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Mass Spectrometry

Mass Spectrometry. Week 2 Lab S117 Fall 2011. Overview. Post-Lab: Percent Alcohol in Wine TurnItIn.com Working with data sets Analyzing data sets Analysis of experimental procedure Pre-lab: Mass Spectrometry Fragmentation Protein Analysis Practical matters of lab meeting.

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Mass Spectrometry

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  1. Mass Spectrometry Week 2 Lab S117 Fall 2011

  2. Overview • Post-Lab: Percent Alcohol in Wine • TurnItIn.com • Working with data sets • Analyzing data sets • Analysis of experimental procedure • Pre-lab: Mass Spectrometry • Fragmentation • Protein Analysis • Practical matters of lab meeting

  3. Assignments • Sign up for turnitin.com • Formal reports due electronically up to time of lab • One minute24 hours late = 20pt penalty • Give copy of lab notebook to AI in lab

  4. Excel • Working with data sets • Statistical Analysis • Certainty and uncertainty • Measurements • Experimental limit • Reproducibility

  5. Experimental Design • Practical Weaknesses? • Propagation of Error • Was the distillation useful?

  6. Pre-Lab: Mass Spectrometry Analysis of isotopes!

  7. Nominal Mass and Exact Mass • Nominal Mass: Low resolution • 35Cl = 35 Da • 37Cl = 37 Da • CO = N2 = 28 Da • Exact Mass: High Resolution • CO = 27.9949 Da • N2 = 28.0061 Da

  8. Analysis of Molecules • Parent peak (molecular ion)and fragment peaks • Analysis of fragment patterns useful • Application to Cl2 isovaleraldehyde caffeine

  9. Analysis of Biomolecules • Complex molecules • Molecular mass in kiloDaltons (kDa) • Nobel Prizes for MS of proteins

  10. Proteins Made of Amino Acids

  11. Mass Spec and Mutations • Sickle Cell Anemia • Point Mutation • How can MS be used to identify Mutations?

  12. Identification of Proteins • Comparison of 2D electrophoresis

  13. Identification of Proteins Trypsin Digest MS Library Mass Spec Purified Protein Mix of Peptides Mass Data of mixture Identity of Protein Example 1 ASR LVMK QQK D ASRLVMKQQKD Example 2 ASRLVKMQKQD

  14. Practical Matters • Meet in chemistry computer lab 006 • Mass Spec lab tours • Dry lab: No special clothing requirements! • Remember formal report and lab notebook

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