HCT plus.indd

The next-generation High Capacity Trap
Enabling Life Science Tools Based on Mass Spectrometry™
High-information content LC-MSn
from your biological samples
Aiming for high productivity and success in LC-MS, your requirements are:
n
• Maximum LC-MS information from your samples.
• Ultrasensitive measurements.
• Smooth data handling and mining.
• Intelligent data interpretation and visualization.
Protein Identification
Protein Characterization
De-novo Sequencing
Differential Proteomics
Metabolite Profiling
Performed by the new
High Capacity Trap
The HCTplus LC-MSn system perfectly answers these needs. Improved HCT (High Capacity
Ion Trap) technology, in combination with a new detector design, offers dramatically
improved performance for proteomics, metabolite profiling and other demanding
LC-MSn applications:
• Ultra-fast acquisition of MS and MS/MS data across the fastest of HPLC
peaks in real-time LC-MSn.
• Low attomole/femtogram sensitivity.
• Superb quality of single mass spectra in combination with
unique scan modes.
Sensitivity
• High in-scan dynamic range for identification of low
abundance compounds in biological samples.
All embedded in a software environment tailored
for high productivity.
Speed
Spectral Information
Dynamic Range
You can have it all at once
The HCTplus provides unique
performance concerning speed,
sensitivity, spectral information
and dynamic range.
ultimate
speed
New, fast scan rates enable real-time
LC-MSn across the fastest of HPLC peaks.
The breakthrough UltraScan™ mode provides full scan MS and
MS/MS spectra at a speed of 26,000 amu/sec with better than
unit resolution.
The Fast Enhanced Mode allows reliable charge state
recognition of charges up to 3+, with a mass accuracy better
than any quadrupole ion trap.
PeptideScan™ combines these two scan modes for fast
peptide separations. Full scan MS are acquired in the Fast
Enhanced scan, while AutoMS/MS scans are performed
using the faster UltraScan.
This ultra-fast data acquisition delivers more
productivity for your application:
• High-quality data even from trace components,
as more MS/MS spectra are averaged across an LC
peak.
• On-line, real-time LC-MSn from complex
samples, as there is enough time for full scan MS
and data-dependent MS/MS spectra acquisition
within a single LC-MS run.
10 MS and MS/MS
per second
3.35
3.40
3.45
4
6
8
2626.85
1500
2000
2500
10
12
14Time [min]
MS (8.67 min), #846
721.85
MS/MS (878.6), #847
586.06
516.17
1002.54
357.90 659.41
1172.49
558.21
1512.52
887.41
1285.58
458.90 736.56
1000
Time [min]
TIC
878.55
500
3.55
LC peak widths just 3 sec
2
1399.44
3.50
High acquisition speed
With real-life LC-MS acquisition settings, the UltraScan
mode generates about 100 scans across a typical LC
peak (10 seconds width).
400
600
837.17
780.92 925.79 1055.18
800
1000
1412.55
1200
1400
m/z
m/z
MS/MS (721.9), #848
1067.41
615.88
375.89
741.17
892.39
480.06
500
1230.32
1500
2000
m/z
873.48
1080.48
333.00
500
1000
1500
700 01
700.01
2000
2500
m/z
MS/MS (586.1), #850
783.20
357.66
985.25
550.87
869.39 1181.15
229.57
500
2500
MS/MS (707.1), #849
760.51
604.51
2712.04
1401.48
1000
1000
2438.25
1500
2000
2500
m/z
Protein identification in 15 minutes
Using capillary LC on a monolithic column, this mixture of
13 digested proteins was separated within 15 minutes.
While each peptide peak is only a few seconds wide, the
HCTplus provides a full m/z scan plus 4 data-dependent
MS/MS scans within 2.8 seconds – yielding enough information for a safe identification of all 13 proteins.
supreme
sensitivity
Substantially improved ion storage capacity provides
highest sensitivity for protein and metabolite research.
The new trap geometry of the HCTplus enables a 10-fold higher ion storage capacity
over older trap designs – providing the highest sensitivity of all ion traps in both full
scan MS and MS/MS.
This sensitivity directly translates into
• More certainty in protein identification and characterization through enhanced
sequence coverage.
• Discovery of low abundance peptides and proteins, like phosphopeptides.
• More confidence in identification of trace metabolites.
• Better quantitation results.
Moreover, the supreme sensitivity in full scan MS mode is the prerequisite
for on-the-fly data-dependent acquisition of LC-MS/MS data even on
trace compounds.
Enolase (50 amol)
Full scan MS - 250 fg Alprazolam
EIC trace 309
b
y
I
E
G
S
T
L
L
T
S
G
I
K
Q
813.39
MS/MS (644.9)
726.430
8.0
9.0
10.0
Time [min]
1270.51
Sub-picogram sensitivity for metabolites
The HCTplus brings full-scan ion trap sensitivities
for small molecules into the femtogram level for
the first time.
512.33
834.42
613.12
347.23 455.13 564.17 676.21
342.12 476.13
947.53
777.33
300
2
4
6
8
10
500
600
700
800
900
1000
1142.37
1100
1200
1300 m/z
Protein identification from 50 attomole
The HCTplus gives you enhanced sequence
coverage for protein database searches from
biological samples down to the sub-femtomole
range. Here, a 50 amol Enolase sample still
yielded an unambiguous MOWSE score, with a
sequence coverage of more than 10%.
MS/MS - 50fg Alprazolam
EIC 274; 281 m/z ± MS2 (309m/z)
0
400
1075.40
12 Time [min]
excellent
spectral information
Unrivaled quality of MS and MS/MS spectra – in terms of mass
resolution, sensitivity and accuracy throughout a wide mass range.
• Superior mass resolution at high scan speed and in reproducible single scans.
• Best mass accuracy of all ion traps for fast protein database searches and successful de novo sequencing.
• Superior mass range: The standard mass range reaches up to m/z 3000 for both MS and MS/MS,
which is a very desirable feature for proteomics. The extended mass range up to m/z 6000 is
essential for AP-MALDI applications.
• Unparalleled MSn capabilities for detailed structural investigations: Manual MSn up
to MS11, AutoMSn up to MS5.
(M-4H)4-
334.1
423.4
424,4
335.1
334.0
423.0
335.0
(M-5H)5-
424.0
425.0
(M-3H)3572,1
274.5
573,1
275.5
274.2
275.0
200
300
(M-2H)2-
572.0 573.0 574.0
(M-6H)6-
870,8
400
500
600
700
800
m/z
Reproducible mass resolution from single full scan: Determination of
negative multiply charge states of a textile dye containing sulfonic groups
Standard mass range up to 3000 m/z,
with superior resolution throughout the
whole mass range: MS/MS spectrum of
Insulin chain B
Mass error <0.09 Da
MS/MS (813.5)
High certainty for protein identification
1 fmol enolase digest separated by nanoLC resulted in 14 identified
peptides, with deviation of measured from expected masses less than
0.09 Da for all peptides.
broad
dynamic range
Intelligent acquisition controls generate high quality
spectra from both the highest and lowest level compounds.
SmartICC*, our new, patented dynamic ion charge control, ensures high trapping
efficiency in any situation: in full scan MS and data-dependent MS/MS, with
high and low-level compounds, and with any mass distribution. An intensive
ion beam from the orthogonal ESI source keeps the accumulation time low;
no time is wasted by pre-scans.
This leads to a superior in-scan dynamic range of about 103, enabling
the simultaneous measurement of major and minor components in
real-life biological samples.
* U.S. Patent No. 6,600,154
Albumin
Enolase (pig)
BSA
Detection of a low level protein in the presence of a
high level protein: Database search result of a mixture of
5 fmol enolase in presence of 1 pmol BSA: Both proteins are
easily identified.
Area
15000000
10000000
5000000
Area
0
Area/106
0
2
4
6
8
10
12
14
16
18
20
750000
300
500000
250
250000
200
150
0
100
y = 0,5733x + 1,2639
R2 = 0,999367
50
0
0
50
100
150
200
250
300
350
400
450
500
Conc. [pg/µl]
0
0,2
0,4
0,6
0,8
1
Conc. [pg/µl]
Quantitation of Reserpine provides a linear regression with
excellent fit over more than 4 decades (100 fg-2500 pg abs.)
Compass
TM
Unified software environment for all
Bruker Daltonics life science instruments.
The HCTplus features Compass,
Bruker Daltonics‘ new unified software environment for all life science
instruments. Compass provides full
system control and data handling by an
intuitive and easy-to-use graphical user
interface (GUI).
Compatibility & Integration
Full compatibility to other Daltonics
instruments like the micrOTOF
ESI-TOF includes similar method setup
and data processing.
Compass integrates the HCTplus with
all popular HPLC and nanoLC systems
to match your separation needs.
A broad selection of UV detectors,
DAD and ELSD is supported
to provide additional spectral
information.
Compass environment for the esquire and HCT ion trap series
Integrates the sucessful modules of the esquire software suite with a new, global method
management and an additional module for work in regulatory compliance.
Regulatory Compliance
Compass Security Pack™ provides all necessary functions for
work in compliance with FDA and EU regulations (21CFR part
11/Annex 11):
• electronic signatures
• audit trailing
• user management
• result history
• system protection during breaks
From sample
Method Setup
The LC-MS sample table integrates all information about
samples and methods. Advanced LC-MS experiments
like UV-and/or mass-directed fraction collection,
multidimensional chromatography or column switching
are supported.
Data Acquisition
The intuitive esquireControl™ gives quick and easy
method setup, featuring unique SmartSuite™ controls
for automated system optimization and increased information from precious samples.
Data Processing
The DataAnalysis™ package stands for smart ways of
interactive and automated LC-MSn data browsing and
mining – including applications-specific automation
and customization of tasks.
Sophisticated Data Interpretation Tools
Powerful applications-specific modules enable automated and interactive data interpretation:
• MetaboliteTools™ software for identification of drug
metabolites. Also applicable for impurity analysis and
QC applications.
• BioTools™/RapiDeNovo™ software for protein
data interpretation, including database searches,
characterization (PTM and other mutations) and de
novo sequencing.
• ProteinScape™ database system for proteome project
management, including automated feedback experiments
via WARP™ and WARP-LC™.
• QuantAnalysis™ - advanced quantitation module.
• LibraryEditor™ and LibrarySearch™ modules for work
with MS and MSn libraries, including user-defined and
NIST libraries. Intelligent filtering of search results
provided.
to knowledge
Comprehensive proteomics platform
The PROTEINEER environment of sample preparation
systems, advanced mass spectrometry and bioinformatics
embeds the HCTplus into an automated proteomics workflow.
Various scenarios are supported to reflect your individual
needs concerning analytical strategy, throughput, and
depth of information.
The HCTplus is seamlessly integrated in various analytical scenarios for protein
identification, characterization (including mutation analysis and de novo sequencing) and quantitation.
Database System for Proteome Project Management
Bruker Daltonics‘ powerful ProteinScape database solution
serves not only for data archiving and management but also
for PROTEINEER workflow control WARP.
ProteinScape‘s built-in GelViewer provides direct
links from gel spots to related data like MS spectra
and search results.
metabolitetools
Complete metabolite ID
and prediction package
Sample
Powerful MetaboliteTools software puts
all of the HCTplus performance features at your
fingertip for metabolite work.
Based on a given drug structure, MetaboliteTools applies
biotransformation rules to predict possible metabolites, and
extracts the relevant information on existing metabolites from
LC-MS data of in-vitro and in-vivo samples.
Reference
eXpose
MetaboliteTools compares sample and reference, extracting the
relevant LC-MS information.
MetaboliteTools supports various analytical scenarios from automated high-throughput
screening approaches to detailed structural confirmation and elucidation.
Technical specifications
Superior HCTplus technology
• Non-linear quadrupole high capacity trap with patented
SmartICC directing optimal ion storage.
• Unparalleled combination of resolution, scan speed, and
mass range in MS and MSn.
• Best mass accuracy of all ion traps.
• Best MS and MS/MS sensitivity of all ion traps.
• Fast Polarity Switching for on-the-fly acquisition of
positive and negative ions.
• SmartFrag™ creates reproducible MS/MS spectra for
successful MS/MS library search.
Scan speed and resolution (MS and MS/MS)
Scan Mode
Res
m/z
u/sec
UltraScan™
2+ ions
3000
26,000
Fast enhanced
3+ ions
3000
8,100
Maximum
4+ ions
3000
800
6000
27,000
3000
3000
8,100
26,000
Extended
PeptideScan™
MS
MS/MS
3+ ions
2+ ions
Scan modes
• Full scan MS and MS/MS in all scan modes for sensitive
and fast analysis of unknown compounds.
• Neutral loss scans e.g. for determination of protein
modifications.
• Multiple reaction monitoring (MRM) with MS/MS
and MS3.
• Manual MSn up to MS11 in all scan modes.
• AutoMSn tree experiments with up to 30 MS precursor
ions and 5 fragment ions per stage.
• Triple Play experiments without time consuming
“charge-state determination-scan“.
Data-dependent experiments
• All types of data-dependent experiments, including
preferred mass list for automated feedback experiments
from MetaboliteTools and PROTEINEER.
Chromatography Choices
Integrated LC-MS control provided for:
• Agilent 1100 HPLC series, Capillary LC, nanoLC
• Waters Alliance HPLC series, CapLC, PDA
• LC Packings nanoLC systems (1D and 2D)
• Gilson 215 Autosampler/Fraction Collector
• Spark Endurance Autosampler
• CTC PAL Autosampler
Source Options
• ESI, APCI, APPI
• Online/Offline NanoElectrospray
• AP/MALDI
• Smart CE-MS coupling with grounded needle
Compass™ Software Features
Integrated LC-MS control and data processing, including
HyStar™, esquireControl™, DataAnalysis™, QuantAnalysis™,
LibrarySearch™ modules
Software options:
• MetaboliteTools™ software for metabolite identification
• BioTools™/RapiDeNovo™ software for protein data
interpretation
• ProteinScape™ database system for proteome project
management
• ACD Labs MS Manager for structural interpretation and
classification
• Compass Security Pack for work in regulated environments
www.bdal.de
www.bdal.com
Bruker Daltonik GmbH
Bremen · Germany
Phone +49 (421) 22 05 - 0
Fax
+49 (421) 22 05 -104
E-Mail
[email protected]
Bruker Daltonics Inc.
Billerica, MA · USA
Phone +1 (978) 663-3660
Fax
+1 (978) 667-5993
E-Mail
[email protected]
Please ask for our worldwide sales and service contacts.