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Title: Applications of mass spectrometry to biomolecular and physical studies
Author: Ghosh, D.
Awarding Body: University College of Swansea
Current Institution: Swansea University
Date of Award: 1991
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The primary objective of this investigation was to evaluate the utility of fast atom bombardment (FAB) and electron impact (EI) tandem mass spectrometry (MS/MS) for the analysis of biomolecules. In this thesis, an introductory overview is followed by descriptions of the mass spectrometric examination of a number of cyclic nucleotides and mononucleotides, together with the differentiation of isomeric ribomononucleotides by FAB and collision-induced dissociation mass-analysed kinetic energy spectrometry (CID-MIKES). The techniques thus developed were then utilized to identify putative cyclic nucleotides extracted from plant tissue. In addition, tandem mass spectrometric analysis was also used to verify successful derivatization of cyclic nucleotides, specifically the lipophilic dibutyryl derivatives necessary for pharmacological studies of the effects of cyclic nucleotides. Further to the successful application of mass spectrometric analysis to qualitative biochemistry, it was applied to quantitative study of cyclic CMP phosphodiesterase. By selecting the protonated molecular ions of both substrate and product, then measuring the peak heights of diagnostic fragments in the daughter ion spectra of both spiked and unspiked samples, it was possible to calculate kinetic parameters for the reaction, these parameters showing excellent correlation with those determined by conventional radioisotopic assay. The feasibility of a dynamic FAB system for the continuous assay of these enzymes was also examined. The stereochemistry of epimeric steroids was investigated by EI MS/MS. Configuration of hydroxy steroids of cholestane, cholane and androstane series were shown to be distinguishable on the basis of the CID MIKE spectra of metastable (M - H2O)+. ions as well as by comparisons of translational energy release values. Glassy carbons (phenol-formaldehyde resins) which are important materials in electrochemistry were analysed by negative ion FAB and compared with graphite samples. Carbon clusters up to C-/15 were observed in both materials with little difference discernible between their respective mass spectra.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available