Mass- Spectrometry-and- Proteomics课件.pptVIP

Mass Spectrometry and Proteomics Paolo Lecchi, PhD Dept. of Pharmacology George Washington University October 13, 2003 * 。 WHAT IS A “MASS SPECTROMETER ”...? MS * 。 The black box problem…... ESI MALDI LC/MS LC-MSMS ESI ESI Ion trap TOF QTOF quad. ESI CRIMS EI APCI FAB FT-ICR qQTOF SELDI …many black boxes ! * 。 “A MASS SPECTROMETER MEASURES THE MW….” “...A MS ANALYSIS GIVES THE MASS-TO-CHARGE RATIO (m/z) OF IONS…IN GAS PHASE”. * 。 Data Processing ION SOURCE ANALYZER ion separation vacuum Detector Pumping system Sample introduction DIRECT INTRODUCTION (solid, liquid, gas) SEPARATION TECHNIQUE (HPLC, CE, GC) MALDI, FAB, EI, ELECTROSPRAY TOF, quadrupole, ION TRAP * 。 ESI Liquid flow Q or Ion Trap analyzer ESI is a solution technique that gives a continuous stream of ions, best for quadrupoles, ion traps, etc. + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + MALDI 3 nS LASER PULSE Sample (solid) on target at high voltage/ high vacuum MALDI is a solid-state technique that gives ions in pulses, best suited to time-of-flight MS. TOF analyzer Atmosphere Low vac. High vac. High vacuum * 。 ….MALDI or Electrospray ? MALDI is limited to solid state, ESI to liquid ESI is better for the analysis of complex mixture as it is directly interfaced to a separation techniques (i.e. HPLC or CE) Until recently only ESI was available for high quality tandem-MS MALDI is easier to use and maintain MALDI is more “flexible” (MW from 200 to 400,000 Da) * 。 MALDI-TOF spectra of apomyoglobin INSTRUMENT: Kratos Axima-CFR Sample: 1 pmole apomyoglobin (horse skeletal muscle) * 。 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 m/z 0 20 40 60 80 100 Rela