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Agilent’s New 7000B Quadrupole GC/MS/MS:

Evolving technology for higher performance tools

Terry L. Sheehan, Ph.D.GC/MS/MS Product ManagerAgilent TechnologiesSanta Clara, CA

Triple Quadrupole MS/MS(also called tandem quadrupole)

is changing the way scientiststhink about GC/MS

Transmission Quadrupole MS & MS/MSSensitivity and Selectivity Scale

Least Sensitive Most Sensitive

Fast 12,500 u/s (5975C)Fast 6,250 u/s (7000)

Typical < 2,000 u/s

Targets & Non-targetsTargets & Non-targets

*Multiple Reaction Monitoring Also called SRM for Selected Reaction Monitoring (similar to SIM – Selected Ion Monitoring)

Dwell 20-50 ms

Targets onlyDwell 1-50 ms

< <

Transmission Quadrupole MS & MS/MS Sensitivity and Selectivity Scale

Least Sensitive Most Sensitive

Fast 12,500 u/s (5975C)Fast 6,250 u/s (7000)

Typical < 2,000 u/sDwell 20-50 ms

Least Selective Most Selective

Unit mass resolutionUnit mass + AMDIS

deconvolution

Targets & Non-targetsTargets & Non-targets

Targets only

Targets only per DBL

Dwell 1-50 ms

< <

< <

Summary of Relative Performance Factors

MDL = Sanalyte

Nneutral + Nmatrix + NGC “bleed”

Quant Error = Smatrix

Sanalyte Smatrix+

GC/MSD Scan

GC/MSD SIM

GC/MSD Scan-DRS

GC/TQ MS/MS

S from analyte + +++ + +++S from matrix -- -- - 0N from neutrals = = = Ultra lowN from matrix --- --- - 0N from "bleed" Low Low Lower 0MDL (clean) Very Low Lowest Lower LowestMDL (very dirty) Low Low Lower LowestQuant Error Low Low Lower Lowest

But scan is essential for non-targets (unknowns)!

Fundamental Benefits of MS/MS for GC

Consistently High Sensitivity for Target Analytes in Complex Matrices

Higher Selectivity to Identify Target Analytes in Complex Matrices

MS/MS Eliminates Scan and SIM Interferences

Single Quad MSselectivity proportional to

spectral resolutionno selectivity against ions

with same m/zinterference

analyte

unit mass resolution


analyte

Product 1Product 3

Product 2



with same m/zinterference

analyte


Product Ion Spectrum


analyte

Product 1Product 3

Product 2



with same m/z

Triple Quad MSHigh probability that one or more of the

product ions will be a unique dissociation product of the precursor only

interference

analyte

interference


Product Ion Spectrum

MS/MS Ensures Lowest Detection LimitsEI: spectrum of analyte can also include ions from matrix, column bleed, gases, etc.

Q1 SIM isolate precursor before CID

chemical noise from these other ions is eliminated

MS/MS Ensures Lowest Detection LimitsEI: spectrum of analyte can also include ions from matrix, column bleed, gases, etc.


Product 1Product 3

Product 2

CID +Q2 SIM

Lower m/z Product Ions measured against zero chemical noise for consistently lower LOQ and LODs


SIM Limited by Unit Mass Resolution

GC/MS Single Quad SIM

Interfering matrix peaks = chemical noise

(mainlib) Eicosane20 40 60 80 100 120 140 160 180 200 220 240 260 280

0

50

100

29

43

57

71

85

99113 127 141 155 169 183 197 211 225 238 253 282

(mainlib) Benzene, hexachloro-20 40 60 80 100 120 140 160 180 200 220 240 260 280 300

0

50

100

2435

47

53

71

8395

107

118

130

142

153 165

177

190

214

237

249

284

Cl

Cl

Cl

Cl

Cl

Cl

in diesel fuel

C20 Eicosane Hexachlorobenzene




(mainlib) Eicosane20 40 60 80 100 120 140 160 180 200 220 240 260 280

0

50

100

29

43

57

71

85

99113 127 141 155 169 183 197 211 225 238 253 282

(mainlib) Benzene, hexachloro-20 40 60 80 100 120 140 160 180 200 220 240 260 280 300

0

50

100

2435

47

53

71

8395

107

118

130

142

153 165

177

190

214

237

249

284

Cl

Cl

Cl

Cl

Cl

Cl

in diesel fuel




At unit mass resolution SIM, HCB is buried by the more intense peaks from C20 to C22 hydrocarbons

in diesel fuel

MS/MS Succeeds Where MS Fails

GC/MS Triple Quad SRM


A chromatographer’s dream: single peak on flat baseline


in diesel fuel

Pesticide Analysis: Comparing SIM to MS/MS

Apple

Orange

Spinach

Ginseng

Cabbage

SIM ‐ EIC (246)

XX

Pesticide Analysis: Comparing SIM to MS/MS

Apple

Orange

Spinach

Ginseng

Cabbage

SIM ‐ EIC (246) MRM (2 transitions)

A c q u is it io n T im e (m in )1 1 .8 1 1 .9 1 2 1 2 .1 1 2 .2 1 2 .3

Cou

nts 4x1 0

-0 .1

0

0 .1

0 .2

0 .3

0 .4

0 .5

0 .6

0 .7

0 .8

0 .9

1

1 .1

2 4 6 .0 -> 1 7 6 .1 , 2 4 6 .0 -> 1 7 5 .1

R a t io = 2 3 .5

A c q u isitio n T im e (m in )1 1 .8 1 1 .9 1 2 1 2 .1 1 2 .2 1 2 .3

Cou

nts 3x1 0

0

1

2

3

4

5

6

7

8

2 4 6 .0 -> 1 7 6 .1 , 2 4 6 .0 -> 1 7 5 .1

R a t io = 2 3 .6


Cou

nts 4x 1 0

-0 .1

0

0 .1

0 .2

0 .3

0 .4

0 .5

0 .6

0 .7

0 .8

0 .9

1

1 .1

1 .2

2 4 6 .0 -> 1 7 6 .1 , 2 4 6 .0 -> 1 7 5 .1

R a t io = 2 4 .0


Cou

nts 3x1 0

0

1

2

3

4

5

6

7

8

2 4 6 .0 -> 1 7 6 .1 , 2 4 6 .0 -> 1 7 5 .1

R a t io = 2 2 .9

Acquisition Time (min)11.8 11.9 12 12.1 12.2 12.3

Cou

nts 4x10

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

246.0 -> 176.1 , 246.0 -> 175.1

Ratio=23.3

S/N = 448

S/N = 456

S/N = 260

S/N = 446

S/N = 241

XX

7000B MS/MS: High Performance Components

New inert, high temp, extraction EI source

New Ramped Iris Triple-Axis Detector

Gold plated quartz, hyperbolic quadrupoles

Split Flow Turbo

High pressure hexapole collision cell with Patent Pending Helium

Quench Gas

Hinged access to source and detector

The best technologies from the industry leading 5975C Series GC/MSD

– Heated monolithic gold plated quartz quadrupole

– Reliable, inert, high performance ion source

– High Signal/Low Noise Triple-Axis Detector

– PCI/NCI ion source

…and the 6400 Series LC/MS/MS

– Linear acceleration enhanced hexapole collision cell

– MassHunter software

7000B MS/MS: Hot (200ºC) Quartz Analyzers

Gold plated quartz, hyperbolic quadrupoles

High temperature analyzers stay clean even with highly concentrated, complex matrices

Proprietary Heated Quartz Quadrupole

• High temperature (200ºC)operation results in greater reliability with “dirty” samples

– MS/MS methods typically have more “dirty” samples and benefit even more from the higher temp quad analyzer

– Virtually eliminates the need to clean quads as a maintenance issue

• More stable tunes and methodsover a longer period of time in real world sample environments

Hyperbolic, Gold-coated, Quartz - Proprietary!

• Why quartz?– Low coefficient of linear expansion (expansivity; α)

• dL = α L0 dt [t = ºC; dL = change in length (m); L0 = length at 0 ºC]

– α varies greatly for different materials (m/m K x 10-6)• Stainless steel 304 = 17.3; Stainless steel 316 = 16.0• Inconel (Ni alloy) = 12.6; Monel (Ni/Cu alloy) = 13.5• Glass pyrex = 4.0; Quartz, fused = 0.59

– For change from 25 to 150 ºC, which material expands the LEAST?

• But what is the “analytical value?”• Low thermal expansion means dimensions/alignments do not change• After maintenance, the quartz quad can be baked at 200 ºC • Under operation, the quartz quad can be run at 150, 180 or even 200 ºC• Heat is the “friend” of GC/MS methods – true for the source and analyzer!

More Information on the Agilent Website

Detailed Video

7000B MS/MS: Collision Cell with Helium Quench

High pressure hexapole collision cell with Patent Pending Helium

Quench Gas

Highly efficient dissociation of precursor ions

Maximum transitions per second without product ion carryover (cross-talk)

Reduction of metastable helium from EI source

Hexapole Cell: Better Transmission Characteristics

00.20.40.60.8

11.2

0 500 1000 1500

Transmission

m/z

Mass Range TransmissionQuadrupole Hexapole Octapole

GC/MSm/z=1050

Quadrupoles are the best mass filters (analyzers)Hexapoles and octapole are the best transmission devices

500 Transitions/sec: Why Is This Important?

• Narrow GC peaks (width <1-2 s)• Sufficient data points (15) to perform acceptable/accurate

quantification• Many compounds to monitor in single run (multiresidue)

when coeluting peaks are quite frequent• Regulated environment where QC checks are required to

ensure data validity– Confirmatory transitions according to the 96/23/CE directive

Measured Collision Cell Clearing Time

1 msec

No product ion signal = no cross-talk at 500 transitions per second

Collision Cell Process: Typical Description

Collision Cell

Precursor Ions In

Product Ions Outcollision induced dissociation

1 ml/min N2Collision Gas

Source Detector

Quad Analyzer Quad Analyzer

This is a good description for LC/MS/MS, but it is not complete for GC/MS/MS

Collision Cell Process: Full Description for GC/MS

Collision Cell

Precursor Ions InHe* +

Product Ions Out + He*

In GC/MS, neutral noise is buried in much higher chemical noise.

In GC/MS/MS, chemical noise is greatly reduced so neutral noise is a critical source of noise.

collision induced dissociation

A high population of highly energetic helium metastables are produced in an EI source; since metastable helium is not charged, it can pass through mass analyzer field into the collision cell and through to the HED-EM


At the detector, metastable helium generates neutral noise.

Source Detector


Collision Cell Process with Helium Quench Gas

Collision Cell

Precursor Ions InHe* +

collision induced dissociation


Transmission of metastable helium to the detector is greatly reduced; the Triple-Axis detector further reduces neutral noise for ultra-low neutral noise.

Source Detector


He Buffer Gas

He* + He → 2 He + heat

Product Ions Out

+ He*

Benefit from “Helium Quench”

3x10

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

5.5

6

6.5

7

7.5

8

+ MRM (283.79999 -> 213.90039) 1pgHCB_std_cond_UnkGain_2090EMV_He20pct_04.d Noise (RMS) = 6.03; SNR (6.465min) = 1294.0

6.465

Counts vs. Acquisition Time (min)4.1 4.2 4.3 4.4 4.5 4.6 4.7 4.8 4.9 5 5.1 5.2 5.3 5.4 5.5 5.6 5.7 5.8 5.9 6 6.1 6.2 6.3 6.4 6.5 6.6 6.7 6.8 6.9 7

3x10

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

5.5

6

6.5

7

7.5


6.470

4.628 6.301

Counts vs. Acquisition Time (min)4.1 4.2 4.3 4.4 4.5 4.6 4.7 4.8 4.9 5 5.1 5.2 5.3 5.4 5.5 5.6 5.7 5.8 5.9 6 6.1 6.2 6.3 6.4 6.5 6.6 6.7 6.8 6.9

Helium Quench Gas ON

Helium Quench Gas OFF

tested with hexachlorobenzeneSame

Signal

SameSignal

S/N 341:1

S/N 1294:1

Noise Decreased2x10

0.5

0.6

0.7

0.8

0.9

1

1.1

1.2

1.3

1.4

1.5

1.6

1.7

1.8

1.9

2


Counts vs. Acquisition Time (min)4.7 4.75 4.8 4.85 4.9 4.95 5 5.05 5.1 5.15 5.2 5.25 5.3 5.35 5.4 5.45 5.5 5.55 5.6 5.65 5.7 5.75 5.8 5.85 5.9 5.95 6 6.05 6.1 6.15 6.2

In MS apps, neutral noise is not visible – it is buried by more intense chemical noise.Since MS/MS eliminates chemical noise, neutral noise is visible and mustbe minimized.

7000B MS/MS: New Higher Sensitivity Source

New inert, high temp, extraction EI source

Increase signal for precursor ions using an extractor source design

Lab-Proven Inert Source

• Solid, inert materials, not a coating that can be damaged by cleaning

• Stay-Clean (350ºC) designextends maintenance intervals with dirty samples

• Source tune parameters in the Autotune file

• Dual filaments reduces maintenance intervals

• 1000’s of satisfied MSD usersPartition between source and analyzer compartments support dual stage pumping

7000B Extraction SourceExternal View - Assembled

At first look, it does not look any different than the 7000A source

Like the 7000A source, the 7000B extraction source is more open than the MSD source – looks similar to the CI source

7000B Extraction SourceExpanded View of Components

And there is new connection to the lens #11

Component #7 is a new insulator

Design uses combination of “push” (repeller) and “pull” (extractor lens) to increase ion intensity

7000B Increased Signal

Extractor Source w/Ramped Iris Detector

Standard Source & Detector mode

Extractor source cutaway

Push-Pull Principle:Repeller and Extractor

Why Is the New Extractor Source More Valuable for

Triple Quad MS/MS than

Single Quad MSD?

Extractor Source Increases Signal for All IonsEI: spectrum of analyte can also include ions from matrix, column bleed, gases, etc.

Single Quad MSChemical Noise Limited

Analyte Signal IncreasedChemical Noise IncreasedS/N does not improve

Extractor Source Increases Signal for All IonsEI: spectrum of analyte can also include ions from matrix, column bleed, gases, etc.


Product 1Product 3

Product 2

CID +Q2 SIM


MS/MS still eliminates chemical noise

Why Is the New Extractor Source Valuable for MS/MS?

Single Quad MSChemical Noise Limited

Analyte Signal IncreasedChemical Noise IncreasedS/N does not improve

Triple Quad MS/MSProduct Ion Signal Limited

Product Ion Signal IncreasedChemical Noise Does NOT IncreaseS/N (and LOD) are improved

Unique Entrance Lens Eliminates Pre-filter

The last lens, which extends into the heated monolithic quadrupole, optimizes coupling of the source to the analyzer. Voltage on this lens is dynamically ramped synchronously with analyzer voltage to focus maximum ion signal for every m/z into the center of the quadrupole field.

Improved lens

7000B MS/MS: New Ramped-Iris Detector

New Ramped Iris Triple-Axis Detector

Ramped-iris improves transfer of ion from the exit of the second analyzer into the Triple-Axis detector

High Performance Triple-Axis Detector with NewDynamically Ramped Iris

• Maximum reduction of neutral noise

– The combination of helium quench gas and the Triple-Axis design delivers ultra low noise

• Long life and high linearity– Triple channel electron multiplier

• “Gain Normalization” corrects tune file for detector aging

– Consistent long term method sensitivity

• Design proven in the 5975C MSD

hyperbolic quartztransmission

quadrupoleanalyzer

triple channelelectron multiplier

high energy dynode

steering rod

ion beamX

Y

OffsetZshield for

secondary particles

triple channelelectron multiplier

high energy dynode

steering rod

ion beamXX

YY

OffsetZ

OffsetZshield for

secondary particles

• Improved high mass peak shape• Improved mid to high mass sensitivity

Mass Iris (V) Signal Increase

69 -1 1.0X

131 -8 1.1X

264 -13 1.5X

414 -20 2.5X

502 -30 2.8X

614 -35 4.0X

Ramped-Iris Triple-Axis Detector

7000B GC/MS/MS Specifications

OFN = octafluoronaphthaleneBZP = benzophenone

EI SRM > 500:1 for 100 fg OFNPCI SRM (CH4) > 50:1 for 100 fg BZP

EI MRM Pk Area RSD < 8% for 100 fg OFN

EI Scan > 300:1 for 1 pg OFNNCI SIM (CH4) > 200:1 for 10 fg OFN

Installation Checkout Specifications

Typical Precision

Typical Sensitivity

ColumnBackflush:

Many Wonderful Advantages for GC/MS/MS of

Complex Matrices

What Real Value Does High Performance EPC Have If Matrix Carryover Destroys Rt Precision?

m in5 . 3 5 . 4 5 . 5 5 . 6 5 . 7 5 . 8 5 . 9 6 6 . 1 6 . 2

N o r m .

2 0 0 0

4 0 0 0

6 0 0 0

8 0 0 0

1 0 0 0 0

1 2 0 0 0

1 4 0 0 0 10 Runs without BackflushingRetention times shift ~4-5 secduring 10 runs

m in5 . 3 5 . 4 5 . 5 5 . 6 5 . 7 5 . 8 5 . 9 6 6 . 1

N or m .

0

2 0 0 0

4 0 0 0

6 0 0 0

8 0 0 0

1 0 0 0 0

1 2 0 0 0

1 4 0 0 0

1 6 0 0 0

10 Runs with Backflushing

Productivity: CFT Backflush

• More analyses per hour . . . – Shorter analysis cycle time (time from injection to injection) – Reduced requirement for sample preparation

• with consistently better results . . . – Quantitative: accuracy, precision, detection limits, linear range– Qualitative: response ratios, confirmatory retention times, library

searches (eliminate false negatives and false positives)

• while using less operator time for each analysis.– Less time performing maintenance on column and ion source

Multiple Backflush ConfigurationsMultiple Backflush Devices

• Backflush from the start of the column– Backflush guard column only

• Backflush from the end of the column– Backflush entire column

• Backflush from the middle of the column– Backflush a percentage of the column– Example: at tee between two 15 m columns

Purged UltimateUnionNew Pressure

Controlled Tee Kit

Backflush – Mid-Column (Backflush Mode)

Vent

Pressure / Flow

Controller

15-m HP-5ms (0.25mm id x0.25um)

15-m HP-5ms (0.25mm id x0.25um)

5975C MSD or 7000B MS/MS

7890A GC

Split/Splitless Injection Port

Decrease inlet pressure during backflush

Pressure Controlled Tee

Backflush with Purged Ultimate Union: Lipid Peroxidation Products in Blood

App Note 5989-9359EN

4-hydroxy-2,3-nonenal (HNE), an indicator of oxidative stress, and its metabolite, 1,4-dihydroxynonene (DHN)

PCI with NH3

Late eluting peaks were quickly contaminating the source

Backflush with Purged Ultimate Union: Loss of Response without Backflush

without backflush

with backflush


Backflush with Purged Ultimate Union: Loss of Response without Backflush

without backflush

with backflush

These were the first backflush results; the system is getting clearer


Pesticide Analysis with New Agilent Jet Stream Technology

Break for Questions

For questions, at break please dial *1 on your phone, or type onto the Q&A box at any time duringthe presentation.

December 12, 2008

MassHunter Software

MassHunter

Quantand

Qualmodules

MassHunter

Quantand

Qualmodules

MassHunter SoftwareCommon Workflow for all Mass Spec. applications

LC/MS Acquisition ModuleTriple Quad LC/MS, LC/TOF, LC/QTOF

LC/MS Acquisition ModuleTriple Quad LC/MS, LC/TOF, LC/QTOF

GC/MS Acquisition ModuleTriple Quad GC/MS

GC/MS Acquisition ModuleTriple Quad GC/MS

MassHunter formatMassHunter format

Translator

Translator

ChemStationSingle Quad LC/MSChemStation

Single Quad LC/MS

ChemStationSingle Quad GC/MS (MSD)

ChemStationSingle Quad GC/MS (MSD)

MassHunter Data Analysis ApplicationsOne software for all your Agilent MS applications

•Target Compound Analysis• High throughput quantitation

•Quantitative Analysis •Qualitative Analysis•Discovery & Method Development• Find features• Identify compounds

Quantitative Analysis View

Results Information


Compound Information

Results Information


Compound Information

Results Information

Calibration Information

Results Information:All Results for batch in one view

• Easy visualization for all results– Customize columns for desired data

• Easy Navigation– Compound and Calibration views change when new compound is selected– Reduce data by filtering on sample type, compound, time segment, and outlier

Results Information:Detailed compound view

•Detailed compound information is easily seen

Results Information:Highlight outliers

•Compounds with results outside specified ranges are highlighted• 30 metrics are available• Custom Calculation is possible• More to be added

– i.e. Peak metrics

Results Information :Simple Metrics – Plot Columns

•Ability to see trends in a batch•Ability to diagnose quantitation problems easily• Missing ISTD amount• Retention time drift

Compound Information:All relevant information in one view

CompoundIntegration

QualifierInformation

SpectrumInformation

ISTDIntegration

ISTD QualifierInformation

ISTD SpectrumInformation

Calibration Curve Views

Can include ISTD, QC, and CC data points on curvePoint information is available as pop-up

QC point

Calibration Curve Assistant

Display all possible Calibration Curve fit options•Review R2, Standard error, or Max % Residual•Can sort, filter, or browse options

Calibration Curve Assistant

Filter possible calibration curves based on requirements

Compare selected calibration fit (black) with proposed (blue).•Linear•All points•Ignore origin

Reporting and Data Transfer

Use of XML results format• Easy data exchange

– LIMS data exchange– Copy/Paste views from MassHunter

Microsoft Excel reporting engine• Powerful Features

– Excel Macros and formulas can be created for custom result manipulation

• Flexible Reports and Layouts– Custom Layouts and graphics

properties.• Preconfigured report template

examples

Productivity of GC/MS/MS

• Sample complexity forces the change from MS to MS/MS– Could not meet required LOD/LOQ in real samples

– Could not avoid interferences from matrix ions (isobaric ions)

Productivity of GC/MS/MS

• Sample complexity forces the change from MS to MS/MS– Could not meet required LOD/LOQ in real samples– Could not avoid interferences from matrix ions (isobaric ions)

• Practical aspects of productivity– MS/MS selectivity for less sample preparation– MS/MS sensitivity for less injected on-column

• In turn, less maintenance– MS/MS selectivity for faster analysis cycles

• MS/MS selectivity lessens the requirement for column selectivity• Less bakeout or backflush may be needed

– Caution: selectivity is a type of ‘blindness’ and can be misleading

– MS/MS selectivity for faster and easier peak integration• What can be easier than a flat baseline and a single peak

Pesticide Analysis with New Agilent Jet Stream Technology

Break for Questions

For questions, at break please dial *1 on your phone, or type onto the Q&A box at any time duringthe presentation.

December 12, 2008

Thank You for Attending Today’s eSeminar

Page 73

Terry L. Sheehan, Ph.D.GC/MS/MS Product Manager

Agilent Technologies, Santa Clara, CA

agilent’s new 7000b quadrupole gc/ms/ms · agilent’s new 7000b quadrupole gc/ms/ms: ... apple...

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