Extending the limits of high resolution quantitative and qualitative analysis · 2013-07-29 ·...

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Extending the limits of high resolution quantitative and qualitative analysis AB SCIEX TRIPLETOF 5600 + SYSTEM AB SCIEX TRIPLETOF 5600 + SYSTEM

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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