Metallomics is the study of the metallome, interactions, and functional connections of metal ions and other metal species with genes, proteins, metabolites, and other biomolecules in biological systems. Its application can handle a variety of sample, from biomedical to environmental. Elemental speciation is a fundamental component of the concept of Metallomics. It permits to investigate the chemical form of metals and metalloids containing species.;Speciation analyses with metallomics approaches can be performed best by high performance liquid chromatography (HPLC) coupled to inductively coupled plasma mass spectrometry (ICP-MS). HPLC represents a suitable separation technique and a variety of different chromatographic techniques, from conventional to capillary, can be applied to obtain good baseline separated peaks corresponding to different species. ICP-MS is the state-of-the-art instrument when elemental mass spectrometry for the detection of ultra trace levels (parts per quadrillion) takes place. ICP-MS is renowned for its phenomenal sensitivity and selectivity as elemental analyzer. Chromatography can be applied to molecular mass spectrometry as well. In the presented works electrospray ionization is the technique of choice. Nano liquid chromatography-Chip-electrospray ionization-ion trap mass spectrometry (NanoLC-Chip-ESI-ITMS) is an excellent tool for the research performed.;The aforementioned instrumentations have been applied to environmental applications in the first part of this dissertation. Chemical warfare agents (CWAs) are present in the environment (sea and soil) because discarded due over production during World War I and II. Those CWAs easily degrade, especially in aquatic surrounding, and give birth to chemical warfare agent degradation products (CWADPs) whose toxicity is not well know. In the following pages a complete study about some of these CWADPs and their speciation and toxicity is reported.;The second part of the dissertation analysis a more complex matrix with metallomics approaches. The combination of subarachnoid hemorrhage (SAH) with cerebral vasospasm (CV) leads to severe debilitation or death of an estimated one million people worldwide every year. A biomarker that would predict CV after a SAH has yet to be found.;The focus of this study is to explore differences in protein phosphorylation in cerebral spinal fluid (CSF) among healthy patients, SAH patients and SAH-CV patients. A significant difference in the phosphorproteom could be one step towards the discovery of a diagnostic marker that may predict CV after SAH. The significance of phosphorylated proteins as a marker is manifested in the constitutive nature of intracellular signaling involved in the pathological events seen post SAH.;Capillary liquid chromatography (cap-LC) coupled to inductively coupled plasma mass spectrometry (ICPMS) and nanoliquid chromatography-PhosphoCHIP/ion trap mass spectrometry (nanoLC-P-CHIP/ITMS) are used to identify and measure protein phosphorylation changes in the CSF of the aforementioned three groups. ICPMS represents a suitable method for screening phosphorous (31P), while nanoLC-P-CHIP/ITMS can be used to identify phosphoproteins and match these with protein database entries.
展开▼
