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. Square Quadrupole Electrospray with Neutral High-Pressure Low- Quadrupole Ion Source S-Lens Blocker Octopole Cell Pressure Cell 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 Reagent 2 Heated Inlet Heated Inlet 3 The highest resolution, most versatile Orbitrap instrument ever 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 46642.586 47+ charge state of yeast enolase showing resolution of >100,000 FWHM at m/z 1,000 100 993.9770 80 100 R=110221 994.0401 R=107426 60 90 993.9134 R=109564 80 994.0827 40 R=107785 Abundance [%] 70 993.8706 20 R=107582 Relative 60 994.1249 R=106063 0 993.8284 50 R=113310 994.1670 46620 46650 R=107915 Mass, Da 993.7860 40 R=106257 Relative Abundance [%] Deconvoluted spectrum of yeast enolase 30 (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 A new standard of performance for comprehensive proteomics Orbitrap Elite LTQ Orbitrap Velos The superior resolution and spectral quality, as well as the higher scan 1000 873 933 803 speed of the Orbitrap Elite instrument, increase proteome coverage in 800 713 complex samples even with very low sample amounts. Here, analysis 694 600 of the E. coli proteome labeled with Thermo Scientific Pierce Tandem 563 Mass Tags demonstrates significant improvement over the already 400 outstanding performance of the previous-generation Thermo Scientific 200 LTQ Orbitrap Velos hybrid MS. Number of Identified Proteins 0 20 ng 80 ng 200 ng Top-down analysis SGRGKGGKGLGKGGAKRHRKVLRDNI ETD QGITKPAIRRLARRGGVKRISGLIYEET HCD RGVLKVFLENVIRDAVTYTEHAKRKTV TAMDVVYALKRQGRTLYGFGG HCD ETD Resolution Resolution ~ 200,000 FWHM at m/z 400 ~ 200,000 FWHM at m/z 400 100 843.9602 100 600.3526 R=158115 R=185866 z=2 Resolution z=2 Resolution 80 80 664.3820 > 100,000 FWHM at m/z 1,000 537.9891 R=171040 > 100,000 FWHM at m/z 1,000 R=189201 z=2 60 60 z=3 787.4543 1093.5867 R=154169 R=125517 40 762.4287 40 z=1 z=4 R=158642 1001.0440 1341.7392 385.7177 z=2 R=115639 R=106010 1620.8704 20 R=197410 z=2 20 z=5 R=100972 z=2 z=3 Relative Abundance [%] 0 0 500 1000 1500 2000 500 1000 1500 2000 m/z m/z 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. 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 386.25510 Buspirone lipids and metabolites in complex biological matrices R=251601 MS Spectrum R > 250,000 700000 RT: 1.95 - 6.06 y 600000 3.87 100 500000 Intensit 90 400000 80 300000 387.25830 e Buspirone 70 5 full MS scans 200000 R=252404 4 MS/MS scans 60 100000 388.26111 R=213904 Abundanc 50 0 386.5 387.0 387.5 388.0 40 m/z Relative 30 20 10 0 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 Time (min) 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 A total solution for metabolism and metabolomics research High-resolution, accurate-mass full-scan MS spectrum of an oxidative metabolite of buspirone 512.2684 150,000 0.50 ppm 1,800,000 402.2502 1,600,000 534.2498 100,000 R=246701 1,400,000 50,000 1,200,000 0 1,000,000 400 y [arbitrary units] it ns 800,000 te 494.2578 In 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™ of endogenous metabolites. The SIEVE™ software finds components that have software integrates acquisition, processing and reporting for metabolite identifica- reproducible differences across multiple sample groups. Optimized component tion from LC/MSn analyses and offers in-depth data mining tools for expected detection dramatically decreases false positive identifications caused by chemical and unexpected metabolites. The Mass Frontier™ software provides chemically background and redundant signals. The SIEVE software searches local and intelligent annotation of MSn 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.