Post on 28-Sep-2020
More Capabilities, New Possibilities in Mass Spectrometry New Jersey, Oct 12, 2011
Iain Mylchreest, Ph.D
San Jose, CA
2
Increasing App’s in Life Sciences Research and Applied Markets
Expanding Application
Base
Metabolite ID/Profiling,pK • Accelerated drug
development and compound assessment
Proteomics • Biomarker discovery • Pathway analysis • Protein identification
BioPharma • Biologics discovery and
development
Metabolomics/Lipidomics • Comprehensive study of
metabolites for disease and therapeutic assessment
Environmental • Water, soil analysis
Clinical/Toxicology • Disease markers • Pain management • Drugs of abuse
Food Safety • Inspection, nutrition • Chemical contaminant,
microbial detection
3
Dionex: Leading Chromatography Provider
Ion Chromatography Systems
• Leading technology and market presence, especially in environmental and food safety
Liquid Chromatography Systems
• UHPLC-ready across entire platform, comprehensive columns offering
Chromatography Data Systems
• Gold standard performance and complementary to our leading LIMS
Complements Our Leading Mass Spectrometry Offering
• Deep applications expertise for compelling, high productivity, integrated solutions
Leading technology and performance across entire portfolio
4
Highest Performance LC Portfolio on the Market
For all applications & configurations, standalone HPLC to MS front end
Transcend UHPLC Solutions For multiplexed workflows
UHPLC Systems 1000 to 1250 Bar
Standard Systems Range of detectors
Dual LC Systems Two systems in one
Nano LC Solutions Flexibility and ease-of-use
5
• Proteomics • Biomarker Discovery • Metabolism • Metabolomics • Lipidomics • PTM Analysis
• Proteomics • Metabolomics • Bioanalysis • Clinical / Toxicology • Environmental • Food Safety
The Industry’s Leading Portfolio of MS Solutions
Triple Quads
Quantitative Qualitative
Non-targeted Analysis
Targeted Analysis Ion Traps
Hybrid Orbitraps ESI-Orbitraps
High Resolution Accurate Mass
High Resolution Accurate Mass Fast Scanning
Fast Scanning Fast Scanning MS and MSn
Research Research Applied Markets Applied Markets
6
2000 2003 2005 2008 1999 2011
Dallas Proof-of-the-principle for the Orbitrap mass analyzer
Denver Exactive benchtop LC-Orbitrap launch
Long Beach Resolving power >100,000 demonstrated on concept instrument
Orbitrap: Continually Extending MS Performance Chicago Orbitrap with external accumulation developed LTQ launch
San Antonio First Commercial LTQ Orbitrap launch
Philadelphia LTQ Orbitrap Velos
2009
Denver Orbitrap Velos Pro Orbitrap Elite Q Exactive
Original 1999 Orbitrap patent US 5,886,346
ASMS 2011: The biggest launches in Orbitrap history
7
Orbitrap: Unmatched Resolution vs. TOFs 400,000
300,000
200,000
100,000
FWHM
2000 m/z 1000 m/z 200 m/z
C B A
Protein + Peptide Fragments Peptides Drugs Thermo S C I E N T I F I C
Routine Orbitrap Resolution
8
• Metabolite profiling, peptide sequencing, toxicology
• Sensitivity equal to high performance triple quads
• Identify, quantify and confirm in one analysis
Strengthening Our Leading Position in Mass Spectrometry
Diverse Technology Introductions
Extending Ion Trap Leadership: Velos Pro
OT Elite Quantum Leap in Orbitrap Performance
Revolutionary Q Exactive The “Quanfirmation” Instrument
June 2011 ASMS Launches
• Metabolite ID, peptide sequencing
• Fastest and most sensitive ion trap on market
• Ideal hybrid front-end • New fragmentation capability
• Proteomics, detailed structural analysis
• High field Orbitrap most significant advance since first Orbitrap
• Resolution and scan speed 4X current Orbitrap
9
Velos Pro: Raising the Bar in Ion Trap Performance
• Fastest: first ion trap with 66,000 amu/sec. scan speed, 2X improvement
• New high dynamic range detection system dramatically enhances peptide/small molecule quantitation
• New Generation II ion optics for improved robustness
• Unique Trap HCD fragmentation, gives triple quad like fragments for improved molecule structural identification
Ultimate in ion trap speed, sensitivity and flexibility
The Fastest, Most Sensitive Ion Trap in the World
10
Orbitrap Elite: The New Performance Standard
• New 7 Hz HCD capability • New secondary detection system for
enhanced dynamic range • Multiple fragmentation techniques for
comprehensive peptide and protein ID; unambiguous small molecule identification
• Ultimate in top down protein analysis A quantum leap in ultra high resolution and accurate mass analysis
Unprecedented mass resolution
Technology Breakthroughs: New High Field Orbitrap Resolving power of >240,000 4X scan speed improvement Advanced signal processing
11
Benefits of New High Field Orbitrap Analyzer
Ultra high resolution and fragmentation flexibility for complex analysis
High-field Orbitrap
+
• Smaller size delivers 2X improved resolution and scan speed • Advanced signal processing gives 2X improved resolution and scan speed • With multiple fragmentation capabilities, significantly enhanced performance
12
IgG Analysis: Orbitrap Elite
gG e5_ 0u_ 5 _ 5e _tube e s 0_ e g ad e t _ t # 60 6 6 08T: FTMS + p ESI sid=45.00 Full ms [1000.00-4000.00]
2000 2100 2200 2300 2400 2500 2600 2700 2800 2900 3000 3100 3200m/z
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Rel
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2815.46
2764.32
2712.06 2865.55
2667.31
2923.732621.38
2576.95
2981.072533.99
2492.48
2452.32 3043.83
2413.38
3102.662336.652269.39
3171.552021.95 2201.192113.94 3242.36
Orbitrap Elite @15000 R
13
gG e5_ 0u_ 5 _ 5e _tube e s 0_ e g ad e t _ t # 60 6 6 08T: FTMS + p ESI sid=45.00 Full ms [1000.00-4000.00]
2700 2720 2740 2760 2780 2800 2820 2840m/z
0
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Rel
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2764.32
2812.502712.06
2818.46
2767.26
2717.81
2809.50 2821.412770.14
2758.432709.18 2720.76
2824.332772.92
2723.442827.19
2776.04 2806.652725.972706.35 2778.93 2830.23
2755.61 2781.592731.80 2833.482783.65 2837.78
2737.41 2791.22 2844.442742.66
2804.11
Orbitrap Elite: Raw Spectrum zoom-in
Glycoforms – Δm = 162 Da
14
OT Elite IgG ReSpect Deconvoluted Spectrum
15
Q Exactive: Redefining Qual/Quan Workflows
• New high performance quadrupole parent ion selection capability
• New S Lens ion optics 3X sensitivity • Targeted & non-targeted screening • 3 workflows in one analytical run • New data dependent scanning • Multiple ion filling capabilities > speed
Exceptional MS/MS sensitivity, speed and data quality
Trace-level metabolites, contaminants, peptides/proteins in complex mixtures
QuanfirmationTM: Simultaneously identify, quantify
and confirm in one analytical run Resolving power of >140,000
16
It is a simple instrument! (Makarov)
C-trap
Central electrode
Outer electrodes
HCD collision cell
Bent Flatapole
Z-lens
Quadrupole mass filter
Injection Flatapole
S-Lens assembly
Capillary Figures and design courtesy J. Rykl, Thermo Fisher Scientific, 2011
17
Exactive: Speed
Peakwidth (FWHM) = 0.8 sec Scans/peak = 21
1.75 1.80 1.85 1.90 1.95 Time (min)
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1.75 1.80 1.85 1.90 1.95 Time (min)
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Peakwidth (FWHM) = 1 sec Scans/peak = 11
Resolution Setting: 35,000 Resolution Setting: 70,000
18
RT: 0.00 - 18.00 SM: 7G
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18Time (min)
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1009.91 10.525.924.22 8.704.633.21 9.57 NL: 4.11E5
m/z= 746.4801-746.4875 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 3.05E5m/z= 371.2193-371.2231 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 1.01E7m/z= 404.0137-404.0177 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 8.83E6m/z= 202.0424-202.0444 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 3.62E5m/z= 353.2206-353.2242 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 6.24E6m/z= 253.0296-253.0322 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 1.48E6m/z= 292.0251-292.0281 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 8.15E6m/z= 409.1350-409.1390 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01
RT: 0.00 - 18.00 SM: 7G
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18Time (min)
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1007.20
3.72
13.54
10.525.57
4.662.16
1.33
7.71 9.995.79 9.52 NL: 2.41E4m/z= 301.1163-301.1193 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 3.74E6m/z= 368.1872-368.1908 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 6.99E5m/z= 210.1114-210.1136 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 3.54E6m/z= 200.1173-200.1193 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 3.12E6m/z= 308.0025-308.0055 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 6.64E6m/z= 328.1920-328.1952 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 8.89E6m/z= 202.0844-202.0864 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 1.72E6m/z= 732.4645-732.4719 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01
RT: 0.00 - 18.00 SM: 7G
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18Time (min)
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1003.72 9.577.802.57 8.002.84
9.19
NL: 9.38E6m/z= 256.0583-256.0609 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 1.60E7m/z= 528.0754-528.0806 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 3.97E6m/z= 249.0180-249.0204 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 1.96E6m/z= 226.0880-226.0902 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 6.12E6m/z= 163.0528-163.0544 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 3.91E5m/z= 369.2155-369.2191 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 1.21E5m/z= 237.1004-237.1028 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 6.12E6m/z= 163.0528-163.0544 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01
RT: 0.00 - 18.00 SM: 7G
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18Time (min)
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1006.013.14 8.007.422.57 10.406.92 NL: 1.07E7
m/z= 336.1012-336.1046 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 2.60E6m/z= 230.0524-230.0548 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 8.78E5m/z= 266.0722-266.0748 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 5.59E6m/z= 330.1084-330.1118 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 2.21E6m/z= 222.1226-222.1248 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 1.41E6m/z= 435.0463-435.0507 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 4.35E6m/z= 353.1062-353.1098 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 4.78E6m/z= 297.0541-297.0571 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01
RT: 0.00 - 18.00 SM: 7G
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18Time (min)
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1009.638.808.002.73 7.20 9.197.625.83 NL: 8.44E6
m/z= 406.0700-406.0740 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 9.34E6m/z= 311.0378-311.0410 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 5.41E6m/z= 388.1291-388.1329 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 4.38E6m/z= 203.1129-203.1149 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 1.62E7m/z= 233.0231-233.0255 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 2.79E5m/z= 392.1585-392.1625 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 1.15E7m/z= 312.1149-312.1181 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01NL: 1.02E7m/z= 320.1064-320.1096 F: FTMS + p ESI Full ms [120.00-1000.00] MS cal6_01
RT: 0.00 - 18.00 SM: 7G
0 1 2 3 4 5 6 7 8 9 10 11 12 13Time (min)
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1007.32 7.895.79 8.80 9.693.78
RT: 0.00 - 18.00 SM: 7G
0 1 2 3 4 5 6 7 8 9 10 11 12Time (min)
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1003.943.28 4.17
2.79
1
2.67 7.535.69
Q Exactive in Small Molecule Analysis Quanfirmation” of 60 Pesticides
Three workflows in a single analytical run
•UHPLC and Q Exactive compatability @70,000 resolution
•Full scan and targeted data dependent MS/MS analysis
•Screen, identify, confirm and Quantify in one run
19
RT: 0.00 - 18.00
0 5 10 15Time (min)
0
1000
1000
1000
100
Rel
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0
1000
1007.32
7.68
7.99
2.64
10.01
9.70
NL: 2.39E7m/z= 233.0231-233.0255 F: FTMS + p ESI Full ms [100.00-1000.00] MS cal5_02
NL: 6.40E6m/z= 404.1221-404.1261 F: FTMS + p ESI Full ms [100.00-1000.00] MS cal5_02
NL: 2.18E6m/z= 343.0382-343.0416 F: FTMS + p ESI Full ms [100.00-1000.00] MS cal5_02
NL: 5.99E6m/z= 163.0528-163.0544 F: FTMS + p ESI Full ms [100.00-1000.00] MS cal5_02
NL: 1.02E6m/z= 746.4801-746.4875 F: FTMS + p ESI Full ms [100.00-1000.00] MS cal5_02
NL: 5.03E6m/z= 528.0754-528.0806 F: FTMS + p ESI Full ms [100.00-1000.00] MS cal5_02
RT: 7.50 - 7.90
7.55 7.60 7.65 7.70 7.75 7.80 7.85 7.90Time (min)
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Rel
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7.69 NL: 3.88E6TIC F: FTMS + p ESI Full ms2 404.02@hcd40.00 [50.00-430.00] MS cal5_02
cal5_02 #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
10
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Rel
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329.0806
344.1041
372.0992
316.1092183.0560 301.0852216.0665134.0606 172.0399287.0824273.0899129.0454 187.0509107.0499 242.032682.0298 226.8040 401.816871.2217 386.1146 420.7813
Azoxystrobin MH+ = 404.1242
Quanfirmation of Azoxystrobin 1.5ppb
Boscalid
Diuron
Azoxystrobin
Methomyl
Indoxacarb
Spinosad D
Extracted Chromatograms Accurate mass Identification
Identification Triggered MS/MS Experiments
MS/MS spectrum Confirmation
20
Q Exactive Quanfirmation– Azoxystrobin 1.5ppb
Quantitation Comparable to Best Triple Quads
21
More Possibilities, Powered by Orbitrap
Exactive • Mass range m/z 50-4000 • Mass accuracy: <2ppm • Max resolution: 100,000
LTQ Orbitrap XL • Mass range m/z 200-4000 • Mass accuracy <1ppm • Mass Resolution >100,000
Q-Exactive • Mass range m/z 50-4000 • Mass accuracy: <2ppm • Max resolution: 140,000
Orbitrap Velos Pro • Mass range m/z 200-4000 • Mass accuracy <1ppm • Mass Resolution >100,000
Orbitrap Elite • Mass range m/z 200-4000 • Mass accuracy <1ppm • Mass Resolution
>240,000
Perf
orm
ance
Screening Research Affordable Flexibility
22
New Software to Support Workflows
New leading edge software for challenging MS workflows
MetQuest 1.0 • For HR/AM qual/quan metabolic screening • Unbiased full-scan data reveals unexpected metabolites
Tracefinder 1.1 • Automate environmental, food safety & clinical workflows • Supports Triple Quad, Ion Trap and Exactive experiments
Xcalibur 2.1 • Core software control for all Thermo Scientific MS systems • Now supports TLX/multiplexing and Turboflow automation
Exactfinder 1.0 • Integrated HR/AM qual/quan workflow analysis • Targeted and non targeted screening
23
Novel MS Technologies for Proteins: Applications Growth Drivers
Advances in Hardware, Software and Reagents Move Applications to Higher Throughput
Technology improves – applications growth follows
Quantitative Qualitative Quantitative /Multiplexed
Structural accuracy at high sensitivity results in complete protein sequence determination including PTMs
High sensitivity, accuracy, precision multiple protein quantitation in biological media
High sensitivity, accuracy, precision multiple protein quantitation in biological media – many experiments in one run
Hardware: VelosPro OrbitrapElite
Hardware: TSQVantage QExactive
Hardware: VelosPro ObitrapElite
Software Software Software
Reagents Reagents Reagents
24
New Proteomics Tools
Proteome Discoverer 1.3 • Comprehensive mass informatics platform for data mining • Integrate biological information from multiple sources
SIEVE 2.0 • For label-free, semiquantitative, differential expression • Expanded to support metabolomic & lipodomic workflows
Pinpoint 1.2 • Enhances speed and confidence in targeted quantitation. • Supports multiple experimental approaches including HRAM
Thermo Scientific Pierce Enrichment Kits • Enrich, capture and identify proteins, peptides, kinases • Peptide Calibration kits and libraries, ActivX kinase probes
Easy-nLC 1000 • Split-free, nano-flow LC optimized for biomolecule separation • Effortless ultra high pressure operation (1000 bar, 15,000 psi)
Leading edge tools for proteomics research
25
Summary
• Dionex acquisition: comprehensive HPLC portfolio complements leading MS technologies
• Velos Pro Ion Trap: leadership extended with high speed quantitative capabilities
• High field Orbitrap Elite: with >240,000 resolution, the most significant advancement since first Orbitrap
• Q Exactive: Revolutionary “Quanfirmation:” screen, identify, confirm and quantify in one run
• Workflow based software: comprehensive new suites for all applications