MALDI-TOF-MS原理.ppt

An Introduction to the ProteinChip? Reader Matrix-Assisted Laser Desorption/Ionization (MALDI) Samples introduced to the MS on a passive probe Desorption/ionization method for non-volatile compounds analyte embedded in a solid-state matrix crystal Matrix - energy absorbing compound responsible for converting laser energy to thermal energy, which facilitates the desorption/ionization process Discovered in late 1980s by groups in Germany and Japan KOICHI TANAKA SHARES 2002 NOBEL PRIZE FOR APPLICATION OF LASER DESORPTION TO LARGE BIOMOLECULES Surface Enhanced Laser Desorption/Ionization (SELDI) Desorption/ionization method of non-volatile compounds in which the sample presenting surface plays an active role in the extraction, presentation, structural modification, amplification, and/or ionization of a given sample Invented by T. William Hutchens and Tai-Tung Yip in early 1990s Matching EAM with Sample Important EAM characteristics High molar absorptivity at laser wavelength Co-crystallizes well with sample Yields peaks that do not interfere with sample spectrum Does not promote multiple protonation Commonly used EAMs CHCA (α-cyano-4-hydroxycinnamic acid) Matrix ions generally 600 Da Promotes multiply protonated protein ions SPA (sinapinic acid) Higher m/z matrix ions Less multiply protonated protein ions EAM-1 The Time-Of-Flight Mass Spectrometer Time-Of-Flight Equation Ion Populations and Interpretation of Mass Spectra Parent molecular ion: formed by adding a charged species (usually a proton) (M+H)+ Pseudomolecular ion: formed by adding multiple protons; an ion formed by adding single or multiple non-proton charging species; an ion formed by clustering of parent molecular species; or a combination of the latter (M+2H)+2, (M+Na)+, (2M+H)+, (2M-H+2Na)+ Interpretation of Mass Spectra M/z: the mass-to-charge ratio Time-Of-Flight Fundamentals Resolution Resolution is the ability to create separate signals from ions of similar mass Analogous to chromatographic efficienc