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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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mass spectrometry

Mass Spectrometry

Week 2 Lab

S117 Fall 2011

overview
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
assignments
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
excel
Excel
  • Working with data sets
  • Statistical Analysis
  • Certainty and uncertainty
    • Measurements
    • Experimental limit
    • Reproducibility
experimental design
Experimental Design
  • Practical Weaknesses?
  • Propagation of Error
  • Was the distillation useful?
pre lab mass spectrometry
Pre-Lab: Mass Spectrometry

Analysis of isotopes!

nominal mass and exact mass
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
analysis of molecules
Analysis of Molecules
  • Parent peak (molecular ion)and fragment peaks
  • Analysis of fragment patterns useful
  • Application to Cl2

isovaleraldehyde

caffeine

analysis of biomolecules
Analysis of Biomolecules
  • Complex molecules
  • Molecular mass in kiloDaltons (kDa)
  • Nobel Prizes for MS of proteins
mass spec and mutations
Mass Spec and Mutations
  • Sickle Cell Anemia
  • Point Mutation
  • How can MS be used to identify Mutations?
identification of proteins
Identification of Proteins
  • Comparison of 2D electrophoresis
identification of proteins1
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

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