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    VAPORIZATION OF BIOLOGICAL MACROMOLECULES USING INTENSE, ULTRAFAST LASERS: MECHANISM AND APPLICATION TO PROTEIN CONFORMATION

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    Genre
    Thesis/Dissertation
    Date
    2011
    Author
    Brady, John Joseph
    Advisor
    Levis, Robert J.
    Committee member
    Owens, Kevin Glenn, 1960-
    Strongin, Daniel R.
    Matsika, Spiridoula
    Department
    Chemistry
    Subject
    Chemistry
    Electrospray
    Femtosecond
    Laser
    Lems
    Mass Spectrometry
    Nonresonant
    Permanent link to this record
    http://hdl.handle.net/20.500.12613/846
    
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    DOI
    http://dx.doi.org/10.34944/dspace/828
    Abstract
    This dissertation details the design and implementation of a state-of-the-art ambient trace analysis technique known as laser electrospray mass spectrometry. This novel technique utilizes an intense, nonresonant femtosecond laser pulse to transfer nonvolatile, fragile molecules into the gas phase from various substrates. The vaporized analyte is subsequently captured, solvated and ionized in an electrospray plume enabling mass analysis. Laser electrospray mass spectrometry is capable of analyzing samples in the liquid or solid states, mass spectral imaging of adsorbed molecules and detecting low vapor pressure analytes remotely. Experiments with biomolecules and pharmaceuticals, such as vitamin B12 and oxycodone, have demonstrated that the nonresonant femtosecond laser pulse allows for coupling into and vaporization of all molecules. This implies that sample preparation (elution, mixing with matrix and choosing samples with a particular electronic or vibrational transition) is not necessary, thus creating a universal mass analysis technique. Investigations using low vapor pressure molecules, such as lipids and proteins, led to the discovery that unfragmented molecules are transferred into the gas phase via a nonthermal mechanism. The laser electrospray mass spectrometry technique has allowed for the nonresonant femtosecond laser vaporization and mass analysis of trace amounts of a nitro-based explosive from a metal surface. The vaporization of unfragmented explosive molecules from a surface facilitates the identification of the explosive, reducing the probability of false positives and false negatives. In addition, this "soft" vaporization of molecules using nonresonant femtosecond laser pulses allows for protein to be transferred from the condensed phase into the gas phase without altering the molecule's structure, enabling ex vivo conformational analysis and possible disease typing.
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