scimax the applications book - bruker.com · 9 2xr maldi imaging maldi imaging rat brain 6 mm 0 1/4...

The Applications Book

Innovation with Integrity MRMS

2

Magnetic Resonance Mass Spectrometry (MRMS) is the panicle of mass spec in terms of mass accuracy, resolving power and flexibility.

scimaX MRMS opens new analytical doors, is reliable and easy to use, pro-viding answers no other instrument can obtain.

Maxwell magnet technology does not require liquid cryogen fills, ever, enabling a smaller footprint that can fit in any standard laboratory.

MRMS

Enabling High Field Performance at 7T

3

MRMS

Welcome Letter

Dear Mass Spec Customer,

Thank you for your interest in Bruker’s scimaX. Powered by MRMS (Magnetic Resonance Mass Spectrometry) and building on a tradition of record setting magnet designs (1.2 GHz NMR, 18T MRI and 21T ICR), Bruker’s novel Maxwell magnet means no liquid cryogen fills – ever.

At the heart of scimaX is the ParaCell with 2xR detection, enabling high field performance at 7T. Having a maximum resolving power in excess of 20M, MRMS brings access to large scale research projects that can benefit population health, global energy, and the environment.

We are happy to provide this synopsis of data acquired on the scimaX MRMS. With instruments in 35 countries, our global MRMS team members are there for you. Please reach out to your local representative to lean how we can integrate this technology into your lab.

Christopher J. Thompson, Ph.D., Business Manager MRMS, Bruker Scientific LLC, Billerica, MA, USA

4

Professor Joe Loo, University of California, Los Angeles, USA

“The development of the scimaX ® will surely be a game-changer for ultra-high resolving power mass spectrometry.”

Professor Eugene Nikolaev, ParaCell lnventor, Russian Academy of Sciences, Moscow

“Continuing the tradition of Bruker innovation, the ParaCell is a new enabling technology for solariX XR. This radical concept is a departure from traditional ICR cell strategies and provides uncommon broadband ion stability resulting in resolution orders of magnitude above other detection schemes. This power enables the user to effortlessly obtain the extreme resolving power needed to probe isotopic fine structure or highly complex mixtures.”

5

Overview of Applications

MALDI Imaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

Metabolomics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

Native and Intact Proteins . . . . . . . . . . . . . . . . . . . . . . . . . 26

Peptide MS, MS/MS and MSn . . . . . . . . . . . . . . . . . . . . . . 38

Petroleomics and Environmental . . . . . . . . . . . . . . . . . . . . 46

Unknowns: Isotopic Fine Structure . . . . . . . . . . . . . . . . . . 58

Mass Accuracy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72

Technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75

Resources . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83

6

MALDI ImagingRequirements for MALDI Imaging

scimaX is the ultimate MALDI Imaging system for analyzing small to medium

molecules, m/z 100-1500. Its unrivaled eXtreme Resolution capability and sub-ppm

mass accuracy, over a wide mass range, can differentiate images that are only mDa

apart and are prerequisite for Isotopic Fine Structure (IFS) analysis and molecular

formula confirmation.

Realize the full benefit of eXtreme Resolution. Above, a 3-color image from rat testis showing distributions of three ions at nominal m/z 848 and differing only by less than 20 mDa.

7

MALDI ImagingCustomer Insights

Professor Dr. Per Andrén, Mass Spectrometry Imaging, Department of Pharmaceutical Biosciences, University of Uppsala, Sweden and Director of the National Resource for Mass Spectrometry Imaging (NRMSI).

“The solariX is never resting. This is why I am so eager to buy a second one. lf I had two, I could obtain my data a lot faster, and also complete more collaborations.”

Professor Richard R. Drake, Ph.D., MUSC Proteomics Center, Medical University of South Carolina

“We have developed and continue to evolve new glycan and glycoprotein methods for MALDI imaging. Bruker has been a strong partner in facilitating and supporting these efforts with their innovative instrumentation and software solutions.”

Professor Jonathan Sweedler, Ph.D., University of Illinois at Urbana Champaign

“If I had to pick one instrument to save in a fire (and did not have to worry about its size or weight), it would be the solariX XR .”

8

MALDI ImagingThe workflow

Sample cohort

Mount sections

Apply matrix

Acquire data

View/analyze data

9

2xR MALDI Imaging

MALDI ImagingRat brain

6 mm

0 1/4 1/2 3/4 1

m/z 828.4931 m/z 828.5163 m/z 828.5521 m/z 828.5711 m/z 828.5796

20

40

a.u.

0

828.50 828.52 828.55 828.57

86 mDa

9 mDa

Compared to XR Imaging, 2xR offers the choice to acquire mass resolution in almost half the time without compromising or to acquire at double mass resolution without increasing acquisition time. Here, data is acquired in less than 7 hours at mass resolution of 800,000.

Imaging of rat brain lateral resolution: 50 µm R: 820,000 (2ω) @ m/z 273

10

2xR MALDI Imaging


810.596

Intensity (arb. unit) 788.61383 m/z ± 5.500 mDa

760.585767.497

788.614

772.524782.567 769.560

0% 62%

4 cm

0 1/4 1/2 3/4 1

scimaX Imaging of a rat brain with standard 2xR detection provides fast imaging speed while retaining eXtreme mass resolution.

Imaging of rat brain (sagital cut) Lateral resolution: 70 µm R: 250,000 @ m/z 400

11

2xR MALDI Imaging


m/z 820.524

m/z 844.523

m/z 810.578

m/z 826.571

m/z 734.570

m/z 830.507

m/z 713.454

0% 62%

Direction of measurement:4 cm

0 1/2 3/4 11/4

scimaX Imaging of a rat brain with standard 2xR detection provides fast imaging speed while retaining eXtreme mass resolution

Imaging of rat brain (coronal cut) Lateral resolution: 70 µm R: 250,000 @ m/z 400

12

MetabolomicsRequirements

Whether Metabolomics, Phenomics or any other complex sample analysis, large scale sample

evaluation is now possible with scimaX and the MRMS aXelerate workflow. The eXtreme Resolution

allows for direct sample analysis and enables true high sample throughput complementary to

established NMR based solutions. From the largest unknown to the smallest, scimaX ensures confident

molecular formula assignment at any level.

13

MetabolomicsCustomer Insights

Professor Philippe Schmitt-Kopplin, Ph.D., Analytical BioGeoChemistry, Helmholtz Zentrum München, Germany

“We set up new discovery approaches to describe the compositional space of any complex system in biology and geochemistry. MRMS eXtreme Resolution enables us to address next generation metabotyping, i.e. simultaneous rapid description of hundreds of known and thousands of new metabolites relevant for dynamic biological/chemical processes. MRMS in combination with MetaboScape will also enable other researchers to shed light to this new exiting research field of this yet dark metabolome.”

Professor Arthur S. Edison, Ph.D., and GRA Eminent Scholar, Departments of Biochemistry and Molecular Biology and Genetics Institute of Bioinformatics, Complex Carbohydrate Research Center, University of Georgia, Athens, GA

“Virtually every metabolomics project we have going right now will benefit from this new instrumentation ... “

Professor Jeremy Nicholson, Ph.D., Director of the Australian National Phenome Center, ProVice Chancellor for Health, Murdoch University

“The performance of our new solariX 7 Tesla MRMS system has met and exceeded all our expectations across a variety of high end metabolic phenotyping challenges in molecular profiling, structure elucidation and imaging – and it is highly user friendly – every laboratory should have one!”

14

MetabolomicsMRMS aXelerate Workflow

SmartFormula Statistical Analysis

T-ReX® 2D Feature

extractions

Putative Structure

Annotations

Rapid, LC-free MS followed by statistical analysis provides a sensitive, high-throughput workflow for modern phenotyping and metabotyping.

• Accelerate throughput (> 200 samples/day)

• Complementary to established NMR based solutions

• Simultaneous analysis of known and unknown metabolites

• Access compounds not readily detectable by LCMS analysis

• 3-tiered confidence in annotation

15

Metabolomics: FIAFIA and CASI, plasma, ESI(+)

High-throughput clinical metabolomics

2.0

3.0

Inte

nsit

y

x 1010

1.0

200 300 400 500 600m/z

700 800

CASI

4.0

6.0

Inte

nsit

y

x 109

2.0

200 300 400 500 600m/z

700 800

Broadband

Flow injection analysis (FIA) enables high-throughput sample analysis by eliminating the time-consuming LC step. This also provides increased chemical coverage as the experiment is not bound to the chemistry of the LC column. The combination of these provides a great cost savings to any lab.

16

Metabolomics: FIAFIA and CASI, plasma, ESI(+)

High-throughput clinical metabolomics

266.00 266.10 266.20266.05 266.15 266.25 266.30m/z

CASI

266.00 266.10 266.20266.05 266.15 266.25 266.30m/z

Broadband

The continuous accumulation of selected ions (CASI) is an enhanced dynamic range experiment that provides increased sensitivity of low intensity species. This sensitivity also brings great utility through the ability to utilize isotopic fine structure for the unambiguous identification of unknown metabolites.

17

Metabolomics: LCMSDarjeeling tea, ESI(+), @ 1 Hz

Isotopic fine structure on an LC timescale

0.8

1.0

Inte

nsit

y

x 109

0.2

0.4

0.6

0

BCP

200

300

400

Inte

nsit

y [m

AU

]

100

010 2 3 4 5

Time (min)6 87

UV(254 nm)

scimaX’s eXtreme Resolution is also compatible to LC analysis. An example is shown for the analysis of metabolites in Darjeeling tea. The following pages show the use of isotopic fine structure for the identification of compounds during a short gradient LC run.

18



Compound 8

Detection of Gallocatechin A+2 isotopic pattern

80

100

Inte

nsit

y [%

]

20

40

60

0200 300 400 500 600 700

m/z

1.25

1.50

Inte

nsit

y [%

]

0.50

0.25

0.75

1.00

0309.084 309.086 309.088 309.090

m/z

Gallocatechin

1.25

Inte

nsit

y [%

]

0.50

0.25

0.75

1.00

0

18O13C2

Compound 8 in the chromatogram is confidently assigned to Gallocatechin. The 18O isotope is baseline resolved and the peak fidelity yield the correct number of Oxygen atoms in the molecule.

Mass error: 175 ppb

19



Compound 10

A+2 isotopic patternDetection of caffeine

80

100

Inte

nsit

y [%

]

20

40

60

0160 180 200 220 240 280 300260

m/z

Caffeine

Inte

nsit

y [%

]

197.088 197.092 197.096 m/z

0.2

0.1

0.3

0.4

0

Inte

nsit

y [%

]

0.2

0.1

0.3

0.4

0

18O

13C15N

13C2

Compound 10 in the chromatogram is confidently assigned to Caffeine. In this example, the A+2 isotope distribution clearly shows three distinct IFS species: 13C1

15N1, 18O1 and 13C2

Mass error: 47 ppb

20



Compound 11

A+2 isotopic patternDetection of Epigallocatechin gallate

Epigallocatechin gallate

80

100

Inte

nsit

y [%

]

20

40

60

0200 300 400 500 600 700

m/z

11.

Inte

nsit

y [%

]

461.092 461.098461.094 461.100461.096

11. 1+ 461.098928

1+ 461.096433

461.102m/z

1.0

0.5

1.5

2.0

2.5

3.0

0

11.1+

461.098991

461.096575In

tens

ity

[%]

18O13C2

1.0

0.5

1.5

2.0

2.5

0

IFS is again present in Compound 11 assigned as epigallocatechin gallate.

Mass error: 51 ppb

21

Metabolomics: LCMSUrine, ESI(+), @ 1 Hz


5.0

6.0

Inte

nsit

y

x 108

2.0

1.0

3.0

4.0

0

BPC

80

60

100

120

Inte

nsit

y [m

AU

]

40

20

010 2 3 4 5

Time (min)

UV (254 nm)

Global health concerns are the focus of many groups across the globe. The analysis of complex samples such as blood and urine are routinely analyzed by LCMS. This example shows the LCMS separation of Urine at 1Hz acquisition rate.

22

Metabolomics: LCMSUrine, ESI(+), @ 1 Hz


Compound 6

Hippuric acid is assigned as compound 6 through the detection of IFS. The combination of high mass accuracy and resolving power provides an unambiguous molecular formula assignment.

Mass error: 227 ppb

A+2 isotopic patternDetection of Hippuric acid

Hippuric acid

80

100

Inte

nsit

y [%

]

20

40

60

0150 200 250 300 350 400

m/z

Inte

nsit

y [%

]

182.068 182.070 182.072 182.074

1+ 182.069765

1+ 182.072229

m/z

0

1+ 182.072226

182.069762

Inte

nsit

y [%

]

18O

13C2

0.2

0.2

0.4

0.4

0.6

0.6

0

23

Metabolomics: LCMSUrine, ESI(+), @ 1Hz


Compound 7

A+2 isotopic patternDetection of Phenylacetylglutamine

Phenylacetylglutamine

80

100

Inte

nsit

y [%

]

20

40

60

0200 300 400 500 600 700

m/z

Inte

nsit

y [%

]

267.116 267.120 267.124 267.128

1+ 267.122528

1+ 267.124993

1+ 267.118673

1+ 265.118263

1+ 529.229164

m/z

1+ 267.125017

267.122554

267.118708

Inte

nsit

y [%

]

18O

13C15N

13C2

0.2

0.2

0.4

0.4

0.6

0.6

0.8

0.8

0

0

15N and 18O are used to assign the molecular formula of Phenylacetylglutamine to compound 7.

Mass error: 77 ppb

24

Metabolomics: LCMSGreen tea, ESI(+), @ 0.6 Hz


2.5

3.5

3.0

4.0

Inte

nsit

y

x 108

x 108

1.0

0.5

1.5

2.0

0

BPC

0.5

1.5

1.0

Inte

nsit

y

062 73 84 95 10

Time (min)

EIC of 773.213

Green tea is analyzed with an acquisition rate of 0.6 Hz. The extracted ion chromatogram of m/z 773.214 is shown in the lower panel.

25

Metabolomics: LCMSGreen tea, ESI(+), @ 0.6 Hz


x 108x 106

x 106

Compound at RT 6.6 minA+2 isotopic pattern

C33H41O21

1.00

1.25

Inte

nsit

y [%

]

0.25

0.50

0.75

0200 400 600 800 1000 1200

m/z

Inte

nsit

y [%

]

775.216 775.218 775.220 775.222

1+ 775.217730

1+ 775.220233

m/z

1+ 775.220316

775.217625

Inte

nsit

y [%

]

18O

13C2

1.0

2.0

2.0

4.0

3.0

6.0

4.0

0

0

With an acquisition rate of 0.6 Hz, IFS is observed at m/z 775 for the chromatographic peak at 6.6 minutes. The molecular formula for Quercitin-3-O-glycosyl-rhamnosyl-galactosid is assigned easily with a mass resolving power of 550,000.

R: 550k @ m/z 773 mSigma: 37.3Mass error: 27 ppb

26

Native and Intact ProteinsRequirements for Native MS

The critical requirement for native MS is an optimal pressure gradient between the electrospray

source and the mass analyzer. The ion path on the scimaX MRMS enables the transmission of intact

biomolecular complexes into the ParaCell, allowing for analysis of protein and protein complexes

with isotopic resolution.

Now fragile fragment-protein, protein-substrate, multi-protein biomolecular complexes, membrane

proteins with nanodiscs, mAbs, and native protein top-down analysis can all be analyzed on a standard

(unmodified) scimaX.

27

Professor Joe Loo, University of California, Los Angeles, USA

“Native mass spectrometry and top-down proteomics are starting to impact studies in structural biology and medicine –and Bruker has all of the tools necessary for these growing technologies.”

Dr. Sally Ann Poulsen, Professor of Chemical Biology at the Griffith Institute for Drug Discovery (GRIDD), Griffith University, Brisbane, Queensland, Australia

“The MRMS method means you avoid wasting valuable time on compounds that will not advance drug discovery”

Professor Ronald Quinn, Griffith Institute for Drug Discovery, Brisbane, Australia

“Our first experiments worked beautifully. Bruker MRMS retains weak non-covalent complexes and also easily handles screening of pools of compounds. The high resolution is perfect to resolve the complex mixtures.”

Native and Intact ProteinsCustomer Perspectives

28

Native and Intact ProteinsThe workflow

Buffer exchange into ammonium acetate(50 mM)

Infuse with syringe pump or automation (4 µl/min)

Acquire (7 sec/sample) with standard ESI source

29

ProteinsUbiquitin, ESI(+)

High resolution protein analysis

2.5

3.0

Inte

nsit

y

x 108

1.0

0.5

1.5

2.0

0600 700 800 900 1000 1100 1200 1300 1400

m/z

scimaX is not limited to the realm of small molecules. Its gentle source design is made to efficiently transfer intact proteins and other large molecular species, without any modifications to the instrument.

R: 2,000,000 @ m/z 779 transient: 5.86 s, 2ω, AMP (Kilgour)

30

ProteinsUbiquitin 10+ Charge State, ESI(+)

Baseline resolved isotopic distribution

2.5

3.0

Inte

nsit

y

x 108

1.0

0.5

1.5

2.0

0779.2 779.6779.4 779.8 780.0 780.4780.2 780.6

m/z

Inte

nsit

y

x 108

1.0

0.5

0779.792 779.794

m/z

The eXtreme Resolution of scimaX produces baseline resolved isotopic peaks that provide confident molecular weight calculation via SNAP deconvolution.

R: 2,100,000

31

Protein FragmentationCID of Ubiquitin 10+ Charge State, ESI(+)

87% sequence coverage

2.5

Inte

nsit

y

x 109

1.0

0.5

1.5

2.0

0200 600400 800 1000 1200

m/z

Rich MS/MS fragmentation provides sequence coverage that benefit from both the high mass accuracy and resolving power, to clearly separate the dense fragment ions.


32

Protein FragmentationCID of Ubiquitin 10+ Charge State, ESI(+)


1.2

Inte

nsit

y

x 108

0.4

0.2

0.6

0.8

1.0

0530 550540 560 570 580

m/z

549.40

Inte

nsit

y

x 107

1.5

1.0

0.5

0549.20 549.30

m/z

Looking closer reveals isotopically resolved fragment ions. R: 1,100,000 @ m/z 400 transient: 1.47 s, 2ω, AMP (Kilgour)

33

Protein FragmentationECD of Ubiquitin 10+ Charge State, ESI(+)


100

Inte

nsit

y [%

]

40

20

60

80

0200 600400 800 1000 1200 1400 1600 1800 2000 2200

m/z

One of scimaX’s many dissociation techniques, electron capture dissociation (ECD), provides complimentary fragmentation to traditional CID.


34

Protein FragmentationECD of Ubiquitin 10+ Charge State, ESI(+)


10501040 1060 1070 1080m/z

8+1136.5700774

1039

Inte

nsit

y [%

]

3

5

6

2

4

1

01036 1037 1038

m/z

Inte

nsit

y [%

]

4

2

6

8

0

With SNAP, complex data is quickly and confidently deconvoluted to provide rich MS/MS information.

R: 1,100,000 @ m/z 400transient: 1.47 s, 2ω, AMP (Kilgour)

35

5 nM sensitivity at normal ESI flow

0.6

0.8

1.0

Inte

nsit

y

x 108

0.2

0.4

0

CA II

CA II

CA II+ acetate

CA II+ acetate

CA II+ substrate

CA II (native) - 5 nM

Ratio drug to CA I: 12.0CA II: 5 nMacetazolamide: 60 nM

CA II + substrate+ acetate

x 104

0.5

1.5

1.0

Inte

nsit

y

0

29050 2925029150 2935029100 2930029200 29400 29450m/z

Fragment Based Drug DiscoveryNative CA II and Acetazolamide, ESI(+)

Using the standard scimaX ESI source for native MS, nanomolar levels of protein are detected with isotopic resolution, in a robust manner suitable for high throughput screening.

R: 70,000 @ m/z 2,620Flow rate: 2 ul/min

36

Rapidly determine binding constants

Inte

nsit

y

x 108

x 107

0.5

4.0

2.0

1.0

6.0

0

0

CA IICA II

+ acetate

CA II+ substrate

CA II: 5 nM

CA II: 5 nM acetacolamide: 20 nM




CA II + substrate+ acetate

2645 26652655 26752650 26702660m/z

x 108

0.5

1.0

0

x 108

x 108

0.5

0.5

1.0

1.0

0

0

Fragment Based Drug DiscoveryNative CA II and Acetazolamide, ESI(+)

The robust source enables native protein-ligand screening of weak and strong complexes with the right balance of desolva-tion and high sensitivity.

R: 70,000 @ m/z 2,620Flow rate: 2ul/min

Spe

cific

bin

ding

0.10.20.30.40.50.60.70.80.9

0

Specific binding data

0 100 500300 700200 600400 800 900nM Ligand

37

Competitive binding of isoforms

0.6

0.8

1.0

Inte

nsit

y

x 104

0.2

0.4

0

CA I

CA I

CA II

CA II

CA I+ substrate

No acetazolamide added

acetazolamide at 400 nM

CA II + substrate

x 104

0.5

1.0

Inte

nsit

y

0

28800 2920029000 2940028900 2930029100m/z

Fragment Based Drug DiscoveryNative CAI and II with Acetazolamide, ESI(+)

Direct observation ligand specificity for protein isoforms is possible on scimaX, reducing need for multiple assays in your screening cascade.

R: 70,000 @ m/z 2,620Flow rate: 2 ul/min

38

Peptide MS, MS/MS and MSn

39

PeptidesPepMix, MALDI(+)

High accuracy and resolution

1000500 1500 2000 2500 3000m/z

Inte

nsit

y [%

]

2

1

3

4

5

x108

0

R: 500,000 @ m/z 400transient: 1.47 s, 2ωRMS mass error of calibration: 303 ppb

x108

2469 2471

Inte

nsit

y [%

]

3

2

4

1

02463 2465 2467

m/z

40

PeptidesACTH (1-39), MALDI(+)


45304520 4540 45804550 45904560 46004570 4610m/z

Inte

nsit

y [%

]

2

1

3

4

5

6

x107

0

R: 165,000 @ m/z 4,541transient: 5.94 s, 2ω

41

PeptidesBSA digest (10 fmol), MALDI(+)


1000800 1200 20001400 22001600 24001800m/z

Inte

nsit

y [%

]

1.0

0.5

1.5

x108

0

R: 500,000 @ m/z 400transient: 1.47 s, 2ω

42

MSn

Lincomycin

Full structural information: MSn

Inte

nsit

y

2.0

3.0

4.0x 108

1.0

0

MS3

100 150 200m/z250 350 450300 400

2.0

3.0

4.0x 108

1.0

0

MS4

scimaX provides complex structural characterization via MSn, using a combination of CID and SORI-CID. MS2 is achieved in the front-end collision cell. The remaining stages of MSMS are done in the ParaCell with SORI-CID.

MS20.50

0.75

1.00

1.25

x 109

0.25

0

43

MS2

Lincomycin

Full structural information: MS2

Inte

nsit

y

100 150 200m/z250 350 450300 400

1.0

x 109

0.5

0

The first stage of MS/MS is done by isolation with the mass selective quadrupole, followed by CID fragmentation in the collision cell. For MS2 only analysis, fragment ions are then sent to the ParaCell for detection.

Meas. m/z Ion Formula err [ppm] mSigma

126.12771 C8H16N 0.15 2.2

172.13317 C9H18NO2 0.21 8.1

216.08661 C9H14NO5 0.19 14.5

234.09717 C9H16NO6 0.20 14.8

264.08996 C10H18NO5S 0.22 12.2

299.19649 C15H27N2O4 0.15 9

317.20705 C15H29N2O5 0.14 13.1

341.20706 C17H29N2O5 0.11 16.8

359.21763 C17H31N2O6 0.08 5.8

371.19987 C18H31N2O4S 0.09 93.3

389.21045 C18H33N2O5S 0.05 8

407.22103 C18H35N2O6S 0 8.4

44

Inte

nsit

y [%

]

2.0

1.0

3.0

x108

0

MS3

Lincomycin


Subsequent stages of MS/MS are performed by isolation and fragmentation in the ParaCell. Ions are fragmentated through the process of sustained off-resonance irradiation collision-induced dissociation, SORI-CID.


126.12769 C8H16N 0.28 48.8

172.13315 C9H18NO2 0.30 49.2

299.19644 C15H27N2O4 0.32 n.a.

341.20699 C17H29N2O5 0.33 n.a.

359.21754 C17H31N2O6 0.34 n.a.

100 150 200m/z

250 350300 400

45

MS4

Lincomycin


Inte

nsit

y

60 70 80m/z

90 110 130100 120

2.0

3.0x 108

1.0

0

The remaining stages of MSn are all achieved with SORI-CID. Even by the fourth stage, rich fragment ions produced with sub-ppm mass accuracy.


54.03382 C3H4N 0.03 n.a.

55.04165 C3H5N 0.06 n.a.

55.05422 C4H7 0.05 n.a.

56.04947 C3H6N 0.05 n.a.

58.06512 C3H8N 0.05 16.2

65.03857 C5H5 0.06 52.5

67.04164 C4H5N 0.09 n.a.

67.05422 C5H7 0.09 n.a.

68.04947 C4H6N 0.08 n.a.

69.06987 C5H9 0.09 29.3

70.06512 C4H8N 0.10 24.2

80.04947 C5H6N 0.11 n.a.

82.06512 C5H8N 0.11 29.8

83.07294 C5H9N 0.12 n.a.

83.08552 C6H11 0.12 30.3

84.08077 C5H10N 0.12 n.a.

93.06986 C7H9 0.14 59.2

95.08552 C7H11 0.12 n.a.

96.08076 C6H10N 0.13 27.5

97.08859 C6H11N 0.13 n.a.

97.10116 C7H13 0.12 n.a.

98.09641 C6H12N 0.14 n.a.

124.11206 C8H14N 0.13 41.3

126.12771 C8H16N 0.14 34.0

46

Petroleomics and EnvironmentalRequirements

When knowing every molecule matters, scimaX empowers 7T systems with high field performance

for any complex mixture.

scimaX powered by 2xR technology is easily matching or surpassing the resolving power of many

conventional high field MRMS systems. Now such high field systems are no longer mandatory for

crude oils, bio-fuels, DOM/NOM or any complex mixture

47

Carlos Afonso, University of RouenPierre Giusti, TOTALC2MC – Complex Matrices Molecular Characterization, Joint Laboratory

“With our Bruker MRMS in Rouen the analysis of highly Complex Mixtures has been pushed further than ever and could also be made on a routine basis in the framework of the C2MC joint lab.”

Petroleomics and EnvironmentalCustomer Insights

Janne Jänis, University of Eastern Finland – JoensuuLaboratory of Bio-organic Chemistry

“Most of our research at UEF heavily relies on the use of Bruker MRMS technology to dig deep into chemistry of bio-oils, natural extracts and highly complex environmental samples”

48

Petroleomics and EnvironmentalProcessing workflow

Export mass spectrum

Internal recalibration

Mass spectrum

Class distribution plot

m/z

DBE vs. C plotMass error plot

Carbon number distribution plot

DBE distribution plot

PetroOrg software Molecular Formula calculation

49

R: 1,400,000 @ m/z 400Transient length: 2.96s2ω detection, AMP100 scans

EnvironmentalSRFA, ESI(-)

Environmental analysis

300200 400 800500 900600 700m/z

Inte

nsit

y [%

]

2

1

3

4

5

6

7

x108

0

50

R: 1,400,000 @ m/z 400Transient length: 2.96s2ω detection, AMP100 scans


Van Krevelen plots

0.4 0.40.2 0.20 00.6 0.60.8 0.81.0 1.0O/C O/C

H/C

H/C

0.2 0.2

0.4 0.4

0.6 0.6

0.8 0.8

1.0 1.0

1.2 1.2

1.4 1.4

1.6 1.6

1.8 1.8

2.0 2.0

0 0

Ox class NOx class

High

Low

% T

otal

High

Low

% T

otal

51

Molecular formulae: ca. 15960 (incl. 13C peaks) R: 1,400,000 @ m/z 400Transient length: 2.96s2ω detection, AMP100 scansAverage mass error: 124 ppb


Class plot of Ox Class

Mass error plot of Ox class

01 02 03 04 05 06 07 08 09 010

011

012

013

014

015

016

017

018

019

020

021

022

023

024

025

026

Rel

. abu

ndan

ce

2

1

3

4

5

6

7

0

350m/z

250150 450 550 850750650

Err

or [p

pm]

Mass error of calculated molecumar formulae

-0.6

-0.4

-0.2

0

0.2

0.6

0.4

0.8

-0.8

52

R: 1,750,000 @ m/z 400 transient: 2.93 s, 2ω, AMP (Kilgour) 3,000 scans

PetroleomicsCrude oil, APPI(+)

Complex mixture analysis via APPI

300200 400 800500 900 1000

API gravity: 24.0Sulphur: 1.52%Asphaltenes: 5.0 %Sodium: 5 ppmNickel: 42 ppmVanadium: 139 ppm

600 700m/z

Inte

nsit

y

0.4

0.2

0.6

0.8

1.0

x109

0

53

Crude oil sample and their compound classes can be analyzed in detail concerning carbon distribution and aromaticity.

R: 1,750,000 @ m/z 400 transient: 2.93 s, 2ω, AMP (Kilgour) 3,000 scans

PetroleomicsCrude oil, APPI(+)

DBE vs. carbon number

Carbon number0 10 20 30 40 50 6070 80

DB

E

510152025

3530

40

0

HC-R

Carbon number0 10 20 30 40 50 6070 80

DB

E

510152025

3530

40

0

S2-R

Carbon number0 10 20 30 40 50 6070 80

DB

E

510152025

3530

40

0

N1-R

Carbon number0 10 20 30 40 50 6070 80

DB

E

510152025

3530

40

0

N1-R

Carbon number0 10 20 30 40 50 6070 80

DB

E

510152025

3530

40

0

S3-R

Carbon number0 10 20 30 40 50 6070 80

DB

E

510152025

3530

40

0

N1S1-R

Carbon number0 10 20 30 40 50 6070 80

DB

E

510152025

3530

40

0

O1-R

Carbon number0 10 20 30 40 50 6070 80

DB

E

510152025

3530

40

0

O1-R

Carbon number0 10 20 30 40 50 6070 80

DB

E

510152025

3530

40

0

O1S2-R

Carbon number0 10 20 30 40 50 6070 80

DB

E

510152025

3530

40

0

S1-R

Carbon number0 10 20 30 40 50 6070 80

DB

E5

10152025

3530

40

0

O1S1-R

Carbon number0 10 20 30 40 50 6070 80

DB

E

510152025

3530

40

0

N1S2-R

54

PetroleomicsCrude oil, LDI(+)

Complex mixture analysis via LDI

300200 400 500 600 700m/z

Many samples require solvents that are not amicable to ESI. The dual ESI/MALDI source of scimaX allows for easy laser desorption ionization for these types of samples.

R: 1,900,000Transient length: 2.96s2ω detection, AMP300 scans

m/z398.10 398.15 398.20 398.25 398.30 398.35

Inte

nsit

y

x108

0

0.25

0.75

0.50

1.00

1.25

55

BroadbandR: 1,900,000 @ m/z 401

CASIR: 3,200,000 @ m/z 401

PetroleomicsCrude oil, CASI-LDI(+)

Enhanced dynamic range by CASI

Carbon number

Carbon number

Carbon number

Carbon number

Carbon number

Carbon number

0

0

0

0

0

0

10

10

10

10

10

10

20

20

20

20

20

20

30

30

30

30

30

30

40

40

40

40

40

40

50

50

50

50

50

50

60

60

60

60

60

60

70

70

70

70

70

70

80

80

80

80

80

80

DB

ED

BE

DB

ED

BE

DB

ED

BE

5

5

5

5

5

5

10

10

10

10

10

10

15

15

15

15

15

15

20

20

20

20

20

20

25

25

25

25

25

25

35

35

35

35

35

35

30

30

30

30

30

30

40

40

40

40

40

40

0

0

0

0

0

0

HC

HC-R

N1

N1-R

S1

S1-R

56


Inte

nsit

y

x 108

Broadband

CASI

x 108

0.5

1.0

1.5

3.0

1.0

2.0

2.0

Inte

nsit

y

0

0401.10 401.30401.20401.15 401.35401.25

m/z


The CASI spectrum detected 54 peaks and assigned 50 molecular formula annotations with average error of 39 ppb in 1 Da window (m/z 401).

BroadbandR: 1,900,000 @ m/z 401

CASIR: 3,200,000 @ m/z 401

57


Inte

nsit

y

Broadband

CASI

x 107

x 107 x 106

1.0

1.0

1.0

3.0

3.0

3.0

5.0

5.0

2.0

2.0

2.0

4.05.0

4.0

4.0

6.0

6.0

Inte

nsit

y

0

0

0

401.2075

401.2172

401.2100

401.2176

401.2125 401.2150 401.2175 401.2200 401.2200 401.2225 401.2250 401.2300401.2275m/z


The insert shows two peaks baseline resolved that are only 0.27 mDa apart. This difference is less than the mass of an electron (0.59 mDa).

BroadbandR: 1,900,000 @ m/z 401

CASIR: 3,200,000 @ m/z 401

58

Isotopic Fine Structure

15N 33S 13C

59

Isotopic Fine StructureRequirements for IFS

Improvement in resolving power is more than simply a larger number. Rather improvements are

gained only when new information is obtained. scimaX is the panicle of resolving power through

routine isotopic fine structure analysis.

Example: A+1 peak of C24H26N6O5S1R=50,000

R=100,000

R=250,000

R=500,000

Monoisotopic

A+1

C24H26N6O5S1

A+1 isotope

A+2

A+3A+2

Resolve unique elemental signatures

511 512 513 514

512.180 512.190

13C

2H15N 33S

512.170512.160

m/z

m/z

60

“This S-atom-driven approach afforded an efficient chemical assignment of S-containing metabolites, suggesting its potential application for screening not only S but also other heteroatom-containing metabolites in MS-based metabolomics.”

Isotopic Fine StructureCustomer Insights

The IFS approach enables the ability to rapidly identify sulfur containing metabolites and calculate

single sum formulas for each, increasing both the speed and accuracy of the workflow. The incredible

power of this workflow is that with extreme resolving power and sum formula determination, rapid

screening for other heteroatom (N and O) containing metabolites is also possible.

In 2013, Prof. Kazuki Saito, of the RIKEN Plant Science Center (group photo shown), was

acknowledged with an award for one of the top downloaded papers. Prof. Kazuki Saito has been

selected as a highly cited Researcher in 2014 and 2015 by Thomson Reuters in the Plant and Animal

Science field and won the 2016 Japanese Society of Plant Physiologists award.

32.0 32.4 32.6 32.8 33.0 33.433.2 33.6 33.832.2m/z

61

Isotopic Fine StructureSubstance P, ESI(+)

Unambiguous molecular formula determination

449.8 450.4450.0 450.6450.2 450.8 451.0 451.2

Inte

nsit

yIn

tens

ity

m/z

2.0

2.0

3.0

3.0

x 109

x 109

1.0

1.0

0

0

A+1

A+2

A+3

Substance P

A+4

scimaX reveals nature’s signature through the ability to see isotopic fine structure, providing unambiguous molecular formula determination. ParaCell technology allows you to work beyond the molecular realm.

R: 3,200,000 @ m/z 449transient: 5.86 s, 2ω,AMP (Kilgour)

62

Isotopic Fine StructureSubstance P, ESI(+)

Unambiguous molecular formula determination

450.248 450.580 450.9150 451.2500450.250 450.584 450.9200 451.2550450.588

Sim

ulat

ion

Exp

erim

ent

m/z

A+1 A+2 A+3

Substance P

A+4

The molecular formula of Substance P is clearly shown by resolving the heavy isotopes for all the elements present in the molecule.

R: 3,200,000 @ m/z 449transient: 5.86 s, 2ω, AMP (Kilgour)

63

Isotopic Fidelity and Ion PopulationLincomycin, ESI(+)

Maintains isotopic fidelity at high ion population

100 400200 500300 600 700 800

Inte

nsit

y

m/z

2.0

2.0

1.0

3.0

4.0

3.0

x 1010

x 109

x 1010

1.0

1.0

0.5

0

0

0

S/N: 75169 accumulation time: 100 msmSigma: 5.7




The high dynamic range of the ParaCell maintains peak fidelity even as the number of ions varies.

R: 650k @ m/z 407Calibrated on Lincomycin 12C peak

2.0x 109

1.0

0

64



A+1 pattern

As the number of ions increases there is no change in the observed isotopic fine structure of Lincomycin.

R: 650k @ m/z 407Calibrated on Lincomycin 12C peak

Part of A+1 pattern

408.218 408.2170 408.2180 408.2190 408.2200 408.2210408.220 408.222 408.224 408.226 408.228

Inte

nsit

y

m/z m/z

6.0

4.0

x 109

2.0

0

S/N: 15833

S/N: 7644

S/N: 2293

S/N: 966

0.80.6

x 109

0.40.2

0

2.0

2.0

4.0

4.0

3.0

3.0

x 108

x 108

1.0

1.0

0

0

2.0

3.0x 109

1.0

0

2.0

3.0x 107

1.0

0

100 ms

50 ms

20 ms

10 ms

simulation

0.80.6

x 108

0.40.2

0

1.0

1.0

1.0

1.5

1.5

1.5

x 108

x 107

x 107

0.5

0.5

0.5

0

0

0

65



No change of IFS with high ion load in analyzer. R: 650k @ m/z 407Calibrated on Lincomycin 12C peak

A+2 pattern Part of A+2 pattern

409.216 409.220 409.222 409.224 409.226 409.228 409.230409.220 409.224 409.228 409.232

Inte

nsit

y

m/z m/z

1.5

1.0

x 109

0.5

0

2.01.5

x 108

1.00.5

0

2.01.5

x 108

1.00.5

0

4.0

6.0x 108

2.0

0

100 ms

50 ms

20 ms

10 ms

simulation

0.8

0.8

0.6

0.6

x 108

x 108

0.4

0.4

0.2

0.2

0

0

0.80.6

x 108

0.40.2

0

4.0

2.0

2.0

5.0

3.0

3.0

x 108

x 107

x 107

3.0

1.02.0

1.0

1.0

0

0

0

S/N: 3664

S/N: 1517

S/N: 486

S/N: 218

66

Isotopic Fidelity and Ion PopulationLincomycin, ESI(+), 10 ms


10 ms accumulation timemSigma: 2.7 R: 650k @ m/z 407Calibrated on Lincomycin 12C peak

408.2150 408.2200 408.2250 408.2300 409.215 409.220 409.225 409.230 409.235

Inte

nsit

y

m/z m/z

3.0

4.0

2.0

x 108

1.0

0

4.0

5.0

7.0

6.0

8.0

9.0

x 107

3.0

1.0

2.0

0

1+ 409.21683

1+ 408.22442

13C

13C2

18O

13C33S

13C15N 13C2H

34S

15N 33S 2H

Experimental tea mix with Lincomycin

Simulated C18H35N2O6S spectrum

67




408.2150 408.2200 408.2250 408.2300 409.215 409.220 409.225 409.230

Inte

nsit

y

m/z m/z

6.0

5.0

8.0

9.0

7.0

4.0

3.0

x 108

2.0

1.0

0

2.0

x 108

1.5

0.5

1.0

0

1+ 409.21687

1+ 408.22439

13C

13C2

18O

13C33S

13C15N13C2H

34S

15N 33S 2H



A+1 pattern A+2 pattern

68




408.2150 408.2200 408.2250 408.2300 409.215 409.220 409.225 409.230

Inte

nsit

y

m/z m/z

3.0

2.5

2.0

1.5

x 109

1.0

0.5

0

6.0

5.0

4.0

3.0

2.0

1.0

0

x 108

1+ 409.216871+

408.22442

13C

13C2

18O

13C33S

13C15N 13C2H

34S

15N 33S 2H




69




408.2150 408.2200 408.2250 408.2300 409.215 409.220 409.225 409.230

Inte

nsit

y

m/z m/z

1.50

1.25

1.00

0.75

0.5

0.25

0

x 109

6.0

5.0

4.0

3.0

2.0

1.0

0

x 109 1+ 409.216951+

408.22451

13C

13C2

18O

13C33S

13C15N 13C2H

34S

15N33S 2H




70

Isotopic Fidelity and Resolving PowerLincomycin, ESI(+)

Maintains isotopic fidelity at high resolving power

Unlike other MS instrumentation, scimaX has the ability to change resolving power from low resolution to eXtreme resolution. Isotopic peak fidelity is maintained across all settings.

R: 42k to 1300k @ m/z 407Calibrated on C12H18F12N3O6P3

150 200 250 300 350 400 450 500 550 600

Inte

nsit

y

m/z

2.0

x 1010

1.0

0

x 1010

1.0

0.5

0x 1010

1.0

2.0

1.0

0

6x 109

x 109

x 109

4

2

0

0

0

R: 330 k

R: 650 k

R: 1.3 M

R: 170 k

R: 85 k

R: 42 k

71

Isotopic Fidelity and Resolving PowerLincomycin, ESI(+)

Maintains isotopic fidelity at high resolving power

Isotopic peak fidelity is maintained even above 1M resolving power (RP), while high mass accuracy and peak fidelity is maintained at low RP.

R: 42k to 1300k @ m/z 407Calibrated on C12H18F12N3O6P3

Inte

nsit

y 4.0

x 109

2.0

0

x 109

0.5

0

R: 650 k

2.0

x 109

1.0

0

4.0

x 108

2.0

0

R: 330 k

x 1010

1.0

x 109

0.50.5

00

R: 1.3 M

408.20 409.210 409.220 409.230408.21 408.22 408.23 408.24m/z m/z

6.0

4.0

x 109

2.0

0

1.0

x 109

0.5

0

simulation

2.03.0

1.00

x 109 x 108

1.0

1.5

0.5

0

R: 170 k

x 109

1.0

x 108

0

0.5

0

R: 85 k

2.03.04.0

1.00

x 108 x 108

0.5

1.0

0

R: 42 k

72

Mass AccuracyRoutine Parts Per Billion

303.195 303.200 303.205 303.210 303.215m/z

73

R: 1,550,000 @ m/z 430RMS mass error: 76 ppbTransient length: 2.96 s 2ω detectionabsorption mode

Calibration Mass AccuracyESI(+), NaTFA

ppb mass accuracy

400200 600 1400800 1600 18001000 1200m/z

Inte

nsit

y

0.4

0.2

0.6

0.8

1.0

x109

0

74

R: 1,760,000 @ m/z 384RMS mass error: 63 ppbTransient length: 2.96 s 2ω detectionabsorption mode

Calibration Mass AccuracyESI(-), NaTFA

ppb mass accuracy

400200 600 1400800 1000 1200m/z

Inte

nsit

y [%

]

1.0

0.5

1.5

2.0

x109

0

75

Technology

76

scimaX MRMSTechnology

2xR ParaCell Detector(n)ECD, SORI-CID, ExD

Analytical Quadrupole CASIDual Stage Ion Funnel

Collision Cell (n)ETD, CID

Dual ESI/MALDI Ion Source

77

Maxwell Magnet TechnologyNo Liquid Cryogens – Ever

scimaX MRMS introduces Maxwell magnet technology to the field of mass spectrometry.

This revolutionary design does not require liquid cryogens fills – ever.

This eliminates the need of monthly and yearly cryogen fills. It also eliminates the cost and

hassle of obtaining cryogens.

Additionally, the reduced footprint allows for the placement of scimaX into any standard

laboratory. Eliminating the need for quench ducts and simplifying the installation to that of any

traditional mass spectrometry instrument.

All of the performance without the headaches of the past.

• 7T magnet

• Ultra-Stable Field

• Very compact design

• Small stray field

• No Liquid Nitrogen

• No Liquid Helium fill needed

• Biennial cold-head exchange

• No Quench duct required293 K

40 K

4 K

78

scimaX is powered by Magnetic Resonance Mass Spectrometry (MRMS) technology which provides superior eXtreme Resolution and mass accuracy, enables routine Isotopic Fine Structure (IFS) analysis for a broad mass range and results in unmatched confidence for compound identification.

scimaX MRMSUnique Feature List

Appendix: Specifications for scimaX

Analytic Performance

Mass Range 100–10,000 m/z

Quadrupole Isolation Mass Range 100–6000 m/z

Quadrupole Isolation Efficiency > 60%

Mass Accuracy (m/z 100–1500)With internal calibrant: < 600 ppb average error With external calibrant: < 1500 ppb average error

Resolving Power @ m/z 400

(Lincomycin)

≥ 450,000 @ 1 Hz for LC experiments20,000,000 maximum resolving power

Isotopic Fine Structure (IFS)

@ m/z 400 (Lincomycin)

Separation of 13C, 15N, 18O, 34S isotopic peaks in M+1 or M+2 isotopic cluster, e.g.:

SensitivityESI (Ubiquitin): S/N > 20:1 for < 100 amol (consumed)MALDI (GluFib): S/N ≥ 20:1 @ m/z 1570.68 for 250 amol on target

MS/MS Efficiency (CID in collision

cell, LHRH, [M+2H]2+> 50% conversion from isolated precursor

Multistage MS (MS3 guaranteed)

(LHRH)LHRH MS/MS (collision cell) -> MS/MS/MS (ParaCell)

Negative Ions Functionality shown on Fibrinopeptide B

Fragmentation Techniques

• In Source CID

• CID in collision cell

• (n)ECD and EID in ParaCell; ECD sensitivity for c5 fragment of Substance P @ m/z 624 S/N > 10:1 for 5 fmol consumed

• SORI

• (n)ETD

MS/MS CapabilitiesAutomated selection, isolation and MS/MS utilizing CID, (n)ECD or (n)ETD

34S

13C15N 13C33S

18O

13C2

79


System Properties

Vacuum System 6 stage differentially pumped vacuum system

• 2 rough pumps (28 m3/h and 5 m3/h) and 4 turbo pumps (each 260 l/s)

• Continuous sample introduction at atmospheric pressure while maintaining ultra-high vacuum (UHV) analyzer conditions

Dual ESI/MALDI source • Apollo II Electrospray Source, flow rate: 1 μL/min – 1 mL/min

• Highly sensitive MALDI source in conjunction with microtiter plate formatted MALDI targets and 1 kHz SmartBeam-II laser

• Fast software controlled switchover between MALDI and ESI operation or simultaneous ESI/MALDI operation

• Dual ion funnel for high ion transmission efficiency over broad mass range

• In Source Collision Induced Dissociation (IS-CID) control

ETD Electron Transfer Dissociation (ETD)

• Robust third generation design

• Provides extensive fragmentation for top-down studies

• Superb tool for studying labile PTMs

Analytic Quadrupole • Facile Q-CID for top-down and LC-MS/MS

• Supports CASI for enrichment of low abundant species

Ion Optics Optimized ion optics to guide the ions, enabling

• Sensitive transmission of broad mass range

• Easy and quick workflow and sample changes

Magnet 7T conduction cooled magnet

• Very stable and homogenous 7T magnetic field for ultimate MRMS performance

• Very compact design with small stray field

• No liquid nitrogen

• No liquid helium fill needed

• No quench duct required

Mass Analyzer Proprietary ParaCell MRMS detector with patented 2 R magnetron control technology

• Dynamic Field shape and shimming enables eXtreme Resolution applications with speed

• Ultrahigh performance low noise 2 R preamplifier for improved sensitivity

• Allows for high resolution, in-cell isolation

• Integrated computer controlled indirectly heated cathode for Electron Capture Dissociation (ECD) and Electron Induced Dissociation (EID) of trapped ions

Acquisition Electronics Nanobay-E featuring

• Fully computer controlled data acquisition

• Data streaming, enabling eXtreme broad-band Resolution

• Flexible and fast data acquisition

Size • Floor standing, L 2.30m x B 0.93m x H 2.04 m

Weight • Mass spectrometer ~ 1450 kg, Accessories ~ 255 kg

80

Magnetic Resonance Mass Spectrometry (MRMS) system

A. Dual ESI/MALDI sourceMass spectrometer with an electrospray ionization (ESI) source allowing for positive and negative ion detection.

• The needle of the near-orthogonal ESI source must be on ground potential.• Ions from the ESI source need to be deflected orthogonally into the mass spectrometer for minimum neutral

gas background.• Atmospheric Pressure Ionization (API) spray chamber shall be compatible with a wide range of ionization

sources including ESI, APPI, APCI, CaptiveSpray, and GC-APCI.• The source shall be compatible with a direct insertion probe (DIP) solution for facile introduction of volatile

compounds.• Fast polarity switching shall be possible in less than a second.

A mass spectrometric source setup including• An ion source with dual ion funnel system enabling simultaneous transfer of a broad m/z-range by monolithic

rf design eliminating compound specific tuning.• A lens system enabling switching from soft ion transfer to in-source-CID experiments.

Mass spectrometer with a combined ESI/MALDI source allowing for positive and negative ion detection.

Switching between ESI and MALDI operation must be fully automatic and SW controlled, the switchover time shall be fast and not exceed 1 minute.

• Simultaneous acquisition of ions originating from ESI and MALDI shall be possible.• Mixing of ions generated by ESI into the MALDI ions from the e.g. tissue sample shall allow for internal

calibration leading to a further increase of mass stability and accuracy.

Solid-state laser capable of at least 1 kHz repetition rate with dynamical shaped beam profile for optimized MALDI performance.

Technology using nth (n≥2) harmonics detection enabling fast MALDI imaging acquisitions while maintaining resolution required for Isotopic Fine Structure (IFS) investigations.

Co-registration for MALDI Imaging data within the suppliers MS software including visualization of mass spectrometry imaging data.

Unique Feature Listapplicable toBDAL #188807050 (for regions with 50Hz power requirements) or BDAL #188807060 (for regions with 60Hz power requirements)


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B. Mass SpectrometerSupport of a technique for the selective enrichment of low abundant species, including

• Quadrupole mass filter for Continuous Accumulation of Selected Ions (CASI) to increase the dynamic range and spectral quality over a selected mass range.

• Continuous Accumulation of Selected Ions (CASI) providing added sensitivity and mass resolution for target m/z ranges.

Transmission of a wide m/z range (~100 – 10,000 m/z)

Provision of various fragmentation techniques• CID in source and collision cell• (n)ECD in MRMS analyzer• SORI-CID in MRMS analyzer• (n)ETD• EID

C. Mass Analyzer TechniqueA mass spectrometric analyzer with the following properties:

• Differential pumping system with ≥ 6 differential pumping stages to achieve ultimate vacuum at the analyzer and best mass spectrometric performance characteristics.

• Fourier Transform Mass Spectrometer (FTMS) based on Magnetic Resonance Mass spectrometric (MRMS) detection system enabling routine Isotopic Fine Structure (IFS) analysis for a broad mass range.

An MRMS system featuring a conduction cooled 7T magnet:• No filling of liquid cryogens needed• No quench line needed

A mass spectrometric detection device as specified below:• Cylindrical MRMS detection cell equipped with ECD capabilities• Capability of mass isolation in the detector cell• MRMS detector with magnetron control features• Exploitation of nth (n≥2) harmonics detection to enable routine Isotopic Fine Structure analysis (IFS) with speed• Mass accuracy (internally calibrated) of 600 ppb or better• Ultimate resolving power in excess of 20 Million at m/z 400• Allowing for LC experiments with an acquisition speed of 1 Hz while maintaining mass resolution of

≥ 400,000 at m/z 400 or ≥ 800,000 at m/z 200.• Allowing for MALDI experiments with an acquisition speed of 1 Hz while maintaining mass resolution of

≥ 400,000 at m/z 400.

D. SoftwareFlexibility to control HPLC systems of various vendors (Agilent, Bruker, Thermo, Waters) within the origi-nal MS vendor software offered (START/STOP signal is insufficient)

Absorption Mode Processing (AMP) enabling• Faster acquisitions for LC and MALDI based applications• Enhanced mass resolution for Isotopic Fine Structure (IFS) interrogation

SW for elemental formula determination from isotopic profile evaluation

E. Further requirementsPossibility of remote service diagnostics via secured Internet connection

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Electro-Spray Ion Source: Apollo II Ion Source with Ion Funnel • Title: "Ionization chamber for atmospheric pressure ionization mass spectrometry" Patent granted: US731502082 • Title: "Ion guide for mass spectrometers" Patents granted: CA2463433C; CA2747956C; EP1465234B; US7495212B2; US7459693 B2; US785l752B2;

US8222597B2 • Title: "Ion funnel with improved ion screening" Patents granted: GB2402261B; US7064321B2

High Precision Multipole Rod Ion Guides • Title: "Multiple rod systems produced by wire erosion" Patents granted: DE102004037B4; GB2416915B; US7351963B2

Electron Capture Dissociation in an ICR cell • Title: "Method and device for irradiating ions in an ion cyclotron resonance trap with photons and electrons" Patents granted: DE10213652B4; GB2390937B; US6803S69B2

ParaCell XR and 2xR Detector • Title: "Ion cyclotron resonance measuring cells with harmonic trapping potential" Patents granted: DE102009050039B4; US8704173B2 • Title: "Correction of asymmetric electric fields in ion cyclotron resonance cells" Patents granted: US8859953B2; US8766174Bl • Title: "ICR cell operating with a duplexer" Patents granted: DE102014226498B4; US9620349B2 • Title: "Introduction of ions into ion cyclotron resonance cells" Patents granted: EP 285809061; US935583082

Isotopic Fine Structure MRMS • Title: "Determination of elemental composition of substances from ultrahigh-resolved isotopic fine structure

mass spectra Patent granted: US9111735Bl

Smart Beam MALDI source • Title: "Laser system for the ionization of a sample by matrix-assisted laser desorption in mass spectrometric

analysis" Patents granted: GB2421352B; US7235781B2 • Title: "A method and system for providing a choice of spatial intensity distributions of laser radiation in

MALDI mass spectroscopy" Patents granted: GB2423187B; US7385192B2

Electron Transfer Dissociation • Title: "Ion fragmentation by electron transfer in ion traps" Patents granted: GB2423865 B; US7456397B2

Appendix: Patents


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Additional Resources

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MRMS LiteratureResources for researchers

MALDI Imaging

High-resolution climate reconstruction using MRMS MALDI Imaging

Designer drug analysis by forensic MALDI techniques

Interrogation of the Spatial Metabolome of Ginkgo biloba with high-resolution MALDI and LDI Mass Spectrometry Imaging

Integrating Ultra-High Speed MALDI-TOF and MALDI MRMS Imaging For Spatial Proteomics

Single Cell Lipid Analysis using Mass Spectrometry

MALDI FTMS Imaging Mass Spectrometry of N-glycans as Tissue Biomarkers of Cancer

MALDI Imaging Success Stories

Microbial Imaging Mass Spectrometry With Fourier Transform Mass Spectrometry

Metabolomics

Fast profiling of sphingolipids alteration in hypertension by MRMS

MRMS aXelerate for targeted metabolomics profiling of myxobacterial extracts

Rapid elucidation of carotenoids in microalgae formulations by MRMS aXelerate

A strategy using isotopic fine structure to reveal potential biomarkers showing the effects of traditional Chinese medicines on Alzheimer disease in rats

Analysis of metabolic changes in murine hair follicles treated with Procyanidin-B2 rich nutraceuticals by DI-MRMS

MRMS aXelerate – rapidly detected micropollutants and plant response metabolites in poplar leaves

Automated MALDI Magnetic Resonance Mass Spectrometry (MRMS) for biomarker identification in large clinical sample sets

Unambiguous Natural Product ID

Electron Induced Dissociation for the Differentiation of Isomeric Metabolites of Diclofenac

Ultrafast Statistical Profiling of Bacterial Metabolite Extracts

Definitive Elemental Formula Determination Debuts with SmartFormula-XR

Native and Glycans

Glycan Sequencing by Electronic Excitation Dissociation Tandem Mass Spectrometry

Comparison of CID and EID Mass Spectrum of Glycosides from solarix XR

Top Down Analysis of Histone H4

Expanded Collisional Energy Files for Automated Top Down and Bottom up Analysis on solariX

MALDI In-Source Decay for Top-Down Analysis of Proteins using Fourier Transform Mass Spectrometry

Petroleomics

Characterization of Petroleum Samples via Thermal Analysis Coupled to APCI FTMS

Reproducibility of Crude Oil Characterization by Flow Injection APPI-FT-ICR Mass Spectrometry

LDI FT-ICR MS as a Tool for Statistical Analysis of Crude Oils

solarix XR: Analysis of Complex Mixtures

Analysis of peat bog after solid phase extraction by FTMS

Analysis of sulfur-rich crude oil and bitumen by FTMS

Analysis of Gas Oil by GC/APCI FTMS

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MRMS LiteratureResources for researchers

Customer Insights

Prof. Richard Drake, MUSC, Translating Cancer Biomarkers into the Clinic with MALDI Mass Spectrometry Imaging

Prof. Ying Ge, University of Wisconsin, Top-Down Proteomics: An Emerging Technique Driven By Cutting-Edge Mass Spectrometer

Prof. Jonathan Sweedler, University of Illinois at Urbana Champaign, How Mass Spectrometry Techniques are Propelling the Advancements of Single Cell Biology

Prof. Sally-Ann Poulsen, Griffith University, Innovating in medicinal chemistry using Fragment Based Drug Discovery combined with Native Mass Spectrometry

Prof. Per Andren, Uppsala University, MALDI Imaging in Neuroscience, Revealing the Biomolecular Map of the Brain

Solutions

Isotopic Fine Structure, Beyond the Molecular Realm: Unambiguous Elemental Formula Determination

Label-Free Molecular Imaging, Discover, localize and quantify biochemical changes and molecular markers

MRMS aXelerate: Phenomics, Metabolomics and beyond

Metabolomics, Novel solutions for Metabolomics, Lipidomics and high-throughput Phenomics

Native MS Solutions, scimaX MRMS: Perfectly designed for Native MS

Please visit Bruker’s Literature Room for the most recent publications and poster halls:

https://www.bruker.com/products/mass-spectrometry-and-separations/literature/literature-room-mass-spec.html

https://www.bruker.com/products/mass-spectrometry-and-separations/literature/literature-room-mass-spec.html

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18T MRMSPushing the boundaries of science

scimaX represents a revolutionary step in

extreme resolution mass spec, bringing high field

performance to 7T. However, there remain heroic

global problems that require a solution that can

address these issues that go beyond the current

standard. The net effect of increasing magnetic

field is not incremental, it is transformative. The

cumulative effect of increased field strength takes

the performance of scimaX to a level beyond

currently available production systems. For these

applications, Bruker offers an 18T magnet that

will define a new class of experimentation.

Please contact your local Bruker representative to

learn more.

Technical Specifications (preliminary)

Magnet Type B-C 180/11 USR-TT

Magnetic Field 18T

Cold Head Dual stage

Magnet Dewar Asymmetric, ca. 85cm to magnetic center

Stray Field (5G, axial/radial)

<3.6m/ <3.6m; in normal operation <2.5m/ <2.5m

Ion Source ESI/MALDI dual source

Ion Opticsrf and dc ion guides, quadrupole, collision cell, ExD

Detector ParaCell MRMS

Foot print ca. 3.4 m x 1.6 m

Height ca. 2.6m; ceiling ≥2.9m

Total Weight ca. 7.5 t

Certification CE

Maintenancemagnet every 2nd year, no field ramping required; mass spectrometer cart yearly

Liquid Helium Volume ca. 1000 Liter

Liquid Helium Refill Volume

None between biennial maintenance; ca. 10 Liter LHe at maintenance

18T MRMS design study: In normal operation the 5G line extends from the magnetic center less than 2.5 m in z (magnetic axis) direction and less than 2.5 m radial.

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Notes

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Bruker Daltonik GmbH

Bremen · GermanyPhone +49 (0)421-2205-0

Bruker Scientific LLC

Billerica, MA · USA Phone +1 (978) 663-3660

[email protected] - www.bruker.com

For Research Use Only. Not for Use in Clinical Diagnostic Procedures.

Pushing science to the max

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