Direct Infusion Lipidomics: Profiling the Lipidome of a Composite Tailings Reclamation Site

Abstract

<p>The comprehensive analysis of intact lipids (called lipidomics) can provide information about the presence of microbial communities in an ecosystem and assist in understanding the biogeochemistry in that system. In previous work we had developed a method to determine the profiles of eight phospholipid classes in a soil microorganism by direct-infusion electrospray mass spectroscopy using tandem mass spectrometry. The work done in this study encompasses first the optimization of previous methodology for use with water and sediment samples containing low concentration of phospholipids and large amounts of organic contaminants and secondly the application of this method to the analysis of phospholipids within composite tailings and recycled process water using a triple quadrupole mass spectrometer to determine the intact lipids in the bacterial community. The results are presented illustrating the phosphatidylcholine (PC) and phosphatidylethanolamine (PE) lipids present in composite tailing samples and recycled process water. This thesis begins with the optimization of a direct infusion mass spectrometry method, which allowed the analysis of intact phospholipids within both water and sediment samples. This method allows for high through-put analysis using both the separation afforded by neutral loss and precursor ion scanning modes and a database containing all possible adduct masses to identify and quantify unknown phospholipids. This method was then applied to water and sediment samples obtained from the Syncrude Sandhill Fen composite tailings site. This analysis discovered multiple differences within the water samples attributed to changes both in well temperature and the ongoing reclamation projects resulting in the change in phospholipid profiles. This thesis also outlines the shortcomings of the direct infusion lipidomics method when used for the analysis of complex samples such as composite tailings sediment samples. In summary, this thesis has demonstrated that direct infusion lipidomics can be successfully applied to the analysis of water samples and yield statistically significant differences within the microbial lipidome.</p>