In spite of their importance in many systems, liquid surfaces have been explored at the microscopic level to a much lesser extent than solids. Most surface analysis must take place in vacuum, a major drawback for liquids. The technique of time-of-flight scattering and recoil spectrometry (TOF-SARS) has been applied to molecular liquid surfaces for the first time. The apparatus borrows key elements from previous TOF-SARS experiments on solids and from molecular beam scattering (MBS) and features excellent surface specificity and the ability to detect all elements. A high-vacuum time-of-flight spectrometer was developed for the purpose of measuring the surface atomic concentration of atoms in low-vapor pressure liquid samples, and hence to infer preferred surface orientations. The TOF-SARS experiment involves surface bombardment with inert gas ions in the 1-3 keV energy range. During the interaction surface atoms may either (a) induce scattering of primary ions or (b) recoil from the surface. A binary collision model describes the kinematics and dynamics of the interactions well, allowing prediction of velocities and probabilities of particles leaving the surface. Particles that reach a detector along a ~1.1 m flight path are separated by velocity, and signals are collected as a histogram, revealing relative measured intensities that are converted to ratios of accessible surface atoms. Comparing the measured atomic ratios with computer-simulated accessible atomic ratios for various possible orientations gives insight into preferred surface orientation. A number of systems were explored m this work: liquids including a complementary pair of molecules having distinct 'head-tails' structures; glycerol as a highly H-bonded system, and a room-temperature molten salt. Preliminary results reveal that surface molecules appear in most cases to adopt some preferred orientation at the interface. The TOF-SARS technique was able to distinguish 'head' from 'tail' in molecules exhibiting that structure, suggesting only part of the head was accessible. In glycerol, all but two possible orientations were ruled out but the symmetrical nature of the molecule prohibits definitive assignment. The ionic liquid was found to have the cation and anion sharing the surface population roughly equally, and a preferred orientation for the substituted aromatic anion was discovered. / Graduation date: 2000

Graduation date: 1995

The work discussed in this dissertation is aimed at creating novel approaches to chemical imaging that ultimately allow for submicron resolution. This goal has been approached from two direction using laser based desorption and coupling it with an AFM using apertureless tip-enhanced laser ablation/ionization. The second direction was through the development a new approach to thermal desorption based mass spectrometry experiments by using a proximal probe to spatially desorb the surface and ionizing the plume of neutrals using a secondary ionization source at atmospheric pressure. The thermal desorption approach allows for the easy scaling of the technique all the way from the millimeter to the nanometer regime. In the nanometer regime an AFM platform with silicon based heating AFM probes is used to locally desorb material from nanometer sized craters. The final work in this thesis focused on trying to improve laser based desorption through a secondary ionization of the neutrals plume by capturing the laser desorption plume into a liquid and then electrospaying the solution into a MS. The added benefit of being able to capture the desorption plume into a liquid is the ability to carry out post sampling processing of the captured analyte via high performance liquid chromatography. The ability to clean up a sample via HPLC also allow for the detection of isobaric compounds as well as trace level materials which otherwise would be obscured by matrix effects in complicated sample matrixes like tissues. This application of laser desorption with a secondary ionization by capture into a liquid could be envisioned to be applied to AFM based laser desorption techniques where boosting the ionization efficiency is crucial for signal detection.

Thesis (M. Phil.)--University of Hong Kong, 2006. / Title proper from title frame. Also available in printed format.

Thesis (Ph. D)--Ohio State University, 2006. / Available online via OhioLINK's ETD Center; full text release delayed at author's request until 2007 Aug 16

M.S. / Environmental Science / A technique to measure arsenobetaine and arsenocholine in fish tissue by fast atom bombardment mass spectrometry was developed with particular attention to quantitative analysis. Experiments were performed which demonstrate analysis of the compounds desorbed directly from thin layer silica chromatography matrices, quantitative analysis of arsenobetaine in real fish samples, and accurate mass measurement of arsenobetaine in normal FAB/MS using peaks from the glycerol matrix as mass references. Improvements to the technique to quantitatively measure these important arsenic metabolites are suggested including optimization of the extraction/isolation procedures and use of isotopically labelled internal standards or surrogates for more accurate measurements.

Ph.D. / Organic Chemistry / The use of electron impact mass spectrometry for isotopic analysis is an established technique; however, its application to problems involving biologically produced molecules enriched in 13C has been severely limited. This limitation is shown to arise because of data reduction techniques which are, for practical purposes, incapable of dealing with the complex situation of multiple enrichment sites, limited isotope abundance, and complex spectral regions. Two approaches for detailed isotopic analysis of EIMS intensity are presented and illustrated using model data, literature data, and original data from biosynthetically enriched compounds. The first approach which allows for sequential variation of any one parameter is shown to be of value in experimental design through construction of ion cluster contour diagrams which summarize the variation of isotopic distribution, molecular size, and uncertainty in measurement with total isotopic content. The second approach differs in concept from previous methods and is especially promising for application to problems involving multiple sites of enrichment, limited isotopic abundance, and complex spectral regions. The key concept of this approach is that the roots of the polynomials constructed from observed spectral intensities are analytically related to the isotopic content and the isotopic distribution of the ions giving rise to the intensities.

Separations coupled to mass spectrometry (MS) are widely used for large-scale protein identification in order to reduce the adverse effects of analyte ion suppression, increase the dynamic range, and as a deconvolution technique for complex datasets typical of cellular protein complements. In this work, matrix assisted laser desorption-ionization is coupled with ion mobility (IM) separation for the analysis of biological molecules. The utility of liquid-phase separations coupled to MS lies in the orthogonality of the two separation dimensions for all analytes. The data presented in this work illustrates that IM-MS relies on the correlation between separation dimensions for different classes (either structural or chemical) of analyte ions to obtain a useful separation. For example, for a series of peptide ions of increasing mass-to-charge (m/z) a plot drift time in the IM drift cell vs. m/z increases in a near-linear fashion, but DNA or lipids having similar m/z values will have very different IM drift time-m/z relationships, thus drift time vs. m/z can be used as a qualitative tool for compound class identification. In addition, IM-MS is applied to the analysis of large peptide datasets in order to determine the peak capacity of the method for bottom-up experiments in proteomics, and it is found that IM separation increases the peak capacity of an MS-only experiment by a factor of 5-10. The population density of the appearance area for peptides is further characterized in terms of the gas-phase structural propensities for tryptic peptide ions. It is found that a small percentage (~3%) of peptide sequences form extended (i.e., helical or β-sheet type) structures in the gas-phase, thus influencing the overall appearance area for peptide ions. Furthermore, the ability of IM-MS to screen for the presence of phosphopeptides is characterized, and it is found that post translationally modified peptides populate the bottom one-half to one-third of the total appearance area for peptide ions. In general, the data presented in this work indicates that IM-MS offers dynamic range and deconvolution capabilities comparable to liquid-phase separation techniques coupled to MS on a time scale (ms) that is fully compatible to current MS, including TOF-MS, technology.

Mass spectrometry has shown its unique potential for studying the structure of proteins. Associated with various specific techniques (H/D exchange, ion mobility, gas-phase spectroscopy, multidimensional mass analysis), it has demonstrated to be an essential tool allowing primary structures to be analyzed and providing a lot of information about high order conformations. This work assesses the capabilities of these emerging mass spectrometry methods, and especially the gas-phase H/D exchange technique, for the structural analysis of nucleic acids. Gas-phase H/D exchange was first used to study single stranded oligonucleotides. The exchange reactions were performed with CD3OD in the collision cell of a 9.4 T FT-ICR MS. In these experimental conditions and in integrating the experimental and theoretical results, gas-phase H/D exchange was shown to be controlled by hydrogen accessibility and not by the chemical nature of the heteroatom bearing the exchangeable hydrogen. This allowed the presence of one structure or several conformers that possess different exchange properties to be detected. Moreover, when several structures were observed, increasing the internal energy of the ions at the entrance of the H/D exchange cell gave access to a qualitative estimation of the relative height of the isomerization barriers compared to the H/D exchange ones. Ion mobility experiments confirmed independently the H/D exchange results. Comparing the ion activation experiments for H/D exchange and for ion mobility revealed that the most compact conformer displays the fastest H/D exchange. This observation showed that H/D exchange and ion mobility provide us with complementary information because accessibility and macromolecule compactness are not univocally associated. Two other methods having independent principles of operations were sequentially combined. The fragmentation of a totally deuterated dinucleotide in exchangeable positions demonstrated the coexistence of several fragmentation channels. The latter were classified according to the involvement of non-labile or labile protons in the fragmentation process. Double resonance experiments were also performed and demonstrated the existence of consecutive fragmentation mechanisms. The involvement of labile, and therefore exchangeable protons in the fragmentation mechanism casts doubt on the use of tandem mass spectrometry to localize incorporated deuteriums in oligonucleotides. Finally, an exploratory work on the gas-phase H/D exchange of non-covalent complexes is presented.

A new laser ablation mechanism of liquid water based on recent insights into its hydrogen bond dynamics has been studied and several applications of the ablation demonstrated. The mechanism, termed as Impulsive Heat Deposition through Vibrational Excitations (IHDVE), is based on the ability of the hydrogen bond network of water to rapidly thermalize vibrational O-H stretch excitations on a time scale of several picoseconds even for excitation intensities that are large enough to bring excited volumes far into the supercritical region. In this way, by using vibrationally resonant picosecond infrared laser pulses with sufficient energy, it is possible to drive ultrafast phase transitions in the excited water volume leading to a rapid and efficient ablation process of water and water rich targets with minimum perturbation of solute molecules of interest. The physics behind the IHDVE ablation process is outlined and the benefits of the IHDVE ablation are demonstrated for two important applications of tissue cutting and mass spectrometry of biomolecules. Finally, the development of two high power infrared laser systems suitable for the practical implementation of IHDVE is presented.