Many plant species around the world are known to contain various psychoactive compounds. Due to their effects when consumed, many of these plants are used as a part of religious and ritualistic practices in many different cultures. As with any psychoactive compounds, these plants have the potential to be used in a recreational manner. In the United States, plant based drugs of abuse, such as marijuana, have become commonly abused substances. Although marijuana is currently regulated by the federal government, many of the plant materials containing potential drugs of abuse are not, and can be purchased legally from various online sources. The goals of this research were to develop methods for the analysis of a wide variety of plant based drugs of abuse by Direct Analysis in Real Time-Mass Spectrometry (DART-MS) and to apply the methods in an effort to differentiate between multiple strains of a single seed species. DART is an ambient ionization technique that allows for rapid analysis of samples while eliminating the need for sample preparation considerations for many applications. Analytes of interest can be detected within the complex plant matrix of ground up seeds, with no need for further extraction or isolation of the analytes. For this study, fourteen different seed samples, including twelve different species, reported to have psychoactive effects on the user were obtained and analyzed. Physical examination was performed, in which average measurements were obtained to describe the length, width, thickness, and mass of each seed species, followed by analytical analysis by DART-MS. The seeds were prepared for analysis by DART-MS by grinding to expose the middle of the seed containing the analytes of interest, and embedding the powder onto QuickStrip(TM) cards (IonSense, Inc.). To optimize the method for analysis, three different DART carrier gas temperatures (250°C, 300°C, and 350°C) were investigated for each seed sample by considering the signal to noise ratio, ion abundance, and presence of the analyte of interest at each source temperature using a single quadrupole mass spectrometer. The analytes detected were then subjected to MS(n) fragmentation in a quadrupole ion trap to confirm the identity of the analytes being detected. Fragmentation patterns were then compared to fragmentation patterns reported in the literature through methods such as chemical ionization, atmospheric pressure chemical ionization, and electrospray ionization. Thirteen of the fourteen seed samples were known to contain compounds with psychoactive properties. One of the species contained no known hallucinogenic compounds, however it was reported to have psychoactive effects when ingested or smoked. Protocols were developed for each sample and the identification of the analytes of interest was successful in twelve of the fourteen samples. DART-MS is a powerful technique for the detection and identification of a variety of plant based drugs of abuse, including tetrahydrocannabinol, lysergic acid amide, and numerous others. The ability to rapidly analyze a large number of samples makes DART-MS a technique with great potential in forensic laboratory settings, such as forensic drug analysis, where case backlog is often an area of concern. The majority of the samples explored in this study are not considered common substances of abuse. However, as their abuse is becoming more common, the high throughput nature of the analytical methods and techniques discussed will become increasingly important.

Gasoline is a substance commonly encountered in forensic settings. Unfortunately, gasoline is an easily obtainable ignitable liquid that arsonists commonly use to initiate or expedite the spread of an intentionally set fire. Fires claim the lives of many people each year in addition to causing widespread property damage. Many fire scene investigations result in charges of arson, which has the legal connotation of a committed crime. For this reason, extensive analysis and investigation must be undertaken before any suspected arson scene is deemed an actual case of arson. Although ignitable liquids, including gasoline, may be present at the scene of a fire, it does not necessarily mean they were intentionally used as accelerants. An accelerant is a fuel used to initiate a fire. These realities, in addition to several other factors, demonstrate why a rapid, reliable, gasoline analysis method is crucial to forensic applications. In this thesis, direct analysis in real time – mass spectrometry (DART-MS) is evaluated as a potential method that could better identify, distinguish and classify gasoline brands from one another. Techniques such as DART-MS could enable forensic laboratories to better identify questioned gasoline samples. Many ignitable liquids share similar chemical properties, and forensically relevant evidence is often obtained from a crime scene in less than favorable conditions. Fire debris can encompass various materials, including burnt carpet, flooring, items of furniture and clothing, among others. If gasoline was used as an accelerant, it may be present in trace amounts after the termination of the fire. Materials submitted for laboratory analysis may be substrates with compositions that have components similar to those found in some ignitable liquids. These are just a few of the potential obstacles that could be encountered with analyzing fire debris in a forensic setting. Traditionally, gas chromatography – mass spectrometry (GC-MS) methods are utilized for gasoline analysis in the criminal laboratory setting. While traditional GC-MS methods are sensitive and able to classify samples as gasoline, they are time consuming in terms of both sample preparation and analysis. Additionally, they do not generate differential mass spectral data based on the brand of gasoline. Conversely, gasoline analysis in this research, utilizing the DART-MS method, demonstrated that five different brands of gasoline could be distinguished from one another both by visual examination of mass spectra and with methods of chemometric analysis. Advantageously, the DART-MS method, an ambient ionization technique, requires little sample preparation and a rapid sample analysis time, which could drastically increase the throughput of standard sample analysis with further method development. The goals and objectives of this research were to optimize the DART-MS parameters for gasoline analysis, determine if DART-MS analysis could distinguish gasoline by brand, develop chemometric models to appropriately classify gasoline samples, and finally lay groundwork for future studies that could further develop a more efficient and discriminating DART-MS gasoline analysis method for forensic casework. Each brand of gasoline was observed to have a chemical attribute signature (CAS) consisting of not only low-mass ions, but also a variety of high-mass ions not usually observed with gasoline samples analyzed by GC-MS. Although variables including season, storage time, dilution and age of the gasoline were observed to contribute to the resulting mass spectral data, once the mass spectra are better understood, they could offer even more discriminating power between samples than simple analysis of the gasoline brand. In this research, DART-MS parameters were first optimized for gasoline analysis. Subsequently, the five acquired brands of gasoline: Shell, Sunoco, Irving, Cumberland Farms and Gulf, were analyzed both undiluted (or neat) and diluted utilizing the DART-MS analysis method. GC-MS data was generated and analyzed to show comparisons. After analyzing the data generated by both approaches, it was apparent that the DART-MS method could generate CASs based on the gasoline brand and offer a degree of differentiation that traditional GC-MS does not. Additional chemometric analyses utilizing principle component analysis (PCA) and the construction of models with Analyze IQ Lab software verified that the gasoline brands were distinguishable when samples were analyzed with this ambient ionization method. PCA plots of the neat gasoline demonstrated clustering based on brand. Additionally, models constructed from training samples generated from DART-MS analysis of the various brands were able to accurately classify gasoline samples as "yes" or "no" when a test set of gasoline was compared to all five brands. The lowest associated testing error rate for some of these models was 0%. However, additional analysis with greater sample sizes needs to be further carried out to more accurately evaluate this method of gasoline analysis and classification.

Human breast milk is a biofluid produced by a woman’s body during pregnancy. Breast milk contains necessary nutrition to a growing infant as well as xenobiotics--including drugs of abuse-- consumed by the woman which diffuse into the breast milk from the bloodstream. Since breast milk is recommended to be part of all infants’ diets, being able to detect any toxic components--such as drugs--in the matrix is critical. However, despite the ease and noninvasive nature of collection, human breast milk is a difficult matrix to analyze due to its high fat and protein content. Thus far, no literature has been published on the analysis of breast milk through direct analysis in real time mass spectrometry (DART-MS). Adapting DART-MS to detect drugs of abuse in human breast milk will allow for quick and timely identification of drugs present in an individual’s breast milk, as well as aid in research regarding the potential harmful effects of drugs--both licit and illicit--on an infant who is breastfeeding. Forensically, this method could potentially allow toxicologists to use breast milk as a matrix to determine if drugs played a role in a woman’s or breastfed child’s death. Using both C18 biocompatible solid-phase microextraction (BIO-SPME) fibers and QuickStrip™ cards, a DART-MS method was developed to be able to detect drugs of abuse in human breast milk. Four drugs of abuse (cocaine, codeine, morphine, and delta-9-tetrahydrocannabinol (Δ9-THC))--all of which are either commonly abused during the postpartum period or are of particular danger to breastfeeding women--were chosen to be studied. The drugs of abuse were extracted from either whole or pre-filtered human breast milk using either liquid-liquid extraction or C18 BIO-SPME fibers and detected with DART-MS using parameters suggested by IonSense, Incorporation (Inc.). Mass spectral results indicated that macromolecules in whole breast milk did not hinder extraction or detection and that a larger amount of the analytes were ionized/desorbed when using the BIO-SPME fibers. Thus, a BIO-SPME method adopted from IonSense, Inc. utilizing C18 fibers and SPME DART-MS parameters (with temperature and rail time adjustments) can be used to quickly detect cocaine, codeine, morphine, and Δ9-THC in human breast milk, indicating that this method may be used for the detection of other drugs of abuse in breast milk. In addition, BIO-SPME fibers can be used to quantify the concentration of cocaine in breast milk between a range of 50 and 200 nanograms per milliliter as demonstrated by a matrix-matched calibration curve created using various concentrations of cocaine. Despite its benefits, the BIO-SPME and DART method cannot be used on samples containing more than one drug of abuse (based upon the drug concentrations utilized in this study) due to competitive adsorption and competitive ionization, respectively, as not all drugs could be detected when this method was applied to breast milk samples containing numerous drugs.

Les lichens, organismes symbiotiques associant un champignon et un partenaire photosynthétique (algue verte et/ou cyanobactérie), sont caractérisés par la biosynthèse de métabolites secondaires uniques dotés de bioactivités variées. Pour valoriser au mieux cette ressource privilégiée, des méthodes innovantes de spectrométrie de masse ont été développées dans le but de minimiser la préparation de l’échantillon et la durée des analyses. Deux techniques de spectrométrie de masse ont été évaluées en ce sens : le DART-MS et le LDI-MS. L’apport de chacune de ces deux méthodes a pu être établi sur un large panel de lichens, représentant une part importante de l’espace chimique couvert par ces organismes. Il a été démontré que des profils chimiques complets pouvaient être obtenus respectivement à partir de thalles lichéniques et d’extraits acétoniques totaux. Compte tenu de la très large utilisation de la CCM pour l’analyse chimique de lichens, les possibilités offertes par le couplage de la CCM à l’ionisation electrospray ont également été explorées. Une seconde partie de ces travaux avait pour but de cartographier la distribution des métabolites secondaires au sein du thalle lichénique. À ces fins, des analyses d’imagerie LDI ont été réalisées sur une coupe transversale d’un lichen crustacé modèle : Ophioparma ventosa. Ce lichen a été étudié en phytochimie pour identifier six napthopyranones à partir des apothécies dont quatre nouvelles structures. Les principaux métabolites de ce lichen ont pu être imagés par LDI-MSI avec une résolution spatiale de 50 μm environ. Une corrélation entre la distribution des molécules et leur rôle écologique présumé permet d’avancer des hypothèses d’écologie chimique. Des approches conjointes reliant histolocalisation et étude génétique des partenaires de la symbiose ont été entreprises. La recherche des gènes de la biosynthèse de la mycosporine sérinol chez les symbiontes isolés de Lichina pygmaea par microdissection capture laser a été initiée en ce sens. D’autres approches innovantes comme l’analyse cristallographique par diffraction de poudre par les rayons X sont également abordées dans ce document articulé autour de six publications issues de ce travail et de deux articles en cours de soumission. / Lichens are self-sustaining symbiotic partnerships comprising a fungus associated with a green alga and/or a cyanobacteria. This consortium produces unique secondary metabolites that are endowed with various biological activities. To harness this privileged chemodiversity, innovative mass spectrometry techniques were developed in the course of this study to accelerate the dereplicative holdup through both a minimal sample preparation and a decrease of the time of analysis. Two approaches were considered during this work: DART-MS and LDI-MS and their adequacy for lichen dereplication was assessed on a vast array of samples encompassing a wide range of metabolites. Both of them facilitated complete chemical profiles, respectively from unprocessed lichen material and crude acetone extracts. Since TLC still enjoys a wide-spread popularity among lichenologists, the advantages offered by TLC-ESI-MS hyphenation were evaluated as well. A second part of this manuscript focused on the histolocalization of lichen metabolites. For this purpose, LDI mass spectrometry imaging studies were undertaken on the crustose lichen Ophioparma ventosa. The phytochemical investigation of this species afforded the isolation of six naphthopyranones from its apothecia, four of them being new molecules. LDI-MSI revealed the distribution patterns of all the main metabolites of this lichen, reaching a spatial resolution of 50 μm. Most interestingly, the distribution pattern of imaged metabolites within the thallus is highly organized and is related to their ecological relevance. Joint strategies combining histolocalization and genetic investigation of lichen symbionts separated using laser capture microdissection were also considered. As such, an investigation of the biosynthesis of mycosporine serinol within Lichina pygmaea dissociated symbionts was initiated. Further analytical strategies such as X-ray powder diffraction are introduced in this thesis that contains six publications and two drafts to be submitted.

Improvised explosive devices (IEDs) continue to be a national threat to the safety and security of the public. Research in explosives analysis for intact and post-blast samples continue to be a topic in which practitioners are constantly improving and searching for faster methods and techniques to analyze these sample types. The key role crime laboratories play in analyzing these sample types can have limitations, such as increasing turnaround times and backlogs. This concern additionally plays a role in the safety of the public if an unknown individual has not been discovered. Current analytical instrumentation in which explosives are analyzed includes Gas Chromatography – Mass Spectrometry (GC-MS), Liquid Chromatography – Mass Spectrometry (LC-MS), and Ion Mobility Spectrometry (IMS). Each instrument has benefits in the analytical results obtained. Direct Analysis in Real Time - Mass Spectrometry (DART-MS) has shown a significant promise as an analytical approach that can help remedy the time an explosive sample is analyzed, while additionally providing discriminating analytical results. Previous research has shown that DART-MS is capable of analyzing explosives, including smokeless powder. A limitation currently in the area of smokeless powder analysis with DART-MS is the application of utilizing this method and technology to realistic casework that may be encountered in forensic laboratories. Intact and post-blast explosive samples encountered in forensic laboratories arrive in various states and conditions. For example, the severity of the blast and environmental factors may play a role in the detection of smokeless powder on these sample types. To provide objective information and additional research, studies were conducted with mixture samples of smokeless powder and potential matrices that may be encountered in real world case samples. Faster processing time, in addition to the discrimination of smokeless powder, was the ultimate goal of this research. Due to the complexity of the mass spectra that may be generated from sample mixtures, an extraction technique coupled with DART-MS was investigated. A liquid-liquid extraction (LLE) method and dynamic headspace concentration using Carbopack™ X coated wire mesh were tested for the effectiveness of separating the analytes of interest of smokeless powder from various matrix interferences. Hodgdon Hornady LEVERevolution (HHL) smokeless powder, Pennzoil 10W-40 (P10W40) motor oil, and residue from metal end caps (China SLK brand) and black steel pipe nipples (Schedule 40) were used during the course of the matrix interference study. The method of applying dynamic headspace concentration using Carbopack™ X coated wire mesh and analysis by DART-MS provides an effective alternative to obtaining mass spectral data in a shorter amount of time, compared to techniques currently used in forensic laboratories. Effective separation was not achieved using the various LLE methods tested. Further testing would be required in order to evaluate the feasibility of implementing the technique as a sample preparation approach prior to analysis by DART-MS.

Improvised explosive devices (IEDs) are charged devices often used by terrorists and criminals to create public panic. When the general public is targeted by an act of terrorism, people who are not injured or killed in the explosion remain in fear until the perpetrator(s) has been apprehended. Methods that can provide investigators and first responders with prompt investigative information are required in such cases. However, information is generally not provided quickly, in part because of time-consuming techniques employed in many forensic laboratories. As a result, case report turnaround time is longer. Direct analysis in real time - mass spectrometry (DART-MS) is a promising analytical technique that can address this challenge in the Forensic Science community by permitting rapid trace analysis of energetic materials. The builder of an IED will often charge the device with materials that are readily available. The most common materials employed in the construction of IEDs are black and smokeless powder. However, other materials may include ammonia- or peroxide-based materials such as common household detergents. Smokeless powder is a propellant that is readily available to civilians. They are typically used for reloading ammunition and are sold in large quantities each year in the United States. Some states have stricter regulations than others but typically a firearms license is all that’s required to possess smokeless powder. Smokeless powder is considered a low explosive which is capable of causing an explosion if a sufficient quantity is deflagrated inside a confined container. The most commonly employed confirmatory techniques for the analysis of smokeless powder are gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-mass spectrometry (LC-MS). These methods often require extensive and time-consuming sample preparation procedures to prepare the powders for analysis. In addition to lengthy sample preparation procedures, GC-MS and LC-MS often require chromatographic separations that can range anywhere from 5 to 30 minutes or longer per sample. Ion mobility spectrometry (IMS) is widely used for the field analysis of smokeless powder and can provide faster results in comparison to GC-MS or LC-MS. However, identification is limited to drift time and no structural information is provided unless coupled to a mass spectrometer. In an effort to accelerate the speed of collection and characterization of smokeless powder, an analytical approach that utilizes novel wire mesh coated with CarbopackTM X, dynamic headspace concentration and DART-MS was evaluated to determine if the approach could generate information rich chemical attribute signatures (CAS) for smokeless powder. CarbopackTM X is a graphitized carbon material that has been employed for the collection of various volatile and semi-volatile organic compounds. The goal of using CarbopackTM X coated wire mesh was to increase the collection efficiency of smokeless powder in comparison to traditional swabbing and swiping methods. DART is an ambient ionization technique that permits analysis of a variety of samples in seconds with minimal to no sample preparation and offers several advantages over conventional methods. Heating time, heating temperature and flow rate for dynamic headspace concentration were optimized using Hodgdon Lil’ Gun smokeless powder. DART-MS was compared to GC-MS and validated using the National Institute of Standards and Technology reference material 8107 (NIST RM 8107) smokeless powder standard. Additives and energetic materials from unburnt and burnt smokeless powders were rapidly and efficiently captured by the CarbopackTM X coated wire mesh and successfully detected and identified using DART-MS. The DART source temperature was evaluated with the goal of providing the most efficient desorption of the analytes adsorbed onto the wire mesh. For this to be a robust approach in forensic analysis, chemometric analysis employing predictive models was used to simplify the data and increase the confidence of assigning a mass spectrum to a particular powder. Predictive models were constructed using the machine learning techniques available in Analyze IQ Lab and evaluated for their performance in classifying three smokeless powders: Alliant Reloder 19, Hodgdon LEVERevolution and Winchester Ball 296. The models were able to accurately predict the presence or absence of these three powders from burnt residues with error rates that were less than 4%. This approach has demonstrated the capability of generating comparable data and sensitivity in a significantly shorter amount of time in comparison to GC-MS. In addition, DART-MS also permits the detection of targeted analytes that are not amenable to GC-MS. The speed and efficiency associated with both the sample preparation technique and DART-MS, and the ability to employ chemometric analysis to the generated data demonstrate an attractive and viable alternative to conventional techniques for smokeless powder analysis.

Molecular ionization is owed much of its development from the early implementation of electron ionization (EI). Although dramatically increasing the library of compounds discovered, an inherent problem with EI was the low abundance of molecular ions detected due to high fragmentation leading to the difficult task of the correct chemical identification after mass spectrometry (MS). These problems stimulated the research into new ionization methods which sought to "soften" the ionization process. In the late 1980s the advancements of ionization techniques was thought to have reached its pinnacle with both electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI). Both ionization techniques allowed for "soft" ionization of large molecular weight and/or labile compounds for intact characterization by MS. Albeit pervasive, neither ESI nor MALDI can be viewed as "magic bullet" ionization techniques. Both techniques require sample preparation which often included native sample destruction, and operation of these techniques took place in sealed enclosures and often, reduced pressure conditions. New open-air ionization techniques termed "ambient MS" enable direct analysis of samples of various physical states, sizes and shapes. One particular technique named Direct Analysis In Real Time (DART) has been steadily growing as one of the ambient tools of choice to ionize small molecular weight (< 1000 Da) molecules with a wide range of polarities. Although there is a large list of reported applications using DART as an ionization source, there have not been many studies investigating the fundamental properties of DART desorption and ionization mechanisms. The work presented in this thesis is aimed to provide in depth findings on the physicochemical phenomena during open-air DART desorption and ionization MS and current application developments. A review of recent ambient plasma-based desorption/ionization techniques for analytical MS is presented in Chapter 1. Chapter 2 presents the first investigations into the atmospheric pressure ion transport phenomena during DART analysis. Chapter 3 provides a comparison on the internal energy deposition processes during DART and pneumatically assisted-ESI. Chapter 4 investigates the complex spatially-dependent sampling sensitivity, dynamic range and ion suppression effects present in most DART experiments. New implementations and applications with DART are shown in Chapters 5 and 6. In Chapter 5, DART is coupled to multiplexed drift tube ion mobility spectrometry as a potential fieldable platform for the detection of toxic industrial chemicals and chemical warfare agents simulants. In Chapter 6, transmission-mode DART is shown to be an effective method for reproducible sampling from materials which allow for gas to flow through it. Also, Chapter 6 provides a description of a MS imaging platform coupling infrared laser ablation and DART-like phenomena. Finally, in Chapter 7 I will provide perspective on the work completed with DART and the tasks and goals that future studies should focus on.

One of the most fundamental challenges in analytical mass spectrometry (MS) is the efficient conversion of neutral molecules into intact gas-phase ions. In this thesis, I investigate the capabilities of various new and established ionization techniques including (a) the Array of Micromachined UltraSonic Electrosprays (AMUSE), (b) Direct Analysis in Real Time (DART) and (c) Electrospray Ionization (ESI) for bioanalytical and biomedical analysis purposes. The AMUSE is a MicroElectroMechanical System (MEMS)-based device that was created as an alternative, and more sensitive approach for ion generation in an array format. In the AMUSE, the processes of droplet formation and DC droplet charging are separated allowing ionization of liquid samples using low charging voltages and a wide variety of solvents. Our analytical characterization work with the AMUSE showed that ion generation with this device was indeed possible, and that incorporation of a Venturi device increased signal stability and sensitivity due to enhanced droplet desolvation and increased ion transfer efficiency. A detailed investigation to determine the optimal source parameters for ionization of aqueous solutions of model compounds including reserpine, leucine enkephalin and cytochrome C was carried out and it was found that ionization was possible even without the application of a DC charging potential. Subsequent experiments using the thermometer ion method to characterize the AMUSE from a more fundamental point of view, showed that AMUSE ions are lower in internal energy than ESI ions, opening interesting possibilities for the mass spectrometric study of labile species. Furthermore, it was found that it was possible to manipulate the internal energy of the ion population by varying the parameters that most strongly affect desolvation and focusing. Our studies with DART were directed at investigating its analytical potential for application to the identification of active ingredients (AIs) in low quality combination medicines and counterfeit antimalarials that are commonly sold in regions of the world (particularly Southeast Asia) where drug resistant malaria is endemic as their use may engender increased resistance against the few remaining effective antimalarials.