Agilent LC/MS Nathan Miller · LC/MSD TOF 1994, 2000 LC/MS 1980 - 2011 2003 Agilent’s > 40 year...

Agilent LC/MS

Overview

Nathan Miller

LC/MS Product Specialist

Over 37,000 GC/MS, 4,500 ICP-MS and 6,500 LC/MS systems through June 2012 Agilent (and previously, as HP) has sold more quadrupole MS systems than any other mfr.

1971, 5930A

2005, 5975A GC/MS 1971 - 2011

LC/MSD, LC/MSD Ion Trap

LC/MSD TOF

1994, 2000

LC/MS 1980 - 2011

2003

Agilent’s > 40 year heritage in mass spectrometry 2008, 7000A

LC/QQQ

LC/QTOF

2006

1976,

5992A

1994, 4500 Series

ICP-MS 1994 - 2011

2012 8800 QQQ

2000, 7500 Series

Direct Liquid Injection

1980, 5985 SQ

Thermospray, 5987

Particle Beam, 5988

ESI on 5989

MS Engine

2011

7200A

Preparative LC Capillary &

Nano LC

Standard LC Ultra High Pressure

LC

High Sensitivity

HPLC-Chip/MS

Agilent High Performance Infinity Series LC:

Best in class “Front End” systems for MS

The industry’s most comprehensive LC portfolio to configure an

integrated LC/MS system

MassHunter Workstation One software for all Agilent Mass Specs

Minimize the learning and optimize the use of software in your

lab across different mass spec instrument platforms

• Control of Agilent 6000 Series QQQ, TOF and Q-TOF

• Control of most 1260 and 1290 HPLC modules, 7100 CE

• Control for GC QQQ and Q-TOF

• Data Processing for QQQ, TOF, Q-TOF, SQ*) and GC Ion Trap*)

• Control & data processing for ICP/MS

LC/MS GC/MS ICP-MS *) after conversion to MassHunter format

7/30/2013

LCMS Overview Jan 2013

Agilent 6000 Series LC/MS portfolio

6200 series TOF

6100 series Quad

6500 series Q-TOF

6400 series QQQ

High Resolution

Low Resolution

Sin

gle

Stag

e Tan

dem

Stage

Qualitative

Quantitative

ChemStation

Easy Access

G2725AA

6120B – 2,000 m/z

6130B – 3,000 m/z

6150B – 1,400 m/z

MassHunter

Data Translator

Single Quad

Control Software

Single Quad

Data Analysis

Software

Single Quad LC/MS – Proven Solutions

Analytical Studio

Reviewer, G3772AA

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MultiMode HPLC Chip

APCI ESI

APPI AJS

nanoESI

Single Quad Supports Wide Range of Ion Sources seven Agilent ion sources and third party sources

Plus Direct Sampling

Sources from Ion Sense:

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Agilent Jet Stream

Gas Dynamics View

The super-heated sheath gas collimates

the nebulizer spray and creates a

dramatically “brighter source”

Nozzle voltage

Resistive sampling

capillary

Nebulizing gas

Super-heated sheath gas

Heated drying gas

Enhanced efficiency nebulizer

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iFunnel Technology

7/30/2013


iFunnel Technology Revolutionizes Ion Sampling

Agilent Jet Stream

• Thermal confinement of

ESI plume

• Efficient desolvation to

create gas phase ions

• Creates an ion rich zone

Hexabore Capillary

• 6 capillary inlets

• Samples ~10 times

more ion rich gas from the

source

• Captures the majority of

the gas from the source

region

Dual Ion Funnel

• Removes the gas but

captures the ions

• Removes neutral noise

• Extends turbo pump life

Captures up to 10 x more ions

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6230B Accurate Mass TOF LC/MS

True Hi-Def TOF Technology:

NO PERFORMANCE COMPROMISE!

• Unmatched Mass Accuracy

< 1 ppm MS

• >22,000 Resolution

• 105 Dynamic Range

• Up to 30 spectra/second

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Formulas for TOF performance

Resolving power =

(m/z)/w1/2

1000/0.025 = 40,000

922/0.022 = 41,909

Mass accuracy ( rel. mass error)

(1000.000-1000.001)/1000)x106=1 ppm

Therefore: 1 ppm @ 1000 m/z is 0.001Da

(1 mDa)

921.5 922.0 922.5 923.0 923.5 924.0 924.5 m/z, amu

923.0121

W1/2 = 0.022

610actual

measured - actual accuracy Mass

+TOF MS: 5.474 to 5.547 min from steroid2TOFap42.wiff Agilent, subtracted (4.696 to ... Max. 1.2e5 counts.

120 140 160 180 200 220 240 260 280 300 320 340 360 380 400m/z, amu

0.00

1.00e4

2.00e4

3.00e4

4.00e4

5.00e4

6.00e4

7.00e4

8.00e4

9.00e4

1.00e5

1.10e5

1.16e5

In

te

ns

ity

, c

ou

nts

289.2159

271.2051

269.1893

287.1997

Epitestosterone

Resolving power:

6890

actual 289.2162

meas 289.2159

-1.06 ppm

922.0092

6000 Overview Diploma Training 2006

Agilent Restricted

Page 14

Rapid Analysis of Pharmaceutical Compounds

0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.45 0.50 0.55

0.60 Time, min

0.0

5.0e4

1.0e5

1.5e5

2.0e5

2.5e5

3.0e5

3.5e5

4.0e5

4.5e5

5.0e5

5.5e5

6.0e5

6.5e5

7.0e5

Inte

nsity

, cp

s

0.34s

0.36s

0.36s 0.42s

0.36s

Atenolol

Metoprolol

Primidone

Verapamil

Beclomethasone-

dipropionate

TOF - MS with Peak Capacity of 40 Cycle in 39 seconds

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Page 14

A big leap forward in 2006

Over 15 Years in LC/MS

atmospheric sampling

and patented orthogonal

geometry - Result in

industry leading sensitivity

and robustness

Rough Pump

Octopole 1

Turbo 1

Turbo 1

Turbo 1

Quad Mass Filter (Q1)

Collision Cell

Lens 1 and 2


10KV Detector

Ion Pulser

Turbo 2

Octopole 1

DC Quad

Rough Pump

Turbo 1

Turbo 1

Turbo 1


Collision Cell Lens 1 and 2

Octopole 2

Rough Pump

Octopole 1

Turbo 1

Turbo 1

Turbo 1


Lens 1 and 2 10KV

Detector

Collision cell incorporates axial

acceleration for high speed MS/MS

analysis

6500 Series Q-TOF 6400 Series QQQ

6100 Series SQ

Agilent 6400 Triple Quadrupole Product Line

A New Agilent LC

Triple Quad

Launched

Every Year

For last

4 years

6460

Triple

Quad

6430

Triple

Quad

6420

Triple

Quad

6490

Triple

Quad

6460

Triple

Quad

Perf

orm

an

ce

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Page 16

What is MRM? (Multiple Reaction Monitoring)

170 210 250 290

210

222

268 280 165

Spectrum with

background

ions (from ESI)

Q1 lets only

target ion 210

pass through

190 210

210

Collision cell

breaks ion 210

apart

150 170 190 210

210 158

191

Q3 monitors only

characteristic

fragments 158

and 191 from ion

210 for quant and

qual.

160

158

190

191

no chemical background

Ions

For Research Use Only.

Single quads vs. Triple quads in complex matrix

m/z 306 → m/z 261

SIM m/z 306

MS-MS

Triple quads enjoy the benefit of filtering out

the noise as observed in SIM

Triggered MRM (tMRM) Acquisition:

Simultaneous Quantitation and Confirmation

Dynamic

MRM MRM Triggered

MRM

Triple Quad MRM

Acquisition

Data

Dependent

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Page 19

Triggered MRM (tMRM) Fast, Sensitive Compound Confirmation to Prevent False Positives

Threshold

Additional MRMs

used

for compound

confirmation

Primary cycle (below threshold)

Triggered cycle (above threshold)

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Page 20

tMRM Product Ion Spectrum A Re-Constructed Product Ion Spectrum that is Library Searchable

x103

8

7

6

5

4

3

2

1

0 80 100 120 140 160 180 200 220 240 260 280 300 320 340

334.2 171.1

145.0

132.1

117.0

105.1 91.1 76.0

tMRM Product Ion

Spectrum

Tebufenpyrad

x103 8

7

6

5

4

3

2

1

0 80

76.0

x103 8

7

6

5

4

3

2

1

0 80 100

91.1

x103 8

7

6

5

4

3

2

1

0 80 100 120

105.1

x103 8

7

6

5

4

3

2

1

0 80 100 120

117.0

x103 8

7

6

5

4

3

2

1

0 100 140

132.1

x103 8

7

6

5

4

3

2

1

0 80 100 120

119.0

x103 8

7

6

5

4

3

2

1

0 100 140 180

145.0 x103

8

7

6

5

4

3

2

1

0 100 140 180

147.0

x103 8

7

6

5

4

3

2

1

0 80 100 120 140 160 180

171.1

334 > 76 334 > 91 334 > 105 334 > 117 334 > 119

334 > 132 334 > 145 334 > 147 334 > 171

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Page 21

6500 QTOF Product Line Excellent Value and Performance

6530 QTOF LC/MS Agilent JetStream for greatest sensitivity

Robust, easy to use

Wide dynamic range for qual/quan applications

6540 UHD QTOF LC/MS Sub 1-ppm mass accuracy for highest confidence

compound identification

Versatile system with highest resolution

Fast scan speed to match UHPLC separations

6550 iFunnel QTOF LC/MS Agilent iFunnel for highest sensitivity

Perfect for most demanding applications

Wide dynamic range for qual/quan applications

Fastest scan speed to match UHPLC separations

7/30/2013


Page 22

6550 iFunnel Q-TOF

LC/MS System

ASMS 2010

Sensitivity

• Dramatically improved quantitative capabilities

• New Qual/Quan Workflows

• Superior metabolite and protein detection

• Non-targeted compound screening

Comprehensive Performance Enhancements

• Mass Resolution >40,000

• 50 spectra /sec MS and 33 spectra/sec MS/MS

• 5 orders of linear dynamic range

• <1 ppm MS mass accuracy; <2 ppm MS/MS

• Unrivalled sensitivity

New 6550 iFunnel QTOF

10X Sensitivity Gain Enables Applications

7/30/2013


Page 23

S/N = 49 (RMS)

Low Femtogram Detection Level

Resolution

~ 31,500

Mass error

-0.1 ppm

EIC

25 fg Reserpine

MS

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Page 24

•Increases MS

flexibility by enabling

both GC and LC

compatibility

•Analysis of wider

range of compounds

•Proven QTOF LC/MS

platform

•Proven 7890 GC

performance

G3212A - GC APCI Interface

7/30/2013


Page 25

GC Transfer line / APCI Source Design

Heated transfer line

Transfer line heat shield

Bayonet lock coupling

Corona needle assembly

Capillary MS inlet

7/30/2013


Page 26

Simplify Your Startup

LC/MS Application

Kits

Method

Standards

On Site Training

LC Column

Databases and

Libraries

7/30/2013


Page 27

RapidFire® Mass Spectrometry

Fast autosampler & SPE system

• Replaces LC in LC-MS

• Integrated, automated, micro-scale solid-phase

extraction

• Integrates with standard ESI MS instruments

• Compatible with most biological matrices

• cycle time: 5–10s/sample

The RapidFire System:

• Fits existing workflows

• Low operational costs

• Improved data quality

• Strong correlation with traditional methods

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Page 28

RapidFire™ Mass Spectrometry (RF-MS)

96 or 384 well

Plate Handler

Microfluidic Sample

Clean-up System Monitor to track

MSMS data

Monitor

to track

RapidFire System

Agilent 6410

QqQ Mass Spectrometer

7/30/2013


Page 29

THANK YOU!

7/30/2013


Page 30

Agilent LC/MS Nathan Miller · LC/MSD TOF 1994, 2000 LC/MS 1980 - 2011 2003 Agilent’s > 40 year...

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