Accurate Mass Spectrometry Lab.ppt

Accurate Mass Spectrometry Lab February 1, 2008 Jon Karty What is Resolution? Resolution is the ability to separate ions of nearly equal mass/charge e.g. C6H5Cl and C6H5OF @ 112 m/z C6H5Cl = 112.00798 amu (all 12C, 35Cl, 1H) C6H5OF = 112.03244 amu (all 12C, 16O, 1H, 19F) Resolving power of 4600 required to resolve these two Two definitions Resolution = Δm / m (0.015 / 112.03 = 0.00013 or 1.3*10-4) Resolving power = m / Δm (112.03 / 0.015 = 7,468 or 7.5*10+3) High resolution facilitates high precision measurements High resolution, high accuracy MS can replace elemental analysis for chemical formula confirmation High resolution is considered RP ≥ 5,000 MAT-95 is capable of 60,000 resolving power LCT is capable of 5,000 resolving power MALDI-TOF-TOF has RP of 10,000-15,000 FTMS instruments can have RP 105 to 106 New “-omics” from MS Metabolomics Application of accurate mass spectrometry to identify small molecule metabolites in biological systems Sertraline’s metabolite has significantly reduced SSRI activity Fluoxetine’s metabolite has same activity, but longer half-life Accurate mass spectrometry determined chemical formula of THG (designer steroid) Petroleomics Application of high resolution, accurate mass spectrometry to the study of fossil fuels Resolving Power Example Calibration Calibration equation determines relationship between observed signals and actual m/z ratios External Calibration: mass spectrum of calibration compounds is acquired in a different experiment than the analyte compound Instrument drift can introduce errors Power supply voltages, temperature in the lab, etc. Internal Calibration: mass spectrum of calibration compounds are recorded at the same time as analyte compound Provides most accurate data Requirement to observe both calibrant and analyte ions in same experiment can be a significant challenge LockSpray on LCT is a unique solution to internal calibration problem Mass Accuracy Mass spectrometer accuracy often reported as a relative