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Title: Separation and identification of biological molecules using chromatographic and mass spectrometric techniques
Author: Martin, R. L.
Awarding Body: University of Wales Swansea
Current Institution: Swansea University
Date of Award: 1999
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Chapter One reviews the historical developments and current technologies of mass spectrometry and chromatography. Chapter Two introduces proteins and peptides, amino acid properties, the elucidation of amino acid sequences and mass spectrometric methods of analysis. The effect of trifluoroacetic acid on electrospray response is discussed and a system designed to compensate for its detrimental effects is demonstrated. Tryptic digests of a number of peptides and proteins are analysed to determined the efficiency and efficacy of the system, resulting in the confirmation of a post-translational modification. Chapter Three discusses the principles of matrix assisted laser desorption ionization time-of-flight mass spectrometry. An investigation is undertaken into the feasibility of C-terminal sequencing of peptides and proteins using on-plate enzyme digests and MALDI-TOF-MS. The net result is a turnaround time of around 30 minutes including sample preparation, digestion time and analysis of the resultant fragments. The identification of the C-terminal sequence of modified hirudin is performed. Chapter Four describes an investigation into protein characterisation using immobilised enzymes and MALDI-TOF mass spectrometry. The system is evaluated using small peptides and a relatively large protein, Cdc4. The average digestion time has been reduced from the 18-24 hours usually required to perform conventional solution-phase techniques to only 2-3 minutes. Chapter Five details an examination of double-ionization energies of alkynes, namely 2-butyne, 2-pentyne, 2-hexyne and 3-hexyne, using double-charge-transfer spectrometry. The principles and background of the technique are explained, followed by a description of experimental methods. In addition, a theoretical investigation is performed using ab initio calculations and a multiple-scattering Xα method. Experimental data is presented in conjunction with a comparison of theoretically obtained results.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available