Thesis submitted to the Division of Science and Engineering. Bibliography: leaves 154-174.

Risk assessment of environmental exposure to chronic, trace concentrations of contaminants presents an analytical challenge to interpret data in a biologically meaningful way. Biomarkers are compounds that can provide integrated information concerning the effects of contaminants on biochemical processes. The metal-binding protein, metallothionein (MT), is a biomarker of toxic heavy metals, such as cadmium (Cd), since these metals bind to MT in vivo and induce transcription of MT genes with subsequent MT protein synthesis. A high performance liquid chromatography (HPLC)-thermospray microatomization-atomic absorption spectroscopy (AAS) method was developed for detecting Cd proteins from two invertebrate models; freshwater mussels (Pyganodon grandis) from a whole lake ecosystem exposure to Cd (Experimental Lakes Area, Canada), and zebra mussels (Dreissena polymorpha) recently introduced to North America. Methods for coupling gel filtration HPLC to AAS or inductively coupled-mass spectrometry (ICP-MS) were developed to provide sensitive and selective information on metal-binding proteins in freshwater mussel extracts. Sensitive metal detection by these methods necessitated precautions to minimize HPLC column-protein exchange of metals. These interactions were relevant to the determination of low concentrations of mussel MT-like proteins (MLPs) by these metal-specific detection systems. Saturation methods employing Cd as a metallic marker for the quantification of characterized MTs were adapted for freshwater mussels exposed to low environmental Cd concentrations. Characterization of Cd-saturated mussel extracts by HPLC-ICP-MS demonstrated the presence of copper and zinc, metals physiologically bound to MT, in the principal metal-binding fraction of mussel extracts. Experimental results also indicated that mussel MLPs are not as heat-stable as mammalian MT. Zebra mussels were chosen as a bioindicator species for obtaining a standard of freshwater MT biomarker for toxi

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