HCT plus.indd
Transcription
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.