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Thermo Scientific Orbitrap Elite Hybrid Mass Spectrometer High-Field Orbitrap Creating new possibilities Ultrahigh resolution • Faster scanning Higher sensitivity • Complementary fragmentation The highest resolution, most versatile Orbitrap instrument ever The Thermo Scientific Orbitrap Elite hybrid mass spectrometer raises the bar for MS and MSn performance and versatility by combining our premier dual-pressure linear ion trap mass spectrometer with a novel high-field Orbitrap™ mass analyzer. The system delivers greater than 240,000 resolution (FWHM), increased sensitivity, high scan speeds and a larger dynamic range. High-field Orbitrap mass analyzer for Higher sensitivity for complex samples ultrahigh resolution Innovations in the pre-amplifier electronics improve sensitivity. Paired The high-field Orbitrap mass analyzer geometry and advanced signal with the ultrahigh resolution, they provide more comprehensive data processing technologies enable resolution of >240,000 FWHM, and a deeper look into complex samples. a 4x increase over previous technology. This ultrahigh resolution is especially useful when analyzing samples of high complexity Fragmentation options increase versatility and targeting analytes of low abundances in applications such as In addition to traditional collision-induced dissociation (CID), the proteomics, metabolomics and lipidomics. Orbitrap Elite mass spectrometer features higher-energy collisional dissociation (HCD) that provides superior fragmentation and Faster scanning for UHPLC compatibility higher-quality MS/MS spectra for many types of samples. Optional The high-field Orbitrap geometry and advanced signal processing also electron transfer dissociation (ETD) preserves labile post-translational provide an up to 4x improvement in scan speed, making the Orbitrap modifications of proteins. Intelligent data-dependent decision tree Elite system more compatible with narrow chromatographic peaks (DDDT) logic can combine multiple fragmentation techniques in a from UHPLC separations. single analysis to significantly improve proteome coverage. Electrospray Ion Source Square Quadrupole with Neutral High-Pressure LowBlocker S-Lens Octopole Cell Pressure Cell Quadrupole Mass Filter C-Trap HCD Collision Cell Transfer Multipole Reagent Ion Source e- New Detection Electronics ETD Option New High-Field Orbitrap Mass Analyzer Reagent 1 Heated Inlet Reagent 2 Heated Inlet 3 The Orbitrap Elite mass spectrometer can analyze complex samples at a greater depth than ever before, helping scientists meet the most demanding challenges in proteomics, metabolomics, lipidomics and metabolism. A quantum leap in Orbitrap MS performance: Key features of the Orbitrap Elite system: • High-field Orbitrap mass analyzer for higher mass resolution and higher scan speed • Advanced signal processing for higher resolution • Improved ion optics for efficient ion injection into the Orbitrap • New Orbitrap image current pre-amplifier for higher sensitivity • Velos Pro linear ion trap generation II ion optics with neutral blocking technology for increased robustness A new standard of performance for comprehensive proteomics Orbitrap-based mass spectrometers are an essential tool in many proteomics labs. The enhanced power of the Orbitrap Elite instrument enables new standards of performance for multiple proteomics applications. Top-down analysis Protein ID Maximum resolution of > 240,000 FWHM and complementary Faster scanning for more data points across a peak, combined with high fragmentation modes facilitate better top-down protein identification and resolution and superior spectral quality, yields more protein identifications in characterization. a single analysis. PTM analysis Quantitative proteomics Available electron transfer dissociation (ETD) preserves labile side chains while Superior resolution and selectivity of the Orbitrap mass analyzer for both full- creating fragmentation throughout the peptide backbone for better analysis of scan MS and MSn enhance relative quantitation analyses such as those using post-translational modifications (PTMs). stable isotope labeling or isobaric mass tags. Intact protein analysis 47+ charge state of yeast enolase showing resolution of >100,000 FWHM at m/z 1,000 993.9134 R=109564 Relative Abundance [%] 80 994.0827 R=107785 993.8706 R=107582 60 50 40 993.8284 R=113310 993.7860 R=106257 994.1249 R=106063 994.1670 R=107915 30 Relative Abundance [%] 994.0401 R=107426 90 46642.586 80 993.9770 R=110221 100 70 100 60 40 20 0 46620 46650 Mass, Da Deconvoluted spectrum of yeast enolase (monoisotopic mass 46642.214 Da). 20 10 0 993.3 993.4 993.5 993.6 993.7 993.8 993.9 994.0 994.1 994.2 994.3 994.4 994.5 994.6 994.7 994.8 m/z Analysis of intact yeast enolase (46.64 kDa) on the Orbitrap Elite hybrid MS demonstrates the superior resolution that makes it the new standard for proteomics applications. 5 Orbitrap Elite LTQ Orbitrap Velos 1000 Number of Identified Proteins The superior resolution and spectral quality, as well as the higher scan speed of the Orbitrap Elite instrument, increase proteome coverage in complex samples even with very low sample amounts. Here, analysis of the E. coli proteome labeled with Thermo Scientific Pierce Tandem Mass Tags demonstrates significant improvement over the already outstanding performance of the previous-generation Thermo Scientific LTQ Orbitrap Velos hybrid MS. 800 933 873 803 713 694 600 563 400 200 0 20 ng 80 ng 200 ng Top-down analysis S G R G K G G K G L G K G G A K R H R K V L R D N I ETD Q G I T K P A I R R L A R R G G V K R I S G L I Y E E T HCD R G V L K V F L E N V I R D A V T Y T E H A K R K T V T A M D V V Y A L K R Q G R T L Y G F G G HCD Relative Abundance [%] 100 843.9602 R=158115 z=2 80 60 40 20 0 385.7177 R=197410 z=2 762.4287 R=158642 z=2 500 ETD Resolution ~ 200,000 FWHM at m/z 400 Resolution > 100,000 FWHM at m/z 1,000 80 60 600.3526 R=185866 z=2 537.9891 R=189201 z=3 40 1001.0440 R=115639 z=2 1000 m/z 100 Resolution ~ 200,000 FWHM at m/z 400 Resolution > 100,000 FWHM at m/z 1,000 664.3820 R=171040 z=2 787.4543 R=154169 z=1 1093.5867 R=125517 z=4 20 1500 2000 0 500 1000 m/z 1341.7392 R=106010 z=5 1620.8704 R=100972 z=3 1500 Analysis of histone H4 demonstrates the complementary nature of the HCD and ETD fragmentation techniques in the Orbitrap Elite instrument. Analysis using HCD and ETD in a single run significantly improved sequence coverage over each of the techniques used separately. 2000 A total solution for metabolism and metabolomics research The Orbitrap Elite system offers a maximum resolution of >240,000 FWHM that allows complete visibility into sample components and distinguishes them from complex matrix components. Improved sensitivity ensures detection of more metabolites in biological samples. High-quality MSn spectral trees provide extensive structure information for metabolite identification and location of modification sites. Improved robustness enables true 24/7 operation. Uncompromised performance • Resolve isobaric species and isotope fine structures for isotope tracing experiments • Fast scan speed and high resolution improve precision and confidence for quantitation experiments •MSn de novo structural elucidation for identification of novel metabolites on a UHPLC time scale 0.1 ppm • Lower maintenance, and higher productivity when analyzing low-level lipids and metabolites in complex biological matrices Buspirone MS Spectrum R > 250,000 386.25510 R=251601 700000 Intensity RT: 1.95 - 6.06 3.87 100 90 500000 400000 80 Relative Abundance 600000 300000 Buspirone 5 full MS scans 4 MS/MS scans 70 60 387.25830 R=252404 200000 100000 388.26111 R=213904 0 50 386.5 387.0 40 387.5 388.0 m/z 30 20 10 0 2.0 2.5 3.0 3.5 4.0 Time (min) 4.5 5.0 5.5 6.0 Even when acquiring MS and MS/MS data, the increased scan speed of the Orbitrap Elite system provides excellent coverage over narrow LC peaks. This analysis of buspirone demonstrates acquisition of full-scan data at a quarter of a million resolution at one Hz. In total, nine scans were acquired during the elution of the peak: five full MS scans and four MS/MS scans. 7 High-resolution, accurate-mass full-scan MS spectrum of an oxidative metabolite of buspirone 512.2684 1,800,000 Intensity [arbitrary units] 1,600,000 534.2498 1,400,000 150,000 0.50 ppm 100,000 402.2502 R=246701 50,000 1,200,000 0 1,000,000 400 800,000 494.2578 600,000 550.2151 464.2835 400,000 412.2851 200,000 0 200 300 400 500 600 700 800 900 1000 m/z The Orbitrap Elite system provides accurate mass determination even in complex bile matrices. Here, an oxidative metabolite of buspirone eluting in the window of several bile matrix components was still detected with an absolute mass deviation of 0.5 ppm. Total solution for Metabolism and Metabolomics Metabolism Metabolomics Thermo Scientific MetWorks and Thermo Scientific Mass Frontier software can Thermo Scientific SIEVE and Mass Frontier software utilize the ultrahigh- utilize the ultrahigh-resolution accurate-mass data from the Orbitrap Elite system resolution accurate-mass data from the Orbitrap Elite for comprehensive analysis for streamlined xenobiotic metabolite profiling and identification. The MetWorks™ software integrates acquisition, processing and reporting for metabolite identifican tion from LC/MS analyses and offers in-depth data mining tools for expected and unexpected metabolites. The Mass Frontier™ software provides chemically n of endogenous metabolites. The SIEVE™ software finds components that have reproducible differences across multiple sample groups. Optimized component detection dramatically decreases false positive identifications caused by chemical background and redundant signals. The SIEVE software searches local and intelligent annotation of MS spectral trees with automated fragment prediction. web-based databases for putative identification. Mass Frontier software provides Fragment Ion Search (FISh) in Mass Frontier software allows for automated confirmation of known compounds through interpretation of MSn spectra, as well localization of biotransformations. as de novo structural elucidation for unknown compounds. Application-specific software turning data into information Thermo Scientific Xcalibur Data System Stable operating platform Xcalibur™ software is the versatile, easy-to-use data system that controls all Thermo Scientific MS systems. The home page of the Xcalibur software offers easy navigation through the process of instrument setup, sequence setup, and data acquisition. Xtract Xtract software deconvolutes isotopically resolved data for intact protein analyses and complex MS/MS spectra acquired in top-down analyses. Thermo Scientific Proteome Discoverer Software Mass informatics platform for protein scientists Proteome Discoverer™ software is a workflow-based proteomics data processing software for in-depth data mining of complex LC-MSn data sets. With the ability to exploit data from different dissociation techniques (CID, HCD, ETD, IRMPD and ECD), Proteome Discoverer software provides extra certainty for peptide and protein identifications. Optional inclusion of multiple search algorithms increases analytical flexibility, and results can now be merged into a single report for easier interpretation. Thermo Scientific ProSightPC Software ProSightPC™ software was developed to address the specific requirements of a top-down and middle-down proteomics strategy. The fragmentation spectra of multiply charged, intact proteins are exceedingly complicated. The software processes high-resolution, accurate-mass data from the Orbitrap Elite MS to produce a list of neutral fragment masses and performs comparisons to proteome databases to identify and characterize proteins. Thermo Scientific ProteinCenter Software Revealing meaningful biological information ProteinCenter™ software is a web-based data interpretation tool that enables scientists to compare and interpret proteomic data sets and extract meaningful biological information quickly and easily. It provides access to a single database consolidated from more than 20 public databases, including all historical data. Thermo Scientific Pinpoint Software Facilitating quantitative proteomics Pinpoint™ software facilitates the transition from early-stage biomarker discovery to larger-scale, quantitative verification of putative biomarkers and general quantitative proteomics. Pinpoint software simplifies the creation of targeted quantitative assays. It allows researchers to leverage previously acquired data from discovery experiments. Pinpoint software largely automates the development of preliminary methods. It enables acquisition and analysis of preliminary data, which is in turn used to optimize the method. SimGlycan™ Software Predicting glycan and glycopeptide structure SimGlycan software analyzes mass spectrometric data to predict the structure of glycans and glycopeptides. It compares experimental MS/MS data against its own database of theoretical fragments and generates a list of probable candidate structures. Each candidate structure is scored to reflect how closely it matches the experimental data. The software also provides additional biological information for probable glycans and identified glycopeptides. Thermo Scientific MetWorks Drug Metabolism Software Simplify the interpretation of complex metabolism data MetWorks software simultaneously searches multiple modifications of one or more parent drugs and interprets simple to complex isotope patterns, or unexpected or low-abundance metabolites. Thermo Scientific Mass Frontier Spectral Interpretation Software Confident path from spectra to structure Mass Frontier software allows confident structural elucidation through chemically intelligent spectral annotation, state-of-the-art fragmentation prediction, and unparalleled spectral and fragmentation mechanism knowledge management. Thermo Scientific SIEVE Differential Expression Software Analysis of differential expression based on comparison of LC-MS datasets SIEVE software provides label-free, semi-quantitative differential analysis of proteins, peptides, or metabolites from the comparison of multiple LC-MS data sets. It is a statistically rigorous tool for analyzing data from metabolism or biomarker discovery experiments. www.thermoscientific.com/OrbitrapElite BR30232_E 06/11G ©2011 Thermo Fisher Scientific Inc. All rights reserved. SimGlycan is a registered trademark of Premier Biosoft International Corp. Tandem Mass Tags is a registered trademark of Proteome Sciences plc. Mass Frontier is a trademark of by HighChem Ltd.. ProSightPC is a trademark of the University of Illinois at Urbana-Champaign. All other trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries. Specifications, terms and pricing are subject to change. Not all products are available in all countries. Please consult your local sales representative for details. 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