gc/ms on the move: steady steps, long strides and ... ms...quantum leaps agenda part 1: overview of...

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

E:/_Jennifer/NPT Videos/EMF2.exe

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


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


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


Utilized Agilent GC/QTOF and

LC/QTOF to characterize the

fragmentation pathways of selective

androgen receptor modulator drug

candidates

http://www.google.com/url?sa=i&rct=j&q=german+sports+university+cologne&source=images&cd=&cad=rja&docid=fK8789twrBMxPM&tbnid=ZZl1H5Y_XAjbsM:&ved=0CAUQjRw&url=http://www.markify.com/trademarks/ctm/deutsche+sporthochsule+k%C3%B6ln+german+sport+university+cologne/006419667&ei=OnWRUdPjKIzQigKT14CgCw&bvm=bv.46471029,d.cGE&psig=AFQjCNEa5te4I6STPNrdc3J1Wkg1c2ODoQ&ust=1368573614765344

7200 GC/Q-TOF Success Stories Shanghai Institute of Organic Chemistry

Rapid Screening of Pesticides in Vegetables


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


Utilized Agilent GC/QTOF to

characterize the targets of putative

antifungal drugs.

http://www.google.com/url?sa=i&rct=j&q=stanford+university+logo&source=images&cd=&cad=rja&docid=WxBa2TAQwiDy-M&tbnid=PPV5AGdywi3hqM:&ved=0CAUQjRw&url=http://www.water-energy-food.org/en/calendar/view__478/connecting-the-dots-the-food-energy-water-and-climate-nexus-stanford-university.html&ei=YniRUcTRDcGyiQLlhoH4BQ&bvm=bv.46471029,d.cGE&psig=AFQjCNHvFLEG3jkmsVZrJWexchkIlAOAMw&ust=1368574428585296

Examples of GC/Q-TOF Applications


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


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


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


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


Chromatographic Deconvolution


• 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


organic acids 16%

amino acids 21%

carbohydrates 18%

lipids 11%

nucleic acid metabolism

11%

other 23%

Statistical Evaluation 129Sv1 strain – control vs treated


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


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)


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)


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


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


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


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


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


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


• 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




• 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



• EI and CI modes for identification


• Pathway analysis in Cytoscape

Characterization and Classification of Heroin from

Illicit Heroin Seizures by GC/Q-TOF Poster WP 123



• 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


• 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


• 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


• 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


• 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


Determination of Odor Compounds in Surface

Water by SPME and GC/Q-TOF MS Poster MP 634


The Role of GC/Q-TOF in Exposomics Poster TP 305


gc/ms on the move: steady steps, long strides and ... ms...quantum leaps agenda part 1: overview of...

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