Metallomics is the study of the metallome, interactions, and the functional connections of metal ions and other metal species with genes, proteins, metabolites, and other biomolecules in biological systems. It has been applied to a variety of samples from biological to environmental. Elemental analysis along with elemental and molecular speciation is a major component of metallomics experiments.
Molecular speciation analysis is performed best with high performance liquid chromatography coupled to inductively coupled plasma mass spectrometry (ICPMS) in conjunction with molecular mass spectrometric techniques. A wide range of liquid chromatography techniques including capillary and conventional to hydrophilic interaction liquid chromatography to strong cation exchange chromatography can be applied for the separation of biological samples. ICPMS is the gold standard for elemental analysis at trace levels (ppt for most elements) and it is known for its sensitivity and selectivity. Liquid chromatography can also be combined with molecular mass spectrometry as well to offer excellent molecular identification of biomolecules. In this work, LC-MALDI-TOF/TOF and LC-ESI-LTQ-MS where both used for protein identification.
The first part of this dissertation focuses on novel ways to separate and identify phosphorothioated oligonucleotides. Hydrophilic interaction liquid chromatography was applied for a novel separation technique and coupled to both ICPMS and ESI-LTQ-MS for their identification and quantification. Both methods offer low detection limits and a fast and easy way for identification.
The second part of this dissertation focuses on metallomics and proteomics techniques for the analysis of a more complex matrix of cerebral spinal fluid (CSF). Subarachnoid hemorrhage (SAH) and its sequela cerebral vasospasm (CV), kill or seriously debilitate an estimated 1.2 million people of all ages, ethnic groups and gender annually. The ability of these techniques to help identify biomarkers has the potential to save lives but no biomarkers for CV have been identified yet.
The focus of the first project was to explore CSF for selenoproteins that could play a role in the etiology of CV. Both elemental and molecular along with multidimensional separation techniques were applied for this exploration. The focus of the second project was to identify proteins that had significant differences between three samples types with common proteomics techniques that include multidimensional separation techniques and two different molecular mass spectrometric techniques.