1 / 29

LC-MS

LC-MS. Lecture 7. LC-MS. LC. Interface. Mass Analyzer. Detector. Extraction of The analyte from The solvent. Ion evaporation or ionization. Fragmentation. LC – Separation of the mixture of analytes Interface – Separation of the analyte from the solvent

teresa
Download Presentation

LC-MS

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  1. LC-MS Lecture 7

  2. LC-MS LC Interface Mass Analyzer Detector Extraction of The analyte from The solvent Ion evaporation or ionization. Fragmentation • LC – Separation of the mixture of analytes • Interface – Separation of the analyte from the solvent • MA (mass analyzer) – separation of the analyte molecular ion and fragments according to their mass to charge ratio

  3. HPLC Liquid phase operation 25 - 50 deg. C No mass range limitations Inorganic buffers 1 ml/min eluent flow is equivalent to 500 ml/min of gas MS Vacuum operation 200 - 300 deg. C Up to 4000 Da for quadrupole MS Requires volatile buffers Accepts 10 ml/min gas flow PROBLEMS IN COMBINING HPLC AND MS

  4. Interface • Historical • Moving belt • Thermospray • Common interfaces • Electrospray • APCI • Special interfaces • Particle beam LC/MS • Continuous flow FAB • Atmospheric pressure photon ionization • MALDI

  5. Formation of gas phase ions from solution phase Basic Electrospray Schematic N 760 Torr 10-5 Torr 10-2 – 10-4 Torr 10-5 Torr 1 Torr Ion optics Richard B. Cole, Electrospray Ionization Mass Spectrometry, Fundamentals Instrumentation and Applications, Wiley Interscience, 1997

  6. API-Electrospray Interface (HP)

  7. Orthogonal ESI schematic (Waters)

  8. ZQ interface (Waters)

  9. ELECTROSPRAY • Factors to consider • Ionic strength • Surface tension of the solvent • Volatility of the solvent • Character of the analyte ions in solution: solvated, ion paired, etc • Mobile phase composition and amount of water • pH of the mobile phase

  10. APCI INTERFACE

  11. APCI IONIZATION CHAMBER

  12. Fragmentation

  13. PROTEIN SPECTRUM (multiple charge) (M+2)/(z+2) (M+1)/(z+1) M/z

  14. MASS ANALYZERS • Quadrupole • Ion Trap • Time-of-Flight • Magnetic Sector • ICR (Ion-cyclotron resonance)

  15. Resolution of MA

  16. Comparison of MA

  17. Quadrupole MA Quadrupole MA – Mass Filter Only ions with the specific M/z could pass between rods at set values of DC and RF voltages

  18. Quadrupole MA

  19. Electromagnetic field in Quadrupole

  20. Ion Trap

  21. Ion trajectory in Ion-Trap

  22. Time-of-Flight

  23. Bipolar TOF

  24. Reflectron Time-of-Flight

  25. MS-MS

  26. TRIPLE QUADRUPOLE

  27. MS-MS (triple quad – TOF)

  28. Magnetic Sector

  29. Double focusing Magnetic sector

More Related