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MSQ Plus North America Sales Training Chapter 2 – MS Modules

MSQ Plus North America Sales Training Chapter 2 – MS Modules. Beta Training Version A 5-Jan-2012. MSQPlus beta Sales Training. Program Goals. Transfer general knowledge of the MSQPlus, and the competitive landscape, our market position in order to better sell the product.

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MSQ Plus North America Sales Training Chapter 2 – MS Modules

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  1. MSQ Plus North America Sales TrainingChapter 2 – MS Modules Beta Training Version A 5-Jan-2012

  2. MSQPlus beta Sales Training Program Goals • Transfer general knowledge of the MSQPlus, and the competitive landscape, our market position in order to better sell the product. • Provide feedback on beta training modules to optimize them prior to launch to the NA Sales Team for chromatography/Advanced Detection. • Impact on MSQPlus order receipts within the 1st quarter of 2012 • Resource within your region for MSQPlus sales

  3. MSQPlus beta Sales Training Attending

  4. Agenda for today’s MSQPlus training program • Feedback and Responses to Session #1 Training • Functional modules common to all MS systems

  5. Feedback Session #1 slide 12 - Basic MS session is too busy and hard to find main points.

  6. + + + + + + + + + + + + + + + + + + + + + Session #1 Slide #12 Mass Spectrometer: Basic Schematic LC Sample Inlet Data System Ionization Source Mass Analyzer(s) Detector + + Creation of Ions Mixture of Solvent and Compounds Separation of Ions Detection of Ions Mass Spectra

  7. Feedback I would eventually like to see a “cheat sheet” of questions that we could use on sales calls with customers so that we can qualify them and understand what MS would suit their application best.

  8. Cheat Sheet Questions – What MS is best for you? • Cheat Sheet Questions: • Clean samples or matrix samples? • IC/Low mass? • Ease of use? Already using Chromeleon? • Known or unknown samples? • Proteins/high mass/high resolution/nano flow? • Rapid separation?

  9. Feedback • I would like a consolidated definition of MS terms – like a glossary: • Atmospheric Pressure Ionization: ESI, APCI • LC/MS vs GC/MS • API vs EI as it refers to a mass spectrum & library searching • CID • MRM, SRM • Adduct • Fragmentation • *most reps like the customers we will sell to do not understand these terms. The reps need to be able to explain them and understand them. • Check-out today’s training session and the MS Apps Guide & Glossary of MS terms sent earlier today….

  10. Feedback • I would like an Introduction to MS Components • Review a schematic of the source components • Discuss how LC/MS works. ESI vs APCI • Discuss CID and how it works. • Discuss +/- ion switching and hi/lo cone voltage • You may want to throw a slide in that discusses how GC/MS works • Check-out today’s training session….

  11. Feedback • Would like a review on How to Interpret Mass Spectra • Discussion of API spectra. • Discussion about positive and negative ion spectra. • Interpretation of spectral results. • Comparison of GC/MS spectra to LC/MS spectra (eg. GCMS Libraries, no commercial libraries for LCMS, generation spectral libraries with LCMS CID data). • Check-out today’s training session…. • Also check out Fred McLafferty’s, Interpretation of Mass Spectra or his ASMS Short Couse

  12. Feedback • I need more on the advantages & limitations of using MS in the determinations of unknowns. • Single state MS systems can provide molecular weight and some structural information – useful in unknown determinations. • Single stage MS systems have their niche primarily in analyte confirmation, low cost, and ease of use. And for us, ICMS. • MS/MS high resolution instruments provide more specific information to help identify unknowns.

  13. Feedback • I need more on the advantage of using MS/MS over MS only detection. • MS/MS instruments can be more sensitive relative to determinations of an analyte in dirty or high matrix samples. • MS/MS instruments can provide more specific, product ion information, better enabling sample confirmation. • MS/MS high resolution instruments provide more specific information to help identify analytes. • MS/MS systems can provide faster sample analysis due to reduced sample prep requirements in high matrix samples. • Compared to MS only systems, MS/MS instruments are generally higher cost to purchase & operate, require more sophisticated users.

  14. Feedback I would like more information on how the MSQPlus is used. The LCMS Applications Guide consolidates most MSQPlus applications. Environmental – Water Testing Industrial – Food & Beverage, Forensic, Chemicals

  15. Agenda for the NA MSQPlus training program • Comparison of Detectors: General, MS only, MS/MS • Functional modules common to all MS systems • MS Features and their importance to customers • MSQPlus Strengths, Weaknesses, Opportunity, and Threats • Decision Tree to better understand when to pursue an MS only order or an MS/MS order

  16. MSQPlus beta Sales Training By the end of this module you will know…. • The Basic Modules of all Mass Spec Instruments. • The Importance of the Basic MS Modules to Customers.

  17. MS Basic Modules Overview Understanding this information will help you identify the proper MS product for your customer.

  18. How does a Mass Spectrometer Work? • Basic Functional Modules of all Mass Spectrometers – Overview. • Inlet systems: Chromatography & Infusion • Ionization: Creating gas phase ions • Ion Extraction & Transfer: Getting ions into the mass filter • Mass Filtering: Separation of ions based on m/z • Detection: Counting ions • Vacuum system: Ion collisions • MSQ Data System

  19. + + + + + + + + + + + + + + + + + + + + + Mass Spectrometer: Basic Schematic LC Sample Inlet Data System Ionization Source Mass Analyzer(s) Detector + + Creation of Ions Mixture of Solvent and Compounds Separation of Ions Detection of Ions Mass Spectra

  20. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Vacuum System Data System Basic Functional Modules of an MS General Block Diagram Functional modules play important roles in… Basic Operation Method Development Maintenance Trouble-shooting

  21. How does a Mass Spectrometer Work? • Basic Functional Modules of all Mass Spectrometers • Inlet systems: Chromatography & Infusion • Ionization: Creating gas phase ions • Ion Extraction & Transfer: Getting ions into the mass filter • Mass Filtering: Separation of ions based on m/z • Detection: Counting ions • Vacuum system: Ion collisions • MSQ Data System

  22. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap Inlets deliver sample to the Mass Spectrometer Liquid Chromatography Inlet Systems Separates mixtures of non-volatile organic and in-organic compounds What is API? Atmospheric Pressure Ionization Includes ESI & APCI Used to combine an LC with an MS Utilizes Atmospheric Pressure Ionization (API) API produces mostly Molecular Ions

  23. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap Inlets deliver sample to the Mass SpectrometerCONTINUED Gas Chromatography Inlets Separates mixtures of volatile and semi-volatile organic compounds Requires Electron Impact (EI) or Chemical Ionization (CI) EI produces mostly Fragments and little or no Molecular Ion

  24. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap Inlets deliver sample to the Mass SpectrometerCONTINUED MSQ Reference Inlet Infuses the MSQ Calibration Solution - No separation. Utilizes Atmospheric Pressure Ionization (API) Produces mostly clusters and adducts that are evenly spaced masses from m/z 23 to m/z 1972

  25. See How Much You Remember – A QuizHOW MASS SPECS WORK - INLETS • What functional modules do all mass specs have? • Name 3 MS inlet systems? • What inlet system is associated with producing fragments of compounds?

  26. How does a Mass Spectrometer Work? • Basic Functional Modules of all Mass Spectrometers • Inlet systems: Chromatography & Infusion • Ionization: Creating gas phase ions • Ion Extraction & Transfer: Getting ions into the mass filter • Mass Filtering: Separation of ions based on m/z • Detection: Counting ions • Vacuum system: Ion collisions • MSQ Data System

  27. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap Mass Specs require Gas Phase Ions Gas Phase Ionization Processes • Atmospheric Pressure Ionization (API) • ESI (Electrospray) • APCI (Atmospheric Chemical Ionization) • Low Pressure Ionization • EI (Electron Impact / Ionization) • CI (Chemical Ionization) • MALDI (Matrix Assisted Laser Desorption Ionization)

  28. MSQPlus Features – API Inlet ESI Probe • FastLoc Probes for ESI & APCI • Easy Fast Change-over (less than 1 minute) • Both probes have all internal connections • High voltage (ESI) and Corona (APCI) • Nebulizing and Sheath Gas • All stainless steel/ceramic for low chemical background • Low cost,replaceable capillaries for low-cost maintenance • Flow rates (workable): 200 to 500µL/min Ionization Ion Extraction & Transfer Mass Filtering APCI Probe Detection Ion Path Roadmap

  29. Inlet MALDI 200,000 Ionization 15,000 Ion Extraction & Transfer 1,000 Mass Filtering Molecular Weight APCI ESI EI/CI Detection Ion Path Roadmap 10 Non polar Very polar Comparison of Ionization ProcessesMass vs. Polarity • ESI can be used over a broad range of polarity and molecular weight • APCI is used for non-polar & smaller compounds

  30. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap Features of ESI What’s ESI look like? High Voltage on Capillary Gas Phase Ions Ready for MS Mobile Phase from HPLC • Evaporates mobile phase – gas phase • Charges mobile phase / analytes – ions • Result: gas phase ions

  31. High Voltage Power Supply Inlet Ion flow + - Ionization Once Rayleigh limit is reached a coulombic explosion happens and releases ions Capillary ± 1.5 - 4 kV Ion Extraction & Transfer Ion containing droplets Off Axis Mass Filtering Detection Ion Path Roadmap Droplets evaporate, Field increases Ions move towards surface Mechanism of Electrospray & Ion Extraction Mobile Phase / Analyte Entrance Skimmer Off Access to Ion Path ± 20 – 190 V

  32. Inlet ±5 kV Ionization Ion Extraction & Transfer Probe Heater Mass Filtering Ion Plume Detection ESI Needle ESI Capillary Ion Path Roadmap MSQ Electrospray Probe & Heater Detail N2 Gas Sheath NebulizingN2 Gas • To create gas phase ions mobile phase is … • Heated • Nebulized • Charged via charge transfer with ESI Needle

  33. ±5 kV Corona Needle APCI Needle(recessed – no voltage) Inlet NebulizingN2 Gas N2 Gas Sheath Ionization Ion Extraction & Transfer Mass Filtering Probe Heater Detection Ion Plume Ion Path Roadmap Ion Plasma MSQ APCI Probe & Heater Detail • To create gas phase ions mobile phase is … • Heated • Nebulized • Charged via Corona Needle

  34. Atmospheric Pressure Ionization Corona Discharge Electrospray Electrospray (ESI) Atmospheric Pressure Chemical Ionization (APCI)

  35. Inlet Filament70ev Ionization Sample Stream Ion Extraction & Transfer High Energy Electrons Mass Filtering + Ions are unstable due to high energy + + + + + Detection Fragmentation of molecular ion occurs Ion Path Roadmap Low Pressure Ionization (Electron Impact - EI) Comparison of EI & ESI

  36. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap See How Much You Remember – A QuizHOW MASS SPECS WORK - IONIZATION • What two characteristics must an analyte have in order to be analyzed via mass spectrometry? • What form of ionization produces an NIST library searchable mass spectrum? • What inlet system is associated with producing fragments of compounds? • In what two ways does an API source enable the analysis of an HPLC sample? • What type of analyte is better suited for ESI?

  37. How does a Mass Spectrometer Work? • Basic Functional Modules of all Mass Spectrometers • Inlet systems: Chromatography & Infusion • Ionization: Creating gas phase ions • Ion Extraction & Transfer: Getting ions into the mass filter • Mass Filtering: Separation of ions based on m/z • Detection: Counting ions • Vacuum system: Ion collisions • MSQ Data System

  38. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap Ion Extraction & Transfer into a Mass Spectrometer The RF Lens transfers ions to the Mass Filter It enables transmission of low mass ions It forms the ion beam & is the region where ion – molecule collisions occur – breaking weak chemical bonds – CID (Collision Induced Dissociation). Ion Extraction MS Detector The Sample Cone may become coated with sample matrix or damaged which may adversely effect Ion Extraction. Ions are attracted to Sample Cone by Voltage & Pressure differences

  39. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap Sample Cone extracts ions from API source region Close-up view of Sample Cone with Obstruction Keep the Sample Cone clear & free of matrix coatings to maintain the most efficient ion extraction

  40. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap RF Lens transfers ions to Mass Filter MSQ RF Lens Assembly Also functions as a Collision Cell – “In-Source” CID Collision energy is a function of Cone Voltage Higher Cone Voltage provides more energy to break weak chemical bonds

  41. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap See How Much You Remember – A QuizHOW MASS SPECS WORK – ION EXTRACTION & TRANSFER What are the processes involved in transfer of ions from the ion source region to mass filter? What parts of the Ion Transfer assembly can be coated with matrix or damaged thus reducing sensitivity? How are ions attracted to the Sample Cone? What is CID? What advantage does the MSQ have over other mass spectrometers?

  42. How does a Mass Spectrometer Work? • Basic Functional Modules of all Mass Spectrometers • Inlet systems: Chromatography & Infusion • Ionization: Creating gas phase ions • IonExtraction & Transfer: Getting ions into the mass filter • Mass Filtering: Separation of ions based on m/z • Detection: Counting ions • Vacuum system: Ion collisions • MSQ Data System

  43. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap Characteristics of Mass Filters • Mass Filters sort ions based up m/z • Measure Exact Mass • Resolve masses • Scan Functions - Mass filter can scan in different ways • A wide range of Mass Filter design & Performance

  44. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap Mass Spectrometers measure Exact Mass m/z 79 m/z 81

  45. Inlet m/m  3044 Low Resolution FWHM  .50 Ionization 522 524 526 1520 1522 1524 1820 1822 1824 Ion Extraction & Transfer Medium Resolution FWHM  .14 m/m  10871 Mass Filtering 522 524 526 1520 1522 1524 1820 1822 1824 Detection High Resolution FWHM  .05 m/m  30440 Ion Path Roadmap 522 524 526 1520 1522 1524 1820 1822 1824 m/z Mass Resolution Comparison (MRFA/UM 1621) The MSQ is a Low Resolution MS – Unit mass resolution

  46. Full Scan MS Scanned “DC Voltage” Sequential m/z are Stable No “DC Voltage” All m/z are Stable MS Filter Rf Lens Focusing Scanning Single Quadrupole

  47. Selected Ion Monitoring (SIM) MS Filter Rf Lens Focusing Selected m/z Single Quadrupole

  48. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap Full Scan enables obtaining a Mass SpectrumSCAN FUNCTIONS – FULL SCAN Mass spectrum obtained from Full Scan Bromide m/z 79 m/z 81 Most significant ions at m/z 79 & 81 m/z 15 m/z 200 Full Scan function: 15 – 250 Daltons in 1.0 sec; + ve; Cone Voltage: 70

  49. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap Full Scan enables Mass Extracted ChromatogramSCAN FUNCTIONS – FULL SCAN Ion Chromatogram peak at 7 min Mass spectrum obtained from Full Scan Extract m/z 81 Plot signal vs. Time m/z 81 m/z 79 Base Peak ion m/z 81 m/z 15 m/z 200 Key point: Mass Chromatogram overlay enables correlation of mass to a chromatographic peak.

  50. Inlet Ionization Ion Extraction & Transfer Mass Filtering Detection Ion Path Roadmap SIM Scans enable sensitive Mass ChromatogramsSCAN FUNCTIONS – SIM SCAN SIM mass chromatogram at m/z 79

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