warnings could result in serious injury.
This safety information is intended to supplement federal, state or provincial, and local
environmental health and safety (EHS) regulations. The information provided covers instrumentrelated safety with regard to the operation of the mass spectrometer. It does not cover every
safety procedure that should be practised. Ultimately, you and your organization are responsible
for compliance with federal, state or provincial, and local EHS regulations and for maintaining a
safe laboratory environment.
For more information, refer to the appropriate laboratory reference material and standard
operating procedures.
Symbols and Conventions
The following conventions are used throughout the guide.
DANGER! Danger signifies an action which leads to severe injury or death.
WARNING! Personal Injury Hazard: A warning indicates an operation that could
cause personal injury if precautions are not followed.
WARNING! Electric Shock Hazard: This symbol indicates a warning of electrical
shock hazard. Read the warning and follow all precautions before performing any
operation described in the guide. Failure to do so can result in serious injury.
WARNING! Burn Hazard: This symbol indicates a warning of potential burns
from hot surfaces. Read the warning and follow all precautions before performing
any operation described in the guide. Failure to do so can result in serious injury.
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WARNING! Biohazard: This symbol indicates a warning of biohazardous
materials. Read the warning and follow all precautions before performing any
operation described in the guide. Failure to do so can result in serious injury.
Caution: A caution indicates an operation that could cause damage to the instrument or
loss of data if precautions are not followed.
Tip! Provides useful information that helps apply the techniques and procedures in
the text for a specific need, and provides shortcuts, but is not essential to the completion
of a procedure.
i
Note: A note emphasizes significant information in a procedure or description.
Qualified Personnel
After installing the system, the FSE (Field Service Employee) uses the Customer Familiarization
Checklist to train the customer on system operation, cleaning, and basic maintenance. Only
qualified AB SCIEX personnel shall install and service the equipment. Only personnel qualified
by AB SCIEX shall operate and maintain the equipment. Contact an AB SCIEX FSE for more
information.
Equipment Use and Modification
Use the system indoors in a laboratory that complies with the environmental conditions
recommended in the system Site Planning Guide. If the system is used in an environment or in a
manner not prescribed by AB SCIEX, the protection provided by the equipment can be impaired.
Unauthorized modification or operation of the system may cause personal injury and equipment
damage, and may void the warranty. Erroneous data may be generated if the system is operating
outside the recommended environmental conditions or with unauthorized modifications. Contact
an AB SCIEX representative for more information on servicing the system.
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Shipping Crate Labels and Indicators
Table 1 Labels and Indicators on the Crate
External Labels and
Indicators
Definition
Action
TIP N TELL
Blue beads in the arrow
Write on the Bill of Lading and
indicate that the container was check for damage. Any claims
tipped or mishandled.
for tipping require a notation.
or
Shock Indicator
The indicator is broken if the
container has suffered a shock
greater than the level marked
on the indicator.
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Write on the Bill of Lading and
check for damage. Any claims
for shock damage require a
notation.
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Safety and Regulatory Information
Regulatory Compliance
This system complies with the standards and regulations listed in this section. Applicable labels
have been affixed to the system.
Australia and New Zealand
•
Electromagnetic Interference—AS/NZ CISPR 11 (Class A)
•
Safety—AS/NZ 61010-1
Canada
•
Electromagnetic Interference—CAN/CSA CISPR11-04. This ISM device complies
with Canadian ICES-001.
•
Safety—CAN/CSA C22.2 No. 61010-1-04, CAN/CSA C22.2 No. 61010-2-061:04
Europe
•
•
•
Electromagnetic Compatibility—Electromagnetic Compatibility Directive
2004/108/EC, as implemented in these standards:
•
EN 55011 (Class A)
•
EN 61326-1
Safety—Low Voltage Directives 2006/95/EC as implemented in these standards:
•
EN 61010-1
•
EN 61010-2-061
WEEE—Waste, Electrical, and Electronic Equipment Directive 2002/96/EEC, as
implemented in EN 40519
United States
•
Electromagnetic Interference, FCC Part 15, Class A—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 (Federal Communications Commission Compliance) 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 and, if not
installed and used in accordance with the operator’s manual, can cause harmful
interference to radio communications. Operation of this equipment in a residential
area is likely to cause harmful interference in which case you will be required to
correct the interference, at your own expense. Changes or modifications not
expressly approved by the manufacturer could void your authority to operate the
equipment.
•
Safety—UL 610101-1-04
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International
•
Electromagnetic Compatibility—IEC 61326-1; CEI/IEC CISPR 11
•
Safety—IEC 61010-1; IEC 61010-2-061
For more information, refer to the Declaration of Conformance included with the system.
Symbols and Labels on the Mass Spectrometer
Table 2 Labels on the Mass Spectrometer
External Labels
Definition
High Voltage
WARNING: NO USER SERVICEABLE PARTS
INSIDE. REFER SERVICING TO QUALIFIED
PERSONNEL.
WARNING: To avoid risk of injury, ensure turbo pump
clamps are mounted with manufacturer’s specified
torque setting. Please contact your Factory
Authorized Service Representative for assistance
prior to replacement.
EN61326—1:2006 CLASS A, GROUP 1, ISM
EQUIPMENT
This ISM device complies with Canadian ICES-001.
Cet appareil ISM est conforme à la norme NMB-001
du Canada.
FCC Compliance. This device complies with Part 15
of the FCC Rules. Operation is subject to the
following conditions: (1) this device may not cause
harmful interference, and (2) this device must accept
any interference received, including interference that
may cause undesired operation.
0211-3199
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Table 2 Labels on the Mass Spectrometer (Continued)
External Labels
Definition
Do not dispose of equipment as unsorted municipal
waste (WEEE).
FOR RESEARCH USE ONLY. NOT FOR USE
IN DIAGNOSTIC PROCEDURES.
This mass spectrometer is for research use only. It is
not intended for use in diagnostic procedures.
This instrument contains the following technology:
LINAC® Collision Cell Curtain Gas™ Interface
WARNING: Hot Surface Hazard.
Operator Guide
Follow operating instructions (mandatory)
Alternating Current
A
Amperes (current)
High voltage. Electrical Shock Hazard
On (Mains supply)
Off (Mains supply)
Protective Earth (ground)
V
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Volts (voltage)
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Table 2 Labels on the Mass Spectrometer (Continued)
External Labels
Definition
V-A
Volts - Amperes (power)
W
Watts (power)
Occupational Health and Safety Symbols
This section describes some occupational health and safety symbols found in the laboratory
environment.
Table 3 Chemical Hazard Symbols
Safety Symbol
Description
Biohazard
Corrosive or Caustic Chemical Hazard
Explosion Hazard
Oxidizing Chemical Hazard
Poison Hazard
Reactive Chemical Hazard
Toxic Chemical Hazard
Table 4 Mechanical Hazard Symbols
Safety Symbol
Description
Automated Machinery Hazard
Crushing Hazard — From Above
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Table 4 Mechanical Hazard Symbols (Continued)
Safety Symbol
Description
Crushing Hazard — From Side
Fire Hazard
Hot Surface Hazard
Laser Radiation Hazard
Lifting Hazard
Magnetic Hazard
Puncture Hazard
Table 5 Pressurized Gas Hazard Warning Symbols
Safety Symbol
Description
Pressurized Gas Hazard
Mains Supply
WARNING! Electrical Shock Hazard: Use only qualified personnel for the
installation of all electrical supplies and fixtures, and make sure that all
installations adhere to local regulations.
The mass spectrometer power consumption is 2400 VA (50 Hz or 60 Hz) at 240 VAC.
An external line transformer is not needed for the mass spectrometer or roughing pump.
Caution: Potential Instrument Damage: Do not unpack or connect any components. The
AB SCIEX FSE will unpack, connect, and configure the system for the proper operating
voltage.
For information on system electrical specifications, refer to the Site Planning Guide.
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Protective Earth Conductor
The mains supply should include a correctly installed protective earth conductor that must be
installed or checked by a qualified electrician before connecting the mass spectrometer.
WARNING! Electrical Shock Hazard: Do not intentionally interrupt the protective
conductor. Any interruption of the protective earth conductor is likely to make the
installation dangerous.
Laboratory Ventilation
The venting of fumes and disposal of waste must be in accordance with all federal, state,
provincial, and local health and safety regulations. The system shall be used indoors in a
laboratory that complies with the environmental conditions recommended in the Site Planning
Guide for the system. The source exhaust system must be vented either to an external fume
hood or to an external exhaust system as recommended in the Site Planning Guide for the
system.
Environmental Conditions
Use qualified personnel for the installation of electrical mains, heating, ventilation, and plumbing
supplies and fixtures. Make sure that all installations follow local bylaws and biohazard
regulations. For more information about the required environmental conditions for the system,
refer to the Site Planning Guide for the mass spectrometer.
DANGER! Explosion Hazard: Do not operate the system in an environment
containing explosive gases. The instrument is not designed for operation in an
explosive environment.
WARNING! Asphyxiation Hazard: Take extreme care to vent exhaust gases
properly. The use of instruments without adequate ventilation to outside air may
constitute a health hazard. In addition, certain procedures required during the
operation of the instrument may cause gases to be discharged into the exhaust
stream; under these conditions, inadequate ventilation may result in serious
injury.
WARNING! Radiation Hazard, Biohazard, Toxic Chemical Hazard: Make
sure the mass spectrometer is connected to the local exhaust system and
ducted to control hazardous emissions. The system should only be used
in a well-ventilated laboratory environment in compliance with local
regulations and with appropriate air exchange for the work performed.
Note: In the USA, OSHA 29 CFR Part 1910-1450 requires 4 to12 air changes
per hour in laboratories.
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Safety and Regulatory Information
WARNING! Biohazard: For biohazardous material use, always apply local
regulations for hazard assessment, control, and handling. This instrument or any
part is not intended to act as a biological containment safety cabinet.
Instrument Disposal (Waste Electrical and
Electronic Equipment)
Do not dispose of system components or subassemblies, including computer parts, as unsorted
municipal waste. Follow local municipal waste ordinances for proper disposal provisions to
reduce the environmental impact of WEEE (waste, electrical, and electronic equipment). To
make sure that you safely dispose of this equipment, contact an FSE for instructions.
European Union customers: Contact a local AB SCIEX Customer Service office for
complimentary equipment pick-up and recycling.
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1
System Information
The AB SCIEX TripleTOF® 5600/5600+ system is designed for the qualitative and quantitative
analysis of chemical species. The system includes a mass spectrometer, a DuoSpray™ ion
source, the optional calibrant delivery system (CDS), and a computer running the Analyst® TF
software.
Theory of Operation
Mass spectrometry measures the mass-to-charge ratio of ions to identify unknown compounds,
to quantify known compounds, and to provide information about the structural and chemical
properties of molecules.
The AB SCIEX TripleTOF 5600/5600+ system has a series of quadrupole filters that transmit
ions according to their mass-to-charge (m/z) value. The first quadrupole in this series is the
QJet® ion guide, which is located between the orifice plate and the Q0 region. The QJet ion guide
does not filter ions, but focuses them before they enter the Q0 region. By prefocusing the larger
ion flux created by the wider orifice, the QJet ion guide increases instrument sensitivity and
improves the signal-to-noise ratio. In the Q0 region, the ions are again focused before passing
into the Q1 quadrupole.
The Q1 quadrupole sorts the ions before they enter the Q2 collision cell. In the Q2 collision cell,
the internal energy of the ions is increased though collisions with gas molecules to the point that
molecular bonds break, creating product ions. This technique allows users to design experiments
that measure the m/z of product ions to determine the composition of the parent ions.
After passing through the Q2 collision cell, the ions enter the TOF region for additional mass
analysis, and then enter the detector. In the detector, the ions create a current that is converted
into a voltage pulse. These voltage pulses are counted, and the number of pulses leaving the
detector is directly proportional to the quantity of ions entering the detector. The instrument
monitors the voltage pulses and converts the information into a signal. The signal represents the
ion intensity for a particular m/z value and the instrument displays this formation as a mass
spectrum.
Data Handling
The Analyst TF software requires a computer running the Windows operating system. The
computer with the associated system software works with the system controller and associated
firmware to control the instrument and data acquisition. During system operation, the acquired
data is sent to the Analyst TF software where it can be displayed as either full mass spectra,
intensity of single or multiple ions versus time, or total ion current versus time.
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System Information
System Overview
Figure 1-1 to Figure 1-2 show the mass spectrometer components and connections.
1
2
4
3
7
6
8
5
Figure 1-1
Front and right side
Item Description
For more information...
1
Optional CDS
Refer to the CDS Operator Guide.
2
DuoSpray™ ion source
Refer to DuoSpray™ Ion Source User
Reference on page 83
3
Syringe pump
Refer to Adjust the Integrated Syringe
Pump Position on page 23.
4
Dress panel LEDs
Refer to Dress Panel LEDs on page 20.
5
TDC bulkhead
Contact an AB SCIEX FSE.
6
InfiniBand cable connection for the TDC
card
Contact an AB SCIEX FSE.
7
USB cable connection for the USB-GPIB
card
Contact an AB SCIEX FSE.
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Figure 1-1
Front and right side (Continued)
Item Description
8
For more information...
Serial (RS-232) cable connection for the
syringe pump
1
Contact an AB SCIEX FSE.
11 10
9
2
8
3
4
7
5
12
6
13
14
15
Figure 1-2
Left side view
Item Description
For more information...
1
Gas and vacuum bulkhead
Contact an AB SCIEX FSE.
2
Calibrant control connection
See the CDS Operator Guide.
3
AUX IO connection. The optional LC
system start signal connects to this port.
Contact an AB SCIEX FSE.
4
External control connection. This port is
intended for future use.
Contact an AB SCIEX FSE.
5
Sources connection. Some ion sources
connect to this port.
Contact an AB SCIEX FSE.
6
Curtain Gas™ (nitrogen) supply
connection
Contact an AB SCIEX FSE.
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System Information
Figure 1-2
Left side view (Continued)
Item Description
For more information...
7
Gas 1 and Gas 2 (zero) supply connection Contact an AB SCIEX FSE.
8
Source exhaust gas (zero air or nitrogen)
supply connection
Contact an AB SCIEX FSE.
9
CAD gas (nitrogen) supply connection
Contact an AB SCIEX FSE.
10
Source exhaust waste connection
Contact an AB SCIEX FSE.
11
Roughing pump vacuum connection
Contact an AB SCIEX FSE.
12
AC distribution panel
Contact an AB SCIEX FSE.
13
Instrument power switch
Refer to Start Up the System on page 20.
14
Cover over circuit breaker
Refer to Start Up the System on page 20.
Use the power switch rather than the
circuit breaker to shut down the system.
15
Mains supply cable
Refer to Start Up the System on page 20.
Dress Panel LEDs
Table 1-1 Dress Panel LEDs
Instrument LED
Color
Name
Description
Green
Power
Lit when the mass spectrometer is turned on.
Green
Vacuum Lit when the proper vacuum has been achieved;
and flashing if the system is not at the proper
vacuum level (during pumpdown and venting.)
Red
Fault
Green
Syringe
Pump
Status
Lit when the mass spectrometer encounters a
system fault.
Lit when the syringe pump is running.
Start Up the System
Note: Before operating the instrument, read the safety information in the Safety and
Regulatory Information.
Before the system is turned on, make sure the site requirements specified in the Site Planning
Guide are met. This guide includes information on the mains supply and connections, source
exhaust, compressed air, nitrogen, roughing pump, ventilation, exhaust, and site clearance.
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System Information
Use the following procedures if you need to turn on or shut down the system. You may need to
shut down the system to perform maintenance.
1. Make sure there is clear access to the mass spectrometer AC mains power cord.
The cord must be accessible in order to disconnect the instrument from the AC
mains power supply.
2. Make sure the 4 L drain bottle is connected to the Exhaust Waste connection on the
rear of the instrument and to the laboratory ventilation system.
3. Make sure that the mains supply cable is plugged in to the instrument.
4. Make sure that the mass spectrometer and roughing pump mains supply cable are
plugged into the 200 to 240 V electrical mains supply.
5. Make sure that the Ethernet cable is connected to both the instrument and the
computer.
6. Turn on the roughing pump.
7. Remove the cover on the circuit breaker switch on the left side of the mass
spectrometer, when viewed from the front (refer to Figure 1-2 on page 19), and then
turn on the circuit breaker.
8. Replace the cover over the circuit breaker switch and then tighten the screw holding
the cover until it is finger tight.
9. Turn on the instrument power switch. Refer to Figure 1-2 on page 19.
10. Turn on the computer, if it was turned off.
11. Start the software.
Shut Down the System
1. Complete or stop any ongoing scans. For more information, refer to Stop Sample
Acquisition on page 61.
Caution: Potential Instrument Damage: Turn off the sample flow before you
shut down the mass spectrometer.
2. Turn off the sample flow to the mass spectrometer and disconnect the sample lines
from the peripheral device to the ion source. Leave the source connected for proper
venting.
3. In the Analyst TF software, deactivate the hardware profile, if it is active, and then
close the Analyst TF software.
4. Turn off the instrument power switch on the left side of the instrument (refer to
Figure 1-2 on page 19).
5. Turn off the roughing pump.
6. Wait 15 minutes.
7. Remove the cover on the circuit breaker switch on the left side of the mass
spectrometer (refer to Figure 1-2 on page 19), and then turn off the circuit breaker.
8. Replace the cover over the circuit breaker switch and then tighten the screw holding
the cover until it is finger tight.
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System Information
Use the Integrated Syringe Pump
Edit the Hardware Profile for the Integrated Syringe Pump
Make sure that the serial (RS-232) cable is connected between the computer and mass
spectrometer.
Make sure the syringe pump is seated properly to avoid damaging the syringe. For more
information about creating and editing hardware profiles, refer to Create a Hardware Profile on
page 29.
1. In the Navigation bar, under Configure, double-click Hardware Configuration.
2. Create or edit the hardware profile containing the instrument.
3. In the Devices in current profile box, select the mass spectrometer and click Setup
Device.
4. In the Configuration tab, select Use integrated syringe pump.
Figure 1-3
Mass Spectrometer dialog
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5. Click Configure Pump.
Figure 1-4
Harvard Syringe dialog
6. Select the COM port for the connection to the syringe pump.
7. Click OK until the Hardware Configuration Editor dialog box appears.
8. Activate the hardware profile.
Adjust the Integrated Syringe Pump Position
1. Press the Release button on the right side of the syringe pump to lower the base and
then insert the syringe as shown in Figure 1-5.
Make sure that the end of the syringe is flush against the base and that the shaft of
the syringe rests in the cutout.
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System Information
1
2
Figure 1-5
Lowering the syringe
Item Description
1
Syringe plunger.
2
Release button. Press to raise or lower the base.
2. Adjust the post, shown in Figure 1-6, so that it triggers the automatic syringe stop
before the syringe plunger hits the glass syringe.
1
2
3
Figure 1-6
Safety stop
Item Description
1
Automatic syringe stop. After the post hits the automatic syringe stop, the
syringe pump stops.
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System Information
Figure 1-6
Safety stop (Continued)
Item Description
2
Post. Adjust the height to prevent the syringe plunger from hitting the
syringe during sample infusion.
3
Post lock screw. Tighten the screw after you have adjusted the height of
the post.
3. Turn the side screws as shown in Figure 1-7 to secure the syringe.
Figure 1-7
Syringe pump
4. In the Analyst TF software, on the Navigation bar, double-click Manual Tuning.
5. Click Start Syringe.
6. To stop the syringe pump, click Stop Syringe.
Configure the Integrated Syringe Pump
1. In the Navigation bar, under Acquire, double-click Build Acquisition Methods.
2. In the Acquisition method pane, click the Syringe Pump icon.
The Syringe Pump method properties tab opens in the Acquisition Method Editor
pane.
3. In the Syringe Diameter (mm) field, type the syringe diameter.
4. In the Flow Rate field, type the flow rate.
5. In the Unit list, select the units of flow.
6. Save the file.
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System Information
Reset the Syringe Pump
If the Analyst TF software stops communicating with the syringe pump, you can reset the syringe
pump.
•
Use a paper clip or similar tool to press the reset button, shown in Figure 1-8.
1
Figure 1-8
Syringe pump reset button
Item Description
1
Reset button
Instrument Safe Fluids
These fluids can safely be used with the mass spectrometer:
•
Methanol (0% to 100%)
•
Acetonitrile (0% to 100%)
•
Water
•
Formic acid (0% to 1%)
•
Ammonium acetate (1 mM to 5 mM)
•
Ammonium formate (1 mM to 5 mM)
•
Acetic acid (0% to 1%)
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2
Hardware Profiles and Projects
Sample Workflows
•
Instrument Setup on page 27
•
Sample Acquisition Workflow on page 27
•
Experienced User Workflow on page 28
Table 2-1 Instrument Setup
Step To do this...
Find the
information in...
What does it do?
1
Create a hardware profile. Create a Hardware
Profile on page 29
Each hardware profile must
include a mass spectrometer.
Only devices included in the
active hardware profile can be
used when creating acquisition
methods.
2
Create projects to store
data.
Create Projects and
Subprojects on page 35
Before starting an experiment,
decide where to store the files
related to the experiment. Using
projects and subprojects helps
manage data better and to
compare the results more easily.
3
Optimize the instrument.
Optimize the Instrument This is the process of optimizing
on page 39
the resolution, optimizing
instrument parameters, and
calibrating the instrument to
obtain the best sensitivity and
performance from the mass
spectrometer.
Table 2-2 Sample Acquisition Workflow
Step
1
To do this...
Find the information What does it do?
in...
Create projects to
store data.
Create Projects and
Subprojects on page 35
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Before starting an experiment,
decide where to store the files
related to the experiment. Using
projects and subprojects helps
manage data better and compare
the results more easily.
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Table 2-2 Sample Acquisition Workflow (Continued)
Step
To do this...
Find the information What does it do?
in...
2
Create an acquisition Basic Acquisition
method.
Methods on page 41
To analyze samples, create an
acquisition method for the mass
spectrometer and any LC devices.
An acquisition method indicates
which peripheral devices to use,
when to use them to acquire data,
and the associated parameters.
4
Create and submit a
batch.
Create and Submit a
Batch on page 51
After creating an acquisition
method, to run samples, create an
acquisition batch and submit the
batch to the Acquisition Queue.
5
Acquire data.
Create and Submit a
Batch on page 51
Running samples involves
managing the acquisition queue and
monitoring instrument and device
status. To submit samples and
acquire data, use the Queue
Manager. The Queue Manager
displays queue, batch, and sample
status, and allows users to manage
samples and batches in the queue.
6
Analyze data in
Explore mode.
Analyzing and
Processing Data on
page 65
Users can use many tools in
Explore mode to view and process
the acquired data, such as
customizing graphs with peak labels
and captions, displaying contour
plots, and saving spectra in the
library.
—OR—
Analyze data and
MultiQuant™ software/
print reports using
PeakView® software
companion software.
Use the MultiQuant software or
PeakView software to analyze data.
For more information, refer to the
documentation that comes with the
software.
Table 2-3 Experienced User Workflow
Step
To do this...
Find the information in...
1
Mass calibrate the instrument.
Mass Calibration Tutorial located in Start >
Programs > AB SCIEX > Analyst® TF 1.6
Software > Hardware and Software Guides
2
Optimize the instrument for an
analyte of interest.
Manual Optimization Tutorial located in Start >
Programs > AB SCIEX > Analyst® TF 1.6
Software > Hardware and Software Guides
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Hardware Profiles
A hardware profile tells the software what instrument and peripheral devices to use, and how the
instrument and the devices are configured and connected to the computer.
Each hardware profile must include a mass spectrometer and only peripheral devices included in
the active hardware profile can be used when creating acquisition methods. Before creating an
acquisition method, make sure that all devices used in the method are included in the hardware
profile. In the configuration options for the mass spectrometer, ensure that the syringe pump or
the CDS is enabled if it will be used during acquisition. Enable both the syringe pump and the
CDS if both will be used during acquisition.
The peripheral devices configured in the active hardware profile and selected in the Add/Remove
Device Method dialog appear as icons in the Acquisition Method Browser pane.
For information about setting up the physical connections, refer to the Peripheral Devices Setup
Guide. For a list of the supported peripheral devices, refer to the Software Installation Guide for
the Analyst® TF 1.6 software.
Create a Hardware Profile
The user can set up multiple hardware profiles, but only one profile can be active at any time.
1. In the Navigation bar, under Configure, double-click Hardware Configuration.
2. In the Hardware Configuration Editor dialog (Figure 2-1), click New Profile.
Figure 2-1
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Hardware Configuration Editor dialog
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Hardware Profiles and Projects
3. In the Profile Name field (Figure 2-2), type a name for the profile. For example,
TripleTOF5600+Shimadzu.
Figure 2-2
Create New Hardware Profile dialog
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4. Click Add Device.
In the Available Devices dialog, in the Device Type field, Mass Spectrometer is the
preset value (Figure 2-3).
Figure 2-3
Available Devices dialog
5. In the Devices list, select the Mass Spectrometer TripleTOF 5600 instrument and
then click OK.
6. In the Devices in current profile list, select the instrument.
7. Click Setup Device.
8. (Optional) On the Configuration tab (Figure 2-4), in the Settings for Integrated
Devices section, select the Use calibrant delivery system (CDS) check box.
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Hardware Profiles and Projects
9. (Optional) If using an integrated syringe pump, select Use integrated syringe
pump.
Figure 2-4
Configuration tab
10. Click OK to return to the Create New Hardware Profile dialog.
11. Click Add Device and then add and configure each peripheral device that is used
with the instrument. Refer to Add Peripheral Devices to Hardware Profiles on
page 33.
12. Ensure all changes are accepted and then click OK.
13. To activate the hardware profile, in the Hardware Configuration Editor, click the
hardware profile and then click Activate Profile.
A green check mark appears next to the profile.
Tip! A hardware profile does not have to be deactivated before activating
another. Click a hardware profile and then click Activate Profile. The other
profile is deactivated automatically.
14. Click Close.
15. Next steps: Either create projects and subprojects or optimize the instrument.
•
Create Projects and Subprojects on page 35.
•
Instrument Tuning and Calibrating on page 39.
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Add Peripheral Devices to Hardware Profiles
Peripheral devices must be configured to enable the software to communicate with them.
Configuring the peripheral devices requires two procedures: setting up the physical connections
and configuring the software to communicate with the peripheral devices.
When the software is installed, the driver required for each peripheral device is also installed
(except if the peripheral devices are controlled through AAO devices; the user has to install the
associated driver.) After the peripheral devices are physically connected to the computer, set up
the appropriate configuration information.
1. Open the Hardware Configuration Editor.
2. In the Hardware Profiles list, if required, deactivate the hardware profile.
3. Click Edit Profile.
4. Click Add Device.
5. In the Available Devices dialog, in the Device Type list, select the device.
6. Click OK.
Figure 2-5
Device Type list
7. In the Devices in the current profile list, select the peripheral device
8. Click Setup Device.
A dialog containing configuration values for the peripheral device opens.
Note: The Alias box may also be referred to as the Name box and may be
found on another tab, under Alias.
9. In the Alias field, type a name or other identifier for the device.
•
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If the device uses a serial connection to the acquisition station, in the COM
Port Number list, select the COM port to which the device is connected.
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•
If the device uses Ethernet communication, type the IP address assigned to
the device or use the corresponding host name for the address.
•
If the device uses a GPIB board as a communication interface, do not change
the settings for the GPIB board.
The rest of the preset values for the device are likely appropriate; do not change
them. For information about the Configuration and Communication tabs, refer to the
Help.
10. To restore the device preset values, on the Communication tab, click Set Defaults.
11. To save the instrument configuration, click OK.
12. Repeat step 4 to step 11 for each device.
13. To save the changes to the hardware profile, click OK.
14. To activate the hardware profile, click Activate Profile.
The check mark should turn green. If a red x appears then there is an issue with the
hardware profile activation. For more information, refer to Troubleshoot Hardware
Profile Activation.
Troubleshoot Hardware Profile Activation
If a hardware profile fails to become active, a dialog appears indicating which device in the profile
failed. A failed profile may be due to communications errors.
1. Read the error message generated. Depending on the message, there may be an
issue with a device or how the communication is set up.
2. Verify that the device has power and is turned on.
3. Verify that the COM port assigned to the device is correct.
4. Verify that the communication settings with the peripheral device (for example, dip
switch settings) are set correctly and match the settings on the Communication tab.
5. Turn off the peripheral device.
6. Wait 10 seconds.
7. Turn the device back on.
Wait until all peripheral device power-up activities are complete before trying to
activate the hardware profile again. Some peripheral devices may require 30
seconds or more to complete their power-up activities.
8. Activate the hardware profile.
9. If the issue persists, delete the failing profile and then create a new one.
10. If the issue still persists, contact AB SCIEX technical support.
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Hardware Profiles and Projects
Create Projects and Subprojects
To use a subproject structure within a project, create the subproject structure when the project is
created.
1. Click Tools > Project > Create Project.
2. In the Project name field, type a project name.
3. To use subprojects in this project, select the required folders in the Projects folders
list and then use the arrow buttons to move them to the Subproject folders list
(Figure 2-6).
Figure 2-6
Create New Project/Subproject dialog
Note: Users cannot create a new subproject for a project that was not
originally created with a subproject.
4. (If subprojects are used.) In the Subproject name field, type a name for the first
subproject or use the existing date.
5. (Optional) To copy the template methods from the API Instrument > Acquisition
Methods > template folder into the new folder, select Copy template methods.
6. (Optional) To use this project and subproject folder organization for all new projects,
select the Set configuration as default for new projects check box.
All new projects will be created with this folder configuration.
7. Click OK.
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Create a New Subproject
Subprojects can only be created in a project that has an existing subproject structure.
1. On the Project toolbar, from the Project list, select the project.
2. Click Tools > Project > Create Subproject.
3. In the Subproject name box, type a name for the subproject.
4. Click OK.
Copy a Subproject
Note: The user can copy a subproject from another project that has
existing subprojects. If the copied subprojects contain folders that also exist
in the project folder, then the software uses the project level folders.
1. Click Tools > Project > Copy Subproject.
2. In the Copy Subproject dialog, click Browse to navigate to the subproject source.
3. In the Source Subproject list, select the desired subproject.
4. Click Browse to navigate to the subproject destination field.
5. In the Target Subproject field, type the name for the copied subproject.
6. Do one of the following:
•
To copy all folders and files from the source subproject into the destination
subproject, select the Copy Contents check box.
•
To copy only the folders in the same structure into the destination subproject,
make sure that the Copy Contents check box is cleared.
7. Click Copy.
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Switch Between Projects and Subprojects
•
On the software toolbar (Figure 2-7), from the project list, click the required project or
subproject.
1
Figure 2-7
Project List
Item Description
1
Project list showing a folder, Tutorial, and the Tutorial folders
subfolders.
Installed Project Folders
Three project folders are installed with the software: API Instrument, Default, and Example.
API Instrument Project
The API Instrument project is unique and very important to the proper functioning of the
instrument. The API Instrument project contains the information required for tuning and
calibrating the instrument. This information includes parameter settings files, reference files,
instrument data files that contain calibration and resolution information, and the acquisition
methods used during automatic tuning. The API Instrument project also contains data files for
manual tuning runs that were performed using the Start button rather than the Acquire button.
These data files are saved automatically in the API Instrument project in the Tuning Cache folder
and named with the date and time they were created. The Tuning Cache is automatically purged
when it reaches 2 GB.
Default Project
The Default project contains folders that are present in new projects and serves as a template for
new projects.
Example Project
The Example project contains sample methods and data files. Users can practice working with
the Explore mode using the example data files. The example files are sorted into subfolders by
instrument type and application area.
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Hardware Profiles and Projects
Back up the API Instrument Project
Back up the API Instrument folder regularly and after routine maintenance has been performed.
1. To create the backup, copy the API Instrument project, paste it to a different location,
preferably to another computer, and then rename the folder. Use the date and an
instrument reference if there is more than one instrument when the folder is named.
2. To recover the folder, rename the current API Instrument folder, copy the backup into
the Projects folder and then change its name back to API Instrument.
Table 2-4 Icons on the Toolbar
Icon
Name
Function
New Subproject
Creates a subproject. Subprojects can only be created later in
the process if the project was originally created with subprojects.
Copy Subproject
Copies a Subproject folder. Subprojects can be copied only from
another project that has existing subprojects. If the same folders
exist at both the project and subproject levels, the software uses
the project level folders.
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3
Instrument Tuning and Calibrating
Run the Verify Performance Only option at any time; however, tune the instrument only if a loss
of sensitivity or resolution is noticed. For more information about tuning and calibration, refer to
the Advanced User Guide.
For tuning the system, use the following solutions that come with the installation kit:
For positive mode:
•
For optimizing TOF MS - MSMS high resolution or MSMS High Sensitivity, use the
Tuning Solution.
•
For Q1calibration, use the PPG POS solution.
In negative mode:
•
For optimizing TOF MS - MSMS High Resolution or MSMS High Sensitivity, use
Taurocholic acid.
Note: AB SCIEX recommends that after using the Taurocholic acid, repeat
the channel alignment using the PPG 3000 solution.
•
For Q1calibration, use the PPG 3000 solution.
Required material
•
Tuning solutions that are supplied in the Standards Chemical Kit shipped with the
system. If needed, a new Kit can be ordered from AB SCIEX.
•
Gas-tight syringes (1.0 ml is recommended)
•
PEEK (red) sample tubing
Prerequisites
•
Make sure that a printer is configured.
•
Make sure that the spray is stable and that the proper tuning solution is being used.
Optimize the Instrument
The following procedure shows how to verify the performance of the instrument. For more
information on using the other instrument performance options, refer to the Help.
1. In the Navigation bar, under Tune and Calibrate, double-click Manual Tuning.
2. Run a TOF MS or Product ion scan type and confirm that there is a stable TIC and
that the peaks of interest are present in the spectrum.
3. In the Navigation bar, under Tune and Calibrate, double-click Instrument
Optimization.
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Instrument Tuning and Calibrating
4. Select a tuning solution. Make sure that the tuning solution matches the reference
table.
5. The Verify Performance Only check box is preselected. Click Next.
For this example, leave this option selected. If the report indicates that the
instrument needs tuning, then run Instrument Optimization again and select one or
more scan modes to optimize.
6. Make sure that the ion source and syringe parameters are suitable.
Note: Users can also use the CDS to inject the solution. Make sure the
tuning solution matches the configuration in the reference table. Set the
appropriate flow rate and then click CDS Inject.
Note: Ensure that the correct Calibrant Valve Position is selected in the
Reference Table Editor for the chosen reference table. CDS can select from
up to four different positions, A to D.
7. Click GO.
The Verifying Performance screen appears. After the process has completed, the
Results Summary appears showing the resolution and intensity for each scan mode.
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4
Basic Acquisition Methods
An acquisition method consists of the method for the mass spectrometer and for liquid
chromatography (LC) devices. Users can easily create an acquisition method using the Method
Wizard.
The Acquisition Method Editor can also be used to create acquisition methods and to add a
sequence of periods and experiments for the instrument and devices.
Users can use the SWATH™ acquisition feature, available in both the Method Wizard and the
Acquisition Method Editor, to create SWATH acquisition methods. For more information, refer to
the Advanced User Guide.
Create an Acquisition Method using the Method
Wizard
The acquisition method can be saved in an existing project.
Tip! To copy the Method Wizard template methods into the Acquisition Methods
folder in the project folder, select the Copy method templates check box in the Create
New Project or Subproject dialog. To open this dialog, click Tools > Project > Create
Project or Create Subproject.
1. Make sure that a hardware profile containing the mass spectrometer and peripheral
devices is active.
2. On the software toolbar, make sure that the appropriate project is selected.
3. On the Navigation bar, in Acquire mode, double-click Method Wizard.
The Method Wizard appears.
Tip!
Move the cursor over the interface to view tool tips and procedures.
4. From the Choose MS Method list, select TOF MS (+).
5. From the Choose LC Method list, select the LC method that was created for the
hardware profile.
6. Type a name for the method and then press Enter.
7. Click Next.
8. On the Ion Source Parameters tab, verify the values, editing them if necessary, and
then click Next.
9. On the TOF MS tab, verify the values, editing them if necessary, and then click
Finish.
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Basic Acquisition Methods
Tip! If required, users can further edit the acquisition method using the
Acquisition Method Editor. In Acquire mode, click File > Open and then
open the method that was created using the Method Wizard.
10. Next steps: The newly created acquisition method can now be used to acquire data
for preliminary analysis.
Create Acquisition Methods using the Acquisition Method
Editor
Tip! If users are creating a new acquisition method file from an existing file, some or
all of the peripheral device methods in the acquisition method may be used.
Only devices configured in the active hardware profile appear in the Acquisition Method Browser
pane. Any devices added to the hardware profile must also be added to existing acquisition
methods. For more information about devices, refer to the Peripheral Devices Setup Guide.
1. Make sure that a hardware profile containing the mass spectrometer and peripheral
devices is active.
2. In the Navigation Bar, under Acquire, double-click Build Acquisition Method.
The Method Editor appears with a method template based on the active hardware
profile.
3. In the Acquisition Method Properties tab, select a Synchronization Mode. For
more information about synchronization modes, refer to the Help.
4. In the Acquisition method pane, click Mass Spectrometer.
5. In the MS tab, select a scan type.
6. Type values in the fields as required. For more information refer to About Instrument
Parameters on page 45.
7. In the Advanced MS tab, type values in the fields as required. For more information
refer to About Instrument Parameters on page 45.
8. Click Edit Parameters.
9. On the Source/Gas tab, specify values in the fields as required.
10. On the Compound tab, specify values in the fields as required and then click OK.
11. Click a device icon.
12. Select the parameters for the devices as required.
13. Add any additional periods and experiments. For more information, refer to Add an
Experiment and Add a Period.
14. Click File > Save.
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Basic Acquisition Methods
Add an Experiment
1. Right-click the period where an experiment needs to be added and then click Add
experiment.
An experiment is added below the last experiment in the period.
Note: An experiment or a period cannot be inserted between experiments
or periods. Users can only add an experiment at the end of the period.
2. In the Acquisition Method Editor pane, select the appropriate device or instrument
parameters.
Add a Period
•
In the Acquisition method pane, right-click the Mass Spec icon, and then click Add
period.
A period is added below the last period created.
Note: Users cannot use multiple periods in an IDA experiment.
Copy an Experiment into a Period
Prerequisite: Multi-period method
•
In the Acquisition method pane, press CTRL, and then drag the experiment to the
period.
The experiment is copied below the last experiment in the period.
Copy an Experiment within a Period
Use this procedure to add the same or similar experiments to a period if most or all of the
parameters are the same.
•
Right-click the experiment and then click Copy this experiment.
A copy of the experiment is added below the last experiment created. This is useful
when the same or similar experiments are added to an acquisition method.
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Basic Acquisition Methods
Scan Techniques
The system is a versatile and reliable system for performing liquid chromatography mass
spectrometry analysis on liquid sample streams to identify, quantify, and examine polar
compounds.
The system uses the following mass spectrometry techniques to analyze samples:
•
•
Two modes of single mass spectrometry (MS):
•
Quadrupole-based single mass spectrometry (for Q1 calibration only)
•
Time-of-flight-based single mass spectrometry
Two modes of tandem mass spectrometry (MS/MS):
•
Product ion mass spectrometry
•
Precursor ion mass spectrometry
Single Mass Spectrometry
Single mass spectrometry (MS) is used to analyze charged molecules to find the molecular
weight and amount of detected ions. Individual ions detected by MS can indicate the presence of
a target analyte.
Quadrupole-Based Single Mass Spectrometry
In a quadrupole-based single mass spectrometry (Q1 MS) scan, the system functions as a
traditional quadrupole mass spectrometer. In this mode, the system generates single mass
spectrometric information using the first quadrupole (Q1) section of the instrument.
Time-of-Flight Single Mass Spectrometry
In a time-of-flight single mass spectrometry (TOF MS) scan, the system generates mass
spectrometric information by pulsing ions into a flight tube and recording their precise arrival time
at the detector. Ions with a greater mass-to-charge ratio take longer to travel the flight tube.
Tandem Mass Spectrometry
The technique of MS/MS is well-suited to mixture analysis because the characteristic product ion
spectra can be obtained for each component in a mixture without interference from the other
components, assuming that the product ions have a unique m/z ratio.
Use MS/MS for targeted analysis by monitoring specific precursor/product ions while the sample
is eluting. This type of analysis is more specific than single MS, which only discriminates on the
basis of the mass-to-charge ratio.
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Product Ion Mass Spectrometry
In a product ion scan (Product Ion), the system generates mass spectrometric information by
selecting a particular precursor ion window in Q1, fragmenting in Q2 (a collision cell) and pulsing
the ions (fragment ions) into a flight tube and recording their precise arrival time at the detector.
Product ions can provide information on the molecular structure of the original (precursor) ions.
Precursor Ion Mass Spectrometry
In a precursor ion scan, the system detects precursor ions that generate a specific product ion.
The instrument uses Q1 in mass resolving mode to scan over the mass range of interest, while
the TOF section records product ion spectra for each precursor ion. The Q1 mass spectrum
shows all precursor ions that produce the product ion of interest.
About Spectral Data Acquisition
Spectral data can be acquired in one of the following modes, as shown in Table 4-1. Spectral
Data can only be acquired from Q1 and Precursor Ion scan types.
Table 4-1 Spectral Data Acquisition
Mode
Description
Profile
The preset value is 0.1 Da. Profile data is the data generated by the
instrument and corresponds to the intensity recorded at a series of
evenly spaced discrete mass values. For example, for the mass range
100 Da to 200 Da and step size 0.1, the instrument scans from 100 Da
to 200 Da in 0.1 Da increments (e.g. 100.0, 100.1, 100.2, 100.3… up
to 200.0).
Peak Hopping
The preset value is 1.0 Da. Peak Hopping is a mode of operating a
mass spectrometer in which large steps (approximately 1 Da) are
made. It has the advantage of speed (less data steps are made) but
with the loss of peak shape information.
About Instrument Parameters
The working parameters are the set of instrument parameters currently being used.
•
Ion Source-dependent (Source and gas) parameters: These parameters can
change depending on the ion source used.
•
Compound-dependent parameters: These parameters consist mostly of voltages
in the ion path. Optimal values for compound-dependent parameters vary depending
on the compound being analyzed.
•
Detector parameters: These parameters affect the detector. The Multi-Channel
Plate is the detector in a TOF instrument and consists of four channels for ion
detection. The total of the channels equals the ion intensity. This parameter can be
optimized using Instrument Optimization.
The following figure shows the location of the parameters on the ion optics path.
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1
3
4
2
Figure 4-1
Ion optics path and parameters
Item Parameter
Parameter
type
Use
Scan type
1
Ion Source
Source and
Gas 1 (Gas 1) Gas
The GS1 parameter controls the
All
nebulizer gas. The nebulizer gas helps
generate small droplets of sample flow
and affects spray stability and
sensitivity.
1
Ion Source
Source and
Gas 2 (Gas 2) Gas
The GS2 parameter controls the flow of All
heater gas, which helps evaporate the
solvent to produce gas phase sample
ions.
1
Curtain Gas
(CUR)
Source and
Gas
The CUR parameter controls the gas
between the curtain plate and the
orifice plate. It prevents the
contamination of the ion optics.
1
Temperature
(TEM)
Source and
Gas
The TEM parameter controls the
All
temperature of the turbo gas in the
TurboIonSpray® probe or the
temperature of the probe in the heated
nebulizer (or APCI) probe.
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All
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Figure 4-1
Ion optics path and parameters (Continued)
Item Parameter
Parameter
type
Use
Scan type
1
IonSpray
Voltage
Floating
(ISVF)
Source and
Gas
The ISVF parameter affects the
stability of the spray and hence the
signal sensitivity. This is the voltage
applied to the needle that sprays the
sample.
All
1
Nebulizer
Current (NC)
Source and
Gas
The NC parameter controls the current All
applied to the corona discharge needle
in the APCI probe used in the
TurboIonSpray® ion source when using
the Heated Nebulizer probe. The
discharge ionizes solvent molecules,
which in turn ionize the sample
molecules.
1
IHT (Interface Source and
Heater
Gas
Temperature)
The IHT parameter controls the
temperature of the NanoSpray®
interface heater and is only available if
the NanoSpray ion source and
interface are installed.
All
The optimal heater temperature
depends on the type of sample being
analyzed and the solvent used. If the
heater temperature is too high, the
signal degrades. Typically, heater
temperatures are in the 130 to 180 °C
range. The maximum heater
temperature that can be set is 250 °C,
but this is too high for most
applications.
2
CAD Gas
Compound
The CAD parameter controls the
All
pressure of collision gas in the collision
cell. The collision gas helps to focus
the ions as they pass through the
collision cell; the preset for the CAD
parameter is in fixed mode. For MS/
MS-type scans, the collision gas aids in
fragmenting the precursor ions. When
the precursor ions collide with the
collision gas, they can dissociate to
form product ions.
Use the preset value and optimize for
the compound.
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Figure 4-1
Ion optics path and parameters (Continued)
Item Parameter
Parameter
type
Use
1
Compound
The DP parameter controls the voltage TOF MS/MS
on the orifice, which affects the ability
to decluster ions between the orifice
and QJet® ion guide. It is used to
minimize the solvent clusters that may
remain on the sample ions after they
enter the vacuum chamber, and, if
required, to fragment ions. The higher
the voltage, the higher the energy
imparted to the ions. If the DP
parameter is too high, unwanted
fragmentation may occur.
DP
(Declustering
Potential)
Scan type
Use the preset value and optimize for
the compound.
2
CE (Collision
Energy)
Compound
The CE parameter controls the
Q1, MS/MS
potential difference between Q0 and
Q2 (collision cell). This is the amount of
energy that the precursor ions receive
as they are accelerated into the
collision cell, where they collide with
gas molecules and fragment.
Use the preset value and optimize for
the compound.
2
CES
(Collision
Energy
Spread)
Compound
The CES parameter, in conjunction
Q1, MS/MS
with the Collision Energy (CE),
determines the collision energy applied
to the precursor ion in a Product Ion
scan. The collision energy is ramped
from low to high. For example, in
positive mode, the collision energy will
be ramped from CE – CES to CE +
CES. By entering a CES value,
collision energy spread is automatically
turned on.
Use the preset value and optimize for
the compound.
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Figure 4-1
Ion optics path and parameters (Continued)
Item Parameter
Parameter
type
Use
3
Compound
The amount of time in milliseconds
MS/MS only,
before the ion pulse. The default (11
Enhanced
msec) is calculated based on the TOF
masses and can be adjusted by the
operator. The range is typically 6 msec
to 333 msec.
Ion Release
Delay (IRD)
Scan type
This parameter is optimized using
Instrument Optimization if the
Enhanced Ion option is selected in the
Advanced options. In general, the
default values do not have to be
changed.
3
Ion Release
Width (IRW)
Compound
This is the width, or duration, of the ion MS/MS only,
pulse in milliseconds and is calculated Enhanced
based on the IRD. The range is
typically 5 to 328 msec with a default
value of 10 msec.
This parameter is optimized using
Instrument Optimization if the
Enhanced Ion option is selected in the
Advanced options. In general, the
default values do not have to be
changed.
4
MCP (CEM)
Detector
The CEM parameter controls the
voltage applied to the detector. The
voltage affects the detector response.
All
Acquisition Method Editor Icons
Table 4-2 Acquisition Method Editor Icons
Icon
Name
Function
Mass Spec
Shows the MS tab in the Acquisition Method Editor.
Period
Right-click to add an experiment, add an IDA Criteria Level, or
delete the period.
Autosampler
Opens the Autosampler Properties tab.
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Table 4-2 Acquisition Method Editor Icons (Continued)
Icon
Name
Function
Syringe Pump
Opens the Syringe Pump Properties tab.
Column Oven
Opens the Column Oven Properties tab.
Valve
Opens the Valve Properties tab.
DAD
Opens the DAD Method Editor. For more information about the
DAD, refer to View DAD Data on page 74.
ADC
Opens the ADC Properties tab. For more information about the
ADC, refer to Show ADC Data on page 67.
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5
Batches
A batch is a collection of information about the samples to be analyzed. Batches tell the software
the order in which to analyze the samples. For information about importing batches, refer to the
Advanced User Guide.
Create and Submit a Batch
Use this workflow to create and submit a batch.
•
Set Queue Options on page 51
•
Add Sets and Samples to a Batch on page 53
•
Set up Sample Calibration on page 54
•
Submit a Sample or a Set of Samples on page 55
•
Stop Sample Acquisition on page 57
Set Queue Options
The queue goes one by one through the list, running each sample with the selected acquisition
method. After all the samples have been acquired, the queue stops and the instrument goes into
the Standby mode. In the Standby mode, the LC pumps and some instrument voltages are turned
off.
The user can change the length of time the queue runs after the last acquisition has finished,
before it puts the instrument into the Standby mode. For more information about the other fields
in the Queue Options dialog, refer to the Help.
1. In the Navigation Bar, click Configure.
2. Click Tools > Settings > Queue Options.
The Queue Options dialog opens.
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Batches
Figure 5-1
Queue Options dialog
3. In the Max Num Waiting Samples field, set the maximum number of samples to a
value that is greater than the number of samples that will be submitted to the queue.
4. In the Max Idle Time field, type the length of time the queue will wait after acquisition
is completed before going into Standby mode. The preset value is 60 minutes.
If using gas cylinders, adjust this time to make sure that the gas in the cylinders is
not depleted.
If using an LC method, before the run is started, make sure that there is enough
solvent in the reservoirs for the primary flow rate for all of the sample runs and the
Max. Idle Time.
5. Select Leave Mass Spec on in Standby to keep the mass spectrometer running
after analysis have been completed. This feature allows the heaters and gasses to
continue running, even after devices have entered idle state, so that the source
housing and front plate are kept free of contaminants.
6. Select Fail Whole Batch in Case of Missing Vial to fail the entire batch when a
missing vial is encountered. If this option is not selected, only the current sample will
fail and the queue will continue to the next sample.
7. Select Fail Whole Batch if Auto Calibration Fails to stop the batch if auto
calibration fails.
8. Select Keep Calibration Data File to keep the calibration data file in a subfolder in
the Data folder of the project from which samples are being submitted.
9. Select Continue Batch when TDC Missing Sync found to continue acquiring the
entire batch when a TDC Missing Sync signal is encountered. If this check box is not
selected, the current sample will fail and the queue will not proceed to the next
sample when this signal is encountered.
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Batches
Add Sets and Samples to a Batch
A set can consist of a single sample or multiple samples.
1. In the Navigation Bar, under Acquire, double-click Build Acquisition Batch.
Figure 5-2
Batch Editor
2. In the Sample tab, in the Set list, type a name.
3. Click Add Set.
4. Click Add Samples to add samples to the new set.
Figure 5-3
Add Sample dialog
5. In the Sample name section, in the Prefix field, type a name for the samples in this
set.
6. To add incremental numbering to the end of the sample name, select the Sample
number check box.
7. If the Sample number check box is selected, in the Number of digits field, type the
number of digits to include in the sample name.
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Batches
For example, if 3 is typed, the sample names would be samplename001,
samplename002, samplename003.
8. In the Data file section, in the Prefix field, type a name for the data file that will store
the sample information.
9. Select the Set name check box to use the set name as part of the data file name.
10. Select the Auto Increment check box to increment the data file names
automatically.
Note: AB SCIEX recommends using a new .wiff file for each sample.
11. In the Sub Folder field, type a name.
The folder is stored in the Data folder for the current project. If the Sub folder field is
left blank, the data file is stored in the Data folder and a subfolder is not created.
12. In the New samples section, in the Number field, type the number of new samples
to be added.
13. Click OK.
The sample table fills with the sample names and data file names.
Tip! Fill Down and Auto Increment options are available in the right-click
menu after a single column heading or several rows in a column are
selected.
14. In the Sample tab, in the Acquisition section, select a method from the list.
15. In the Vial Position column, indicate the positions of the vials.
Tip! It is also possible to automatically fill in the samples from the
Locations tab by clicking on the first and last vial within a set with the Shift
key held down. These vials appear as red circles. On the Locations tab,
multiple injections from the same vial can be done by holding down the Ctrl
key while clicking the vial location. The red circle turns green.
16. To set sample locations, do one of the following:
•
Set Sample Locations in the Batch Editor on page 56.
•
Select Vial Positions using the Locations Tab (Optional) on page 57.
Set up Sample Calibration
The software can automatically schedule and execute the external auto calibration while samples
are being acquired in batch mode. This ensures good mass accuracy is maintained throughout
the acquisition.
If the CDS is not configured, calibration is done using an autosampler and users must supply the
calibration method (*.dam) and the vial position of the calibrant sample.
1. In the Batch Editor, click the Calibrate tab.
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2. In the Calibrate Every _ Samples field, type the number of samples to be acquired
between calibration samples.
3. From the Calibrant Reference Table, select a table from the list of all calibrant
reference tables available for the current polarity. Ensure that the selected reference
table has the correct Calibrant Valve Position.
4. Set the CDS Inject Flow Rate.
When the batch is submitted, the calibration samples are inserted into the queue.
Each set starts with a calibration sample. The calibration method is named Cal_ plus
the acquisition method name (for example, Cal_TOF.dam). If the CDS is configured,
the software automatically creates a calibration method that matches the acquisition
method that is used for the next sample in the queue. Calibration data is saved to a
separate data file for each calibration sample. The calibration data file is saved in the
subfolder Cal Data and named with Cal plus the time stamp and calibration sample
index (for example, Cal200906261038341.wiff) if the Keep Calibration Data File was
selected in the Queue Options dialog.
Note: Re-ordering or deleting samples in the queue may result in
calibration samples being lost or no longer matching the following method.
Submit a Sample or a Set of Samples
1. In the Batch Editor, click the Submit tab.
2. If the Submit Status section contains a message about the status of the batch, do
one of the following:
•
If the message indicates that the batch is ready for submission, proceed to
step 3.
•
If the message indicates that the batch is not ready for submission, make the
changes as indicated by the message.
3. Click Submit.
The Acquisition dialog appears.
4. Save the file.
Change Sample Order
The order of the samples can be edited before they are submitted to the Queue.
•
In the Submit tab, double-click any of the numbers on the far left of the table (a very
faint square box is visible), and then drag them to the new location
Acquire Data
The system should not be in Tune mode when sample acquisition is started. Also, if the system
has been previously run that day and has not yet been set to Standby, sample acquisition will
start automatically.
1. In the Navigation Bar, click Acquire.
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2. Click View > Sample Queue.
The Queue Manager opens with all submitted samples.
2
3
1
Figure 5-4
Queue Manager
Item Description
1
The Tune icon should not be pressed in.
2
Queue status.
3
Queue Server should be in Normal mode. For more information, refer to
Queue States on page 59.
3. Click Acquire > Start Sample.
4. Next steps: After the data has been acquired, it can be analyzed. Depending on the
application, use the MultiQuant™ software, the MetabolitePilot™ software, or the
PeakView® software. For more information, refer to the documentation that comes
with the software.
Set Sample Locations in the Batch Editor
If an autosampler is used in the acquisition method, then the vial positions of the samples must
be defined in the acquisition batch. Define the location in the Sample tab or in the Locations tab.
For more information about creating batches, refer to Add Sets and Samples to a Batch on
page 53.
Note: Depending on the autosampler being used, it may not be necessary to
type details in additional columns.
1. In the Sample tab, in the Set list, select the set for which sample locations need to
be specified.
2. For each sample in the set, do the following if applicable:
•
In the Rack Code column, select the rack type.
•
In the Rack Position column, select the position of the rack in the
autosampler.
•
In the Plate Code column, select the plate type.
•
In the Plate Position column, select the position of the plate on the rack.
•
In the Vial Position column, type the position of the vial in the plate or tray.
3. Save the file.
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Select Vial Positions using the Locations Tab
(Optional)
1. In the Batch Editor, click the Locations tab.
2. In the Set list, select the set for which sample locations need to be specified.
3. In the Autosampler list, select the autosampler.
The appropriate number of rack spaces for the autosampler is shown in the graphic
rack display.
4. In the space associated with the rack, right-click and then select the rack type.
The plates or trays are shown in the rack.
5. Double-click one of the rectangles, or select a rectangle and then click Plate/
Autosampler View.
The circles depicting the wells or vials for the plate or tray appear.
6. To select whether samples are marked by row or column, click Row/Column
Selection.
If the button shows a red horizontal line, the Batch Editor marks the samples by row.
If the button shows a red vertical line, the Batch Editor marks the samples by
column.
7. Click the sample wells or vials in the order to be analyzed. Click a selected well or
vial again to clear it.
8. Save the file.
Tip! To auto fill in the samples, hold down the Shift key and then click the
first and last vial within a set. To perform multiple injections from the same
vial, hold down the Ctrl key and then click the vial location. The red circle
changes to a green circle.
Stop Sample Acquisition
The following procedure shows one way to stop sample acquisition. For more information on the
other ways to stop sample acquisition, refer to Table 5-4. When a sample acquisition is stopped,
the current scan finishes before the acquisition is stopped.
1. In the Queue Manager, click the sample in the queue after the point where
acquisition should stop.
2. Click Acquire > Stop Sample.
The queue stops after the current scan in the selected sample is complete. The
sample status on the Queue Manager (Local) window changes to Terminated, and
all others following in the queue are Waiting.
3. When ready to continue processing the batch, click Acquire > Start Sample.
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Batch and Acquisition Method Editor Tips
Table 5-1 Tips
To do this...
...do this
To change all the values in a column click a column heading and then right-click. From the
simultaneously
menu, use the Auto Increment and Fill Down commands
to change the values in the column.
This also works for multiple cells in the same column.
To change an existing acquisition
method
from the list, select the method and then click Method
Editor. To create a new acquisition method, from the list,
select None and then click Method Editor. Only
experienced users should use this feature.
Do not use this feature if the Use Multiple Methods option
is used.
To select more than one well or vial hold down the Shift key and then click the first and last
at a time
well or vial in the range.
Batch Editor Right-Click Menu
Right-click in the Batch Editor table to access the following options.
Figure 5-5
Batch right-click menu
Menu
Function
Open
Opens a batch file.
Import From
Imports a file.
Save As Batch
Saves the batch with a different name.
Save As a Template
Saves the batch as a template. Used with the Express View
feature.
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Figure 5-5
Batch right-click menu (Continued)
Menu
Function
Hide/Show Column
Hides or shows a column.
Save Column Settings
Saves the batch column settings.
Add Custom Column
Adds a custom column.
Delete Custom Column
Deletes a custom column.
Fill Down
Fills the same data into the selected cells.
Auto Increment
Automatically increments data into the selected cells.
Delete Samples
Deletes the selected row.
Select Autosampler
Selects an autosampler.
Queue States and Device Status
The Queue Manager shows queue, batch, and sample status. Detailed information about a
particular sample in the queue can also be viewed.
Queue States
The current state of the queue is indicated in the Queue Server.
Figure 5-6
Queue Server indicator showing Normal mode
Figure 5-7
Queue Server indicator showing Tune mode
The first icon in Figure 5-6 shows the queue state. The second icon indicates whether the queue
is in Tune mode (for tuning) or Normal mode (for running samples). Table 5-2 shows the various
queue states.
Table 5-2 Queue States
Icons
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State
Definition
Not Ready
In the Not Ready state, the hardware profile is
deactivated and the queue is not accepting any
sample submissions.
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Table 5-2 Queue States (Continued)
Icons
State
Definition
Standby
In the Standby state, the hardware profile has been
activated, but all devices are idle. Pumps are not
running and gases are turned off.
Warming Up
In the Warming Up state, the instrument and
devices are equilibrating, columns are being
conditioned, the autosampler needle is being
washed, and column ovens are reaching
temperature. The period of equilibration is selected
by the operator. From this state, the system can go
to the Ready state.
Ready
In the Ready state, the system is ready to start
running samples and the devices have been
equilibrated and are ready to run. In this state, the
queue can receive samples and will run after
samples are submitted.
Waiting
In the Waiting state, the system will automatically
begin acquisition when the next sample is
submitted.
Prerun
In the Prerun state, the method is being
downloaded to each device and device equilibration
is occurring. This state occurs before the acquisition
of each sample in a batch.
Acquiring
In the Acquiring state, the method is run and data
acquisition occurs.
Paused
In the Paused state, the instrument has been
paused during acquisition.
View Instrument and Device Status Icons
Icons representing the instrument and each device in the active hardware configuration appear
on the status bar in the bottom right corner of the window. The user can view the detailed status
of an LC pump to check if the LC pump pressure is appropriate, or view the detailed status of the
instrument to check the temperature of the ion source.
•
On the status bar, double-click the icon for the device or instrument.
The Instrument Status dialog opens.
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Table 5-3 Instrument and Device Status (showing the instrument icon)
Status
Icon Background color
Description
Idle
Green or yellow
The device is not running. If the background
color is yellow, the device should be equilibrated
before it is ready to run. If the background color
is green, the device is ready to run.
Equilibrating
Green or yellow
The device is equilibrating.
Waiting
Green
The device is waiting for a command from the
software, from another device, or for some
action by the operator.
Running
Green
The device is running a batch.
Aborting
Green
The device is aborting a run.
Downloading
Green
A method is being transferred to the device.
Ready
Green
The device is not running, but is ready to run.
Error
Red
The device has encountered an error that
should be investigated.
Note: For each status the background color can also be red. This situation means that
the device encountered an error while in that status.
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Queue Right-Click Menu
Right-click in the Queue table to access the following options.
Figure 5-8
Queue Manager right-click menu
Menu
Function
Sample Details
Opens the Sample Details dialog.
Reacquire
Acquires a sample again.
Insert Pause
Inserts a pause, in seconds, between two samples.
Delete
Deletes either the batch or the selected samples.
Move Batch
Moves the batch within the queue.
Sort
Sorts by the preselect column.
Column Settings
Changes the column settings.
Icon Quick Reference: Acquire Mode
Table 5-4 Acquire Mode Icons
Icon
Name
Function
View Queue
View the sample queue.
Instrument Queue
View a remote instrument station.
Status for Remote Instrument View the status of a remote instrument.
Start Sample
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Starts the sample in the queue.
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Table 5-4 Acquire Mode Icons (Continued)
Icon
Name
Function
Stop Sample
Stops the sample in the queue.
Abort Sample
Aborts a sample acquisition in the middle of the
processing of that sample.
Stop Queue
Stops the queue before it has completed processing
all the samples.
Pause Sample Now
Inserts a pause in the queue.
Insert Pause before Selected
Sample
Inserts a pause before a specific sample.
Continue Sample
Continues acquiring the sample.
Next Period
Starts a new period.
Extend Period
Extends the current period.
Next Sample
Stops acquiring the current sample and to start
acquiring the next sample.
Equilibrate
Click to select a method to use to equilibrate the
devices. This method should be the same as the
method used with the first sample in the queue.
Standby
Puts the instrument in Standby mode.
Ready
Puts the instrument in Ready mode.
Reserve Instrument for
Tuning
Reserves the instrument for tuning and calibrating.
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Analyzing and Processing Data
6
Use the sample files installed in the Example folder to learn how to view and analyze data using
the most common analysis and processing tools. For more information about the following topics,
refer to the Advanced User Guide.
•
Labelling graphs
•
Overlaying and summing spectra or chromatograms
•
Performing background subtractions
•
Smoothing algorithms
•
Working with smoothed data
•
Working with centroid data
•
Working with contour plots
•
Working with the fragment interpretation tool
•
Working with library databases and library records
Open Data Files
Prerequisite: The Example project is selected.
1. In the Navigation Bar, under Explore, double-click Open Data File.
The Select Sample dialog opens.
2. In the Data Files field, double-click TOF.
3. Select Reserpine MSMS.wiff.
4. In the Samples list, select a sample.
5. Click OK.
The data acquired from the sample is shown. If data is still being acquired, the mass
spectrum, DAD/UV trace, and TIC continue to update automatically.
Tip! To turn off the automatic update on the mass spectrum, right-click
the mass spectrum and then click Show Last Scan. If there is a check
mark beside Show Last Scan, then the spectrum will update in real time.
Navigate Between Samples in a Data File
Prerequisite: The Example project is selected.
Table 6-9 shows the navigation icons used in this procedure. If samples were saved in separate
data files, then open each file individually.
1. In the Navigation Bar, under Explore, double-click Open Data File.
2. In the Data Files list, double-click TOF.
3. Select Reserpine MSMS.wiff.
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4. To skip to the next sample in the data file, click the icon with the arrow pointing to the
right.
5. To skip to a non-sequential sample, click the icon with the arrow curving to the right.
6. In the Select Sample dialog, in the Sample list, select the sample.
7. To go to the previous sample in the data file, click the icon with the arrow pointing to
the left.
Show Experimental Conditions
The experimental conditions used to collect data are stored in the data file with the results. The
information contains the details of the acquisition method used: the MS acquisition method (that
is, the number of periods, experiments and cycles) including instrument parameters, and HPLC
device method (LC pump flow rate). In addition, it also contains the MS resolution and mass
calibration tables used for the sample acquisition. Table 6-1 shows the software functionality
available when the user views the file information.
Note: If data is acquired from more than one sample into the same .wiff file, the file
information pane will not refresh automatically as the user scrolls through the samples.
Close the file information pane and then reopen it to view the details for the next sample
in the .wiff file.
•
Click Explore > Show > Show File Information.
The File Information pane opens below the graph.
Tip! To create an acquisition method from the file information pane, rightclick the file information pane and then click Save Acquisition Method.
Table 6-1 Right-Click Menu for Show File Information Pane
Menu
Function
Copy
Copies the selected data.
Paste
Pastes data.
Select All
Selects all the data in the pane.
Save To File
Saves data in an .rtf file.
Font
Changes the font.
Save Acquisition Method
Saves the acquisition method as .dam file.
Save Acquisition Method to
CompoundDB
Opens the Specify Compound Information dialog. Select
the IDs and molecular weights to be saved in the
compound database.
Delete Pane
Deletes the pane.
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Show Experimental Data in Tables
•
With a data file open, click Explore > Show > Show List Data.
The data is shown in a pane below the graph.
Table 6-2 Right-Click Menu for the Spectral Peak List Tab
Menu
Function
Column Options
Opens the Select Columns for Peak List dialog.
Save As Text
Saves the data as text file.
Delete Pane
Deletes the pane.
Table 6-3 Right-Click Menu for the Chromatographic Peak List Tab
Menu
Function
Analyst Classic
Parameters
Opens the Analyst Classic dialog.
IntelliQuan Parameters
Opens the Intelliquan dialog.
Save As Text
Saves the data as text file.
Delete Pane
Deletes the pane.
Show ADC Data
ADC (analog-to-digital converter) data is acquired from a secondary detector (for example from a
UV detector through an ADC card), and is useful for comparison with mass spectrometer data. To
have ADC data available, acquire the data and the mass spectrometer data simultaneously and
save it in the same file.
Prerequisite: The Example project is selected.
1. In the Navigation Bar, under Explore, double-click Open Data File.
The Select Sample dialog opens.
2. In the Data Files list, navigate to the file to open, and then select and pan a sample.
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3. Click Explore > Show > Show ADC Data.
Figure 6-1
Select ADC Channel dialog
4. In the Channel list, select a channel.
5. Click OK.
The ADC data opens in a new pane below the active pane.
Show Basic Quantitative Data
1. With a data file open, click Explore > Show > Show List Data.
2. In the Peak List tab, right-click and select Show Peaks in Graph.
Peaks appear in two colors.
3. To change the peak finding algorithm settings, right-click and then select either
Analyst Classic Parameters or Intelliquan Parameters, which ever is active.
4. (Optional) To remove the colored peaks, right-click in the Peak List tab and then
clear Show Peaks in Graph.
Chromatograms
Table 6-4 Chromatograms
Types of chromatograms Purpose
TIC (Total Ion Chromatogram)
A chromatographic display generated by plotting the intensity
of all ions in a scan against time or scan number.
When a data file is opened, it is preset to appear as a TIC. If
the experiment contains only one scan, it is shown as a
spectrum. For more information about using the available
icons, refer to Table 6-8.
If the MCA check box is selected during acquisition of the data
file, then the data file opens to the mass spectrum. If the MCA
check box is not selected, then the data file opens with the TIC
XIC (Extracted Ion
Chromatogram)
An ion chromatogram created by taking intensity values at a
single, discrete mass value, or a mass range, from a series of
mass spectral scans. It indicates the behavior of a given mass,
or mass range, as a function of time.
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Table 6-4 Chromatograms (Continued)
Types of chromatograms Purpose
BPC (Base Peak
Chromatogram)
Chromatographic plot that shows the intensity of the most
intense ion within a scan versus time or scan number.
TWC (Total Wavelength
Chromatogram)
A chromatographic display created by summing all of the
absorbance values in the acquired wavelength range and then
plotting the values against time. It consists of the summed
absorbances of all ions in a scan plotted against time in a
chromatographic pane.
XWC (Extracted Wavelength
Chromatogram)
A subset of TWC, an XWC shows the absorbance for a single
wavelength or the sum of the absorbance for a range of
wavelengths.
DAD (Diode Array Detector)
A UV detector that monitors the absorption spectrum of eluting
compounds at one or more wavelengths.
Show TICs from a Spectrum
Prerequisite: The Example project is selected.
1. In the Navigation Bar, under Explore, double-click Open Data File.
The Select Sample dialog opens.
2. In the Data Files list, open a file containing spectra.
3. Click Explore > Show > Show TIC.
The TIC opens in a new pane.
Tip!
TIC.
Right-click inside a pane containing a spectrum and then click Show
Show a Spectrum from a TIC
1. In a pane containing a TIC, select a range.
2. Click Explore > Show > Show Spectrum.
The spectrum opens in a new pane.
Tip! Double-click in the TIC pane at a particular time to show the
spectrum.
Generate XICs
The user can generate XICs only from single period, single experiment chromatograms or
spectra. To obtain an XIC from multi-period or multi-experiment data, split the data into separate
panes by clicking the triangle that is under the x-axis. For more information about using the
available icons, refer to Table 6-8.
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Analyzing and Processing Data
There are several methods for extracting ions to generate an XIC, depending on whether
chromatographic or spectral data is used. Table 6-5 contains a summary of methods that can be
used with chromatograms and spectra.
Table 6-5 Summary of XIC Generation Methods
Method
Use with
chromatogram
Use with
spectrum
Extraction
Selected range
No
Yes
The selected range method extracts
ions from a selected area in a
spectrum.
Maximum
No
Yes
The maximum method extracts ions
from a selected area in a spectrum
using the most intense peak in the
selected area. This creates an XIC
using the maximum mass from the
selected spectral range.
Base peak
masses
Yes
No
The base peak masses method can be
used only with BPCs (Base Peak
Chromatograms.) Use the Use Base
Peak Masses command to extract ions
results in an XIC with a different
colored trace for each mass. If the
selection includes multiple peaks, the
resulting XIC will have an equal
number of colored traces representing
each mass.
Yes
The specified masses method extracts
ions from any type of spectrum or
chromatogram. Select up to ten start
and stop masses for which to generate
XICs.
Specified masses Yes
To generate an XIC using a selected range
1. In the Navigation Bar, under Explore, double-click Open Data File.
The Select Sample dialog opens.
2. In the Data Files list, select a file containing spectra.
3. Click OK.
4. To select a range inside the pane, click and hold the left mouse button at the start of
the range and then drag the cursor to the stop point and release.
The selection is highlighted in blue.
5. Click Explore > Extract Ions > Use Range.
An XIC of the specified selection opens in a pane below the spectrum pane. The
experiment information at the top of the pane contains the mass range and the
maximum intensity in counts per second.
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To generate an XIC using the maximum peak
1. In the Navigation Bar, under Explore, double-click Open Data File.
The Select Sample dialog opens.
2. In the Data Files list, select a file containing spectra.
3. Click OK.
4. Select a range.
The selection is highlighted in blue.
5. Click Explore > Extract Ions > Use Maximum.
An XIC of the maximum peak specified selection opens below the spectrum pane.
The experiment information at the top of the pane contains the mass range and the
maximum intensity in counts per second.
To generate an XIC using base peak masses
1. In a BPC, select the peak from which to extract ions.
The selection is highlighted in blue.
2. Click Explore > Extract Ions > Use Base Peak Masses.
An XIC of the specified selection opens below the spectrum pane. The experiment
information at the top of the pane shows the mass range and the maximum intensity
in counts per second.
To extract ions by selecting masses
1. Open a spectrum or chromatogram.
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2. Click Explore > Extract Ions > Use Dialog.
Figure 6-2
Extract Ions dialog
3. Type the values for each XIC to be created. If a stop value is not typed, then the
range is defined by the start value.
•
In the Start field, type the start value (lower value) for the mass range to be
extracted.
•
In the Stop field, type the stop value (higher value) for the mass range to be
extracted.
4. Click OK.
An XIC of the selection opens below the chromatogram pane. The experiment
information at the top of the pane includes the masses and the maximum intensity in
counts per second.
Generate BPCs
BPCs can be generated only from single period, single experiment data. For more information
about using the available icons, refer to Table 6-8.
1. Select an area within a TIC.
The selection is highlighted in blue.
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2. Click Explore > Show > Show Base Peak Chromatogram.
The selections are shown in the Start Time and End Time fields.
Figure 6-3
Base Peak Chromatogram Options dialog
3. In the Mass Tolerance field, type the value to dictate the mass range used to find a
peak. The software finds the peak using a value twice the typed range (± the mass
value).
4. In the Minimum Intensity field, type the intensity below which peaks are ignored by
the algorithm.
5. In the Minimum Mass field, type the mass that determines the beginning of the scan
range.
6. In the Maximum Mass field, type the mass that determines the end of the scan
range.
7. To set the start and end times, select the Use Limited Range check box and do the
following:
•
In the Start Time field, type the time that determines the start of the
experiment.
•
In the End Time field, type the time that determines the end of the experiment.
8. Click OK.
The BPC is generated in a new pane.
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Generate XWCs
The user can extract up to three ranges from a DAD spectrum to generate the XWC. For more
information about using the available icons, refer to Table 6-8.
1. Open a data file that contains a DAD spectrum.
2. Anywhere in the pane, right-click and then click Extract Wavelengths.
Figure 6-4
Extract Wavelengths dialog
3. Type start and stop values.
4. Click OK.
The XWC opens in a pane below the DAD spectrum.
View DAD Data
The user can view DAD data in chromatogram or spectrum form, the same as mass
spectrometer data.
1. Open a data file containing data acquired with a DAD.
2. The TWC, which is analogous to a TIC, opens in a pane below the TIC.
3. In the TWC pane, click a point to select a single point in time, or highlight an area of
the spectrum to select a range of time.
4. Click Explore > Show > Show DAD Spectrum.
The DAD spectrum opens in a pane below the TWC. The y-axis shows absorbance
and the x-axis shows wavelength.
Tip! If the pane with the TWC is closed, click a point anywhere in the TWC
to open it again. Click Explore > Show > Show DAD TWC.
Generate TWCs
A TWC shows total absorbance (mAU) on the y-axis plotted against time on the x-axis. For more
information about using the available icons, refer to Table 6-8.
1. Open a data file that contains a DAD spectrum.
2. Click Explore > Show > Show DAD TWC.
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The TWC opens in a pane below the DAD spectrum.
Tip! Right-click inside the pane containing the DAD spectrum and then
click Show DAD TWC.
Adjust the Threshold
The threshold is an invisible line drawn parallel to the x-axis of a graph that sets a limit below
which the software will not include peaks in a spectrum. The line has a handle, represented by a
blue triangle to the left of the y-axis. Click the blue triangle to view a dotted line that represents
the threshold. The threshold can be raised or lowered, but changing the threshold value does not
change the data. The software does not label any peaks in the region that lies below the
threshold.
1. Open a data file.
2. Adjust the threshold using one of the following steps:
•
To raise the threshold, drag the blue triangle up the y-axis. To lower the
threshold, drag the blue triangle down.
•
Click Explore > Set Threshold. In the Threshold Options dialog that opens,
type the threshold value.
•
Click Explore > Threshold.
The graph updates to show the new threshold. Peak labeling and the peak list are
also updated.
Tip! To view the current threshold value, move the pointer over the
threshold handle.
Table 6-6 Right-Click Menu for Chromatogram Panes
Menu
Function
List Data
Lists the data points and integrates the peaks found in
chromatograms.
Show Spectrum
Generates a new pane.
Show Contour Plot
Shows a color-coded plot of a data set, where the color represents
the intensity of the data at that point. Only certain MS modes are
supported.
Extract Ions
Extracts a specific ion or set of ions from a selected pane and then
generates a new pane containing an extracted chromatograph for the
specific ions.
Show Base Peak
Chromatogram
Generates a new pane containing a base peak chromatogram.
Show ADC Data
Generates a new pane containing the UV data trace, if acquired.
Save to Text File
Generates a text file of the pane, which can be opened in Excel or
other programs.
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Table 6-6 Right-Click Menu for Chromatogram Panes (Continued)
Menu
Function
Save Explore History
The Explore History File records changes to processing parameters,
also called Processing Options, when a .wiff file is processed in
Explore mode. The processing history is stored in a file with an .EPH
(Explore Processing History) extension.
Add Caption
Adds a caption at the cursor point in the pane.
Add User Text
Adds a text box at the cursor point in the pane.
Set Subtract Range
Sets the subtract range in the pane.
Clear Subtract Range
Clears the subtract range in the pane.
Subtract Range Locked
Locks or unlocks the subtract ranges. If the subtract ranges are not
locked then each subtract range can be moved independently. The
subtract ranges are preset to locked.
Delete Pane
Deletes the selected pane.
Table 6-7 Right-Click Menu for Spectra Panes
Menu
Function
List Data
Lists the data points and integrates the peaks found in the spectrum.
Show TIC
Generates a new pane containing the TIC.
Extract Ions
Extracts a specific ion or set of ions from a selected pane and then
generates a new pane containing a chromatograph for the specific
ions.
Save to Text File
Generates a text file of the pane, which can be opened in Excel or
other programs.
Save Explore History
The Explore History File records changes to processing parameters,
also called Processing Options, when a .wiff file is processed in
Explore mode. The processing history is stored in a file with an .EPH
(Explore Processing History) extension.
Add Caption
Adds a caption at the cursor point in the pane.
Add User Text
Adds a text box at the cursor point in the pane.
Show Last Scan
Shows the scan prior to the selection.
Select Peaks For Label
In this dialog, select the parameters to reduce peak labeling.
Re-Calibrate TOF
Opens the TOF Calibration dialog.
Abscissa (Time)
Changes the view to display TOF values on the x-axis.
Delete Pane
Deletes the selected pane.
Add a Record
Add records and compound-related data including spectra to the
library. An active spectrum is required to perform this task.
Search Library
Searches the library without constraints or with previously saved
constraints.
Search With Constraints Searches using the Search Constraints dialog.
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Data Processing
Graphical data can be processed many ways. This section provides information and procedures
for using some of the most commonly used tools.
The user can zoom in on part of a graph to view a particular peak or an area in greater detail in
both spectra and chromatograms. The user can also zoom in repeatedly to view smaller peaks.
Graphs
The same data can be examined in different ways. Data can also be kept for comparison
purposes before performing processing operations such as smoothing or subtraction.
A window contains one or more panes arranged in such a way that all the panes are fully visible
and they do not overlap.
Panes may be of variable or fixed size. Panes are automatically tiled within the window and are
arranged into column and row format. If the size of a window is changed, the panes within the
window change in size to accommodate the new size. A window cannot be sized to the point
where any of the panes would become smaller than its minimum size.
Two or more windows or panes containing similar data can be linked, for example, spectra with
similar mass ranges. As one pane or window is zoomed in, the other pane zooms in
simultaneously.
For example, the user can link an XIC to the BPC from which the XIC was extracted. Zooming in
the BPC also zooms the XIC, so that both chromatograms show the same magnification.
Table 6-8 Graph Options
To do this...
use this menu option...
Copy a graph to a
new window
• Select the graph to copy. Click Explore >
Duplicate Data > In New Window.
Rescale graph to its
original size
• Select the graph. Click Explore > Home Graph.
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...or click this
icon
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Table 6-8 Graph Options (Continued)
To do this...
use this menu option...
Move a pane
• Select the graph. Click Window > Move Pane.
...or click this
icon
• Select the pane or window and then drag it to
the new position. This position can be within the
same window or within another window.
A four-headed arrow is shown when the cursor is
on the boundary of the active window or pane.
• If the pane is at the top or bottom of the
target pane, the pane moves above or
below that pane, respectively.
• If the pane is at the left or right of the target
pane, the pane moves to the left or right of
that pane, respectively.
• If the pane is at any other position, the pane
moves to the target row. The drop shadow
of the pane as the pane is moved around
indicates its new position.
Link panes
1. With the two graphs open, click one to make
that pane active.
2. Click Explore > Link and then click the other
pane.
Remove linking
• Close one of the panes. Click Explore >
Remove Link
Delete a pane
• Select the graph. Click Window > Delete Pane.
Lock a pane
• Select the graph. Click Window > Lock Panes.
Hide a pane
• Select the graph. Click Window > Hide Pane.
Maximize a pane
• Select the graph. Click Window > Maximize
Pane.
Tile panes
• Select the graph. Click Window > Tile all
Panes.
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Zoom in on the y-axis
1. Position the pointer to the left of the y-axis and then drag vertically away from the
starting point.
A box is drawn along the y-axis representing the new scale.
Note: Take care when zooming in on the baseline. Zoom in too low and
the zoom-in box disappears.
2. Release the mouse button to draw the graph to the new scale.
Tip! To return the graph to the original scale, double-click on either axis.
To restore the entire graph to original scale, click Explore > Home Graph.
Zoom in on the x-axis
1. Position the pointer under the x-axis to either side of the area to expands and then
drag away from the starting point in a horizontal direction to expand the area of
interest.
2. Release the mouse button to redraw the graph to the new scale.
Tip! To return the graph to the original scale, double-click on either axis.
To restore the entire graph to original scale, click Explore > Home Graph.
Table 6-9 Explore Quick Reference: Chromatograms and Spectrum
Icon
Name
Function
Open File
Opens files.
Show Next Sample
Navigates to the next sample.
Show Previous Sample
Navigates to the previous sample.
GoTo Sample
Opens the Select Sample dialog.
List Data
Views the data in tables.
Show TIC
Generates a TIC from a spectrum.
Extract Using Dialog
Extracts ions by selecting masses.
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Table 6-9 Explore Quick Reference: Chromatograms and Spectrum (Continued)
Icon
Name
Function
Show Base Peak
Chromatogram
Generates a BPC.
Show Spectrum
Generates a spectrum from a TIC.
Copy Graph to new Window Copies the active graph to a new window.
Baseline Subtract
Opens the Baseline Subtract dialog.
Threshold
Adjusts the threshold.
Noise Filter
Click to use the Noise Filter Options dialog to define
the minimum width of a peak. Signals below this
minimum width are regarded as noise.
Show ADC
Displays ADC data.
Show File Info
Shows the experimental conditions used to collect
the data.
Add arrows
Adds arrows to the x-axis of the active graph.
Remove all arrows
Removes arrows from the x-axis of the active graph.
Offset Graph
Click to compensate for slight differences in the time
during which the ADC data and the mass
spectrometer data were recorded. This is useful
when overlaying graphs for comparison.
Force Peak Labels
Labels all the peaks.
Expand Selection By
Sets the expansion factor for a portion of a graph to
be viewed in greater detail.
Clear ranges
Return the expanded selection to normal view.
Set Selection
Click to type start and stop points for a selection.
This provides more accurate selection than is
possible by highlighting the region using the cursor.
Normalize to Max
Click to scale a graph to maximum, so that the most
intense peak is scaled to full scale, whether or not it
is visible.
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Table 6-9 Explore Quick Reference: Chromatograms and Spectrum (Continued)
Icon
Name
Function
Show History
View a summary of data processing operations
performed on a particular file, such as smoothing,
subtraction, calibration, and noise filtering.
Open Compound Database Opens the compound database.
Set Threshold
Adjusts the threshold.
Show Contour Plot
Shows selected data as either a spectrum graph or
an XIC. Additionally, for data acquired by a DAD, a
contour plot can show selected data as either a DAD
spectrum or an XWC.
Show DAD TWC
Generates a TWC of the DAD.
Show DAD
Generates a DAD.
Extract Wavelength
Extracts up to three wavelength ranges from a DAD
spectrum to view the XWC.
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7
DuoSpray™ Ion Source User Reference
The DuoSpray™ ion source combines a TurboIonSpray® probe and an APCI (atmospheric
pressure chemical ionization) probe in a single ion source housing.
WARNING! Toxic Chemical Hazard: Use the ion source only if you have
knowledge of and training in the proper use, containment, and evacuation of
poisonous or injurious materials used with the ion source. Any poisonous or
injurious materials introduced into the equipment will be present in the ion
source and exhaust output.
Introduction to the Ion Source
Figure 7-1 shows the parts of the ion source.
1
1
9
2
8
7
3
6
4
5
Figure 7-1
Parts of the ion source
Item Description
1
Electrode adjustment nut
2
Corona discharge needle adjustment knob
3
APCI probe
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Figure 7-1
Parts of the ion source (Continued)
Item Description
4
Y-axis adjustment knob for the APCI probe, used to position the probe on the vertical
axis for ion source sensitivity adjustments
5
Corona discharge needle, which ionizes the trace species, or sample gas. Primary ions,
which are formed as a result of the discharge, are converted by collisional processes to
final ion-molecule reaction products.
6
Turbo heater
7
X-axis adjustment knob for the TurboIonSpray probe, used to position the probe on the
horizontal axis for ion source sensitivity adjustments
8
Y-axis adjustment knob for the TurboIonSpray probe, used to position the probe on the
vertical axis for ion source sensitivity adjustments
9
TurboIonSpray probe
Probes
Choose the probe and method most suitable for the compound in the sample stream flow.
Table 7-1 Specifications of the Ion Source
Parameter
TurboIonSpray probe
Ion source temperature Probe temperature from ambient
range
temperature to 750°C, depending on
liquid flow
APCI probe
Probe temperature from
50°C to 750°C,
depending on liquid flow
Liquid chromatography
Interfaces to any liquid chromatography system
Nebulizer gas (Gas 1)
Refer to the Site Planning Guide for the mass spectrometer.
Heater gas (Gas 2)
The TurboIonSpray probe produces ions through ion evaporation. The APCI probe vaporizes the
sample before inducing ionization through atmospheric pressure chemical ionization. This
process is induced by a corona discharge needle as the ions pass through the ion source
housing to the interface region.
All of the data acquired using the ion source is identified with an abbreviation representing the
probe used to acquire the data (TIS for the TurboIonSpray probe, HN for the APCI probe).
TurboIonSpray Probe
The TurboIonSpray probe is suited for LC/MS/MS analyses. The sensitivity that is achieved with
this technique is dependent on both flow rate and analyte. At higher flow rates, ionization
efficiency increases, resulting in improved sensitivity. Compounds with extremely high polarity
and low surface activity usually show the greatest sensitivity increases. The TurboIonSpray
technique is mild enough to be used with labile compounds, such as peptides, proteins, and
thermally labile pharmaceuticals.
When the heater is turned off, the TurboIonSpray probe functions as a conventional IonSpray™
ion source. It also functions with flow rates from 5 µL/min to 3000 µL/min and it vaporizes 100%
aqueous to 100% organic solvents.
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1
2
3
Figure 7-2
Parts of the TurboIonSpray probe
Item Description
1
Electrode adjustment nut (black collar) that adjusts the extension of the electrode tip
2
Bronze retaining ring that fastens the probe to the probe tower on the ion source
housing
3
Electrode tip through which samples are sprayed into the sample inlet area of the ion
source
APCI Probe
The APCI probe is suitable for:
•
Ionization of compounds that do not readily form ions in solution. These are usually
non-polar compounds.
•
Creation of simple APCI spectra for MS/MS experiments.
•
High-throughput analyses of complex and dirty samples. It is less sensitive to ion
suppression effects.
•
Rapid sample introduction by flow injection with or without an LC column
The APCI probe can accept the entire effluent, without splitting, at flow rates from 50 µL/min to
3000 µL/min (through a wide bore column). It can vaporize volatile and labile compounds with
minimal thermal decomposition. The rapid desolvation and vaporization of the droplets and
entrained analyte minimizes thermal decomposition and preserves molecular identity for
ionization by the corona discharge needle. Buffers are readily tolerated by the ion source without
significant contamination and the flash vaporization of the sprayed effluent allows up to 100%
water to be used without difficulty.
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1
Figure 7-3
2
3
Parts of the APCI probe
Item Description
1
Electrode adjustment nut (black collar) that adjusts the extension of electrode tip
2
Bronze retaining ring that fastens the probe to the probe tower on the ion source
housing
3
Electrode tip through which samples are sprayed into the sample inlet area of the ion
source
Gas and Electrical Connections
Gas and high-voltage electrical connections enter through the front plate of the interface and
connect internally through the ion source housing. When the ion source is installed on the mass
spectrometer, all of the electrical and gas connections are made.
Ion Source Latch
A latch disables the high-voltage power supply for the mass spectrometer and the source
exhaust system if:
•
The ion source housing is not installed or is improperly installed.
•
A probe is not installed.
•
The mass spectrometer senses a gas fault.
Source Exhaust System
WARNING! Toxic Chemical Hazard: Be sure to use the source exhaust system to
safely remove sample vapor exhaust from the laboratory environment. For
requirements for the source exhaust system, refer to the AB SCIEX TripleTOF®
5600/5600+ Instruments Site Planning Guide.
A passive pressure exhaust system removes ion source gases through a drain port without
introducing chemical noise. The drain port connects through a drain chamber and a source
exhaust pump to a drain bottle, and from there to a customer-supplied exhaust ventilation
system. For more information on the requirements for the source exhaust system, refer to the
Site Planning Guide for the mass spectrometer.
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WARNING! Toxic Chemical Hazard: Vent the source exhaust system to an
external fume hood or an external vent to prevent hazardous vapors from being
released into the laboratory environment.
A pressure switch mounted on the source exhaust pump measures the pressure in the source
exhaust line. If the pressure in the line rises above the set point while the probes are installed,
the high-voltage power supply is turned off.
Installation
When the ion source is installed, the mass spectrometer recognizes the ion source and displays
the ion source identification in the software.
The interior of the ion source is visible through the tempered glass windows on the side and end
of the ion source housing. The ion source housing is connected to the vacuum interface housing
and is held in position by two source latches.
WARNING! Electrical Shock Hazard: Install the ion source on the mass
spectrometer as the last step in this procedure. High voltage is present when the
ion source is installed on the equipment.
Required tools
• Ion source housing assembly
• Probes
• Ion source hardware kit (Do not discard the empty package. Use it to store the ion source
when not in use.)
Install the Probe in the Ion Source Housing
Install the probes in the ion source housing before installing the ion source. Always remove the
ion source from the mass spectrometer before exchanging probes. Refer to Remove the Ion
Source on page 96.
If the probes are not properly installed in the ion source housing, then high-voltage power for the
mass spectrometer and source exhaust system is turned off.
WARNING! Electrical Shock Hazard: Make sure that the ion source housing is
completely disconnected from the mass spectrometer before proceeding.
WARNING! Electrical Shock Hazard: When installing the ion source, install the
probe before installing the ion source on the mass spectrometer.
Caution: Potential Equipment Damage: Do not let the protruding electrode tip touch any
part of the ion source housing, to avoid damaging the probe.
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1. Adjust the black electrode adjustment nut on the probe to move the electrode tip
inside the electrode tube.
For optimum stability and performance, the electrode tip should extend between 0.5
mm and 1.0 mm from the end of the probe.Insert the probe into the tower.
2. Insert the APCI probe into the tower that is on the left side of the ion source when the
glass window is facing you, inserting the raised plastic post into the groove on the
probe.
3. Gently push down on the probe until the contacts engage with those in the tower.
4. Turn the bronze retaining ring over the probe and push it down to engage its threads
with the threads on the tower.
5. Tighten the ring until it is finger-tight.
6. Insert the TurboIonSpray® probe into the tower on the top of the ion source, inserting
the raised plastic post into the groove on the probe.
7. Gently push down on the probe until the contacts engage with those in the tower.
8. Turn the bronze retaining ring over the probe and push it down to engage its threads
with the threads on the tower.
9. Tighten the ring until it is finger-tight.
Install the Ion Source
1. Make sure that the source latches on the side of the ion source are pointing up in the
12:00 position.
2. Align the ion source with the vacuum interface, making sure that the latches on the
ion source are aligned with the sockets in the vacuum interface.
3. Push the ion source gently against the vacuum interface and then rotate the ion
source latches, shown in Figure 7-1 on page 83, fully downwards to lock the ion
source into place.
Connect the Sample Tubing and Cables for Sample
Introduction with the TurboIonSpray Probe
If you are using the optional CDS, refer to the CDS Operator Guide to install the CDS and
connect the tubing and cables.
WARNING! Toxic Chemical Hazard: Make sure that the sample tubing nut is
tightened properly before operating this equipment. If the sample tubing nut is
not tight, the sample may leak, and you may be exposed to dangerous chemicals.
WARNING! Electrical Shock Hazard: Do not bypass the grounding union
connection. The grounding union provides safety grounding between the mass
spectrometer and the sample introduction device.
1. Insert a 30 cm piece of red PEEK tubing into the sample tubing nut at the top of the
TurboIonSpray probe.
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2. Install the sample tubing nut on the fitting at the top of the TurboIonSpray probe.
3. Tighten the sample tubing nut until it is finger-tight.
4. Connect red PEEK tubing from the sample supply device to the grounding union on
the ion source.
5. Connect the other end of the red PEEK tubing to the grounding union.
Tip! Refer to Figure 7-1 on page 83 for a picture on the parts referenced
in this procedure.
Connect the Sample Tubing and Cables for Sample
Introduction with the APCI Probe
If you are using the optional CDS, refer to the CDS Operator Guide to install the CDS and
connect the tubing and cables.
WARNING! Toxic Chemical Hazard: Make sure that the sample tubing nut is
tightened properly before operating this equipment. If the sample tubing nut is
not tight, the sample may leak, and you may be exposed to dangerous chemicals.
WARNING! Electrical Shock Hazard: Do not bypass the grounding union
connection. The grounding union provides safety grounding between the mass
spectrometer and the sample introduction device.
1. Insert a 30 cm piece of red PEEK tubing into the sample tubing nut at the top of the
APCI probe.
2. Install the sample tubing nut on the fitting at the top of the APCI probe.
3. Tighten the sample tubing nut until it is finger-tight.
4. Connect red PEEK tubing from the sample supply device to the grounding union on
the ion source.
5. Connect the other end of the red PEEK tubing to the grounding union.
Tip! Refer to Figure 7-1 on page 83 for a picture of the parts referenced in
this procedure.
Optimization
Optimize the ion source whenever the analyte, flow rate, or mobile phase composition changes.
Use appropriate analytical procedures and practices to minimize external dead volumes. Prefilter
samples so that the capillary tubing in the sample inlets is not blocked by particles, precipitated
samples, or salts.
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Tip!
It is easier to optimize signal and signal-to-noise with FIA or on-column injections.
Optimize the TurboIonSpray Probe
Optimize performance while injecting a known compound and monitor the signal of the known
ion. Adjust the parameters to maximize the signal-to-noise ratio and signal stability.
Caution: Potential Equipment Damage: If the LC system connected to the mass
spectrometer is not controlled by the Analyst® TF software, then do not leave the mass
spectrometer unattended while in operation. The LC system can flood the ion source
housing when the mass spectrometer goes into Standby mode.
Note: The IonSpray Voltage Floating (ISVF) is always applied to both the
TurboIonSpray probe and the APCI probe simultaneously, and the Temperature (TEM)
is always applied to both the turbo and APCI heaters simultaneously.
Table 7-2 Typical Values for Optimizing the TurboIonSpray Probe
Parameters
LC flow rate
5 µL/min to
50 µL/min
200 µL/min
Operational
range
1000 µL/
min
Probe X-axis position 3 mm to 8 mm
Probe Y-axis position 5 mm to 10 mm
5 µL/min to
3000 µL/min
0 mm to 10 mm
0 mm to 5mm
0 mm to 13 mm
The optimal X-axis position is within 0 mm to 3 mm on either side of the orifice.
The optimal Y-axis position is within 3.0 mm to 7.0 mm of the orifice.
Run the Method
1. Start the Analyst TF software.
2. In the Analyst TF software, in Tune and Calibrate mode, double-click Manual
Tuning.
3. Set the Temperature (TEM) parameter to 450 and let the ion source warm up for 30
minutes or until the ion source housing is warm to the touch.
The 30-minute warm-up stage prevents solvent vapors from condensing in a cold
probe.
4. Start the sample flow.
5. Run the method to be used to optimize the ion source.
Set the Starting Conditions
1. On the Source/Gas tab in the Tune Method Editor, type a starting value for Ion
Source Gas 1 (GS1).
For LC pumps, use a value between 40 and 60 for GS1.
2. Type a starting value for Ion Source Gas 2 (GS2).
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For LC pumps, use a value between 30 and 50 for GS2.
Note: Gas 2 is used with higher flow rates typical with an LC system and in
conjunction with increased temperature.
3. Type a starting value for IonSpray Voltage Floating (ISVF).
4. In the Curtain Gas (CUR) field, type 20.
5. On the Compound tab, in the Declustering Potential (DP) field, type 80.
Adjust the TurboIonSpray Probe Position
At low flow rates, the probe can be adjusted to its lowest Y-axis position. For high flow rates,
position the probe higher than the orifice. The curtain plate orifice should remain clear of solvent
or solvent droplets at all times.
For multiply-charged proteins and peptides introduced at a few microliters per minute, position
the sprayer nozzle higher than the curtain plate orifice.
1. Look through the window of the ion source housing to view the position of the probe.
2. Set the X-axis adjustment knob to 5 and the Y-axis adjustment knob to 5.
3. Infuse or inject the sample.
4. Monitor the signal in the software.
5. Use the X-axis adjustment knob to adjust the probe position in small increments until
the best signal or signal-to-noise ratio is achieved.
The TurboIonSpray probe may optimize slightly to either side of the orifice.
6. Use the Y-axis adjustment knob to adjust the probe position in small increments to
achieve the best signal or signal-to-noise ratio.
Note: The vertical position of the probe depends on flow rate. At low flow
rates, the probe should be closer to the orifice. At higher flow rates, the
probe should be farther away.
WARNING! Toxic Chemical Hazard: Make sure that the electrode tip
extends past the end of the probe, to prevent the escape of hazardous
vapor from the ion source.
7. Adjust the black electrode adjustment nut on the probe to move the electrode tip
relative to the sprayer tube. For optimal performance, the electrode should protrude
0.5 mm to 1.0 mm.
After the probe is optimized, it needs only minor adjustment. If the probe is removed,
or if the analyte, flow rate, or solvent composition changes, repeat the optimizing
procedure after installation.
Optimize the Source/Gas Parameters
Optimize nebulizer gas (Gas 1) for best signal stability and sensitivity. The heater gas (Gas 2)
helps in the evaporation of solvent, which helps to increase the ionization of the sample. Too high
a temperature can cause premature vaporization of the solvent at the TurboIonSpray probe tip,
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especially if the probe is too low, resulting in signal instability and a high chemical background
noise. Similarly, a high heater gas flow could produce a noisy or unstable signal.
1. Adjust GS1 and GS2 in increments of 5 to achieve the best signal or signal-to-noise
ratio.
2. Increase CUR until the signal starts to decrease.
Note: To prevent contamination, use the highest value for CUR possible
without sacrificing sensitivity. Do not set CUR lower than 20.
3. Adjust ISVF in increments of 100 V to achieve the best signal or signal-to-noise ratio.
Note: If the ISVF is too high, a corona discharge can occur. It is visible as
a blue glow at the tip of the TurboIonSpray probe. This will result in
decreased sensitivity and stability of the ion signal.
Optimize the Turbo Heater Temperature
The quantity and type of sample affects the optimal TurboIonSpray probe temperature. At higher
flow rates the optimal temperature increases.
WARNING! Toxic Chemical Hazard: Vent the source exhaust system to an
external fume hood or an external vent to prevent hazardous vapors from being
released into the laboratory environment.
•
Adjust the TEM value in increments of 50°C to achieve the best signal or signal-tonoise ratio.
Optimize for the Compound
•
Adjust the DP value in increments of 10 to achieve the best signal or signal-to-noise
ratio. DP optimizes between 60 to 200.
Optimize the APCI Probe
Optimize performance by injecting a known compound and monitoring the signal of the known
ion. Adjust the parameters to maximize the signal-to-noise ratio.
Table 7-3 Typical Values for Optimizing the APCI Probe
Parameter
Typical value
Operational range
Probe X-axis position
5 mm
Scale 0 mm to 10 mm
Probe Y-axis position
5 mm
Scale 0 mm to 13 mm
The optimal X-axis position is within 0 mm to 2 mm on either side of the orifice.
The optimal probe Y-axis position is within 3.0 mm to 7.0 mm of the orifice.
Run the Method
Caution: Potential Equipment Damage: Warm the ceramic heater slowly to avoid thermal
shock to the heating element.
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Caution: Potential Equipment Damage: If the LC system connected to the mass
spectrometer is not controlled by the Analyst TF software, then do not leave the mass
spectrometer unattended while in operation. The LC system can flood the ion source
housing when the mass spectrometer goes into Standby mode.
1. Start the Analyst TF software.
2. In the Analyst TF software, in Tune and Calibrate mode, double-click Manual
Tuning.
3. Set the Temperature (TEM) parameter to 450 and let the ion source warm up for 30
minutes or until the ion source housing is warm to the touch.
The 30-minute warm-up stage prevents solvent vapors from condensing in a cold
probe.
4. Start the sample flow.
5. Run the method to be used to optimize the ion source.
Set the Starting Conditions
1. On the Source/Gas tab in the Tune Method Editor, in the Ion Source Gas 1 (GS1)
field, type 0.
Note: The value for the GS1 parameter, which is used by the
TurboIonSpray probe, may influence performance of the APCI probe. You
may need to adjust the GS1 parameter value to achieve optimal
performance.
2. In the Ion Source Gas 2 (GS2) field, type 20.
Note: Gas 2 is used as a nebulizer gas for the APCI probe.
3. In the Curtain Gas (CUR) field, type 20.
4. Type a starting value for IonSpray Voltage Floating (ISVF).
5. On the Compound tab, in the Declustering Potential (DP) field, type 80.
Optimize Gas 1, Gas 2, and Curtain Gas™ Flow
1. Adjust GS1 and GS2 in increments of 5 to achieve the best signal or signal-to-noise
ratio.
2. Increase CUR until the signal starts to decrease.
Note: To prevent contamination, use the highest value for CUR possible
without sacrificing sensitivity. Do not set CUR lower than 20.
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Adjust the Corona Discharge Needle
When using the APCI probe, make sure that the corona discharge needle is pointing toward the
orifice.
WARNING! Electrical Shock Hazard: Follow this procedure to avoid contact with
the high voltages applied to the corona discharge needle and the curtain plate.
1. Use the slotted screwdriver to rotate the plastic screw on the top of the needle.
2. Look through the glass window to make sure that the needle is aligned with the tip
facing the orifice.
Adjust the APCI Probe Position
Make sure that the curtain plate orifice remains clear of solvent or solvent drops at all times.
The position of the probe relative to the curtain plate orifice affects sensitivity and signal stability.
For lower flow rates, position the probe closer to the orifice. For higher flow rates, position the
probe farther away from the orifice.
1. Set the Y-axis adjustment knob to 10.
2. Monitor the signal in the Analyst TF software.
3. Use the Y-axis adjustment knob to adjust the probe in small increments until you
achieve the best signal or signal-to-noise ratio.
The APCI probe optimizes toward the orifice plate.
After the probe is optimized, it needs only minor adjustment. If you remove the
probe, or if the analyte, flow rate, or solvent composition changes, repeat the
optimizing procedure after reinstallation.
Note: The position of the TurboIonSpray probe may influence performance
of the APCI probe. You may need to adjust the TurboIonSpray probe
position to achieve optimal performance.
Optimize the IonSpray Voltage Floating
•
In positive mode, start at a value of 5500, and decrease in steps of 500 V; in
negative mode, start at a value of –4500, and increase in steps of 500 V; continue
adjusting until you achieve the best signal or signal-to-noise ratio.
This parameter usually optimizes around 5500 V in positive mode. If you observe no
changes in signal with increasing ISVF, then leave the ISVF at the lowest setting that
provides the best signal or signal-to-noise ratio.
Note: If the ISVF is too high, a corona discharge can occur. It is visible as
a blue glow at the tip of the TurboIonSpray probe. This will result in
decreased sensitivity and stability of the ion signal.
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Optimize the APCI Probe Temperature
The quantity and type of solvent affects the optimal APCI probe temperature. At higher flow
rates, the optimal temperature increases.
WARNING! Toxic Chemical Hazard: Vent the source exhaust system to an
external fume hood or an external vent to prevent hazardous vapors from being
released into the laboratory environment.
•
Adjust the TEM parameter in increments of 50°C to achieve the best signal or signalto-noise ratio.
Optimize for the Compound
•
Adjust the DP value in increments of 10 to achieve the best signal or signal-to-noise
ratio. DP optimizes between 60 to 200.
Maintenance
To determine how often to clean the ion source or perform preventive maintenance, consider the
following:
•
Compounds tested
•
Cleanliness of the preparation methods
•
Amount of time an idle probe contains a sample
•
Overall system run time
These factors can cause changes in mass spectrometer performance, indicating that
maintenance is required.
Perform periodic gas leakage tests and general maintenance inspections to be sure of safe
operation of the system. Clean the ion source regularly to keep it in good working condition.
Caution: Potential Instrument Damage: Use only the recommended cleaning method to
avoid damaging the equipment.
Required tools
• 1/4 inch open-ended wrench
• 9/64 inch Allen key (supplied)
• 5 mm Allen key
• 2.5 mm Allen key
• Phillips screwdriver
• Slotted screwdriver
Clean the Probes
Flush the ion source periodically, regardless of the type of compounds sampled. Do this by
setting up a method in the Analyst TF software specifically for performing a flushing operation.
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1. Switch to a mobile phase that is 50:50 water:acetonitrile or 50:50 water:methanol.
2. Adjust the position of the TurboIonSpray and APCI probes so that they are as far
from the orifice as possible.
3. In the Analyst TF software, set Temperature (TEM) to between 500 and 600, Ion
Source Gas 1 (GS1) and Ion Source Gas 2 (GS2) to at least 40, and Curtain Gas
(CUR) to the highest setting possible.
4. Wait until the TEM setpoint is reached.
5. Infuse or inject mobile phase through the sample tubing and probe at 1 mL/min for
about 10 to 15 minutes.
6. Make sure that the probe and sample tubing are flushed thoroughly.
Remove the Ion Source
Always remove the ion source from the mass spectrometer before you perform any maintenance
on the ion source or exchange probes.
WARNING! Hot Surface Hazard: Surfaces of the ion source become hot during
operation. Let the ion source cool for at least 10 minutes before starting any
maintenance procedures.
1. Stop any ongoing scans.
2. Shut down the sample stream.
3. Using the Analyst TF software, put the mass spectrometer in Standby mode. Refer
to the Analyst TF software Help.
4. Let the ion source cool for at least 10 minutes.
5. Disconnect the sample tubing from the grounding union.
6. Turn the two source latches upward to release the ion source.
7. Pull the ion source gently away from the vacuum interface.
8. Put the ion source on a clean, secure surface.
Remove the Probe
WARNING! Electrical Shock Hazard: Disconnect the source from the mass
spectrometer before starting any maintenance procedures.
1. Remove the ion source from the mass spectrometer. Refer to Remove the Ion
Source.
2. Loosen the 1/8-in. sample tubing nut and remove the sample tubing from the probe.
3. Loosen the bronze retaining ring that fastens the probe to the ion source housing.
4. Gently pull the probe straight up out of the tower. Do not let the tip of the probe touch
anything during removal or storage.
5. Put the probe on a secure, clean surface.
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Clean the Electrode Tube
Clean the electrode tube periodically, or when performance decreases.
This procedure applies to both the TurboIonSpray and APCI probes. Use this procedure to
remove the electrode tube for cleaning. If the electrode tube cannot be cleaned, then use this
procedure to replace it with a new part.
WARNING! Electrical Shock Hazard: Remove the ion source from the mass
spectrometer before starting any maintenance procedures.
1. Remove the ion source from the mass spectrometer. Refer to Remove the Ion
Source on page 96.
2. Remove the probe from the ion source. Refer to Remove the Probe on page 96.
3. Remove the electrode adjustment nut. Hold the probe with the tip pointing
downwards so the spring remains inside the probe as the electrode tube is
withdrawn. Refer to Figure 7-4.
1
8
6
7
2
Figure 7-4
5
3
4
Probe - expanded view
Item Description
1
Electrode adjustment nut
2
PEEK union
3
Spring
4
Bronze retaining ring
5
Electrode tip
6
Sprayer tube
7
Electrode tube
8
1/4-inch retaining nut
4. Pull the PEEK union and the attached electrode tube from the probe. Refer to
Figure 7-4.
5. Use the 1/4 inch open-ended wrench to remove the retaining nut that holds the
electrode tube in the PEEK union.
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6. Remove the electrode tube from the retaining nut.
7. Clean the electrode tube with a 50:50 methanol:water solution, by running the
solution through the electrode tube or by soaking the tube in an ultrasonic bath.
Replace the Electrode
1. Insert the electrode tube into the retaining nut and then into the PEEK union fitting.
Make sure that the electrode tube is inserted as far into the PEEK union fitting as it
will go. If there is a gap between the electrode tube and its seat inside the union
fitting, a dead sample volume may occur.
2. Align the electrode tube with the narrow opening in the sprayer tube, and then insert
the PEEK union fitting and attached electrode tube into the probe. Be careful not to
bend the electrode tube.
3. Make sure that the spring is still inside the probe and then tighten the electrode
adjustment nut.
4. Insert the probe into the tower, taking care not to allow the tip of the probe to touch
any part of the ion source housing.
5. Push down the bronze retaining ring to engage its thread with the thread on the ion
source housing and then tighten the ring.
6. Insert the sample tubing into the sample tubing nut, insert the sample tubing nut into
the fitting at the top of the probe, and then tighten the sample tubing nut until it is
finger-tight.
7. Install the ion source on the mass spectrometer. Refer to Install the Ion Source on
page 88.
8. Adjust the electrode tip to specification. Refer to Adjust the Electrode Tip Extension
on page 98.
Adjust the Electrode Tip Extension
The electrode tip extension should be adjusted for best performance. The optimal setting is
compound-dependent. The distance that the electrode tip extends affects the shape of the spray
cone, and the cone shape affects mass spectrometer sensitivity.
WARNING! Toxic Chemical Hazard: Make sure that the electrode tip extends past
the end of the probe, to prevent the escape of hazardous vapor from the ion
source.
•
Adjust the black electrode adjustment nut on the top of the probe to extend or retract
the electrode tip. The electrode tip should extend between 0.5 mm and 1.0 mm from
the end of the probe as shown in Figure 7-5.
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1
2
0.5 - 1 mm
Figure 7-5
Electrode tip extension adjustment
Item Description
1
Probe
2
Electrode
Remove the Corona Discharge Needle
The corona discharge needle tip may become so corroded that it must be cut off from the corona
discharge needle. If this occurs, replace the corona discharge needle.
WARNING! Electrical Shock Hazard: Remove the ion source from the mass
spectrometer before starting any maintenance procedures.
WARNING! Piercing Hazard: The tip of the needle is extremely sharp. Take care
to handle it safely.
1. Remove the ion source and probe from the mass spectrometer.Refer to Remove the
Ion Source on page 96.
2. Rotate the ion source so that the open side is toward you.
3. Press down on the corona discharge needle adjustment knob on the top of the
tower. The corona discharge needle extends.
4. Holding the corona discharge needle tip between the thumb and forefinger of one
hand and the corona discharge needle with the other hand, rotate the corona
discharge needle tip counter-clockwise to loosen and gently remove the tip.
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Figure 7-6
Corona discharge needle tip at rear of ion source
5. Turn the corona discharge needle adjustment knob counter-clockwise until it is
loose.
6. Gently pull the corona discharge needle adjustment knob to remove the corona
discharge needle from the ceramic sleeve, taking care not to break the sleeve.
7. Remove the corona discharge needle adjustment knob from the corona discharge
needle.
8. Install the corona discharge needle adjustment knob on a new corona discharge
needle and then insert the needle into the sleeve.
9. Tighten the corona discharge needle adjustment knob until the connection is firm.
10. Holding a new tip between the thumb and forefinger of one hand and the corona
discharge needle with the other hand, rotate the corona discharge needle tip
clockwise to install the tip.
11. Install the ion source on the mass spectrometer. Refer to Install the Ion Source on
page 88.
Replace the Sample Tubing
Use the following procedure to replace the sample tubing if it has a blockage.
1. Stop the sample flow and make sure that any remaining gas has been removed
through the source exhaust system.
2. Remove the ion source. Refer to Remove the Ion Source on page 96.
3. Disconnect the sample tubing from the probe and the union.
4. Replace the sample tubing with the same length of tubing used previously.
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5. Install the ion source. Refer to Install the Ion Source on page 88.
6. Resume the sample flow.
Troubleshooting
Table 7-4 Troubleshooting
Symptom
Possible cause
Solution
®
• The probe is not installed.
The Analyst TF software
reports that the mass
spectrometer is in Fault state.
(The mass spectrometer icon on
the Analyst TF software status
• The probe is not connected
bar is red.)
securely.
• Install the probe. Refer to
Install the Probe in the Ion
Source Housing on
page 87.
The heater does not work.
Contact your FSE.
The F3 fuse is blown.
• Remove and replace the
probe. Tighten the probe
connection bronze ring
securely. Refer to Remove
the Probe on page 96 and
Install the Probe in the Ion
Source Housing on
page 87.
The spray does not appear to be The electrode is blocked.
uniform.
Clean or replace the
electrode. Refer to Clean the
Electrode Tube on page 97.
Sensitivity is poor.
Excess high voltage induces
fragmentation before the ions
enter the mass filters.
Optimize ISVF or DP. Refer
to Optimize the
TurboIonSpray Probe on
page 90 or Optimize the
APCI Probe on page 92.
During testing, the ion source
fails to meet specifications.
The mass spectrometer has
not passed the installation
tests.
Perform installation tests on
the mass spectrometer with
the default source.
The test solution was not
prepared correctly.
Confirm that the test
solutions were prepared
correctly.
If the problem cannot be
resolved, contact an FSE.
Background noise is high.
• Temperature (TEM) is too
high.
• Optimize TEM.
• Gas 2 flow rate (GS2) is too • Optimize GS2.
high.
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Table 7-4 Troubleshooting (Continued)
Symptom
Possible cause
Solution
Arcing or sparks occur.
The position of the corona
discharge needle is incorrect.
Turn the corona discharge
needle toward the curtain
plate, and away from the
stream of Gas 2. Refer to
Adjust the Corona Discharge
Needle on page 94.
Consumables
The following tables list the orderable parts for the DuoSpray™ ion source. The parts are
available in the Consumables Kit for the mass spectrometer (PN 1026540).
Table 7-5 Orderable Parts for the Ion Source
Part No. Description
Quantity
016316
PEEK tubing, Red, 1/16 o.d. × 0.005 bore
100 cm
016325
PEEK fitting, Brown, 10-32 × 1/16 inch
5
025388
Electrode, Nebulizer
1
025392
Electrode, TuroIonSpray
1
027471
PEEK Graph-tite fitting, Black, 1/16 inch
2
®
1005601
PEEK tubing kit to connect to TurboIonSpray probe, 30 cm
1
1005602
PEEK tubing kit to connect to APCI probe, 45 cm
1
025348
PEEK union in probe
1
026626
Spring for probe
1
Table 7-6 Spares
Part No. Description
Quantity
1006177
APCI Corona discharge needle tip
1
1006174
APCI Corona discharge needle rod
1
027497
Gold-plated spring for HV connection
1
027013
Spring for corona discharge needle
1
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8
Cleaning and Maintenance
Regularly clean and maintain the instrument for optimal performance. Table 8-1 provides a
recommended schedule for cleaning and maintaining the instrument. Contact your Qualified
Maintenance Person to order consumable parts.
Table 8-1 System Maintenance Tasks
Component
Frequency
Task
For more information, refer to...
Curtain plate
As needed
Clean
Clean the Curtain Plate
QJet®
As needed
Clean
Contact an AB SCIEX FSE.
Q0 and IQ1 lens As needed
Clean
Contact an AB SCIEX FSE.
Instrument
cooling fan filter
Every 6
months
Replace
Contact an AB SCIEX FSE.
Instrument
surfaces
As needed
Clean
Clean the Surfaces
Drain bottle
As needed
Empty
Empty the Drain Bottle
Roughing pump
oil
As needed
Check and fill
Contact an AB SCIEX FSE.
Every 6 to 12
months
Replace
Contact an AB SCIEX FSE.
Electrode
As needed
Inspect and
Clean the Electrode Tube on page 97
clean or replace
ion guide
Corona
As needed
discharge needle
Replace
Remove the Corona Discharge Needle on
page 99
For “As needed” tasks, follow these guidelines:
•
Clean the curtain plate, orifice plate, QJet ion guide, and Q0 region if system
sensitivity degrades.
•
Clean the instrument surfaces after a spill, or when they become dirty.
•
Empty the drain bottle when it becomes full.
Contact an AB SCIEX FSE for maintenance service and support.
Health and Safety Precautions
•
Determine what chemicals may have been used in the instrument prior to service.
Refer to Safety Data Sheets for the health and safety precautions that must be
followed with chemicals.
•
Work in a well-ventilated area.
•
Always wear assigned personal protective equipment, including powder-free nitrile
gloves, safety glasses, and a laboratory coat.
•
Follow required electrical safe work practices.
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Cleaning and Maintenance
•
Avoid ignition sources when working with flammable materials, such as isopropanol,
methanol, and other flammable solvents.
•
Take care in the use and disposal of any chemicals. Potential risk of personal injury if
proper handling and disposing of chemicals are not followed.
•
Avoid skin contact with chemicals during cleaning, and wash hands after use.
•
Comply with all local regulations for the handling of biohazard, toxic, or radioactive
materials.
WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard:
Determine whether instrument decontamination is required prior to
cleaning. Instrument decontamination should be conducted prior to
cleaning if radioactive materials, biological agents, or toxic chemicals
have been used with an instrument.
Caution: Potential Instrument Damage: Rinse off any acid-containing cleaning solvents
with water. Do not use chlorinated solvents because these may damage the QJet ion
guide components
Clean the Surfaces
Clean the external surfaces of the instrument after a spill, or when they become dirty.
•
Using warm, soapy water and a soft cloth, wipe the external surfaces.
Empty the Drain Bottle
Empty the drain bottle when it becomes full.
WARNING! Biohazardous Material: Deposit biohazardous material in
appropriately labelled containers. Potential risk of personal injury if proper
handling and disposing of biohazardous materials are not followed
1. Shut Down the System on page 21.
2. Disconnect the tubes from the top of the drain bottle.
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1
2
3
4
Figure 8-1
Item
Vent connections
Description
1
Roughing pump exhaust connection to vent
2
Source exhaust output connection to vent
3
Vacuum hose
4
Source exhaust drain bottle
3. Unscrew the cap and dispose of the waste.
4. Replace the cap and connect the tubes.
Front-End Cleaning
Clean the instrument front-end using the routine cleaning method, to:
•
Minimize unscheduled instrument downtime.
•
Maintain optimum sensitivity.
•
Avoid more extensive cleaning that requires a service visit.
Symptoms of contamination: Significant loss in sensitivity and increased background noise.
When contamination occurs, perform an initial routine cleaning. Clean up to and including the
front of the orifice plate. If routine cleaning does not resolve issues with sensitivity, a full cleaning
may be necessary.
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Cleaning and Maintenance
This section provides instructions for performing routine cleaning without breaking vacuum and
full cleaning under atmospheric pressure, after venting the instrument.
Note: Follow all applicable local regulations. For health and safety guidelines, refer to
Health and Safety Precautions for more information.
Required tools and materials
• Powder free gloves (nitrile recommended)
• Safety glasses
• Laboratory coat
• Fresh, high quality water (at least 18 Mohm de-ionized water [DI water] or ultra-pure HPLCgrade water). Old water can contain contaminants which can further contaminate the mass
spectrometer.
• HPLC- or LCMS-grade methanol, isopropanol (2-propanol), or acetonitrile
• Cleaning solution. Use one of:
•
•
•
•
100% methanol
100% isopropanol
50:50 acetonitrile:water solution (freshly prepared)
50:50 acetonitrile:water with 0.1% acetic acid solution (freshly prepared)
Caution: Potential Instrument Damage: Do not use chlorinated solvents.
Note: For consumables ordering information and inquiries, call 877-740-2129 (U.S.
only), or visit www.absciex.com.
Table 8-2 Tools and Supplies Available from AB SCIEX
Description
P/N
Small polyester swab (thermally bonded)
1017396
Small lint-free wipe (11 cm x 21 cm); available in Consumables kits
WC018027
Best Practices
WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard:
Determine whether instrument decontamination is required prior to
cleaning. Instrument decontamination should be conducted prior to
cleaning if radioactive materials, biological agents, or toxic chemicals
have been used with an instrument.
•
Always wear clean, powder-free gloves for the cleaning procedures.
•
After cleaning the instrument components and before reassembling them, put on a
clean pair of gloves.
•
Do not use cleaning supplies other than those specified in this procedure.
•
If possible, prepare cleaning solutions just before beginning.
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•
Prepare and store all organic solutions and organic-containing solutions in very
clean glassware only. Never use plastic squirt bottles. Contaminants can leach from
these bottles and further contaminate the mass spectrometer.
•
Allow only the center area of the wipe to contact the instrument surface. Cut edges
can leave fibers behind.
Tip!
Figure 8-2
Wrap the wipe around a thermally-bonded polyester swab.
Example: Folding the wipe
•
Allow the wipe or swab to contact the surface once, and then discard it, to avoid
cross-contamination.
•
Larger parts of the vacuum interface, such as the curtain plate, may require several
cleanings, using multiple wipes.
•
To avoid contaminating the solution, pour the solution on the wipe or swab.
•
Only moisten the wipe or swab slightly when applying water or cleaning solution.
Water, more so than organic solvents, may cause the wipe to deteriorate, leaving
residue on the instrument.
Prepare for Routine Cleaning
In routine cleaning, clean the curtain plate and the front of the orifice plate. Routine cleaning can
be performed while the instrument remains under vacuum.
Note: Instruments with a NanoSpray® ion source may require a full cleaning for best
results. Contact an AB SCIEX FSE.
1. Deactivate the hardware profile.
WARNING! Hot Surface Hazard: Surfaces of the ion source become hot during
operation. Let the ion source cool for at least 10 minutes before starting any
cleaning procedures.
2. Remove the ion source. Be sure to place the ion source in a safe location.
3. Wait at least 20 minutes for the curtain plate and orifice plate to cool.
4. Cover the source drain with the exhaust cover plate (if available), or a similar cover.
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1
Figure 8-3
Source drain on the vacuum interface
Item Description
1
Source drain
Clean the Curtain Plate
Caution: Potential Instrument Damage: Do not rest the curtain plate on the orifice. Make
sure that the conical side faces up.
1. Remove the curtain plate and then place it, conical side up, on a clean, stable
surface.
Figure 8-4
Interface with curtain plate removed
2. Using wipes and water, clean both sides of the curtain plate.
3. Repeat step 2 using the cleaning solution.
4. Using a dampened wipe or small poly swab, clean the aperture.
5. Wait until the curtain plate is dry.
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Cleaning and Maintenance
6. Inspect the curtain plate for solvent stains or lint, removing any residue with a clean,
slightly damp lint-free wipe.
Note: Persistent spotting or filming is an indicator of contaminated solvent.
Clean the Front of the Orifice Plate
Note: If the standard orifice plate has a removable interface heater, do not remove the
heater during cleaning.
1. When cleaning a NanoSpray orifice plate, remove the interface heater and clean it:
i. Wipe the heater with a lint-free wipe dampened with water.
ii. Wipe the heater with a lint-free wipe dampened with cleaning solution.
2. Moisten the lint-free wipe with water and then wipe the front of the orifice plate.
Caution: Potential Instrument Damage: Do not insert a wire or metal brush
into the orifice, to avoid damaging the aperture.
3. Repeat step 2 using the cleaning solution.
4. Wait until the orifice plate is dry.
5. Inspect the orifice plate for solvent stains or lint, removing any residue with a clean,
slightly damp lint-free wipe.
Note: Persistent spotting or filming is an indicator of contaminated solvent.
Put the Instrument Back into Service
1. Install the curtain plate on the front end of the instrument.
2. Remove the protection from the ion source drain.
3. Install the ion source on the mass spectrometer.
4. Tighten the ion source by turning the ion source release latches down into the
locking position. (Refer to the ion source Operator Guide.)
5. Activate the hardware profile.
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9
Basic System Troubleshooting
This appendix contains basic information for troubleshooting basic system issues. Certain
activities may be carried out by the AB SCIEX trained Qualified Maintenance Person in the
laboratory. For advanced troubleshooting, contact an AB SCIEX Field Service Employee (FSE).
Table 9-1 System Issues
Issue
Possible cause
Corrective action
Sensitivity loss
Instrument or ion source
For more information, refer to:
requires tuning and optimizing
• Instrument Tuning and
Calibrating
• DuoSpray™ Ion Source
User Reference on page 83
appendix
• Analyst® software Help
system
Dirty curtain plate
Refer to Clean the Curtain
Plate on page 108 for more
information.
Dirty orifice plate
Contact an AB SCIEX FSE or
your local AB SCIEX trained
Qualified Maintenance Person.
Dirty QJet® ion guide, Q0 or
IQ0
Contact an AB SCIEX FSE or
your local AB SCIEX trained
Qualified Maintenance Person.
Frequent or extreme
contamination of the QJet ion
guide
Curtain Gas™ flow rate is too
low.
Verify, and if applicable,
increase the Curtain Gas™
flow rate.
Low vacuum level
Low roughing pump oil level.
Check the roughing pump oil
level, and add oil if necessary.
Contact an AB SCIEX FSE or
your local AB SCIEX trained
Qualified Maintenance Person.
For sales, technical assistance or service, contact an AB SCIEX FSE or visit the AB SCIEX Web
site at www.absciex.com for contact information.
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A
Recommended Calibration Ions
The following tables list the standards recommended by AB SCIEX for calibrating the AB SCIEX
TripleTOF® 5600/5600+ Instruments. For information about tuning solutions, refer to Chapter 3:
Instrument Tuning and Calibrating.
Table A-1 Q1 PPG Positive Calibration Ions
Masses
59.04914
175.13286
442.3374
674.50484
906.67228
1196.88158
Table A-2 Q1 PPG Negative Calibration Ions
Masses
44.99819
411.25991
585.38549
933.63665
1165.80409
Table A-3 APCI Positive Calibration Solution: TOF MS
TOF MS
Masses
aminoheptanoic acid
146.11756
amino-dPEG 4-acid
266.15981
clomipramine
315.16225
amino-dPEG 6-acid
354.21224
amino-dPEG 8-acid
442.26467
reserpine
609.28066
amino-dPEG 12-acid
618.36953
Hexakis(2,2,3,3-tetrafluoropropoxy) phosphazine 922.0098
Hexakis(1H,1H,5H-octafluoroheptoxy)
phosphazene
1521.97148
Table A-4 APCI Positive Calibration Solution: MSMS (Clomipramine)
MSMS (Clomipramine)
Masses
C3H8N
58.0651
C5H12N
86.0964
C16H14N
220.1121
C14H10NCl
227.0496
C17H17N
235.1356
C15H13NCl
242.0731
C17H17ClN
270.1044
C19H23ClN2
315.16225
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Table A-5 APCI Negative Calibration Solution: TOF MS
TOF MS
Masses
7-aminoheptanoic acid
144.103
amino-dPEG 4-acid
264.14526
sulfinpyrazone fragment
277.09825
amino-dPEG 6-acid
352.19769
sulfinpyrazone
403.11219
amino-dPEG 8-acid
440.25012
amino-dPEG 12-acid
616.35498
amino-dPEG 16-acid
792.45984
Table A-6 APCI Negative Calibration Solution: MSMS (Sulfinpyrazone)
MSMS (Sulfinpyrazone)
Masses
C6H5O
93.0344
C6H5OS
125.0067
C10H8NO
158.06114
C17H13N2O2
277.0983
C23H20N2OS3
403.11219
Table A-7 APCI Negative Calibration Solution: MSMS (Sulfinpyrazone fragment)
MSMS (Sulfinpyrazone fragment)
Masses
C6H5
77.03967
C8H6N
116.0506
C9H8N
130.0662
C10H8NO
158.0611
C11H8N2O2
200.0591
C15H9N2
217.0771
C16H13N2O
249.1033
C17H13N2O2
277.09825
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B
Exact Masses and Chemical Formulas
PPG
Table B-1 contains the exact monoisotopic masses and charged species (positive and negative)
observed with the PPG (polypropylene glycol) calibration solutions. The masses and ions were
calculated using the formula M = H[OC3H6]nOH, while the positive ion MSMS fragments used the
formula, [OC3H6]n(H+). In all calculations, H = 1.007825, O = 15.99491, C = 12.00000, and N =
14.00307.
Note: When performing calibrations with the PPG solutions, use the correct
isotope peak.
Table B-1 PPG Exact Masses
n
Exact Mass (M) (M + NH )+
4
MSMS
(M + NH4)2+
fragments
(M + COOH)–
1
76.05242
94.08624
59.04914
56.06003
121.05061
2
134.09428
152.12810
117.09100
85.08096
179.09247
3
192.13614
210.16996
175.13286
114.10189
237.13433
4
250.17800
268.21182
233.17472
143.12282
295.17619
5
308.21986
326.25368
291.21658
172.14375
353.21805
6
366.26172
384.29554
349.25844
201.16468
411.25991
7
424.30358
442.33740
407.30030
230.18561
469.30177
8
482.34544
500.37926
465.34216
259.20654
527.34363
9
540.38730
558.42112
523.38402
288.22747
585.38549
10 598.42916
616.46298
581.42588
317.24840
643.42735
11 656.47102
674.50484
639.46774
346.26933
701.46921
12 714.51288
732.54670
697.50960
375.29026
759.51107
13 772.55474
790.58856
755.55146
404.31119
817.55293
14 830.59660
848.63042
813.59332
433.33212
875.59479
15 888.63846
906.67228
871.63518
462.35305
933.63665
16 946.68032
964.71414
929.67704
491.37398
991.67851
17 1004.72218
1022.75600
987.71890
520.39491
1049.72037
18 1062.76404
1080.79786
1045.76076
549.41584
1107.76223
19 1120.80590
1138.83972
1103.80262
578.43677
1165.80409
20 1178.84776
1196.88158
1161.84448
607.45770
1223.84595
21 1236.88962
1254.92344
1219.88634
636.47863
1281.88781
22 1294.93148
1312.96530
1277.92820
665.49956
1339.92967
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Reserpine (C33H40N2O9)
Table B-2 Reserpine Exact Masses
Description
Mass
Molecular Ion C33H41N2O9
609.28066
Fragment C23H30NO8
448.19659
Fragment C23H29N2O4
397.21218
Fragment C22H25N2O3
365.18597
Fragment C13H18NO3
236.12812
Fragment C10H11O4
195.06519
Fragment C11H12NO
174.09134
Taurocholic Acid (C26H45NO7S)
Table B-3 Taurocholic Acid Exact Masses
Description
Mass
Molecular Ion C26H44NO7S
514.28440
Fragment C2H3O3S
106.98084
Fragment C2H6NO3S
124.00739
Fragment SO3
79.95736
TOF Calibration Solution
Table B-4 TOF Calibration Solution Exact Masses
Description
Molecular Ion Cs
Mass
+
Molecular Ion Peptide
ALILTLVS
132.90488
829.53933
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