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AB SCIEX TripleTOF 5600 System
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AB SCIEX TripleTOF 5600 System Introduction Technology Overview Conclusions References Instrument Sole Source Specifications Proteomics Applications In Depth Protein Identification Phosphopeptide Identification Relative Quantitation of Peptides Using High Resolution Full Scan TOF MS Protein Quantitation using Multiplexed Isobaric Tagging Reagents Targeted Peptide Quantitation using MRMHR Intact Protein Analysis Conclusions References Lipidomics Applications MS Approaches to Lipidomics Workflows on the TripleTOF System for Lipid Analysis Linking Lipid Discovery to Untargeted Lipid Relative Quantitation and
Statistical Profiling Conclusions References
Introduction A time-of-flight (TOF) mass spectrometer separates ionized molecules, with
the same kinetic energy, based upon their flight time in a vacuum. In
contrast to a quadrupole mass analyzer, the TOF mass analyzer is not a
narrow mass, selective range scanning device. This distinction confers upon
the TOF mass analyzer several advantages. The primary advantages are
increased sensitivity and scanning speed.[1],[2],[3],[4] With recent
improvements in instrument design, TOF mass analyzers are now capable of
mass resolution exceeding >25,000 in both MS and MS/MS modes with mass
accuracy of better than 2 parts per million.2 It is this combination of
features that makes TOF mass spectrometry an ideal approach for the
identification and quantitation of numerous analytes, including
biomolecules and small molecule metabolites. This overview will demonstrate
the capabilities and advantages of a new QqTOF instrument, the TripleTOF
5600 System for qualitative/quantitative workflows carried out
simultaneously for modern proteomics research. The capabilities,
accuracy, and ruggedness of these workflows for the quantitation of
peptides in biological samples will be discussed. For reference purposes,
the use of electrospray ionization (ESI) TOF mass spectrometry will be
compared to an ESI-triple quadrupole mass spectrometer for the accurate
quantitation of such biological targets in complex samples, as previously
introduced by Stolker et al, 2004 in application to the detection and
quantification of small drug compounds.[5] Modern quadrupole-hybrid time-of-flight instruments are designed to consist
of a first quadrupole that can focus all ions (in RF-only mode) or use a
combination of DC and RF to transmit selected ions only into the second
quadrupole (collision cell). Ions transiting the collision cell will
either be fragmented by collision induced dissociation (CID) or will be
transferred without fragmentation into the time-of-flight region for
detection (Chernushevich et al, 1997 & 2001).[6],[7] The benefits of
detecting these ions using a time of flight analyzer are many; Full scan
spectra can be acquired at fast acquisition rate due to the high
sensitivity of the mass analyzer and the resulting full scan MS or MS/MS
spectra have high mass resolution and high mass accuracy. Powerful
information dependant acquisition (IDA) strategies are possible on QqTOF
instruments that enable fast, in-depth qualitative analysis of complex
samples for peptide and protein identification.[8] In addition, while
traditionally considered to be primarily of qualitative importance, new
detector improvements on the TripleTOF 5600 system has moved QqTOF
technology into more quantitative applications with sufficient sensitivity
and dynamic range. [pic] Figure 1. TripleTOF 5600 System Ion Path consisting of the QJET® Ion Guide,
Q0 High Pressure Cell, Q1 resolving quadrupole, and the LINAC® Collision
Cell (5500 platform technology) and unique Accelerator TOF Analyzer
technology featuring high acceleration voltage and a 40 GHz Multichannel
TDC detector.
Technology Overview The TripleTOF System is considered the first accurate mass, high resolution
system of its kind, designed to provide high mass accuracy, high spectral
peak resolution, high sensitivity, and wide dynamic range for quantitation
capabilities similar to a triple quadrupole mass spectrometer. Improved
front-end ion sampling using the QJet® Ion Guide,[9] and accelerated
transmission of the ions through the time-of-flight region can provide
sensitivity at high acquisition speed. Combined with a 40 GHz TDC
multichannel detection system, the TripleTOF 5600 System can achieve 50
high resolution MS/MS in information dependent acquisition mode with a
cycle time of 1 s for deep interrogation of complex protein digest
mixtures. Summary of TripleTOF 5600 System features: Accelerator TOF Mass Analyzer
Up to 40,000 resolution at as little as 10 milliseconds accumulation time
Greater than 25,000 resolution at m/z 100
No loss in sensitivity in full mass scanning at high resolution
Advanced TOF entrance optics enables two resolution modes
High resolution mode > 30 000 resolution
High sensitivity mode > 15 000 resolution
Two-stage reflector for high resolution
30 kHz Accelerator for increase duty cycle
15 kV TOF acceleration for improved sensitivity
40 GHz Multichannel TDC detector
High resolution compact instrument (flight path of ~ 2.5 m)
Computer controlled polarity switching Specifications:
Up to 25 Hz operation in MS mode (acquiring 25 MS scans per second
Up to 100 Hz operation in MS/MS mode (acquiring 100 MS/MS per second)
Up to 50 MS/MS scans in data dependant MS/MS mode (acquired in IDA
mode per second)
Detection System
40 GHz multichannel time-to-digital converter (TDC)
Ion signal handling as fast as every 25 ps in both positive and
negative mode
Fast sampling in the time domain provides sufficient points across the
spectral peaks to ensure high resolution and mass accuracy at low mass
[