The first section of this thesis presents a review of the literature on endocrine hormones, synthetic steroids, and synthetic steroid receptor antagonists. Particular emphasis is placed upon the estrogen, progesterone and glucocorticoid activity of these compounds. The structure, activity and preparation of some progesterone/glucocorticoid receptor antagonists is also reviewed. The second section of this thesis presents a route for the synthesis of 19-aryl substituted androstanes. These compounds have been designed to explore their potential antiprogestational and antiglucocorticoid activity. It is demonstrated that a bulky aromatic functionality may be introduced at the sterically hindered C-19 methyl group, by the nucleophilic attack of an organometallic nucleophile on a 10~-formyl-19-norandrostane, which is suitably protected at positions C-3 and C-17. The subsequent careful manipulation of the C-3 and C-17 functionalities has led to a succesful 11 step synthesis (scheme 4.19) of 17~-hydroxy-17a-(prop-l-yne)-19-(p-N,N- dimethylaminophenyl)androst-5-en-3-one (3.1), which should be of interest for biological evaluation.

The work described in this thesis concentrates on chemically modified heterocyclic conducting polymers based upon pyrrole, thiophene and indole monomers with -particular emphasis placed upon carboxylic acid substituents. A brief description of the electrochemistry of chemically modified poly(pyrroles) is given with an explanation of the problems associated with the chemical modifications. A reasonable understanding of the properties of chemically modified poly(thiophenes) has been achieved. In particular the properties of poly(3-thiopheneacetic acid) have been characterised in both aqueous and nonaqueous solution using standard electrochemical techniques in conjunction with reflectance FFIR studies. Finally the understanding of poly(indole) and poly(5-carboxyindole) electrochemistry has been greatly improved especially with regards to the chemical structure of each polymer, which was elucidated from several reflectance FTIR studies. The characterisation of the electrochemical growth and the aqueous electrochemistry of poly(5-carboxyindole) are the areas in which the greatest advances have been made. The techniques of reflectance FTIR spectroscopy, UV/vis spectroscopy and impedance spectroscopy have aided the study of poly(5-carboxyindole) immensely. More studies need to be performed before a fuller understanding of these polymeric systems is achieved and the final chapter gives suggestions for further work which will add to the information given in this thesis.

Mass spectrometry and tandem mass spectrometry (MS/MS) has been employed to analyse peptides (<600 daltons), synthetic polymers of low molecular weight (<10,000 daltons) and a mixture of polymer additives (300-1200 daltons). Mass spectrometry experiments were performed on a four sector mass spectrometer, a tandem quadrupole mass spectrometer and a time-of-flight instrument. High and low energy collision induced dissociation (CID) spectra were obtained by means of a four sector mass spectrometer and a tandem quadrupole instrument respectively. Surface induced dissociation (SID) spectra of peptides were obtained by means of a four sector mass spectrometer with a modified collision cell in the third field free region. Sequence data were generated by SID from protonated and cationated precursor ions of all four peptides analysed. Furthermore, broad metastable ion peaks were observed in the spectra, arising from fragmentation of precursor ions in the field free region between the electric sector and the magnetic sector of the second mass spectrometer. Field desorption was a good ionisation technique for the generation of molecular weight information from the polymer additives used. High energy eID was found to be more applicable than low energy eID to the structural determination of polymer additives as characteristic ions were observed in the spectra. Mechanisms have been proposed for the generation of some of the fragment ions observed. Ultraviolet-matrix assisted laser desorptionlionisation spectra of synthetic polymers of low molecular weight (<10,000 daltons) were used to calculate molecular weight averages. Furthermore, end group information was also obtained from CID spectra of the same polymer samples. End group and structural information was also obtained from polystyrene and a substituted polystyrene by means of FD-MS/MS.

This thesis describes the development and application of hydrodynamic modulation voltammetric (HMV) techniques coupled to ultramicroelectrodes (UMEs) that possess intrinsically high mass transport rates in quiescent solutions. This study demonstrates that the well defined convective-diffusive conditions of the microjet electrode (MJE) arrangement allows mass transport to be enhanced by almost two orders of magnitude compared to a 25 mm diameter disc-shaped UME. The MJE comprises a nozzle which is used to deliver solution to a UME surface at high velocity. Scanning electrochemical microscopy (SECM) with small UMEs has been used to image the hydrodynamics of the jet system with high precision. Variations in local mass transport for both IrCl63- and Fe(CN)64- oxidation at a range of flow rates has been observed at various positions within the impinging jet and the stagnation zone has been thoroughly characterised under a variety of experimental conditions. Agreement has been found between experiment and theory for voltammetric data recorded with the nozzle and UME aligned in the stagnation zone, for a range of viscous solutions examined. By modulating the mass transport rate to the surface of an UME, in the MJE arrangement, by the introduction of a rotating blade between the end of the nozzle and the UME, it was possible to enhance the current sensitivity of the system. Trace level detection, to 2 x 10-7 mol dm-3 IrCl63- solution, was readily facilitated. This type of HMV experiment has utilised two methods to provide the reference signal for phase-sensitive detection of the current signal, involving either a dual-disc electrode or a single UME coupled to an LED detection system. Both HMV methods have been shown to work well.

In this thesis, the work undertaken in an attempt to gain insight into the location of the binding site of bilirubin is described. The history, properties and biosynthesis of both bilirubin and albumin are reviewed in Chapter One. Chapter Two contains an account of the novel one-step method of labelling bilirubin of high yield and specific activity. In Chapter Three, the effect of light on bilirubin in the presence and absence of albumin, followed by the labelling of the albumin in an attempt to obtain information regarding the nature of the amino acid residues involved in the binding of bilirubin to albumin. Attachment of labelled bilirubin to albumin enabled the possible site on the latter to be followed during degrading procedures. Possible effect of this photo-induced binding on phototherapy is described. Chapter Four contains an account of the synthesis and binding studies of mesobilirubin. Fluorescence and spectroscopic methods are employed to determine the association constants of mesobilirubin and comparison with bilirubin made. The importance of the vinyl groups in bilirubin binding is cited and the possibility of mesobilirubin being a kernicteric pigment is hypothesized.

Liquid polysulfide polymers (LPs) H(SCH2CH20CH20CH2CH2S)nH are of wide commercial importance in the sealants industry. Chemical oxidation of the mercaptan groups, by inorganic and peroxide catalysts, leads to the formation of durable, flexible and adhesive elastomers. Terminal modification of the prepolymers, by substituting a wide range of carbonylcontaining groups for the thiol protons, has led to the creation of photopolymerisable oligomers with certain of the desirable properties mentioned above. UV irradiation of all the modified polysulfides, which exist as viscous oils, resulted in the development of a solid elastomeric material with considerable powers of adhesion. The acylation process involves the addition of a slight excess of an acyl halide to the prepolymer in solution, followed by the addition of an amine to neutralise the HCI formed. This procedure has been adapted to react di- and polyfunctional termini with LPs to produce block copolymer polysulfides. When exposed to UV irradiation, the oligomers slowly photocrosslink, forming an insoluble elastomer. The increase in degree of polymerisation has been successfully monitored by 1H NMR spectroscopy; this was achieved by irradiating the oligomers in NMR tubes containing a thin capillary tube containing a deuterium-lock solvent. These new materials were characterised by IR, 1H and 13C NMR, GPC, viscometry and, particularly electrospray and FAB-mass spectrometry. FAB spectra have also been obtained for the first time for the parent polysulfide as well as the modified forms; these enable characterisation of the polysulfide backbone and its irregularities. Characterisation of these cured products has been made by hardness testing, solvent resistance, flexibility and tensometry. Lap shear measurements have been made of all of the functionalised cured polysulfides; adhesion is greatly enhanced by the addition of an adhesion promoter (optimum level 2%). All the systems slowly photopolymerise without sensitisers or additives, but the addition of radical photoinitiators shortens the cure time considerably. The modified LPs can be combined with commercially available polyurethane systems to make a fast curing photosystem yielding products with excellent adhesion and flexibility. Each of the modified LPs contains a carbonyl group, which has photoactive potential, thus excitation of the carbonyl group leads to an n->π* transition to form a reactive triplet state which then attacks another LP chain to crosslink the polysulfide. Monitoring of 1H NMR linewidths shows a gradual broadening of lines as 120 Hz polymerisation proceeds, reaching a plateau value of ΔH1/2=120 Hz, illustrating the elastomeric properties of the polysulfide. Addition of plasticisers eases the uniform spreading of the oligomers onto sheets before irradiating, which gives a potential application as a surface coating. Photoresist imaging agents are another potential application as the modified oligomers act as negative resists.

The analysis of polymer systems is a crucial field in modern petrochemicals and also other sectors of the chemical industry. This analysis stretches from simple mass measurement, to sample identification and structural studies. Mass spectrometry is one of a number of analytical fields that are used to investigate such samples, but it has been shown to have certain limitations for the routine analysis of polymers. Chapter one discusses the development of mass spectrometry and the progression that has led to the development of the methods of sample ionisation and separation that are most used today. It also discusses the field of polymer analysis and the other analytical techniques that are applied to such investigations in industry. Chapter two details the instrumentation used in this study and also the nature of the main samples that were investigated. Chapter three covers analysis by the technique of matrix-assisted laser desorption / ionisation which is the major mass spectrometric method currently for polymer analysis. Details of work investigating sample preparation arc given leading into a larger discussion of the mass biases that arise in such polymer analysis and a rationalisation of their causes. Chapter four details investigations of polymer analysis using an electrospray source on a Fourier-transform ion cyclotron resonance instrument. The added complexity of charge states is discussed leading to further study of the mass and charge variations given by instrumental conditions. Chapter five presents a small body of work in the field of structural analysis by the technique of collision-induced dissociation. Work with buckministerfullerene and a range of ethoxylate polymers is given to demonstrate the capabilities of such analysis. Chapter six concludes the thesis with a discussion of the topics raised in the previous chapters and how they affect the use of mass spectrometry as a routine tool for the industrial analysis of polymeric systems.

This thesis describes the development of new techniques and new approaches for studying the kinetics and mechanisms of reactions which occur at the interface between two immiscible liquids. New approaches for studying the kinetics of transfer processes at the air/water interface are also described. Scanning electrochemical microscopy (SECM) is employed in the equilibrium perturbation (BP) mode for studying reversible transfer processes at liquid/liquid and air/water interfaces. In this application, an ultramicroelectrode (UME) located in an aqueous phase, at micro metre distances from the interface of interest, is employed to drive a transfer process, initially at equilibrium, in the direction of the aqueous phase by depleting the local aqueous concentration of a target species by electrolysis. The UME current flow depends on the transfer kinetics at the interface. The development of a submarine UME allows SECM to be conducted at the air/water interface, or at a liquid/liquid interface with the electrode in the more dense phase. The kinetics of the extraction/stripping reactions of aqueous copper (II) with an oxime ligand (Acorga PSO) in 1,2-dichloroethane (DCE) and heptane are first investigated. Subsequently, measurements of the kinetics of oxygen transfer across a condensed monolayer of l-octadecanol, as a function of surface area, demonstrate that the accessible free area of the interface primarily governs the rate of oxygen transfer. SECM double potential step chronoamperometry is developed to study irreversible transfer processes at interfaces. Theoretical modelling is applied and tested through experimental measurements on model interfaces. Subsequently, the rate of transfer of bromine from aqueous solutions to DCE and to air is shown to be above the upper limit measurable by the technique, however, a lower limit on the first order transfer rate constant ofO.S cm S-l is assigned. A new technique, termed microelectrochemical measurements at expanding droplets (MEMED) is developed for studying spontaneous reactions at liquid/liquid interfaces. In MEMED, the two liquids are contacted by flowing one (feeder) through a capillary submerged in the second (receptor), resulting in the growth of drops at the capillary tip. The interfacial reaction generates a product or reactant concentration profile, which extends into the receptor phase. This is probed directly using a UME positioned opposite the capillary in the solution, operated in either a potentiometric or an amperometric mode, as a local concentration probe. A numerical model for mass transport in this configuration is developed, and the technique and model are assessed by measuring both bromine transfer from aqueous sulfuric acid solutions to drops of DCE, and bimolecular electron transfer between iridium (IV) chloride in the aqueous solution and ferrocene in the organic phase, which exhibit transport-controlled transfer rates under the conditions employed. The MEMED technique is applied to measure the kinetics of the hydrolysis of triphenylmethyl chloride (TPMCl), at the DCE/water interface, through potentiometric measurement of the chloride ion concentration profile. The reaction is shown to be first-order in TPMCI, occurring interfacially, with a rate constant of 6.5 x 10-5 cm s". Subsequently, the oxidation of methylanisole (MA) (feeder) by aqueous solutions of cerium (IV) (Ce(IV» (receptor), is probed. Under the conditions of this study, the reaction at the interface dominates, with a negligible contribution from the aqueous phase reaction between dissolved MA and Ce(IV).

Streptomyces subrutilus ATCC 27467, when grown on a glucosecontaining soyabean medium, produces both 1-deoxymannonojirimycin (DMJ) and 1-deoxynojirimycin (DNJ) in its culture medium. When 1- or 2-[2H]-D-glucose is used, the deuterium label appears at C6 in both alkaloids and the labelling pattern suggests that the first step in the biosynthesis of both DNJ and DMJ is a glucose to fructose isomerisation. Studies with 5-[2H]-D-glucose and 6,6-[2H2]-D-glucose indicate that oxidation of the 6-position of the glucose/ fructose occurs during the biosynthesis and that mannonojirimycin is the first amino sugar to be formed. Mannonojirimycin can then undergo dehydration and reduction to DMJ. Alternatively, epimerisation of mannonojlrlmycin can occur at C2 to give nojirimycin which is then dehydrated and reduced to DNJ. Studies with another microorganism, B. subtilis var niger ATCC 9372, indicate that a similar biosynthetic pathway is in operation. DMJ, however, is not produced by this microorganism and only low levels of NJ are postulated from enzyme inhibition and deuterium labelling studies. A minor biosynthetic route is also evident from labelling studies with 1-[13C]-D-glucose and 1-[13C]-D, L-glyceraldehyde. It is suggested that the fructose at the beginning of the biosynthesis can split into two C3 trioses which are in equilibrium with each other. These can then recombine to continue in the usual biosynthetic pathway. New chemical routes to 542H]-D-glucose and 2-[2H]-D-glucose are described in Chapter 3 along with a method for introducing deuterium or tritium into DNJ or NJ. Other isotopically enriched glucoses or intermediates have also been prepared using literature methodology. Studies have been undertaken to assess whether S. subrutilus can utilise glucose analogues in the biosynthetic pathway. To this end, new or modified routes to these glucose derivatives have been investigated and some work has focused on the chemical synthesis of DNJ analogues themselves.

The thesis presents a review of the literature on the HPLC resolution of enantiomers using chiral stationary phases and discusses the mechanisms of separation and the utility of these phases. The synthesis and chemical, chiroptical and spectroscopic properties of helicenes are also reviewed and the potential utility of helicene-based for the HPLC resolution of enantiomers is discussed. The work carried out involved the preparation of a hexahelicene-based chiral stationary phase and demonstrations of its utility for the resolution of chiral analytes. This phase was chemically bonded rather than physically coated, in order to make it stable to hydrolysis and solvent stripping, and therefore to permit its employment with a wide range of mobile phases. This thesis describes the synthesis of hexahelicen-7-ylacetic acid methyl ester, including confirmation of the structure of the key synthetic intermediates by spectroscopic analysis, and the investigation of several proceduresfor the resolution of hexahelicen-7-ylacetic acid methyl ester into its enantiomers, one of which enabled around 100 mg amounts of each of (+) and (-) enantiomers of hexahelicen-7-ylacetic acid to be obtained in highly chemically and optically pure form. The thesis also gives an account of several synthetic approaches to covalently bonding of the chiral selector (hexahelicen-7-ylacetic acid) to a modified silica stationary phase. Chiral stationary phases were prepared from each of the enantiomers in sufficient amount to permit the packing of analytical columns of these phases. Chiral solutes containing nitroaryl functionalities were synthesised to investigate the chiral resolving power of these novel phases. Adequate separations were obtained with both chiral stationary phases and, as anticipated, the eluting order of chiral analytes was reversed between the (+) and (-) stationary phases. The work demonstrated the utility of these columns for the resolution of nitroaryl-containing chiral analytes.