Development of novel analytical methods to study the metabolism of coumarin

The research in this thesis revolves around developing analytical methods for the determination of coumann and 7-hydroxycoumann for various applications. The techniques used in this work were, capillary electrophoresis, immunosensing and electrochemistry. Chapter 1 serves as general review of the analysis of coumann and 7-hydroxycoumann, including the many different types of analytical technique which have been used to analyse this drug. Capillary electrophoresis was used as the basis of a method for the determination of 7-hydroxycoumann in unne and serum. This method offered advantages over existing technology, due to the speed of analysis compared to existing methods Capillary electrophoresis was also used to study of the metabolism of coumann by liver microsomes. This work proved very interesting with a novel method being developed for the determination of the mam metabolite of coumann in humans This method was also applied to an mterspecies study on the metabolism of coumann m a vanety of species, while it also found application m tiying to understand some of the different reactions which take place in hver microsomes Electrochemistry was used to study further some microsome reactions with some success. A biosensor was constructed for the determination of 7-hydroxycoumann which used electrochemical detection. An immunosensor based on a competitive electrochemical immunoassay was developed. The antibody-based biosensor employed horseradish peroxidase-labelled anti-7- hydroxycoumann, with the enzyme catalysed reaction involving the reduction of hydrogen peroxide m the presence of a mediator.