Speciation of metals in proteins using gel electrophoresis with laser ablation-ICP-MS
A study into the applicability of laser ablation inductively coupled plasma-mass spectrometry (LA ICP-MS) coupled with native polyacrylamide gel electrophoresis (PAGE) to metal speciation in proteins is described in this thesis. Chapter one of the thesis outlines the various roles played by metal ions in clinical samples. Health risks can arise from anthropogenic metal enrichment and metallodrug therapy is identified as a key source. The use of platinum and gold metallodrugs is discussed in detail. Speciation of metals in clinical samples is examined, concentrating on some of the techniques currently employed and their limitations. Particular attention is paid to the techniques employed in the study. The aim of the study is outlined - to develop an alternative speciation strategy for the study of metals in proteins. The reagents and instrumentation used are described in chapter two along with all experimental procedures. Chapter three describes the method development and determination of the analytical performance of the technique. The technique is used to study platinum speciation of protein samples enriched in vitro, the metal distribution profiles obtained are shown. The technique is next applied to the analysis of in vivo samples obtained from platinum therapy patients and a control source. Chapter four outlines the application of the technique to analysis of gold enriched samples both in vitro and in vivo from chrysotherapy patients. The gold distribution profiles obtained are shown and discussed. Chapter five looks at multi-element distribution and interactions occurring in serum. Elements naturally occurring at trace levels in serum, such as Cu and Fe, are studied in vivo. Other, lower level metals are studied following in vitro enrichment. Chapter six concludes the study. It discusses how the technique was found to be applicable for metal speciation of clinical samples by meeting the criteria laid out at the start of the study. Recommendations for future work are also outlined.