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Mass Spectrometry

Mass Spectrometry. WEB resources: http://www.colby.edu/chemistry/OChem/DEMOS/MassSpec.html http://www-methods.ch.cam.ac.uk/meth/ms/theory/index.html. Ionization. Electron. separate m/z. ionize. Fast Atom Bombardment. Plasma Desorption. Ionization. Thermospray. Electrospray. +. -. -.

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Mass Spectrometry

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  1. Mass Spectrometry WEB resources: http://www.colby.edu/chemistry/OChem/DEMOS/MassSpec.html http://www-methods.ch.cam.ac.uk/meth/ms/theory/index.html Biophysical Tools - Mass Spectrometry

  2. Ionization Electron separate m/z ionize Fast Atom Bombardment Plasma Desorption Biophysical Tools - Mass Spectrometry

  3. Ionization Thermospray Electrospray Biophysical Tools - Mass Spectrometry

  4. + - - + Mass analysis • Quadrupole Mass Filter • 4 parallel rods with mixture of DC and RF electrical voltage • Magnetic sector Force on a moving charge in B field = Centrifugal force: zvB = mv2/r Calculate v from kinetic energy: mv2/2 = z Vacc mass to charge ratio: m/z = r2B2/2Vacc • Electrostatic Analyzer • instead of magnet use electric field: • F = zE Time-of-Flight ions of different m/z and the same energy will have different velocity Biophysical Tools - Mass Spectrometry

  5. Matrix Assisted Laser Desorption accuracy +/- 1 Da on a 10 kDa protein Biophysical Tools - Mass Spectrometry

  6. Ion Cyclotron Resonance • radius of motion is defined by centrifugal force (mv2/r) balancing the magnetic force (zvB) • Angular velocity (cyclotron frequency) defined by m/z ratio Biophysical Tools - Mass Spectrometry

  7. Distribution of charged states in electrospray High resolution neighboring m/z values differing by a single charge: isotope peaks separation: Biophysical Tools - Mass Spectrometry

  8. Combined techniques MS/MS Triple – quadrupole trap Hybrid sector/trap LC/MS Biophysical Tools - Mass Spectrometry

  9. Application: Mass determination Perils of sample preparation Biophysical Tools - Mass Spectrometry

  10. Application: Oligomeric structures Biophysical Tools - Mass Spectrometry

  11. Application: Peptides Biophysical Tools - Mass Spectrometry

  12. Application: Sequencing Biophysical Tools - Mass Spectrometry

  13. Application: postranslational modification Biophysical Tools - Mass Spectrometry

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