by jonathan haltiwanger and teresa leslie. analytical technique to determine the elemental...
Post on 19-Dec-2015
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What is Mass Spectrometry (MS)?
Analytical technique to determine the elemental composition of a sample
Vaporization ionization electric field mass-to-charge (m/z)
MS/MS- Tandem Mass Spectrometry Fragment specific ions to identify resulting fragment ions Data is used to form structure Can find specific compounds in complex mixture
http://upload.wikimedia.org/wikipedia/commons/e/eb/MS_MS.png
Components:
Q- Quadrupole mass analyzer Filters sample ions (m/z)
TOF- Time-of-flight mass analyzer Velocity depends on m/z q- Radio frequency collision B- Magnetic sector E- Electric sector
http://www.ich.ucl.ac.uk/services_and_facilities/lab_services/mass_spectrometry/images/HPLC_ESI_MS_3.gif
MODE TIME!
Precursor ion scan Product ion scan Neutral loss scan Selected reaction monitoring
Fourier Transform MassSpectrometer (FTMS)
How does this apply to us?
Accurate Molecular Weight Measurements Sample confirmation, purity
Reaction Monitoring Monitor enzymes
Amino Acid Sequencing Identification of protein using database “tag”
Oligonucleotide Sequencing Post-Translation Modifications-
Glycosylation, phosphorylation, disulfide bridges
Protein Structure Protein-protein, protein-ligand, protein-DNA
Expanding Newborn Screening
Screens for 30+ diseases Usually rare, inherited metabolic disorders Measures acylcarnitine and amino acid levels
Fast, accurate, cheap, and non-invasive Star-G Project- Information on disorders Examples: Sickle cell disease,
phenylketonuria, maple syrup urine disease
http://www.dshs.state.tx.us/lab/nbsHeelstick.jpg
Phenylketonuria (PKU)
Difficulty breaking down phenylalanine Enzyme phenylalanine hydroxylase absent Leads to mental retardation, eczema,
seizures, and possibly death
http://www.bmb.leeds.ac.uk/illingworth/bioc1010/image017.gif
http://media.wiley.com/CurrentProtocols/HG/hg1701/hg1701-fig-0002-1-full.jpg
Realms of MS/MS
Biotechnology Analysis of proteins, oligonucleotides
Pharmaceutical Drug discovery, drug metabolism
Clinical Neonatal screening, hemoglobin analysis, drug
testing Environmental
Water quality, food contamination Geological
Oil compositionhttp://www.astbury.leeds.ac.uk/facil/MStut/mstutorial.htm
Sources:
Mørtz E, O'Connor PB, Roepstorff P, Kelleher NL, Wood TD, McLafferty FW, Mann M (1996). "Sequence tag identification of intact proteins by matching tanden mass spectral data against sequence data bases". Proc. Natl. Acad. Sci. U.S.A. 93 (16): 8264–7. doi:10.1073/pnas.93.16.8264. PMID 8710858.
Sleno L, Volmer DA (2004). "Ion activation methods for tandem mass spectrometry". Journal of mass spectrometry : JMS 39 (10): 1091–112. doi:10.1002/jms.703. PMID 15481084.
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