The contamination of 316L stainless steel surfaces within an electrospray ionisation source of a mass spectrometer is investigated. An accelerated method of contamination is used. Following initial test method development and investigation of the contamination resulting on the ion transfer components (sample cone, outer cone and extraction cone), flat samples are employed within the ionisation source. This enables characterisation of the contamination composition, morphology and build-up with time. Blood plasma is introduced into the mass spectrometer as it is a widely analysed substance that is known to result in contamination. The contamination from a mixture of human blood plasma, diluted in methanol, and a water/acetonitrile mobile phase is found to contain inorganic NaCl crystals embedded in a matrix of organic residues. The morphology shows self-organising features as the contamination builds. A model is proposed to explain the morphology, involving rapid evaporation of the droplets that impinge on the stainless steel surface. Two types of surface modification are considered for the stainless steel: electrochemically grown films and coatings deposited by vapour deposition. A method for electrochemical film growth is developed, enabling nanoporous films to be formed on the stainless steel in 5 M sulphuric acid at 60°C by square wave pulse polarisation between active or transpassive and passive potentials. The films are characterised using glow discharge optical emission spectroscopy, X-ray photoelectron spectroscopy, scanning electron microscopy, Rutherford backscattering spectroscopy and nuclear reaction analysis. The films are shown to be chromium- and molybdenum-rich relative to the substrate, and to consist mainly of sulphates, oxides and hydroxides. The morphology and composition of the films are discussed in relation to the polarisation conditions and mechanism of film formation. A range of vapour deposited coatings are considered: TiN, TiC, TiB2, Graphit-iC, and diamond-like carbon coatings with Si and N2 dopants and with varying sp2:sp3 ratios. In addition, a hydrophobic coating is deposited on the stainless steel by immersion, in order to provide a significant variation in surface energy. Surface analysis of the coatings is carried out, considering their sp2:sp3 ratios, their electrical conductivities, their water contact angle, and the various components of the surface energy. The contamination build-up on the surface of uncoated 316L stainless steel is compared with that on stainless steel with the various surface modifications. A method for quantification of the build-up of contamination on flat samples is developed using white light interferometry. The surface modifications which result in the slowest contamination build-up with time are then applied to the ion transfer components of the mass spectrometer. The robustness of the mass spectrometric response for the selected coated surfaces is compared with that of the uncoated stainless steel. The electrochemically grown films and two of the doped diamond-like carbon coatings are found to be successful in reducing the build-up of contamination.

Non-targeted metabolite profiling by liquid chromatography-mass spectrometry (LC-MS) was used to determine the metabolite responses of Arabidopsis roots to auxin or ethylene. Crosstalk between these hormones regulates many important physiological processes in plants, including the initiation of lateral root formation and the response to gravity. These occur in part through alterations in the levels of flavonoids, specialized plant metabolites that have been shown to act as negative regulators of auxin transport. However, much remains to be learned about auxin and ethylene responses at the level of the metabolome. LC-MS analysis showed that a number of ions changed in response to both hormones in seedling roots. Although classes of specialized metabolites such as flavonols and glucosinolates change in abundance in response to both auxin and ethylene, there was little overlap with regard to the specific metabolites affected. These data will be integrated with information from transcriptomic and proteomic experiments to develop framework models that connect phytohormones and specialized metabolism with specific physiological processes. Previous studies by imaging techniques have shown that flavonols increase in response to both auxin and ethylene in the root elongation zone, but LC-MS showed that flavonols decreased in abundance in response to these hormones. Therefore a method was developed for targeted metabolite profiling of flavonols in individual root tips by flow injection electrospray mass spectrometry. This method uncovered spatial differences in metabolic profiles that were masked in analyses of whole roots or seedlings, and verified that flavonols increase in response to these hormones in root tips. / Master of Science

La présence de contaminants métalliques dans les produits pétroliers engendre des effets indésirables, comme la dégradation de leur stabilité au stockage. Pour limiter ces perturbations, des additifs sont incorporés aux produits pétroliers dans le but de complexer les cations métalliques en solution. Le sujet de la thèse porte sur le développement d’une méthodologie pour étudier les interactions non covalentes dans une matrice hydrocarburée par spectrométrie de masse électrospray (ESI-MS). Plusieurs approches ont été utilisées pour comparer le pouvoir complexant de différentes molécules vis-à-vis des métaux ciblés, puis identifier les structures chimiques les plus efficaces pour inhiber l’action catalytique des contaminants. Ces travaux mettent en évidence l’apport de l’ESI-MS comme outil de présélection d’additifs, moins coûteux et chronophage que les méthodes actuelles de screening. / The contamination of petroleum products by metallic compounds leads to adverse effects, as the degradation of their stability under storage. To prevent these phenomena, specific molecules are added to petroleum products in order to complex metallic cations in solution. This work focus on the development of an electrospray mass spectrometry (ESI-MS) method to study non covalent interactions in a hydrocarbon medium. Several approaches were employed to compare the complexing ability of different molecules towards targeted metals, then to identify the most efficient chemical structures to inhibit the catalytic action of contaminants. Results highlight the contribution of ESI-MS as a screening tool, less expensive and time-consuming than the methods currently used.

Analytical method validation is the process of establishing that an analytical technique is applicable for a proposed objective. Early in the method development of a new analytical technique an understanding of the instrumental components and procedures is elaborated through scientifically based optimization. The optimization experiments are used to define the operational parameters that yield the maximum performance by the analytical technique for the target analyte before commencing validation studies. This dissertation details method development through experimental investigations instrumental components of LEMS (substrate, laser parameters, and electrospray source conditions). Each instrumental component has a number of induvial parameters which are optimized to yield the maximum laser electrospray mass spectrometry (LEMS) signal intensity for a given analytical problem. LEMS uses a nonresonant, femtosecond (fs) laser to ablate analytes from a surface. Those ablated analytes are then captured by a perpendicular electrospray, ionized, and desolvated to produce ions which travel into the inlet of the mass spectrometer for analysis. Each element of the LEMS experimental setup works in a complementary fashion to generate a mass spectral signal which have specific optimization steps that can dramatically impact the data that can be acquired. The results of the optimization for each instrumental component will then be applied to preliminary method development experiments for the analysis of pharmaceutical compounds from complex formulations biomarker discovery for mice afflicted with a traumatic brain injury.The effect of the laser pulse duration on the ablation mechanism and amount of laser induced conformational changes of aqueous myoglobin was investigated using 55 fs, 56 picosecond (ps), and 10 nanosecond (ns) pulses and laser pulse energies from 0.05 to 1.6 mJ. It was found that the optical properties of the substrates (stainless-steel and quartz) and laser intensity regimes accessible by each pulse duration determined the amount of myoglobin ablated and subsequent mass spectral signal intensity. Laser ablation of myoglobin from both substrates using all laser pulse energies was observed for the 55 fs pulse while the 10 ns pulse required minimum pulse energies of 0.4 and 1.2 mJ for ablation of myoglobin to occur from stainless-steel and quartz, respectively. As the pulse duration increases, thermal processes increase which dictated the relative amount of protein unfolding, number of phosphate adducts, and degree of solvent adduction. Many of the common laser electrospray ionization (ESI) hybrid techniques employ ns pulse durations. However, the amount of ablated myoglobin originating from a ns pulse was observed to be dependent on the amount of energy that was absorbed by the substrate or sample. Experiments to increase the signal intensity while implementing ns laser electrospray mass spectrometry (ns-LEMS) were performed by exploiting the optical properties of nanomaterials as a potential matrix for desorption and detection of myoglobin. To estimate the contribution of the surface plasmon resonance (SPR) to the desorption of myoglobin under the different pulse duration regimes, the addition of an aqueous gold nanostar (GNS) matrix was implemented. GNSs have a SPR maximum of ~750 nm which overlaps strongly with the 780 nm laser wavelength. Gold nanospheres, which have a SPR of ~530 nm, have an absorption overlap 25 times less than that of the nanostars with the 785 nm laser light and therefore were chosen as a control gold nanoparticle matrix. It was observed that protein mixed with solution phase GNSs improved the laser ablation and consequent mass spectral signal intensity of the protein in comparison to both the nanosphere addition and ablation from quartz without nanomaterial addition for the 55 fs, 56 ps, and 10 ns pulses. This dissertation also extends to an investigation of the electrospray source and the roles that the nebulizing gas pressure, electrospray solution flow rate, and needle protrusion from the emitter sheath effects the electrospray analyte signal and stability. Interactions between the electrospray droplets and nebulizing gas were elucidated using an ablation chamber in which laser ablated analytes were carried via the nebulizing gas flow through the nebulizer sheath to interact with the electrospray Taylor cone, jet, and subsequent droplets. The signal intensity and relative standard deviation (RSD) of an infused Victoria blue solution was used to assess conventional ESI optimization experiments while a mixture of Gly-Gly-His, lactose, adenosine, and vitamin B12 was laser ablated within the ablation chamber for the optimization of the remote ablation device. It was found that a needle protrusion flush with the nebulizing sheath wall, 9 psi nebulizing gas pressure, and 9 µL/min ESI flow rate yielded the highest signal intensity for low and high mass analytes when utilizing the ablation chamber. However, the conventional ESI signal and stability was maximized using a needle protrusion of 0.6 mm from the sheath, 18 psi nebulizing gas pressure, and 9 µL/ min ESI flow rate. The last two chapters describe collaborative efforts with GlaxoSmithKline (GSK) and Temple University’s Lewis Katz School of Medicine with the application of LEMS to real world problems. The first of these chapters explores the preliminary method development results for sampling protocols of LEMS in a pathway to measuring the active ingredient in a formulation when differences in concentration are a percent or less for GSK. The results from the method development and optimization experiments in the previous chapters were applied to the GSK pharmaceutical manufacturing paradigm to test product quality in-line and in real-time instead of testing in a lab at the end of the manufacturing process. The LEMS sampling protocols involved ablation of either powder, compressed form, or solution containing powder using laser ablation. The ablated material was then entrained in an electrospray aerosol and transferred into a mass spectrometer for quantitative measurement of the molecules making up the powder, pill, or solution. Measurement time was on the order of seconds so that thousands of samples can be potentially measured in an hour. Future prospective experiments include additional optimization of the solution phase and compressed form sampling methods and, ultimately, the method validation of LEMS for quantifying active ingredients in pharmaceutical formulations. The last chapter seeks to develop new methods to map all biomarkers in traumatic brain injury (TBI) through mass spectrometry imaging (MSI), serum analysis, and protein derivatization assays. In this work, the Ramirez laboratory employs the controlled cortical impact model of experimental TBI in mice, harvests the brain (post injury) and prepares sections for analytical analysis. TBI is a complex injury involving multiple physiological and biochemical alterations to tissue. The potentially thousands of relevant biomarkers spread over a volume of thousands of mm3 makes the spatially resolved chemical analysis of brain a big data problem to which principal component analysis is applied. / Chemistry

Atmospheric aerosols affect climate change by altering the energy balance of the atmosphere, and public health due to their variable chemical composition, size, and shape. While the formation of secondary organic aerosol (SOA) from gas phase precursors is relatively well understood, it does not account for the abundance of SOA observed during field measurements. Recently it has become apparent that in-aerosol aqueous chemical reactions likely provide some of the missing sources of SOA production, and many studies of aqueous phase processes are underway. This work explores the fates of the simplest 2-oxocarboxylic acids, glyoxylic acid (GA) and pyruvic acid (PA), under simulated solar irradiation in the aqueous phase. Field measurements have revealed that mono-, di-, and oxocarboxylic acids are abundant species present in atmospheric waters. Of particular interest are 2-oxocarboxylic acids because their conjugated carbonyl moieties result in significant UV-visible absorption above 300 nm, allowing absorption of sunlight in the lower troposphere, thereby initiating radical photochemistry and leading to formation of SOA. In Chapter 2 of this work, GA is demonstrated to primarily undergo α-cleavage, producing CO, CO2, formic acid, and the key SOA precursor glyoxal. Trace amounts of oxalic acid and tartaric acid are also quantified. Additionally, the dark thermal aging of glyoxylic acid photoproducts, studied by UV-visible and fluorescence spectroscopies, reveals that the optical properties of the solutions are altered radically by the glyoxal produced. The optical properties display periodicity during photolytic-dark cycles, reflecting behavior expected for aerosols during nighttime and daytime cycles. In contrast, Chapter 3 shows that PA photoreacts via a proton-coupled electron transfer (PCET) mechanism that produces CO2 and organic acids of increased complexity with 6 to 8 carbons. A combination of analytical techniques including 1H and 13C NMR; 13C gCOSY NMR; mass spectrometry; chromatography; and isotope substitutions allows the organic products to be identified as: 2,3-dimethyltartaric acid; 2-hydroxy-2-((3-oxobutan-2-yl)oxy)propanoic acid; and the quasi-intermediate 2-(1-carboxy-1-hydroxyethoxy)-2-methyl-3-oxobutanoic acid. In Chapter 4, PA irradiation is also shown to consume dissolved oxygen so fast that solutions become depleted within a few minutes depending on reaction conditions. This fast process directly produces the atmospheric oxidant singlet oxygen, which enhances the oxidizing capacity of the atmosphere. Additionally, PA photochemistry only proceeds under very acidic conditions (pH ≤ 3.5), like those in most atmospheric aerosols. Finally, we require a thorough understanding of the behavior of 2-oxocarboxylic acids at the air-water interface of aerosols because much of the GA and PA present in the atmosphere is produced in the gas phase and needs to partition into the aqueous phase to undergo photoreaction. Therefore, Chapter 5 uses surface sensitive online electrospray ionization mass spectrometry (OESI-MS) to demonstrate that carboxylic acids delivered from the gas phase onto the surface of aqueous microdroplets display enhanced acidities relative to bulk water solutions. This work demonstrates that aqueous photolysis is a very competitive atmospheric fate for both GA and PA. It also shows that these photoreactions are capable of contributing substantially to SOA formation by building chemical complexity and forming oxidants directly.

Desorption Electrospray Ionization Mass spectrometry Imaging (DESI-MSI) is an area of great interest and a promising tool in the field of chemical imaging. It is a powerful, label-free technique, which can determine, map and visualize different molecular compounds on a sample surface. The amount of information acquired in a single DESI-MSI experiment is enormous compared to other techniques, as it can simultaneously detect different compounds with their spatial distribution on the surface. The experiment can be used to produce two-dimensional and three-dimensional images. Chapter 2 focuses on the design and optimization of the setup for performing DESI-MS imaging on various substrates. The proposed setup was tested for its lateral spatial resolution. To provide proof-of-concept of the design, preliminary tests were performed to generate images from commercial thin layer chromatographic plates and photographic paper. Chapter 3 focuses on demonstrating the compatibility of novel microfabricated Thin Layer Chromatography plates (M-TLC plates) for detection with DESI-MSI.