Orbitrap ID-X Tribrid mass spectrometerTransforming small molecule identification and characterization

A leap ahead for small molecule analysesAcquisition and interpretation of mass spectra to characterize and identify small-molecule unknowns is a formidable task. Sample complexities, background interferences, compound structural diversity, and the lack of robust data processing tools can make it difficult to set up analytical methods and interpret data. Overcoming these bottlenecks requires advances in both data acquisition and computational tools.

Equipped with the novel, automated AcquireX intelligent data acquisition workflow including various characterization capabilities using intelligent MSn and a multitude of mass spectral data processing tools, the Thermo Scientific™ Orbitrap ID-X™ Tribrid™ mass spectrometer can help you extract meaningful data faster and with the highest confidence.

Acquisition and interpretation of mass spectra to characterize and identify small-molecule unknowns is a formidable task. Sample complexities, background interferences, compound structural diversity, and the lack of robust data processing tools can make it difficult to set up analytical methods and interpret data. Overcoming these bottlenecks requires advances in both data acquisition and computational tools.

Equipped with the novel, automated AcquireX intelligent data acquisition workflow including various characterization capabilities using intelligent MSn and a multitude of mass spectral data processing tools, the Thermo Scientific™ Orbitrap ID-X™ Tribrid™ mass spectrometer can help you extract meaningful data faster and with the highest confidence.

2

Focus on science, not instrument setup The unique combination of Thermo Scientific™ Tribrid™ mass spectrometer architecture, built-in method editor templates, and ready-to-use experimental parameters enable you to analyze complex samples without extensive expertise or effort. The easy-to-use method templates support a variety of small molecule applications to define data acquisition using combinations of high-resolution accurate-mass (HRAM) and ion trap mass analyzers, and MSn fragmentation using either collision-induced dissociation (CID) or high-energy collision dissociation (HCD) techniques.

Collect more meaningful data, not just more dataMS and MSn data acquisition methods for compound identification have been limited by the relative abundances of compounds of interest relative to the background and/or sample matrices. Duty cycle is often wasted triggering on background. Manually excluding background and creating inclusion lists for relevant features is too laborious. A breakthrough in intelligent automation, AcquireX, increases profiling efficiency by automating inclusion/exclusion list creation for MSn data acquisition so you find more compounds with distinguishable fragmentation patterns.

Meet data analysis requirements for speed, simplicity, and flexibility High-quality MSn data generated by the Orbitrap ID-X Tribrid mass spectrometer reduces compound identification bottlenecks. Confidently identify knowns and unknowns, and elucidate structures of unknowns using spectral library searching, MSn mass spectral tree annotation, in silico fragmentation, and precursor ion fingerprinting. High-certainty identification and characterization of small molecules is now remarkably fast.

Demystifying unknown annotation using high-quality MSn data Confidently identify known unknowns and elucidate structures of unknown unknowns with novel spectral prediction tools and processing routines. New data processing methods leverage custom library searching, in silico fragmentation modeling, and precursor ion fingerprinting to associate high-quality MSn data from unknown precursors with compound structures.

Break your bottlenecks with intelligence and technology— from acquisition to analysisGo beyond today’s small molecule analyses and capture more low-abundance analytes in complex samples using intelligent automation and streamlined mass spectral processing, elucidation, and identification software.

3

Building on proven Tribrid architectureThe Orbitrap ID-X Tribrid mass spectrometer is based on the proven and trusted tribrid architecture that combines the best of quadrupole ion trap and HRAM Thermo Scientific™ Orbitrap™ mass analyzer technology for acquisition of the richest MSn data for every sample run. Dual fragmentation techniques—CID and HCD—are available at any stage of MSn, with subsequent mass analysis in either the ion trap or ultra-high-resolution Orbitrap mass analyzer.

t

4

Active Beam GuidePrevents neutrals and high-velocity clusters from entering the mass resolving quadropole

Optional EASY-IC Ion SourceGenerates internal calibrant ions for real-time mass calibration

t

S-LensOptimizes ion transmission into the mass spectrometer, while minimizing in-source fragmentation

Quadrupole Mass FilterSelects precursor ion with resolution up to 0.4 amu; Yields high ion transmission from 50 to 2000 m/z

Dual-Pressure Linear Ion TrapProvides MSn rates up to 40 Hz and sensitive mass analysis of CID and HCD fragment ions

Ultra-High-Field Orbitrap Mass AnalyzerOffers resolution up to 500,000 FWHM and MSn scan rates up to 30 Hz

Ion-Routing MultipoleEnables parallel analysis; Allows HCD at any MSn stage

5

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Focus on your science, not on instrument setupThe intuitive method editor features a user-friendly interface with predesigned optimized method templates for a wide range of applications.

Effortlessly run advanced experiments

for a variety of applications using validated method

templates Ready-to-use templates with predefined instrument parameters enable generation of high-quality data regardless of analyte chemical structure.

0.86 0.87 0.88 0.89 0.90 0.91 0.92 0.93

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132.0766-1.5ppm

0.92132.0767-0.75ppm

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132.0767-0.75ppm

0.88

132.0767-0.75ppm

0.88

132.0767-0.75ppm

0.89

132.0767-0.75ppm

0.89

132.0767-0.75ppm

0.90

132.0767-0.75ppm

0.90

132.0767-0.75ppm

0.90

132.0767-0.75ppm

0.91

132.0767-0.75ppm

0.91

Background ExclusionIncrease the MSn sampling efficiency of unknown compounds within one replicate individual, or batch analysis.

Exclusion and Component Inclusion Increase the breadth and depth of compound identification and structural elucidation using various ion tree experiments.

Deep Scan with inclusion and exclusion Attain exhaustive characterization using fully-automated, complex data acquisition enabling comprehensive sample interrogation.

Excellent scan-to-scan mass measurement accuracy Unprecedented run-to-run mass measurement accuracy

Cumulative mass measurement accuracy for replicate LC-MS analyses of 24 small molecule standards conducted over 72 hours.

Orbitrap ID-X Tribrid mass spectrometer analysis of creatine (theorectial m/z of 132.0768) LC-MS analysis.

–3

–2

–1

1

3

5

0 200 400

Molecular weight (DA)

600 800 1000

Match experimental design to your challenging samplesThree modes of AcquireX intelligent data acquisition enable rapid profiling of multiple samples or exhaustive interrogation of replicate sample injections. Each mode increases the number of compounds sampled with distinguishable fragmentation spectra to use for sample and study characterization.

MS internal calibration adds confidenceThe Orbitrap ID-X Tribrid mass spectrometer instrument calibration routine ensures high mass measurement accuracy over the mass range used for small molecule analysis, regardless of the MSn level. The result is high confidence in structural analysis and elucidation for low m/z ions within a sample and over extended periods of time.

7

Collect more meaningful data, not just more dataIntelligent AcquireX efficiently acquires data with significantly less manual experimental setup and interpretation expertise required.

8

The three modes of AcquireX data acquisition permit rapid profiling across multiple samples or exhaustive sample interrogation through replicate injections. Initial analysis of a blank and representative sample enables automated creation of an inclusion/exclusion list used to direct DDA sample analysis. Each replicate injection increases the number of compounds sampled with distinguishable fragmentation spectra for sample and study characterization.

INCLUSION LISTINCLUSION LIST

400.253 400.253

468.271EXCLUSION LIST

322.561

418.253

579.457

EXCLUSION LIST

322.561

418.253

468.271

579.457

EXCLUSION LIST

322.561

400.253

418.253

468.271

579.457

EXCLUSION LIST

Blank Full MSSample Full MS

Sample Iterative DDA

Sample Iterative DDA

Sample Iterative DDA

INCLUSION LIST

MS Method

INCLUSION LIST

MS Method

MS Method

9

Detect up to 140% more unique unknown compounds that are truly experimentally related, even at low abundance as shown in this example of three replicate injections of the NIST SRM 1950 plasma extract sample.

The AcquireX Intelligent data acquisition approach samples more compounds with distinguishable product ion spectra. With each iteration and subsequent injection, the same compounds chosen to be fragmented are eliminated, minimizing redundancy.

Injection1

Injection2

Injection3

1 2Total number of DDA injections

3

73% increase

125% increase139% increase

Traditional DDA DDA with AcquireX

915

1578

1013

2282

1061

2534

Obtain more distinguishable fragmentationFragment more unique unknown compounds

The Orbitrap ID-X Tribrid mass spectrometer improves precursor sampling by automatically updating the inclusion/exclusion lists following each replicate. The data-informed AcquireX approach utilizes sequential injections to interrogate more unique sample components through MSn acquisition. AcquireX more than doubled the number of unique compounds with fragmentation spectra in human plasma (NIST SRM1950) in comparison to traditional DDA, resulting in a greater metabolome coverage.

HIG HE R E FFIC IE NCY

10

Go beyond traditional untargeted metabolomics Increase metabolome coverage and confidence in unknown identification.

Insufficient metabolome annotation has limited the biological interpretation of untargeted metabolomics studies. The Orbitrap ID-X mass spectrometer provides a streamlined approach to routinely achieve comprehensive metabolome coverage with confident compound annotation against spectral libraries to provide compound annotations or perform de novo structural elucidation of unknown metabolites. The power of HRAM Orbitrap mass analyzer technology with AcquireX intelligent acquisition of MSn spectra reinvents the way metabolomics is done today.

Number of compounds with fragmentation spectra

Injection 1

Injection 2

Injection 3

No MS/MS

Traditional DDA DDA with AcquireX

192.0656

94.0651

136.0757

174.0551209.0923

m/z

146.0596

146.0596

128.0493

118.0645

77.038675.0229

m/z

HCD MS/MS

CID MS3

The Orbitrap ID-X Tribrid mass spectrometer offers MSn fragmentation and the flexibility of complementary HCD and CID techniques to increase the probability of generating information-rich product-ion spectra across more compound classes. In this example, fragmentation of the metabolite kynurenine illustrates the complementarity of HCD and CID.

Annotation of more high-quality spectra leads to improved metabolome coverage, comprehensive pathway annotation, and functional interpretation of results. Confident annotations of metabolites in the tryptophan degradation pathway enabled detection of changes in the levels of anthranilic acid in a comparison among healthy donors and donors with Crohn’s disease.

HIG HE R QUA LIT Y

11

“ Confident metabolite identification remains a challenging step in the untargeted metabolomics workflow where the data acquired are applied to derive the structure of all metabolites detected. Progress has been made to putatively annotate metabolites using MS or MS/MS data using established UHPLC/MS assays through the use of metabolomics database and mass spectral library searching workflows. The Orbitrap ID-X Tribrid MS delivers complementary information with multiple dissociation techniques and robust MSn data required to deduce structural information and increase confidence in metabolite annotation. With built-in intelligent data acquisition, AcquireX, we now collect more informative data, and not just more data, we can maximize metabolome coverage and increase confidence in the identification on unknowns applying intelligent-DDA approaches.”

— Warwick Dunn, PhD, Professor in Analytical and Clinical Metabolomics, Phenome Centre Birmingham, School of Biosciences and Institute of Metabolism and Systems Research, University of Birmingham, UK

Complimentary MSn fragmentation of kynurenine metabolite Metabolite intermediate kynurenine identified in a Crohn’s disease study

1.13.11.111.13.11.52

Tryptophan

N-formylkynurenine Kynurenine

Alanine

OH

O

NH2

NH

OH

O

NH

H2N

O

Anthranilic acid

1.60 -0.952.55

O

HN2

HO

OH

O

O

H2N

NH

O

O

OHH2N

O

3.5.1.9

3.7.1.3

In pharmaceutical research and development, small molecule characterization plays a critical role in active pharmaceutical ingredient (API) impurity and metabolite identification, degradation profiling, and analysis of extractables and leachables. High-resolution mass spectrometry (HRMS) is a pivotal tool for these applications. The Orbitrap ID-X Tribid mass spectrometer elevates HRMS to a new level. With AcquireX intelligent data acquisition, complex matrix interferences are removed by automatic background exclusion, ensuring the capture of low-abundance analytes and significantly improving identification. Information-rich MSn fragments provide in-depth structural knowledge of compounds, and facilitate high-confidence identification of unknown or unexpected compounds.

Coupled with the powerful spectral library and data processing software, the Orbitrap ID-X Tribrid mass spectrometer provides the ultimate solution for small molecule structural analysis, significantly increasing accuracy, efficiency, and overall productivity in drug impurity and metabolite identification, extractable and leachable analysis, and other related applications.

Abundantly certain in no time Identify and characterize structures of small molecule pharmaceuticals.

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13.3913.5213.29 15.2412.576.336.04 6.52 13.566.755.826.855.365.22 13.61

7.09 12.424.87 12.387.14 14.914.72 14.61 15.4112.347.264.51

7.38 12.2415.774.10 16.0312.093.893.44 10.747.57 10.66 16.617.743.071.211.15 2.43 10.008.88 9.40 17.130.36

14.06

14.22

12.8711.68 13.8910.38

714 716 718 720 722 724 726 728 7300

20

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718.4763

718.9782

725.4829

728.5124726.4994722.2712720.1359 729.5181714.1635724.4760715.7983

723.2744717.9698714.4966

700 705 710 715 720 725 730 7350

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702.4810724.9962722.2719

701.4735

700.4627 728.5131

703.4876

732.8096715.1379 731.1905

0.22 ppm

0.22 ppm

Impurity analysis performed using AcquireX data acquisition

Analysis of the excipient sample, along with an analysis to determine background precursors was performed. Background precursors were automatically incorporated into an exclusion list and imported into the refined method. The exclusion list significantly decreased the number of compounds of interest targeted during the DDA method, resulting in more efficient MS/MS and MS3 data acquisition. The highlighted retention times show the measured elution times for two isomeric impurities.

TIC

MS

XIC m/z 722.2715

13

MS3 spectral analysis enables differentiation of impurity structures Overlaying ion tree spectra with in silico dissociation mechanisms to propose impurity structures

Plausible precursor and product ion structures were evaluated using the Fragments and Mechanisms tool in Thermo Scientific™ Mass Frontier™ 8.0 software. The fragmentation pathways of two plausible structures were evaluated for fragmentation pathways to predict formation of the m/z 583.1873 product ion. Structure A has a direct fragmentation pathway justifying the fragmentation of m/z 704 to form the m/z 583 product ion. Structure B predicts that m/z 583.1873 fragment would originate from the m/z 601 MS/MS product ion instead of the m/z 704 product ion.

HRAM MS/MS and MS3 spectra acquired at the retention times 11.68 and 14.06 minutes. The comparative MS/MS spectra show complex, but conserved fragmentation, reducing the confidence in proposing precursor structures that lead to different elution profiles. Specifically, the MS/MS product ions at m/z 704 and 583 are detected for both m/z 722 precursors, however when the m/z 704 MS/MS product ion is isolated and fragmented, only the structure eluting at 11.1 minutes has an MS3 product ion at m/z 583, indicating different precursor structures.

RT 11.68 min Structure “A”

RT 14.06 minStructure “B”

50 100 150 200 250 300 350 400 450 500 550 600 650 700m/z

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722.2714704.2610133.0883

122.0811

201.1276104.0705

89.0596

343.0728188.0555362.1970

279.1106159.1169504.1413 557.209281.0698

583.1879 686.2501

50 100 150 200 250 300 350 400 450 500 550 600 650 700

704.2604

722.2707

201.1274

133.0883

91.0541122.0811

343.0727188.0554 362.1966279.1107

504.140781.0697 686.2489575.2177583.1868

200 250 300 350 400 450 500 550 600 650 700

686.2532

504.1432

362.1962530.1206201.9671

583.1810334.2036

280.3376207.7233

200 250 300 350 400 450 500 550 600 650 700

504.1428

686.2523

362.1957530.1220343.0721

415.9328

OH

H+

HN

HN+H

HN

HOOH

HO

NH

OH

OH

OH

OO

m/z 722.2717

m/z 722.2717

OO

N

FF

F

O

O

O

S

S

FF

F

N

OH

HN

NH

OH

OH

OO

m/z 704.2612

OO

S

FF

F

NNH

OO

m/z 583.1873

OO

+

S

FF

F

N

OO

HN

HN

HOOH2

HO

m/z 722.2717

N

FF

F

O

O

O

S

OO

HN

HN

HO

HO

m/z 704.2612

N

FF

F

O

O

O

S

OO

Lib Lib

-H2O

+

+

H+

RT 11.68 min Structure “A”

RT 14.06 minStructure “B”

50 100 150 200 250 300 350 400 450 500 550 600 650 700m/z

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722.2714704.2610133.0883

122.0811

201.1276104.0705

89.0596

343.0728188.0555362.1970

279.1106159.1169504.1413 557.209281.0698

583.1879 686.2501

50 100 150 200 250 300 350 400 450 500 550 600 650 700

704.2604

722.2707

201.1274

133.0883

91.0541122.0811

343.0727188.0554 362.1966279.1107

504.140781.0697 686.2489575.2177583.1868

200 250 300 350 400 450 500 550 600 650 700

686.2532

504.1432

362.1962530.1206201.9671

583.1810334.2036

280.3376207.7233

200 250 300 350 400 450 500 550 600 650 700

504.1428

686.2523

362.1957530.1220343.0721

415.9328

OH

H+

HN

HN+H

HN

HOOH

HO

NH

OH

OH

OH

OO

m/z 722.2717

m/z 722.2717

OO

N

FF

F

O

O

O

S

S

FF

F

N

OH

HN

NH

OH

OH

OO

m/z 704.2612

OO

S

FF

F

NNH

OO

m/z 583.1873

OO

+

S

FF

F

N

OO

HN

HN

HOOH2

HO

m/z 722.2717

N

FF

F

O

O

O

S

OO

HN

HN

HO

HO

m/z 704.2612

N

FF

F

O

O

O

S

OO

Lib Lib

-H2O

+

+

H+

RT 11.68 min Structure “A”

RT 14.06 minStructure “B”

50 100 150 200 250 300 350 400 450 500 550 600 650 700m/z

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722.2714704.2610133.0883

122.0811

201.1276104.0705

89.0596

343.0728188.0555362.1970

279.1106159.1169504.1413 557.209281.0698

583.1879 686.2501

50 100 150 200 250 300 350 400 450 500 550 600 650 700

704.2604

722.2707

201.1274

133.0883

91.0541122.0811

343.0727188.0554 362.1966279.1107

504.140781.0697 686.2489575.2177583.1868

200 250 300 350 400 450 500 550 600 650 700

686.2532

504.1432

362.1962530.1206201.9671

583.1810334.2036

280.3376207.7233

200 250 300 350 400 450 500 550 600 650 700

504.1428

686.2523

362.1957530.1220343.0721

415.9328

OH

H+

HN

HN+H

HN

HOOH

HO

NH

OH

OH

OH

OO

m/z 722.2717

m/z 722.2717

OO

N

FF

F

O

O

O

S

S

FF

F

N

OH

HN

NH

OH

OH

OO

m/z 704.2612

OO

S

FF

F

NNH

OO

m/z 583.1873

OO

+

S

FF

F

N

OO

HN

HN

HOOH2

HO

m/z 722.2717

N

FF

F

O

O

O

S

OO

HN

HN

HO

HO

m/z 704.2612

N

FF

F

O

O

O

S

OO

Lib Lib

-H2O

+

+

H+

RT 11.68 min Structure “A”

RT 14.06 minStructure “B”

50 100 150 200 250 300 350 400 450 500 550 600 650 700m/z

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722.2714704.2610133.0883

122.0811

201.1276104.0705

89.0596

343.0728188.0555362.1970

279.1106159.1169504.1413 557.209281.0698

583.1879 686.2501

50 100 150 200 250 300 350 400 450 500 550 600 650 700

704.2604

722.2707

201.1274

133.0883

91.0541122.0811

343.0727188.0554 362.1966279.1107

504.140781.0697 686.2489575.2177583.1868

200 250 300 350 400 450 500 550 600 650 700

686.2532

504.1432

362.1962530.1206201.9671

583.1810334.2036

280.3376207.7233

200 250 300 350 400 450 500 550 600 650 700

504.1428

686.2523

362.1957530.1220343.0721

415.9328

OH

H+

HN

HN+H

HN

HOOH

HO

NH

OH

OH

OH

OO

m/z 722.2717

m/z 722.2717

OO

N

FF

F

O

O

O

S

S

FF

F

N

OH

HN

NH

OH

OH

OO

m/z 704.2612

OO

S

FF

F

NNH

OO

m/z 583.1873

OO

+

S

FF

F

N

OO

HN

HN

HOOH2

HO

m/z 722.2717

N

FF

F

O

O

O

S

OO

HN

HN

HO

HO

m/z 704.2612

N

FF

F

O

O

O

S

OO

Lib Lib

-H2O

+

+

H+

RT 11.68 min Structure “A”

RT 14.06 minStructure “B”

50 100 150 200 250 300 350 400 450 500 550 600 650 700m/z

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722.2714704.2610133.0883

122.0811

201.1276104.0705

89.0596

343.0728188.0555362.1970

279.1106159.1169504.1413 557.209281.0698

583.1879 686.2501

50 100 150 200 250 300 350 400 450 500 550 600 650 700

704.2604

722.2707

201.1274

133.0883

91.0541122.0811

343.0727188.0554 362.1966279.1107

504.140781.0697 686.2489575.2177583.1868

200 250 300 350 400 450 500 550 600 650 700

686.2532

504.1432

362.1962530.1206201.9671

583.1810334.2036

280.3376207.7233

200 250 300 350 400 450 500 550 600 650 700

504.1428

686.2523

362.1957530.1220343.0721

415.9328

OH

H+

HN

HN+H

HN

HOOH

HO

NH

OH

OH

OH

OO

m/z 722.2717

m/z 722.2717

OO

N

FF

F

O

O

O

S

S

FF

F

N

OH

HN

NH

OH

OH

OO

m/z 704.2612

OO

S

FF

F

NNH

OO

m/z 583.1873

OO

+

S

FF

F

N

OO

HN

HN

HOOH2

HO

m/z 722.2717

N

FF

F

O

O

O

S

OO

HN

HN

HO

HO

m/z 704.2612

N

FF

F

O

O

O

S

OO

Lib Lib

-H2O

+

+

H+

RT 11.68 min Structure “A”

RT 14.06 minStructure “B”

50 100 150 200 250 300 350 400 450 500 550 600 650 700m/z

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722.2714704.2610133.0883

122.0811

201.1276104.0705

89.0596

343.0728188.0555362.1970

279.1106159.1169504.1413 557.209281.0698

583.1879 686.2501

50 100 150 200 250 300 350 400 450 500 550 600 650 700

704.2604

722.2707

201.1274

133.0883

91.0541122.0811

343.0727188.0554 362.1966279.1107

504.140781.0697 686.2489575.2177583.1868

200 250 300 350 400 450 500 550 600 650 700

686.2532

504.1432

362.1962530.1206201.9671

583.1810334.2036

280.3376207.7233

200 250 300 350 400 450 500 550 600 650 700

504.1428

686.2523

362.1957530.1220343.0721

415.9328

OH

H+

HN

HN+H

HN

HOOH

HO

NH

OH

OH

OH

OO

m/z 722.2717

m/z 722.2717

OO

N

FF

F

O

O

O

S

S

FF

F

N

OH

HN

NH

OH

OH

OO

m/z 704.2612

OO

S

FF

F

NNH

OO

m/z 583.1873

OO

+

S

FF

F

N

OO

HN

HN

HOOH2

HO

m/z 722.2717

N

FF

F

O

O

O

S

OO

HN

HN

HO

HO

m/z 704.2612

N

FF

F

O

O

O

S

OO

Lib Lib

-H2O

+

+

H+

MS3MS/MS

MS3MS/MS

Demystifying unknowns using high-quality MSn data annotationTake advantage of superior MSn data coupled with selective mass-spectral prediction tools and conquer unknown identification challenges.

High-quality MSn data generated by the Orbitrap ID-X Tribrid mass spectrometer combines precursor ion fingerprinting with substructure analysis to facilitate unknown identification. MSn provides more relevant ion fragment information, helps identify isomeric species, and aides unknown compound structure annotation. By performing an MSn tree/subtree search in Mass Frontier 8.0 software against the mzCloud mass spectral library, the substructures present in an unknown compound can be identified by matching the partial MSn spectral tree data. The partial matches can be rank ordered using mzLogic and/or the FISh score to propose the best candidates.

14

Comparative MS/MS spectra for two flavonoid structural isomers where the only structural difference is associated with an aglycone substructure. The addition of the two sugar rings to the base aglycone substructure results in similar MS/MS spectra, prohibiting correct structural identification despite low mass errors for precursor and MS/MS product ions. Incorporation of neutral-loss triggered MS3 acquisition and subsequent analysis results in clearly defining two isomeric compounds.

287.0546

449.1072

433.1124

449.1074

287.0546

433.1125

241.0492165.0180

213.0543

153.0180

258.0519121.0282

231.0648

269.0441133.0282

197.0594

287.0546

111.0075

153.0180

241.0493161.0231135.0439 287.0547259.0598269.0442

219.0648231.0650116.8602

3.7.1.3

1.13.11.111.13.11.52

3.5.1.9

+

L-Tryptophan

Oxygen

N-Formylkynurenine L-Kynurenine

Formate

0.17 0.120.05

0.29 -0.160.35 -0.16 0.14-0.30

L-Alanine-0.20 -0.270.07

Anthranilate1.60 -0.952.55

H

H H

OH

OH

OH

OH

OH

O

OO

O

O

H2N

H2N

NH2NHHN2

+H

HO+

+

+

H

H2N

O

NH2

NH

+

+

OH

OH

OH

O

O

OH

O O

HO

HO

HO

HO

OHO

O

OH

OH

OH

OH

OH

OO

O

O O

OHO

HO

HO

HO

H

H

H

H

HH

H

H

H

H

OH

OH

O

O

O

O

OO

O O

OO

HO

OH

OH

OHHO

HO

HO

HO

HO

H HO

3.7.1.3

1.13.11.111.13.11.52

3.5.1.9

+

L-Tryptophan

Oxygen

N-Formylkynurenine L-Kynurenine

Formate

0.17 0.120.05

0.29 -0.160.35 -0.16 0.14-0.30

L-Alanine-0.20 -0.270.07

Anthranilate1.60 -0.952.55

H

H H

OH

OH

OH

OH

OH

O

OO

O

O

H2N

H2N

NH2NHHN2

+H

HO+

+

+

H

H2N

O

NH2

NH

+

+

OH

OH

OH

O

O

OH

O O

HO

HO

HO

HO

OHO

O

OH

OH

OH

OH

OH

OO

O

O O

OHO

HO

HO

HO

H

H

H

H

HH

H

H

H

H

OH

OH

O

O

O

O

OO

O O

OO

HO

OH

OH

OHHO

HO

HO

HO

HO

H HO

3.7.1.3

1.13.11.111.13.11.52

3.5.1.9

+

L-Tryptophan

Oxygen

N-Formylkynurenine L-Kynurenine

Formate

0.17 0.120.05

0.29 -0.160.35 -0.16 0.14-0.30

L-Alanine-0.20 -0.270.07

Anthranilate1.60 -0.952.55

H

H H

OH

OH

OH

OH

OH

O

OO

O

O

H2N

H2N

NH2NHHN2

+H

HO+

+

+

H

H2N

O

NH2

NH

+

+

OH

OH

OH

O

O

OH

O O

HO

HO

HO

HO

OHO

O

OH

OH

OH

OH

OH

OO

O

O O

OHO

HO

HO

HO

H

H

H

H

HH

H

H

H

H

OH

OH

O

O

O

O

OO

O O

OO

HO

OH

OH

OHHO

HO

HO

HO

HO

H HO

3.7.1.3

1.13.11.111.13.11.52

3.5.1.9

+

L-Tryptophan

Oxygen

N-Formylkynurenine L-Kynurenine

Formate

0.17 0.120.05

0.29 -0.160.35 -0.16 0.14-0.30

L-Alanine-0.20 -0.270.07

Anthranilate1.60 -0.952.55

H

H H

OH

OH

OH

OH

OH

O

OO

O

O

H2N

H2N

NH2NHHN2

+H

HO+

+

+

H

H2N

O

NH2

NH

+

+

OH

OH

OH

O

O

OH

O O

HO

HO

HO

HO

OHO

O

OH

OH

OH

OH

OH

OO

O

O O

OHO

HO

HO

HO

H

H

H

H

HH

H

H

H

H

OH

OH

O

O

O

O

OO

O O

OO

HO

OH

OH

OHHO

HO

HO

HO

HO

H HO

MS/MS results are indistinguishable without spectra from standards

MS3 results easily distinguish the isomers

Luteolin 7-rutinoside

Kaempferol 3-O-β-rutinoside

3.7.1.3

1.13.11.111.13.11.52

3.5.1.9

+

L-Tryptophan

Oxygen

N-Formylkynurenine L-Kynurenine

Formate

0.17 0.120.05

0.29 -0.160.35 -0.16 0.14-0.30

L-Alanine-0.20 -0.270.07

Anthranilate1.60 -0.952.55

H

H H

OH

OH

OH

OH

OH

O

OO

O

O

H2N

H2N

NH2NHHN2

+H

HO+

+

+

H

H2N

O

NH2

NH

+

+

OH

OH

OH

O

O

OH

O O

HO

HO

HO

HO

OHO

O

OH

OH

OH

OH

OH

OO

O

O O

OHO

HO

HO

HO

H

H

H

H

HH

H

H

H

H

OH

OH

O

O

O

O

OO

O O

OO

HO

OH

OH

OHHO

HO

HO

HO

HO

H HO

3.7.1.3

1.13.11.111.13.11.52

3.5.1.9

+

L-Tryptophan

Oxygen

N-Formylkynurenine L-Kynurenine

Formate

0.17 0.120.05

0.29 -0.160.35 -0.16 0.14-0.30

L-Alanine-0.20 -0.270.07

Anthranilate1.60 -0.952.55

H

H H

OH

OH

OH

OH

OH

O

OO

O

O

H2N

H2N

NH2NHHN2

+H

HO+

+

+

H

H2N

O

NH2

NH

+

+

OH

OH

OH

O

O

OH

O O

HO

HO

HO

HO

OHO

O

OH

OH

OH

OH

OH

OO

O

O O

OHO

HO

HO

HO

H

H

H

H

HH

H

H

H

H

OH

OH

O

O

O

O

OO

O O

OO

HO

OH

OH

OHHO

HO

HO

HO

HO

H HO

Meet your data analysis requirements with speed, simplicity, and flexibility Overcome challenges in small molecule full-scan data analysis and interpretation with Compound Discoverer software.

Thermo Scientific™ Compound Discoverer™ software efficiently extracts high-confidence insights from information-rich, small-molecule HRAM data. It serves as a hub to seamlessly connect you to the tools needed to analyze large amounts of HRAM data productively and confidently.

15

mzCloud™ mass spectral libraryRank search more effectively with the industry-leading online spectra fragmentation library

mzLogic algorithmFast, automated logical analysis eliminates thousands of candidates and hours of work

Mass Frontier spectral interpretation softwareSupports MSn spectral ion tree searching and custom libraries

Leverage MSn data to find best candidate structures

Monitor your system, access your data and system remotely and securely from anywhere in the world. Leverage real-time monitoring of your instrument with automatic email notifications anytime, anywhere. Store data in secure accounts and share with colleagues and collaborators around the world.

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