Extending the limits of high resolution quantitative and qualitative analysis · 2013-07-29 ·...
Transcript of Extending the limits of high resolution quantitative and qualitative analysis · 2013-07-29 ·...
Extending the limits of high resolution quantitative and qualitative analysis
AB SCIEX TRIPLETOF™ 5600+ SYSTEM
AB SCIEX TRIPLETOF™ 5600+ SYSTEM
AB SCIEX has been a leader in the field of mass
spectrometry for more than 25 years. With each
generation of our system solutions, we have
raised the bar for quantitative and qualitative analysis
in pharmaceutical drug development, life science
research, food and environmental safety, forensic
toxicology, and clinical research.
The original TripleTOF™ 5600 system was the first
system to combine comprehensive qualitative
exploration, rapid profiling, and high-resolution
quantitation on a single accurate mass platform.
The next generation, TripleTOF™ 5600+ system adds
innovative acquisition strategies, tools for regulatory
compliance, and compatibility with ion mobility
separation to enable research that was
previously impossible.
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
The AB SCIEX TripleTOF 5600+ extends the market-leading capabilities of the TripleTOF family,
by providing broader applicability in the pharmaceutical industry and enabling a new paradigm in
proteomics research. No other accurate mass MS/MS instrument has the speed and sensitivity to
deliver comprehensive qualitative exploration, rapid profiling, and high-resolution quantitation
in complex matrices – all in a single injection, on a single platform.
The AB SCIEX TripleTOF™ 5600+ SystemHigh resolution quantitative and qualitative analysis
For the first time, pharma, academic and analytical testing scientists can integrate comprehensive
qualitative exploration, rapid profiling, and high-resolution quantitation workflows on a single
platform providing faster and more accurate answers to “what is in the sample, how much is there,
and does it change?”
One system, flexible workflows
Speed and resolution for definitive identification
Select up to 100 precursors and get high resolution MS/MS in a second for fast LC conditions
Identify precursors with confidence with high mass accuracy in MS and MS/MS mode
Target analyte classes with advanced IDA (information-dependent acquisition) workflows
> Multiple mass defect scanning
> Neutral loss scanning
> Exclusion / inclusion lists / isotope pattern matching
Identify unknowns with robust, intelligent multiple collision energy workflows
EXPLORE
Explore complex samples in greater depth:
Select up to 100 precursors – and generate high
resolution and high mass accuracy MS/MS spectra in
a second using powerful IDA (information-dependent
acquisition) algorithms and high-resolution, accurate-
mass MS and MS/MS.
Explore Profile Quantify
QUALITATIVE ANALYSIS
QUANTITATIVEANALYSIS
LC High resolution MS High resolution MS/MS
Quantitative and qualitative information in a single run
Combine high-resolution MS quantification with high-resolution MS/MS ID confirmation
Quantify every compound—even under fast LC conditions
Re-visit data for further interrogation
Powerful new MS/MSALL workflow with SWATH™ Acquisition provides additional
specificity and robustness to the profile workflow
Relative and absolute quantitation with high resolution Scheduled MRMHR
Maximize specificity with MRM quantitation using high resolution fragment ions
Multiplex functions for many analytes with triple quadrupole MRM acquisition rates and collect full scan MS/MS spectra for each precursor with no additional cycle time
Maintain fast cycle time for superior peak definition, even in fast LC separations
Linear dynamic range of 4 orders or greater for quantitative accuracy
PROFILE
Profile samples for quant and qual in a single run:
Quantify virtually every compound in the sample
with high-resolution MS quantification and fast
chromatography; confirm compound identity with high-
resolution, accurate-mass MS/MS; then mine the data
for the information you need.
QUANTIFY
Quantify analytes with high resolution MRM for highest
specificity and confidence: Obtain high-resolution at
high mass and low mass in the same spectrum for
high-resolution MRM quantification. LOQ’s and dynamic
range are equivalent to high-performance triple quad
instruments. Cycle times as low as 10 msec generate
superior peak definition even with fast LC separations.
Quantification requires little optimization, and you
can choose multiple fragments per precursor from the
acquired full scan MS/MS data.
LC
Multiplexed high resolution MRM
MS Quantification
High resolution, accurate mass MS/MS
High resolution, accurate mass quantification
High resolution, accurate mass MS/MS
High resolution, accurate mass MS
Accelerator TOF™ Analyzerimproves resolution and sensitivity
15kV acceleration voltage for higher sensitivity and
resolution
High transparency grids throughout for minimal ion loss
Two-stage ion reflector compensates for energy dispersion
to maximize resolution
40 GHz four channel TDC and detector provide highest sampling
speeds and maintains high resolution – even at
low mass
30kHz accelerator for highest acquisition rate
New entrance optics improves ion beam focusing post LINAC
Innovative technologyBeginning with the best-in-class quantitative performance of the Triple Quad™ 5500 system,
AB SCIEX researchers developed the AcceleratorTOF™ Analyzer to deliver high resolution at the
speed and sensitivity required to maintain the precision and limits-of-quantitation associated
with MRM. The result is the breakthrough AB SCIEX TripleTOF™ 5600+ System —the next generation
in quadrupole TOF technology.
Intelligent implementationImprove productivity and specificity by acquiring information
instead of just data.
Dynamic background subtraction (DBS) minimizes collection
of MS/MS on background ions to increase identification of
low-level analytes in the presence of background noise.
Multiple mass defect triggered IDA (Information Dependent
Acquisition) improves efficiency and reduces the need for
multiple injections to get comprehensive information. Mass
defect can be applied for specific mass ranges, and multiple
mass defects can be applied.
MS/MSALL is a powerful workflow enabled by the speed
and sensitivity of the Triple TOF™ 5600+ system, where high
quality MS/MS spectra are generated for all masses. This is the
foundation of the ground-breaking SWATH™ Acquisition that
is changing the landscape of quantitative proteomics.
Ion mobility enabled, for a new dimension in selectivity
On the TripleTOF 5600+, SelexION™ technology adds a new
dimension to accurate mass selectivity for enhanced quantitative
and qualitative performance.
AB SCIEX SelexION Technology compliments and extends the power
of accurate mass and the Scheduled MRMHR workflow, by removing
interferences at the structural level.
The 5600+ system with SelexION technology is the ultimate
accurate mass tool for the separation of isobaric species, isolation
of challenging co-eluting contaminants and reduction of high
background noise, improving data quality and enhancing selectivity
for challenging samples requiring advanced analytical separations.
Reserpine
Inte
nsi
ty,
cps
m/z, Da
R=29,950 R=34,600
Performance delivered
High-resolution, high-sensitivity data with acquisition speed up to 100 spectra/sec
makes the TripleTOF 5600 system a perfect compliment to ultra-fast chromatography.
The system also provides 4-5 orders of magnitude of linearity to ensure accurate
quantitation in MS and MS/MS modes.
Reserpine and fragments demonstrate high resolution at low mass, ~30,000 at 100 MS/MS per second. Resolution of insulin (6+) is maintained at >43,000 with data
accumulation times of 1 second, 100 milliseconds, and 10 milliseconds.
EasyMass™ Accuracy delivers high mass accuracy without continuous recalibration.
With external calibration, the mass accuracy holds rock-steady at <1ppm over 30
minutes, and RMS=1.69 over 100 hours.
Time (Mins)
Ma
ss A
ccu
racy
(p
pm
)
Log Concentration Ratio
Log
Are
a R
ati
o
Scientists have come to expect industry-leading performance from AB SCIEX mass spectrometers.
The TripleTOF™ 5600+ system delivers with high-resolution, high-sensitivity data, excellent mass
accuracy stability, and fast acquisition rates. If you’re looking for unmatched performance, look
no further than the TripleTOF 5600+ system.
R=36,500
Mass/charge, amu
Inte
nsi
ty,
cps
1 s acquisitionR=43,750
100 ms acquisition
R=44,020
10 ms acquisitionR=44,000
Insulin
Applied powerIn-depth identification and expression analysis
for omics and biomarker studies.
A new standard for protein identification and targeted
quantitation: The TripleTOF™ 5600+ system achieves the highest
acquisition speeds while maintaining maximum resolution and
mass accuracy for unmatched protein identification. The resolution
and speed also enable MRM-like quantitation without significant
method development.
Comparative analysis for metabolomics studies can be
performed on the TripleTOF 5600+ system with greater precision
and confidence than ever before. Profiling of endogenous
metabolites can be achieved with unmatched speed and sensitivity
in a single injection to determine statistical differences between
samples for the rapid determination of putative biomarkers.
Novel workflows for Lipidomic analysis: Lipid profiling on the
TripleTOF 5600+ system can be used to analyze every lipid species
simultaneously. With the new MS/MSALL workflow it is possible to
acquire precursor ion data at every unit mass to obtain high quality
MS/MS data on every lipid class within a sample.
MultiQuant™ Software
Process high-resolution MS
and MS/MS data quickly and
easily for absolute and relative
quantitative information.
LipidView™ Software
Characterize lipid data by
searching parent and fragment
ion masses against a database
of thousands of entries.
MetabolitePilot™ Software
Streamline data analysis for
metabolite identification
with tools for easy structural
elucidation and relative
quantitation of metabolites.
ProteinPilot™ Software
Get more identifications and
quantification information out
of every proteomic sample
using the powerful algorithms
within ProteinPilot Software.
One Touch Productivity
Simple, clear metabolite ID, characterization,
and powerful quantitation for bioanalysis.
Identify and characterize metabolites at new levels of speed,
efficiency and data quality: The TripleTOF 5600+ system is the
ideal platform for rapid assessment of metabolic stability and
in vivo characterization and definitive metabolite ID. With the
ability to perform both TOF MS and MS/MS scanning at rates
suitable for fast LC, you can apply completely generic methodology
to acquire high-quality qualitative and quantitative data
simultaneously.
Quantify drug metabolites with performance equivalent to
MRM on a triple quad. The TripleTOF 5600+ system is the first
accurate-mass, high-resolution MS system with the speed and
sensitivity to deliver triple quad-like quantitation.
Accurate mass for regulated bioanalysis: The TripleTOF 5600+
has a full suite of tools for to support 21 CFR Part 11 compliance
and and uses the Analyst® software platform, recognized as the
industry standard for quantitative LC/MS/MS. With Scheduled
MRMHR, the selectivity of accurate mass is merged with the added
utility of sensitive full-scan data in your quantitative workflows.
Time
m/z
MS/MSALL with SWATH™ AcquisitionMS/MSALL with SWATH™ Acquisition is changing the way scientists
work in quantitative proteomics. Driven by the superior speed and
high-resolution performance of the TripleTOF™ 5600+ system in
both MS and MS/MS mode, this technique provides comprehensive
quantitative MS/MS data on every component in a sample in a
single injection.
SWATH Acquisition requires no sample-specific method
development, yields better dynamic range than methods that
derive quantitation from MS (precursor) data, and delivers limits
of detection that were previously only attainable using targeted
MRM strategies on triple quadrupole systems.
Finally, one of the greatest benefits of using MS/MSALL with
SWATH Acquisition is that for every sample analyzed, the technique
creates a complete digital archive of all possible fragment ions in
that sample. As your research evolves and new hypotheses are
formed, you can re-interrogate a sample without the need to
re-acquire data.
Differential protein expression across samples
DATA ACQUISITION PROTEIN IDENTIFICATION MS PEAK EXTRACTION STATISTICAL ANALYSIS
Inte
nsi
ty
m/z
CID
Time, min
High Resolution XICs
Eksigent NanoLC-Ultra® with cHiPLC®
The most precise quantitation requires the most reproducible nanoflow separations
Quantitative results using MS/MSALL with SWATH™ Acquisition
4 orders of dynamic range
Low detection levels
LCMS reproducibility is a key component of targeted quantitation experiments such
as MS/MSALL with SWATH™ Acquisition. Eksigent nano LC systems using the cHiPLC®
provide high quality, reproducible separations, as illustrated by these 10 replicate
injections of depleted plasma acquired using SWATH Acquisition.
Selected Peptides Abundances
SSAAGNTVIIGGGDATAVAKYCR012W3-phosphoglycerate kinase314 024 copies per cell (log2=18.2)
HLNDQPNADIVTIGDKYBR039WGamma subunit of the F1 sectorof mitochondrial F1F0 ATP synthase28,103 copies (log2=14.8)
NQQLEEDLEESDTKYNL079CMajor isoform of tropomysosin3,000 copies per cell (log2=11.6)
LNTGHEILQIQPRYER024WCarnitine acetyltransferase319 copies per cell (log2=8.3)
Sensitivity and Dynamic Range Equivalent to MRM Assays
18
16
14
12
10
8
6
Cop
ies
per
cell
(log2
)
0 100 200 300 400 500 600
100’000’s
10’000’s
1’000’s
100’s
Time, min
Inte
nsi
ty,
cps
For Research Use Only. Not for use in diagnostic procedures.
© 2012 AB SCIEX. The trademarks mentioned herein are the property of AB Sciex Pte. Ltd. or their respective owners. AB SCIEX™ is being used under license.
5261412-01 05/2012
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