Thermo Scientific Q Exactive Orbitrap LC-MS/MS System

O

O

O

N

260.1645

165.0910

303.2067

N

O

O

O

O

N

260.1645

165.0910

303.2067

N

O

Quan/Qual AnalysesUnmatched Confidence for

Identify • Quantify • Confirm

2|

Quanfirmation

\‚kwän-f r-'m -shän\ n 1 : the ability to confidently identify, quantify, and confirm using a single

mass spectrometer : syn Q EXACTIVE MASS SPECTROMETER

With the Q Exactive LC-MS/MS:

• Proven S-lens ion source provides the enhanced sensitivity needed to find more analytes

• 140,000 resolution at m/z 200 and <1 ppm mass accuracy provide outstanding ID confidence

• Spectrum multiplexing and advanced signal processing ensure UHPLC-compatible data acquisition speed for increased throughput

• Fast scan-to-scan polarity switching in both MS and MS/MS reveals more compounds in a single run

The unsurpassed performance of Orbitrap™ technology has revolutionized

mass spectrometry. The Thermo Scientific Q Exactive benchtop LC-MS/MS

is the next step in that revolution, combining high-performance quadrupole

precursor selection with high-resolution, accurate-mass (HR/AM) Orbitrap

detection for outstanding performance and tremendous versatility.

The Q Exactive™ mass spectrometer is superbly suited to untargeted

or targeted screening with high-confidence confirmation, but is equally

capable of a broad range of qualitative and quantitative applications.

The superior quality of Q Exactive data enables identification and

quantitation of more compounds

with greater confidence.

|3

Orbitrap mass analyzer – unrivaled mass resolution and unmatched spectrum quality

Hyperbolic quadrupole mass filter – precise selection of precursor ions and outstanding ion transmission

Nominally isobaric pesticides carbofuran and formetanate are unresolved or only poorly resolved at the highest resolutions available in Q-TOF instruments. The higher resolutions easily achieved by the Q Exactive instrument clearly resolve analytes from complex matrix interferences without sacrificing sensitivity.

Carbofuran C12H15NO3 M+H+ = 222.1125Formetanate C11H15N3O2 M+H+ = 222.1237

222.0 222.1 222.2m/z

0

10

20

30

40

50

60

70

80

90

100

Rel

ativ

e A

bund

ance

222.1263

222.1463

222.1838

R=17,500

222.0 222.1 222.2m/z

222.1238

222.1124

222.1467

222.1843

R=35,000

222.0 222.1 222.2m/z

222.1238

222.1129

222.1465

222.1854

R=70,000

222.00 222.10 222.20m/z

222.1237

222.1127

222.1466

R=140,000

HCD collision cell – higher fragmentation efficiency for unsurpassed MS/MS spectra quality

C-trap – premier in-spectrum dynamic range

S-lens ion source – excellent ion transmission for enhanced sensitivity

HCD collision cell – higher fragmentation efficiency for unsurpassed MS/MS spectra quality

C-trap – premier in-spectrum dynamic range

S-lens ion source – excellent ion transmission for enhanced sensitivity

HCD collision cell – higher fragmentation efficiency for unsurpassed MS/MS spectra quality

C-trap – premier in-spectrum dynamic range

S-lens ion source – excellent ion transmission for enhanced sensitivity

The flexible quanfirmation capabilities of the Q Exactive instrument make it a versatile

solution for drug discovery and metabolism research.

For DMPK quan/qual screening studies, the Q Exactive instrument provides triple quadrupole-equivalent ability to quantify parent compounds and metabolites, and identity confirmation with MS/MS sensitivity and selectivity superior to Q-TOFs. Thermo Scientific MetQuest software enables laboratories to quantify metabolic stability and confirm putative metabolites from a single injection for better throughput.

For structural identification of metabolites, the Q Exactive mass spectrometer leverages the accuracy and confidence of Orbitrap technology to eliminate interferences. The Q Exactive instrument provides superior MS/MS sensitivity and greater confidence in structural identification. Thermo Scientific Metworks metabolite identification software and Mass Frontier spectral interpretation software assist with fast, confident structural elucidation.

4|

Greater throughput for drug discovery studies

Confident metabolite structural identification

Full MS/MS spectra of busiperone, an anxiolytic drug, and a hydroxylated metabolite of busiperone. Assigned masses of identified fragments were all within 1 ppm of their theoretical values. High resolution, accurate mass, and outstanding spectral quality add Q Exactive confidence to metabolite studies and structural ID.

50 100 150 200 250 300 350 400m/z

0

20

40

60

80

Rel

ativ

e Ab

unda

nce

386.2547

122.0714

150.1026 222.1488265.1909180.1019

50 100 150 200 250 300 350 400m/z

0

20

40

60

80

100

Rel

ativ

e Ab

unda

nce

402.2497

122.0714

150.1026 238.1437 281.1858

BuspironeFull MS2

100 HNN

NN

N N

O

O N

O

O

N N NN

N

O

O

HN

HN+

+

++

N

O

OOH N NH

O

OOH

N N NN

N

O

OOH

+

+

Hydroxylated Metabolite of BuspironeFull MS2

HNN

N+

NN

HN+

Facilitating Confident Metabolic Analyses in Drug Discovery

From acquisition to reporting, MetQuest software automates processing of metabolic stability screening data from the Q Exactive LC–MS/MS.

With the high mass resolution to differentiate analytes from interferences, the scan speed

to keep up with UHPLC, and fast polarity switching to find more endogenous metabolites,

the Q Exactive mass spectrometer is the platform of choice for the large-scale profiling of

complex metabolomics samples. The component elucidator algorithm in Thermo Scientific

SIEVE software removes chemical background signals and groups related adducts, dimers

and isotopes, dramatically improving confidence, reducing the false discovery rate, and

accelerating biomarker discovery.

The Q Exactive mass spectrometer is also well-suited for lipid profiling.

Its outstanding resolution helps unravel complex samples. The Q Exactive

LC-MS/MS is an excellent solution for analysis of fragile labile lipids and can

perform qualitative and quantitative analyses of all classes of lipids.

Metabolites from the same parent compound can have a large variance in ionization efficiency between ionization polarities. With fast scan-to-scan polarity switching in both MS and MS/MS, the Q Exactive instrument maximizes metabolite detection and identification from a single chromatographic run.

Whether you are screening to find the toughest impurities, genotoxic compounds, or leachables and extractables, the quanfirmation capabilities of the Q Exactive mass spectrometer provide a greater measure of confidence.

|5

Profiling All Classes of Lipids

Faster reactive-metabolite screening High-confidence drug impurity analysis

A 12-hour continuous UHPLC-MS analysis of d5 hippuric acid (m/z 185.0969) from rat plasma demonstrates the Q Exactive mass spectrometer’s robust ion statistics and highly stable mass accuracy.

100

100

100

Rela

tive

Abun

danc

e

100

100 95,693,541-1.0% CV

96,475,711-0.2% CV

95,147,394-1.6% CV 95,147,394-1.6% CV

98,394,592+1.8% CV

97,722,945+1.1% CV

4.15 4.20 4.25 4.30 4.35 4.40

Time (min)

7:51 PM(~25 hr post

externalcalibration)

11:14 PM

1:23 AM1:23 AM

5:59 AM(~35 hr post

externalcalibration)

3:50 AM

-0.27 ppm

-0.59 ppm

-0.27 ppm

-0.92 ppm

-0.43 ppm

+0.16 ppm

-0.21 ppm

-0.05 ppm

-0.59 ppm

-0.21 ppm

Mean 185.09681-0.50 ppm

Mean 186.10023-0.18 ppm

Peak Width - 3.6 secMean Scans - 14.5

185.0 185.5 186.00

0

0

0

0

2.0x106185.09685

186.10029

185.09679

186.10022

185.09685

186.10025

185.09673

186.10015

185.09682

186.10022

2.0x106

2.0x106

2.0x106

2.0x106

m/z

0

0

0

0

Mean Area96,686,8371.41% CV

Powerful Engine for Large-Scale Metabolomic Profiling

6|

Proteomics, more than any other field, has benefited from Orbitrap technology. The Q Exactive

mass spectrometer provides robust, easy-to-use high-resolution, accurate-mass (HR/AM)

capabilities for quantitative and qualitative proteomics applications. It is ideally suited for

proteomics labs already benefiting from Orbitrap technology that need to expand their

analytical capacity. And it makes HR/AM confidence available to labs relying on unit resolution

instruments for high-throughput quantitation.

Timed SIM MS quantitative analysis of two peptides in a 1-µg yeast digest demonstrates both outstanding LOD (10 attomoles) and excellent linearity.

Superior sensitivity and spectrum quality of the Q Exactive mass spectrometer yield identifications even for low-copy-number proteins in small amounts of sample.

• Higher-energy collisional disso-ciation (HCD) often yields superior fragmentation and higher-quality MS/MS spectra compared CID.

• Superior spectral quality and a wide dynamic range make it possible to explore deeper in the proteome, identifying more proteins and quantifying proteins other instruments cannot find.

• Unlike Q-TOF technology, Orbitrap technology requires no tradeoff between high sensitivity and mass resolution.

• The quan/qual capabilities of the Q Exactive instrument make it easier than ever to transition from proteomic discovery experiments to targeted protein quantitation and confirmation.

Supporting the Q Exactive LC-MS/MS for proteomics research is a range of software including Thermo Scientific Proteome Discoverer software for protein identification, SIEVE software for differential expression analysis, and Pinpoint software for quantitation and verification.

GLILVGGYGTR*

NGFILDGFPR* Log10 (Load Amount (fmol))

Log1

0 (P

eak

Area

)

y = 1.058x + 6.583R2 = 0.9984

y = 1.112x - 6.148R2 = 0.9990

y = 1.112x - 6.148R2 = 0.9990

-3 -2 -1 0 1 32

9.0

8.0

7.0

6.0

5.0

4.0

3.0

0

100

200

300

400

500

600

30

1.00E

+02

3.00E

+02

1.00E

+03

3.00E

+03

1.00E

+04

3.00E

+04

1.00E

+05

3.00E

+05

1.00E

+06

3.00E

+06

10 ng100 ng1000 ng

Cellular Copy Number of Yeast Protein

Num

ber

of

Ind

enti

fied

Pro

tein

s w

ith

Est

imat

ed C

op

y N

umb

er

b5+-NH3

498.17874y6+-H20

758.45410

y10+

1120.55273y2+

322.18576

y8+

1006.51752y6+

776.46594y9+

1177.57886

y1+

175.11826

y5+

647.42157

y3+

435.26990

y4+

548.35284

200 400 600 800 1000 1200m/z

0

10

20

30

40

50

Inte

nsity

(10^

3 co

unts

)

L G N D D E V I L F R

Peptide of protein YOR020C of 149 copy number-identified from 10 ng yeast digest

MASCOT Ion Score : 90Exp Value: 3.8 E-9

Making High-Resolution, Accurate-Mass Quan/Qual More Accessible for Proteomics Research

|7

The Q Exactive LC-MS/MS is the ideal quan/qual screening platform, with specificity that far

exceeds triple quadrupole, Q-TOF, or Q-Trap levels. Whether screening and quantifying

antibiotics, pesticides or hormones, its data-dependent™, full-scan MS/MS capabilities,

superior 140,000 resolution and mass accuracy ensure the highest possible confidence in every

ID. Thermo Scientific ExactFinder software simplifies every step from method development

through data processing and reporting. Even with challenging experiments like multi-residue

analyses involving hundreds of compounds in multiple classes, the Q Exactive MS and

ExactFinder™ software provide results that stand up to the closest scrutiny.

#1739 RT:7.69 AV:1 NL:7.33E5F: FTMS + p ESI Full ms2 404.02@hcd40.00 [50.00-430.00]

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420m/z

0

50

100

Rela

tive

Abun

danc

e

329.0806

344.1041

372.0992

316.1092183.0560301.0852

216.0665134.0606

172.0399

287.0824

273.0899129.0454

187.0509107.0499

EICs from Full-Scan MS

0 5 10 15Time (min)

0

100

0

100

0

100

0

100

Rela

tive

Abun

danc

e

0

100

0

1007.32

7.68

7.99

2.64

10.01

9.70

Diuron - NL: 2.39E7m/z = 233.0231-233.0255

Azoxystrobin - NL: 6.40E6m/z = 404.1221-404.1261

Boscalid - NL: 2.18E6m/z = 343.0382-343.0416

Methomyl - NL: 5.99E6m/z = 163.0528-163.0544

Spinosad D - NL: 1.02E6m/z = 746.4801-746.4875

Indoxacarb - NL: 5.03E6m/z = 528.0754-528.0806

7.55 7.60 7.65 7.70 7.75 7.80 7.85 7.90Time (min)

0

10

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40

50

60

70

80

90

100

Rela

tive

Abun

danc

e

7.69

AzoxystrobinAzoxystrobin

AzoxystrobinMH+ = 404.12421.5 ppb

Y = -62765.7+2.36363e+007*X+83740.5*X^2R^2 = 0.9991 W: 1/X

0 5 10 15ppb

0

1.0

2.0

3.0

4.0

Area

x108

Quantitation

Confirmation

Multiple MS/MSScans Across Each Peak

Screening and Quantifying Environmental and Food Samples with Greater Confidence

Quanfirmation capabilities of the Q Exactive system provide simultaneous quan/qual analysis of a 60-pesticide mixture. Fast scanning ensures high-quality MS/MS data from narrow UHPLC peaks.

BR41270_03/11C

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www.thermoscientific.com/qexactive

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©2011 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries. Specifications, terms and pricing are subject to change. Not all products are available in all countries. Please consult your local sales representative for details.

SRM is the standard approach to quantitative analysis in clinical research, toxicology, and

doping control. The Q Exactive LC-MS/MS takes quantitative analyses to another level,

providing the strength of SRM with a level of confidence only possible with high-resolution,

accurate-mass Orbitrap technology. The superior Q Exactive quanfirmation capabilities

make it possible to identify, quantify, and confirm endogenous steroids, drugs of abuse,

and pain-management therapeutics with equal ease.

Analysis of testosterone in water and in charcoal-stripped serum demonstrates the excellent sensitivity and ion-ratio stability of the Q Exactive LC-MS/MS.

Testosterone - 109Y = 0.0970157+0.00839124*X R^2 = 0.9980 W: 1/X

0 100 200 300 400 5000.0

1.0

2.0

3.0

4.0

Area

Rat

io

Standard % Difference

10 pg/mL 0.97

7.45

-5.78

100 pg/mL -0.29

-5.35

2.99

50 pg/mL

20 pg/mL

250 pg/mL

500 pg/mL

Sample Amount (pg/mL)

0

50

100SN: 74

RT: 4.13

Testosterone 10 pg/mL in Serum

Sample Fragment m/z 109

ng/dL(Calibrators in

water)

T1 15 15 13 14

T2 17 17 19 20

T3 156 156 156 152

T4 34 34 37 36

T5 8 8 8 7

Fragment m/z 97

ng/dL(Calibrators in

water)

ng/dL(Calibrators in

plasma)

ng/dL(Calibrators in

plasma)

Moving Quantitation Confidence to Another Level in Clinical Research and Forensic Toxicology