L20 - Biological Applications of Mass Spectrometry Lecture...

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Unformatted text preview: Biological Applications of Mass Spectrometry Lecture 20 November 24, 2011 Identifying biological metabolites using mass spectrometry Discovery compound, X Collect urine and bile samples over 24 hrs Centrifuge; supernatant analyzed by LC/MS/MS Identifying biological metabolites using mass spectrometry 1) Precursor Scan Neutral Loss Scan 2) Product ion analysis of potential metabolites 3) Accurate mass: used to differentiate between metabolites having the same nominal mass - Externally calibrated mass spectrometer, using for example, mixture of various molecular-weight polyethylene glycol compounds. Accuracy = 0.05 Da. - Infuse an internal standard of known accurate mass into the LC column eluent prior to electrospray. Calibration slope can be adjusted to compensate for scan-to-scan variations. Accuracy = 0.005 Da. Further structural confirmation? NMR techniques… Philips et al. 2007; Martin et al. 2005; Fasano et al. 2006; Fasano et al. 2009 Nabb et al. 2007 8:2 FTOH F(CF 2 ) 8 CH 2 CH 2 OH (8:2 FTAL) (8:2 FTCA) (PFOA) (PFNA) +GSH/-HF +GSH/-HF (GS-8:2 FTUCA) CYP450 (8:2 FTUAL) (8:2 FTUCA) -HF -HF (GS-8:2 FTUAL) F(CF 2 ) y COO- F(CF 2 ) x CH 2 CH 2 OH x = 6, 8, 10, etc....
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This note was uploaded on 01/05/2012 for the course CHM 410 taught by Professor - during the Fall '11 term at University of Toronto.

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L20 - Biological Applications of Mass Spectrometry Lecture...

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