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Four Reasons to Buy Thermo Scientific LC

Four Reasons to Buy Thermo Scientific LC. Accela UHPLC and Easy-nLC II Andy Altman VP & GM LC LCMS. Technology that delivers quality results, faster & easier. EASY-nLC II Nano-LC. Accela UHPLC. Four Reasons to buy Thermo Scientific LC.

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Four Reasons to Buy Thermo Scientific LC

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  1. Four Reasons to Buy Thermo Scientific LC Accela UHPLC and Easy-nLC II Andy Altman VP & GM LC LCMS

  2. Technology that delivers quality results, faster & easier EASY-nLC IINano-LC Accela UHPLC

  3. Four Reasons to buy Thermo Scientific LC • Maximum Pumping Performance: maximum reproducibility and base resolution using quaternary solvent delivery with innovative technologies for HPLC and UHPLC. • Increased Sampling Flexibility: multiple autosampling technologies specifically designed for UHPLC pressures and minimum carryover. • Easy Split-Free Nano-Flow: effortless, split-free nano LC with unrivalled ease of use and touch screen technology. • The Complete Workflow Solution: one source for solvents, consumables, instrumentation, service, and support.

  4. 1. Maximum Pumping Performance

  5. 1. Maximum Pumping Performance Four solvents, one pump, no waiting • Multiple pumping options: • HPLC flow rates up to 5ml/min at 600 bar • UHPLC flow rates up to 2ml/min at 1250 bar • Rapid method development with quaternary solvent capability. • Extraordinary retention time reproducibility and gradient performance using unique Forced Feedback Control technology. • Stable baselines without a pulse dampener. • Maximum productivity by utilizing low delay volumes to shorten run times. • Increases functionality with optional seal wash capability.

  6. Z-arm with force sensor 1. Maximum Pumping Performance Force Feedback Control • Stable and rapid solvent delivery for all HPLC and UHPLC applications. • Gradients with the highest compositional accuracy. • Highest flow rate accuracy across the entire pressure range. • Stable baselines virtually pulsation free with no pulse dampeners. • Extraordinary retention time reproducibility.

  7. 2.4 2.4 2.2 2.2 2.0 2.0 1.8 1.8 1.6 1.6 1.4 1.4 mAU 1.2 mAU 1.2 1.0 1.0 0.8 0.8 0.6 0.6 0.4 0.4 0.2 0.2 0.0 0.0 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 1.70 1.75 1.80 1.85 1.90 1.95 2.00 Minutes 1. Maximum Pumping Performance FFC vs. no FFC: Baseline Fluctuation No pulse dampener needed to obtain stable baselines when using FFC! Flow rate: 1 mL/min FFC no FFC

  8. 1. Maximum Pumping Performance Improving the Detection of Low Level Drug Impurities 0.04% Impurities Detected

  9. 1. Maximum Pumping Performance Pressure trace overlap shows excellent pressure stability at 1200 bar with clean and dirty matrices. Column: Hypersil Gold PFP 100x2.1mm 1.9 µm Flow rate: 900 uL/min, 1200 bar Gradient: A: water, 0.1% FA, B: methanol, 0.1% FA Detector: Accela PDA 1. Gallic Acid 2. 3,5-dihydroxybenzoic acid 3. Epigallocatechin 4. Caffeine 5. Epicatechin 6. Epigallocatechin Gallate 7. Epicatechin Gallate 8. Anisic acid 9. Catechin Gallate 10. 3,4,5-Trimetheoxycinnamic acid 11. Mycricetin

  10. 1. Maximum Pumping Performance Unbeatable retention time and peak area reproducibility at 1200bar Column: Hypersil Gold PFP 100x2.1mm 1.9 µm Gradient: A: water, 0.1% FA, B: methanol, 0.1% FA Flow rate: 900 µl/min, pressure 1200 bar

  11. 2. Increased Sampling Flexibility Accela Open Autosampler • Maximizes precious bench space with a stackable design. • Integrated Dynamic Load and Wash for near zero carryover. (0.003% chlorhexidine) • Specifically configured to handle UHPLC applications up to 1250bar. • Compact size open architecture for easy access to components. • Maximum flexibility with fast cycles and outstanding reproducibility. • Optional column oven for increased sample to sample reproducibility.

  12. Aspirating sample into holding loop, using front and rear air segments Inject Sample into sample loop, turn valve and load to column Flush valve and syringe at the same time using continous flow solvent pumps 2. Increased Sampling Flexibility Dynamic Load and Wash

  13. 2. Increased Sampling Flexibility Accela Closed Autosampler/Column Oven • 5to 95C settable in 1C increments. • Integrated solvent pre-heater with 3μL delay volume. • Diamond coated 1250bar valve. • Inner and outer coated needle for low carryover • Isothermal mobile preheater, injection valve, sample loop, and column contained in one temperature controlled compartment.

  14. 3. Easy Split-Free Nano-Flow Boosting Productivity with EASY-nLC II Effortless, split-free nano LC Unrivalled ease of use with touch screen Simple to interface and synchronize with any MS Maximum performance with 2D-enabled operation - identifying more proteins than conventional approaches Quality assured – system pre-configured and QC-tested with MS Easy switching between 1- or 2-column configuration Easy maintenance, access, and service

  15. 3. Easy Split-Free Nano-Flow Why split-free nano LC? Increased reproducibility Nanoscale HPLC pumps generate gradients with maximum accuracy at nanoliter flow rates. Increased sensitivity Extremely low delay volumes transfer the entire sample to the mass spectrometer ensuring more ions in the source. Increased system stability Integrated sample temperature control maintains sample integrity. Specialized flow path guarantees actual ‘on-column’ gradient is unaffected by changes in back pressure.

  16. 3. Easy Split-Free Nano-Flow Specialized functionality for Nano applications saves time Automated leak and system back pressure testing pinpoint leaks and blockages for maximum productivity. Unparalleled ease of use with automated maintenance schedules. Unmatched system stability with ‘Idle-flow’ function providing solvent flow at all times with automated pump refill between experiments. ‘Intelligent flow control’ operates according to user-defined pressure. Ensures rapid build-up of pressure and flow. Speeds up equilibration and sample loading steps. Avoids interruptions caused by increases in back pressure. Analysis continues despite increasing back pressure. *Irregular pressure profile due to changing solvent composition

  17. 3. Easy Split-Free Nano-Flow Maximizing Sample Throughput Load up to 384 samples Run unattended up to 3 weeks – 2 x 25 mL solvent bottles Selection of vial and microplate formats -56 vials positions -1 x 96 well MTP and 6 vials -4 x 24 well MTP strips and 6 vials -2 x 48 well MTP strips and 6 vials -1 x 384 well MTP and 6 vials Enclosed refrigerated compartment avoids condensation, maintains sample stability

  18. 3. Easy Split-Free Nano-Flow Stable, pulsation-free gradients from 100 nL/min Factory-configured flow paths minimize swept volume. Maximum chromatographic accuracy with: Split-free gradient mixing Step-controlled nano-flow syringe pumps Flow sensor control of mobile phase delivery

  19. 3. Easy Split-Free Nano-Flow Easy maintenance, access, and service Built-in PC & software Automated maintenance steps Remote access via internet for system interrogation Event log for troubleshooting Modular design for easy component exchange Pulsation-free pumps minimize need for replacement of seals, valves and fittings Buffer bottles Switching valves PC and software Dual nano-flow sensors Nanoliter pumps

  20. 3. Easy Split-Free Nano-Flow EASY-Column: Chromatographic excellence at nanoscale Compatible with any nanoscale HPLC system Optimized for online LC-MS Simple, flexible design Each column supplied with certificate of analysis 653.3 100 90 582.3 80 70 722.3 60 Relative Abundance 50 40 30 732.2 507.8 20 547.3 10 747.70 0 10 15 20 25 30 Time (min) Sample: 100 fmol tryptic BSA digest Precolumn:EASY-Column, 2cm, ID100µm, 5µm, C18-A1 (SC001) Analytical column:EASY-Column, 10cm, ID75µm, 3µm, C18-A2 (SC200) Method: 300 nL/min, 5-35 % acetonitrile in 30 minutes. Typical peak widths (FWHM) ~ 7 sec

  21. 3. Easy Split-Free Nano-Flow EASY Nano Spray Source • Single and easy setup for all online nano-flow applications. • Maximum sensitivity and resolution with extremely stable spray. • Optimized for easy handling and performance with zero dead volume connections. • Fast and flexible route for ion generation.

  22. 3. Easy Split-Free Nano-Flow Excellent Results: Fast separations with typical peak widths ~ 3.5 sec (FWHM) Sample: 100 fmol tryptic BSAdigest Column: EASY-Column C18-A2, 10cm,ID 75μm, 3μm Gradient: 300 nL/min, 5-35% B in 10 min Solvents: A: 0.1% formic acid B: acetonitrile, 0.1% formic acid Detector: Easy-nLC with LCQ

  23. 3. Easy Split-Free Nano-Flow Consolidated sequence and annotation database ProteinCenter™ database Bridge the gap between search engine and publication • Progress from data sets to meaningful biology - in minutes, not months. • Historic tracking of outdated proteins and cross-reference mapping. • Computational sequence enrichment by prediction of trans-membrane domains, signal peptides and protein domains. • Access >10 million non-redundant proteins. • Map accession keys in time and between databases. • Remove redundancy - allellic, sequence variance, splice-isoforms. • Collect protein annotations from all major sources. • Share results – online.

  24. 3. Easy Split-Free Nano-Flow Search engine Excel Literature ProteinCenter™ workflow

  25. 4. The Complete Workflow Solution

  26. 4. The Complete Workflow Solution Accela PDA and UV Detection • LightPipe™ technology produces the highest sensitivity in PDA detection. • Wide wavelength range: 190-800nm. • Maximum resolution with the fiber-optic beam shaper. • Pre-aligned Deuterium and Tungsten lamps. • Reduced flow cell volume improves high speed chromatographic efficiency. (2uL / 10mm pathlength)

  27. 4. The Complete Workflow Solution The right column for your application • Hypersil GOLD • Hypersil GOLD with C18 Selectivity, C8, aQ, PFP, CN, Phenyl, C4, Amino, AX, SAX, Silica • Outstanding peak shape in multiple phases, lengths, and particle size from 1.9 to 5 μm. • 1.9 μm particles improve speed and efficiency.

  28. 4. The Complete Workflow Solution Accela HPLC/UHPLC: most versatile and best MS front-end. • Efficient separations over multiple pressure ranges. • Maximum flexibility with quaternary gradient capability. • Increased productivity with Accela Open Autosampler.

  29. 0.46 100 95 90 85 80 75 70 65 60 0.76 Relative Abundance 0.43 55 50 0.64 0.13 45 0.27 0.31 40 35 30 25 20 15 10 5 0 0.00 0.10 0.20 0.30 0.40 0.50 0.60 0.70 0.80 0.90 1.00 Time (min) 4. The Complete Workflow Solution LC/MS Analysis of β-Blockers in under 1 minute Column: 30 x 2.1mm 1.9 mm Hypersil Gold Mobile phase: A - H2O + 0.1% HCOOH B - MeCN + 0.1% HCOOH Gradient: Time %A %B 0.00 85 15 0.10 75 25 0.50 70 30 1.00 60 40 Flow rate: 1000 l/min Pressure: 310 bar Temperature: 30 ْC Injection: 5 mL Detection: LTQ-Orbitrap Resolution (∆m / m): 15,000 (FWHM) Interface: ESI + Analytes: 1. Atenolol 2. Nadolol 3. Pindolol 4. Timolol 5. Metoprolol 6. Labetalol 7. Propanolol Short column Fast analysis Conventional pressure

  30. 4. The Complete Workflow Solution Low retention time RSD values of complex mixtures Accurate label-free quantification Sample: 100 fmol annexin V 800 fmol gelsolin 300 fmol BSA digestion mixture Column: Easy-Column C18 15 cm, 75 µm ID, 5 µm Gradient: 300 nl/min, 3 - 35% B in 25 minutes Detector: EASY-nLC with LTQ Orbitrap XL FT scan: 390-1500 m/z

  31. Uncompromised Performance with Thermo Chromatography UHPLC • Reduces run times • Accelerates method development • Increases sensitivity and resolution • Reduces operational costs • Increased Flexibility • Increased Productivity • Increased Throughput • Enhanced Data Quality Accela Systems • Unmatched quaternary performance • Eliminates need for pulse dampeners • Reduced carry over with Open AS • Increased sensitivity with unique PDA • Best UHPLC-MS Front End EASY-nLC II • Unrivalled ease of use with touch screen • Maximum performance with state-of-the-art MS systems • Easy maintenance, access, and service • Easy switching between 1- or 2-column configuration

  32. Thank you!

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