The world leader in serving science GOH Lin-Tang, PhD Senior Manager (SEA) Latest Omics Developments based on Orbitrap Mass Spectrometry
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The world leader in serving science
GOH Lin-Tang, PhDSenior Manager (SEA)
Latest Omics Developmentsbased on Orbitrap Mass Spectrometry
2
A Historical Review of (Bio)Mass Spectrometry
3
1999: The First Orbitrap Mass Analyzer Patent
4
2000: The Foundation of a New Mass Analyzer
5
Orbitrap Mass Analyzer: Principle of Operation
)/ln(2/2
),( 222mm RrRrzkzrU
z
φHyper-logarithmic potential distribution:
“ideal Kingdon trap”
r
12
2
RRmz 2
2
RRm
zr qm
kz /
Makarov A. Anal. Chem. 2000, 72, 1156-1162.
Characteristic frequencies:• Frequency of rotation ωφ• Frequency of radial oscillations ωr• Frequency of axial oscillations ωz
6
2003: The Beginnings were Basic and «UnExciting»
Courtesy of A Makarov
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2005: The First Hybrid Orbitrap MS
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Mass Resolution: The Most Direct Approach to Deal with Complexity
70K FWHM
140K FWHM
Protonated AFB1: C17H13O6; m/z = 313.071215
9
Unparallel Discriminating Power: Midazolam Mystery
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2006: Success on the GO!
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2007: Addition of HCD to Complement Ion Trap CID
12
Problem: Quantitativeinformationaboutexpressionlevelofaproteinisessentialtounderstandingitsbiologicalroleinresponsetochangeordisease.
Qualitative Quantitative
Alterationsinexpressioncanrevealameaningfulbiologicalpatternnotapparentinapureidentificationexperiment,whichprovidesonlyalistofdetectedproteins
Addanotherdimensiontoanyexperimentbydeterminingtherelativeabundanceofeachidentifiedprotein
0
1
2
3
4
5
0 500 1000 1500
RelativeAbundance
Minutes
Changeswithtime
Changeswithtreatment
0
0.5
1
1.5
2
2.5
3
Changeswithcellline
Moving Beyond Qualitative Proteomics
13
2008: Enabling Absolute SILAC Experiments
0.5 fmol 150 fmol
Experiment requires: Resolving power Mass accuracy Dynamic Range Parallel MS/MS Scan Speed Sensitivity
Pure FormBackground of Total Cell Lysate
S. Hanke, H. Besir, D. Oesterhelt, M. Mann J. Proteome Res. 2008, 7 (3), 1118–1130.
Linearity of Quantitation of a Protein
45,000 fmol
A B
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2008: Introduction of ETD on Hybrid Orbitrap MS
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EThcD: The Best of Both Worlds
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2011: Fully Featured & Enhanced Hybrid Orbitrap (Elite)
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2011: Introduction of Quadrupole-Orbitrap MS Platform
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2011: A Superior Fully-Benchtop Orbitrap MS Platform
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QqQ MRM vs Orbitrap-based HR-MSMS (PRM)
• Serial monitoring
• Parallel monitoring
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Linearity & CV of PRM
R² = 0.9994
R² = 0.9992
Inte
grat
ed In
tens
ity
Peptide Concentration (fmol)
1E10
1E9
1E8
1E7
1E6
1E5
1E40.005 0.01 0.05 0.1 0.5 1 5 10 50 100
GISNEGQNASIK (2 amu) 2.19% 9.70%
GISNEGQNASIK (0.7 amu) 2.09% 2.03%
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2014: Recent HRMS Comparison Study by US FDA
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Critical Parameter #1: Mass Accuracy
Detection of 48 compounds (antibiotics, toxins, pesticides, drugs etc) in various food matrices.
Within 5 ppm
Within 30 ppm
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Critical Parameter #2: Isotopic Abundance/Pattern
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Superior HRAM Attributes in Complex Matrix Analysis
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Impeccable Mass Stability at High Mass Accuracy
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Animal Feed Matrix Challenge: Orbitrap vs TOF MS
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Quantitative Comparative Study: Orbitrap MS vs QqQ
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Quantitative Comparative Study: Orbitrap MS vs QqQ
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2015: Quadrupole-Orbitrap MS Quantifies like a QqQ
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Assessment of False Negative Detection by QqQ and Q-Orbi
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Orbitrap-based Validated Method: Qual-Quan in Single Run
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US FBI Rapid Screening Method for DoB by LC_Q-Orbitrap
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US FBI Rapid Screening Method for DoB by LC_Q-Orbitrap
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US FBI Rapid Screening Method for DoB by LC_Q-Orbitrap
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Orbitrap MS for Forensic & Clinical Toxicology Screening
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Orbitrap MS for Forensic & Clinical Toxicology Screening
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Doping Control Screening based on Orbitrap MS
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Doping Control Screening based on Orbitrap MS
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HRAM Workflow for Known Screening & Unknown AnalysisSample X
Unknown
Known
LC-HRAM MS System
Reduced putative candidate list
1. High mass accuracy precursor ion measurement2. High mass accuracy isotopic pattern/ratios
3. High mass accuracy MSn fragment data 4. High resolution MSn spectral data (known cpds)
Experimental Data Acquisitions
Known CompoundLibraries / Databases
Target candidate list
Elemental composition
analysisList of molecular formula
Reduced list of molecular formula
Software prediction of molecular structures
Structure elucidation
IP matching
Unknown CompoundAnalysis
TraceFinder
Compound Discoverer
40
Library Searching for “Unknown” Samples – mzCloud™
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Regularly Updated mzCloud™ & Statistics Report
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Visit PlanetOrbitrap.com
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