61st ASMS Conference on
Mass Spectrometry and
Allied Topics
Minneapolis-St Paul, MN,
USA
GC/MS on the Move:
Steady Steps,
Long Strides and
Quantum Leaps
Agenda
Part 1:
Overview of Changes in the GC/MS Product Lines
Elizabeth Almasi, Product Manager, GC/TQ and GC/IT
Part 2:
Update of the 7200 GC/Q-TOF Applications
Sofia Aronova, Ph.D, Application Chemist
Today’s Challenges Economic Backdrop: Tension and Uncertainty
• Operating expenses increasing
• Capital Budgets decreasing
• Asset management and protection a priority Operations
• Regulations, compliance and technology driving need for lower level detection
• Sample complexity increasing
• Many analytes are labile
Technology
Agilent Response Solutions to Assure Business Continuity & Reduce Uncertainty
Reduced operating
expenses:
gas use and power use
Increased productivity:
fast MS vent mode &
Rapid consumables
Parts Finder tool
Improved asset
utilization & protection:
maintenance feedback &
improved GC:MS
communication
Economic
A more facilitated user
experience
Powerfully Intuitive
MassHunter Software
wit GC<-> MS
Integrated Intelligence
Built-in optimization
tools
Analyzer Solutions
For rapid start-up
Execution
Better analyte detection:
Improved performance
specifications
Improved software tools
and databases:
Rapid method
development Optimized
analysis parameters
Better assurance of
sample integrity,
protection of labile
compounds:
Agilent Inert Flow Path
Technical
Innovations in Hardware, Software, Chemistries and Workflows
7890B/5977A GC/MSD
• Performance
• Carbon footprint
• Cost of operation
• Productivity
Extractor EI Source: All About More Ions
Better Signal, Improved Precision Extractor ion source
Non-Extractor 40ppb
solution (SVOC)
Extractor 40ppb
solution (SVOC)
Better Science + Engineering = Better Specs
Higher S/N > 1500:1 for 1 pg OFN
Lower IDL < 10 fg OFN
8 consecutive injections
Tested at factory and at
installation
Faster Scan Rate = 20,000 u/s
Vacuum system Optimized for GC/MS Designed and manufactured to Agilent Quality Standards
EDM – 8 turbo
stages (rotor)
designed to optimize
pumping speed for
light gases
“i” for inverter technology
Energy saving sleep mode
More than 80% of the MS power
usage comes from the foreline
pump.
During Sleep mode power
usage of the pump is reduced
by 40%
3 TwisTorr drag stages (spiral
channels) designed for high
compression ratio
performances
255 L/s helium
220 L/s hydrogen DS42i
Agilent TwisTorr 304 FS New Foreline Pumps
GC/MSD Software MassHunter Qual
MassHunter Quant MH
CS
MassHunter GC/MS Acquisition
And Use Either
MassHunter Data Analysis
OR
MSD ChemStation Data Analysis MSD ChemStation DA
G1701FA
5977 with Both ChemStation &
MassHunter Addressing customer concerns of obsolescence • Full support for the ‘past’ and ‘future’
5977 MSD delivered with ChemStation and MassHunter
Software support agreement provides free
update/upgrades for 1 year
•For the 40,000+ ChemStation Users
Instrument control modeled after ChemStation - with
enhancements
Data backward compatible with previous product
All ChemStation features retained including macros and
macro-based programs such as EnviroQuant and
Deconvolution Reporting Software
For all GC/MSD Users Productivity benefits of MassHunter Quant and Qual
Use MassHunter or ChemStation or both -- at any time
MSD ChemStation OR MassHunter?
MSD ChemStation
Experienced GC/MSD User
• Adding instrument to routine production
• Established workflows
• Macros, Macros, Macros
DRS or Screener DB
EnviroQuant User
• Uses the US EPA workflow, especially
reports and QA/QC criteria.
• MassHunter EnviroQuant targeted for
early 2014
MassHunter New to Agilent or GC/MSD
Superior Quantitation Tools
21CFR11 or DrugQuant
Targeted deconvolution,
Multi disciplinary MS lab
• Already using MassHunter for LC/MS, ICP-MS, & GC/MS
Change as new tools are introduced
• Agile II integrator
Lower Detection Levels,
Higher Productivity:
The 7000C Triple Quad MS
The 7000C performance is powered by these components
Triple-Axis Detector Drastically reduces neutral
noise, ensuring a cleaner signal
and lower detection limits.
Second-generation extractor EI or PCI/NCI
ion source with improved thermal profile Maximizes the number of ions that are successfully
transferred out of the ion body and into the quad
analyzer.
High-temperature, gold-plated
monolithic quartz quadrupole The quartz monolith guarantees perfect
alignment for hyperbolic surfaces throughout
the life of the MS. Gold surfaces stay clean and
maintenance free in high temperatures – up to
200 °C.
MassHunter Software: From Tune to reports, with
efficient workflows
High-capacity
split flow turbo
molecular pump Ensures optimal vacuum
conditions even at high
flow rates.
Pesticides and Environmental
Pollutants Database – The most extensive
and comprehensive database with all pertinent
MS/MS parameters
New MRM
Optimization Efficient, simple,
customizable
Proven specifications
IDL: 4 fg OFN 10fg OFN, Splitless injection
transition 272 m/z =>222
S/N: 7000:1 100fg OFN injected,
transition 272 => 222
MS/MS your way, the best way
Seamless MRM
method
optimization
Ease of use
Efficiency
• The new MRM optimization allows maximum
utilization of the dwell time for the best sensitivity
• Compound based adjustment of the optimization is
possible, unique to Agilent
• New, direct connection between the acquisition
method editor and MRM optimization program
Benefits of the new MRM Optimization
Customizable dwell times
Simple
Efficient
Flexible
0.1ppb
Dichlobenil: 7000C – winter squash
0.1-100ppb; R2=0.9996
5 injections at each level
Quant/Qual ion
ratio stability at
different
concentrations
Self-Cleaning Ion Source (He + H2)
Novel Approach to Hydrogen
• Real Change in How We Think about H2
– The past = ‘digital’ approach of 100% helium or 100% hydrogen
• 100% H2 has limitation: spectral changes, lower sensitivity, surface activity
– Now = optimal blend of He and H2 (EPC controlled)
– Now = optimal amount of H2 to clean the ion source
• So Novel . . . It’s Patented
– Impacts consistency of MS performance
– Impacts frequency of source maintenance
Available on Advanced PAH Analyzer
Self Cleaning Ion Source: PAH results
Integrated Intelligence –
Reducing Time, Energy and Gas use Operational Savings -Time
• FastVent – shorter maintenance cycle
• Better Early Maintenance Feedback
• Fix before failure
• Parts Finder
• More wizards and aids
Energy Savings -Power
• Sleep/Wake functions control:
• GC and MS Heaters
• New Agilent Pumps (5977 only)
Gas Savings
• Conservation of costly helium
Agilent 5977 GC/MSD
Agilent 7000C
Triple Quadrupole GC/MS
Improve Asset Utilization Leverage and Protect Your Investment Better
5977/7000C
7890B
During MS vent… Increases GC carrier gas flows,
Cools MS faster
Upto 40% Faster Vent Times
(280C 100C)
In event of MS Failure… Stops GC Flows
Don’t waste expensive He
Don’t build-up H2
5977/7000C
7890B
GC monitors GC/MS
connection.
If communication lost… GC shuts down various
(GC) thermal zones
Integrated GC-MS Communication
Faster Maintenance, better asset protection, safer H2 use
Integrated Early Maintenance Feedback Improving Operational Efficiency - Time
Select time-
based or
frequency-
based
counters
Choose
maintenance
parameters
that are
relevant to
your own app
Set warning
and Action
limits
Customize
user-defined
EMF
Example:
Check gas
cylinder after x
injections
Integrated Parts Finder Click-click shopping list – Time savings
Set Up Backflush Easily Saves time and keeps system clean
Interviews
customer about
configuration &
method and
makes
recommendation
New Integrated
Backflush MTS
GC Option
7890B Integrated Sleep/Wake Modes Reduce Power/Gas Costs
Set sleep and
wake times
Customize
sleep-wake
protocol:
Example -
Condition GC
before daily
workflow
begins Define sleep
and wake
methods
Reducing Operating Expenses Reducing Dependence on He
Switching to N2 gas overnight and on weekend downtime can save up to 180 days of He per
GC instrument
He Conservation Module
Manage He Use Optimally
• Use He only when needed
• Extend He tank usage
• Reduce operating costs
• Reduce gas supply uncertainty
C o r e C o m p e t e n c y
Proprietary Deactivation Chemistry for
Glass, Steel, Gold, Fused Silica, Glass Wool
Ultra Inert Flow Path:
Protection from Injection to Detection
Flexible Metal Ferrule Design Grooves
reduce bulk stiffness during
compression and improves ID
constriction.
Angle eliminates wall frictional
contact to allow the material to
flow into the ID area for larger
range of constriction.
Tighter ferrule ID
specification and
tolerances (30%)
Wider opening
allows for easier
column insertion
1
2
3
4
Chromatograms for sensitive OP pesticides
UltiMetal Plus on Flexible Metal Ferrule Is deactivation needed for such small surface?
4 x10
0
1
2
4 x10
0
1
2
Counts vs. Acquisition Time (min)
4.5 5 5.5 6 6.5 7 7.5
Untreated FMF Post column Std 500ppb
FMF + UltiMetal Plus Post column Std 500ppb
Acephate
Omethoate
Demeton-S
Acephate
Omethoate Demeton-S
H2 as carrier gas: Advantages and Concerns Driven by the decreased supply and increased cost of He
Safety
Spectra
Sensitivity
Surface
Activity
• Safety Considerations
• Gas Source Requirements
• Hardware considerations
Magnet
Drawout Plate
• Plumbing considerations
• Column selection
• Flow rate
• Method translation -
preserving Rt
Complete H2 Conversion Protocol from Agilent
7000C MS, 5977 MSD and 7890B GC Driving Business Value and Success
Best Sensitivity
Proven GC/MS Reliability
Unmatched Software Options
Integrated Intelligence
Greater Operating Efficiency
Expanded Accessories and Options
Part 2:
Application Update for the 7200 GC/Q-TOF
Sofia Aronova, Ph.D, Application Chemist
GC/MS on the Move
7200 GC/Q-TOF Applications
Sofia Aronova
GC/Q-TOF Application Chemist
September 18, 2013
September 19, 2013 35
7200 Series GC/Q-TOF
September 19, 2013 36
High resolution
Accurate mass
Fast acquisition of full spectra
MS/MS mode
• High resolution/accurate mass product ion spectrum
• Increased selectivity
• Structural elucidation
Ideal tool for solving complex analytical problems
7200 GC/Q-TOF Applications Summary ASMS Posters and Applications
September 19, 2013 37
Metabolomics and
metabolic profiling
Pesticide residues
in food
Flavors and
Fragrances
Natural products
Forensics
Petrochemicals
Environmental
7200 GC/Q-TOF Success Stories German Sports University
Expanding Sports Drug Testing Assays
September 19, 2013 38
Utilized Agilent GC/QTOF and
LC/QTOF to characterize the
fragmentation pathways of selective
androgen receptor modulator drug
candidates
7200 GC/Q-TOF Success Stories Shanghai Institute of Organic Chemistry
Rapid Screening of Pesticides in Vegetables
September 19, 2013 39
Utilized Agilent GC/QTOF to create a
187 pesticide library that was
subsequently used to successfully
screen vegetables obtained from a
local market.
7200 GC/Q-TOF Success Stories Stanford University
Identification of Drug Targets
September 19, 2013 40
Utilized Agilent GC/QTOF to
characterize the targets of putative
antifungal drugs.
Examples of GC/Q-TOF Applications
September 19, 2013 41
Metabolic profiling of yeast sterols using the Agilent 7200 Series GC/Q-TOF system
Metabolomics of Carbon Fixing Mutants of Cyanobacteria by GC/Q-TOF
Metabolomics of Opiate-Induced Changes in Murine Brain by GC/Q-TOF
Untargeted Metabolomic Analysis of UV Stress Response in C. reinhardtii by GC-QTOF
Simultaneous analysis of tryptophan, kynurenines and amino acids using the GC/QTOF in Negative CI mode
Accurate mass retention time locked flavor database by GC/Q-TOF
Analysis of trace levels of sulfur compounds in coffee by the Agilent 7200 GC/Q-TOF system
Olive oil characterization using Agilent GC/Q-TOF MS and Mass Profiler Professional software
Rapid simultaneous screening of multiple pesticide residues in Food matrices
Simultaneous targeted and non-targeted screening for pesticides in vegetables by GC/Q-TOF MS
Analysis of biomarkers in crude oil using the Agilent 7200 GC/Q-TOF
Characterization and classification of heroin from illicit heroin seizures by GC/Q-TOF
Unknowns analysis of natural products using GC/Q-TOF and GC/IonTrap in EI and PCI modes with MS/MS
Determination of odor compounds in surface water by solid phase micro-extraction and GC/Q-TOF
The role of GC/QTOF in exposomics
7200 GC/Q-TOF Applications Summary ASMS Posters and Applications
September 19, 2013 42
Pesticide residues
in food
Flavors and
Fragrances
Natural products
Forensics
Petrochemicals
Metabolomics
Environmental
Metabolomics of Opiate-induced Changes in
Murine Brain by GC/Q-TOF The Problem
September 19, 2013 43
Chronic opiate therapy often results in opiate dependence and
addiction.
Understanding the mechanisms of opiate addiction is important
for developing novel therapeutic strategies.
Untargeted metabolomics approach was utilized to characterize
a murine model of opiate dependence.
Sofia Aronova1 and Stephan Baumann1
Manhong Wu2, Ming Zheng2, David Clark2 and Gary Peltz2
Peyman Sahbaie3
1 Agilent Technologies, Santa Clara, CA 2 Department of Anesthesia, School of Medicine, Stanford University, CA 3 Veterans Affairs Palo Alto Healthcare System, Palo Alto, CA
Experimental Setup
September 19, 2013 44
morphine Saline
(control)
C57BL/6
(morphine-sensitive)
129Sv1
(resistant)
Brainstem tissue collected
Metabolites extracted and derivatized
morphine Saline
(control)
4-day treatment
Data Analysis Workflow
Chromatographic deconvolution and library search
Export compound information as CEF files
Statistical evaluation of the data in MPP
• Data clustering on PCA plot
• Significance and Fold Change Analysis
Confirmation of compound identity using accurate mass
information and structure elucidation tools of MassHunter
MS/MS and positive CI for identification of unknowns
September 19, 2013 45
Chromatographic Deconvolution
September 19, 2013 46
• Deconvolution was able to distinguish about 700 components in each sample.
• For about 100 components the corresponding hits were found in the Fiehn metabolomics RTL library
Metabolite Classes
September 19, 2013 47
organic acids 16%
amino acids 21%
carbohydrates 18%
lipids 11%
nucleic acid metabolism
11%
other 23%
Statistical Evaluation 129Sv1 strain – control vs treated
September 19, 2013 48
Saline
(control)
morphine
PCA plot of 129Sv1 strain mice
(control vs morphine treated)
shows the existence two distinct
clusters
Principal Components Analysis (PCA) plot
Statistical Evaluation 129Sv1 vs C57BL/6
September 19, 2013 49
PCA plot of interstrain
comparison (controls) shows the
existence two distinct clusters
C57BL/6
(morphine-sensitive) 129Sv1
(resistant)
Principal Components Analysis (PCA) plot
Fold Change Analysis C57BL/6 (morphine-sensitive)
September 19, 2013 50
adenosine
morphine Saline
(control) adenosine
C57BL/6
(morphine-sensitive)
The level of adenosine is decreased in morphine-administered mice as
compared to control in C57BL/6 strain (p<0.05)
Fold Change Analysis C57BL/6 (morphine-sensitive)
September 19, 2013 51
C57BL/6
(morphine-sensitive) 129Sv1
(resistant) Citric acid
α-Hydroxyglutaric acid
Glyceric acid
Adenosine 5’-
monophosphate
Adenosine
Cholesterol
N-Acetylaspartylglutamic acid
C57BL/6 (morphine dependent) vs
129Sv1 (resistant)
The level of several compounds significantly differs between the two strains (p<0.05)
Easy Confirmation of Identified Compound Using
Accurate Mass Information
September 19, 2013 52
Adenosine
Molecular Formula Generator (MFG) with Fragment Formula Annotation (FFA) and Library Search allows for easy confirmation of identified compounds
Identification of the Molecular Formula of an
Unknown Using Accurate Mass Information
September 19, 2013 53
Molecular ion is unknown, making it difficult to confirm the correct
annotation even using accurate mass information
Annotated spectrum of an unknown compound at 10.34 min accumulated
in 129Sv1 after morphine administration as compared to untreated control
MI ?
Importance of MS/MS in Unknown Identification Co-eluting compounds
September 19, 2013 54
EI MS/MS, selecting ion 228, CE 5 eV
EI MS
• m/z 129.1022 and 72.0808 are not product ions of m/z 228.0665 used as a precursor
• In fact, abundances of m/z 129.1022 and 72.0808 ions differed between the two strains but abundances of m/z 228.0665 and its product ions did not
Identification of Molecular Formula: PCI Methane
September 19, 2013 55
EI MS
CI MS
• CI helps to identify the molecular ion (MI) and thus empirical formula of the unknown compound
• Multiple adducts for MI as an additional confirmation point
Conclusions
September 19, 2013 56
Metabolomics studies can take advantage of multiple hardware features of 7200 GC/Q-TOF such as: • accurate mass information • full spectrum sensitivity • MS/MS capability • availability and easy switch between EI and CI modes Using multiple software capabilities such as: • chromatographic peak deconvolution and library search
(Unknowns Analysis) • statistical analysis in MPP • automatic fragment formula annotation of the tentative hits (Qual) • and more
Metabolomics of Opiate-induced Changes in
Murine Brain by GC/Q-TOF Poster TP 388
September 19, 2013 57
• Chromatographic deconvolution in MassHunter
• Statistical analysis in MPP
• PCI with accurate mass to identify empirical formula
• MS/MS to find contaminating ions
• Structure elucidation and ID confirmation with MSC
Metabolomics of Carbon Fixing Mutants of
Cyanobacteria by GC/Q-TOF Poster TP 419
September 19, 2013 58
• Chromatographic deconvolution in MassHunter
• Statistical analysis in MPP
• ID confirmation using accurate mass
• Pathway analysis in MPP
Untargeted Metabolomic Analysis of UV Stress
Response in C. reinhardtii by GC-QTOF Poster TP 412
September 19, 2013 59
• Chromatographic deconvolution in MassHunter
• EI and CI modes for identification
• Statistical analysis in MPP
• Pathway analysis in Cytoscape
Characterization and Classification of Heroin from
Illicit Heroin Seizures by GC/Q-TOF Poster WP 123
September 19, 2013 60
• Chromatographic deconvolution in MassHunter
• PCA and hierarchical clustering in MPP
• ID confirmation using accurate mass and MassHunter
tools
• Building Class Prediction model in MPP
Accurate Mass Retention Time Locked Flavor
Database by GC/Q-TOF Poster MP 685
September 19, 2013 61
• Custom accurate mass RTL library
• Evaluation of accurate mass library using olive
oil sample
Rapid Simultaneous Screening of Multiple
Pesticide Residues in Food Matrices Poster ThP 552
September 19, 2013 62
• Exact mass RTL database
• Rapid targeting screening workflow using the exact
mass database
• The database was evaluated using 3 different food
matrices
Simultaneous Targeted and Non-targeted Screening
for Pesticides in Vegetables by GC/Q-TOF Poster TP 605
September 19, 2013 63
• Targeted pesticide screening workflow using
accurate mass database
• Untargeted pesticide screening workflow
• MS/MS confirmation
Unknowns Analysis of Natural Products Using
GC/Q-TOF and GC/IonTrap in EI and PCI Modes with
MS/MS Poster MP 684
September 19, 2013 64
• Workflow for identification of unknowns using full
spectrum acquisition mode
• Structure elucidation of the unknowns acquired in
MS/MS mode with MSC
Simultaneous Analysis of Tryptophan, Kynurenines
and Several Amino Acids using GC/Q-TOF in NCI Poster TP 297
September 19, 2013 65
Determination of Odor Compounds in Surface
Water by SPME and GC/Q-TOF MS Poster MP 634
September 19, 2013 66
The Role of GC/Q-TOF in Exposomics Poster TP 305
September 19, 2013 67