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UHRMS has enormous potential to provide complete molecular characterisation of organic aerosol. However atmospheric aerosol often has complex, mixed organic and inorganic composition and soft ionisation techniques that have been developed to ionise organic aerosol specifically, are susceptible to salt suppression effects and competitive ionisation when applied to complex ambient aerosol. In this thesis, characterisation experiments were conducted on a novel soft ionisation technique called Extractive Electrospray Ionisation (EESI) using mixtures of water-soluble organic compounds (WSOCs) with increasing ammonium sulphate solute concentrations. The WSOCs MS signals in the mixtures were not significantly affected by salt suppression over an atmospherically relevant range of 0-70% inorganic molar percentage relative to organic, recommending EESI-MS as a suitable technique for analysing ambient aerosol from a range of environments. EESI-MS was also used for the first time to detect Secondary Organic Aerosol (SOA) generated in the Cambridge Atmospheric Simulation Chamber (CASC) from the OH radical-initiated oxidation and ozonolysis of Anthropogenic Volatile Organic Compounds (AVOCs) and Biogenic Volatile Organic Compounds (BVOCs). Several oligomer products from complex BVOC and AVOC oxidation, previously observed in chamber studies using similar soft ionisation techniques, were detected using EESI-MS. This thesis also includes the first study, to our knowledge, characterising the detailed molecular composition of organic compounds present in rainwater and aerosol collected during the same rain event, using chip-based direct infusion Nano-ESI and LC-ESI-MS. A Solid Phase Extraction (SPE) method was developed to extract the organic compounds from the rainwater and aerosol samples collected during the AERORAIN field campaign conducted in León, Spain, prior to MS analysis. More mono-aromatic compounds were observed in the aerosol samples whereas more poly-aromatic compounds were observed in the rainwater. HYSPLIT air mass backward trajectories revealed that both air mass origin and seasonality had strong influences on the sample composition. Samples collected during the summer and samples influenced by the Sahara desert contained the most compounds due to increased photochemical activity and extra industrial emissions from North Africa respectively. All samples contained semi-volatile oxygenated organic aerosol and multiple organosulphates were tentatively identified. Continental samples contained low-volatility oxidised organic aerosol and exhibited the most efficient aerosol scavenging and CHO and CHON compounds were preferentially scavenged over CHOS and CHONS compounds. The combination of EESI-MS, Nano-ESI-MS and LC-ESI-MS analysis in this thesis, along with optimised analytical and data processing methods, has provided an in depth characterisation of the composition of complex aerosol, and highlights the importance of seasonality, long range transport and below-cloud scavenging on the compound distribution in ambient samples.

Although the benefits of ICP-MS are well documented, the determination of analytes at low levels in high concentrations of matrix elements has proved difficult. As ICP-MS is a 'flow into' instrument the deposition of salts throughout the system is a common cause of significant loss of signal. The application of desolvation of aluminosilicate samples, to aid in the production of more robust plasma conditions, was investigated to increase the efficiency of the ICP in processing the sample. The performance of the ICP-MS was monitored with different cone arrangements and by running the skimmer cones at elevated temperatures. An alternative to modification of the instrument is to employ chemical modification of the sample and hence the separation of Au and Pt from an aluminosilicate matrix via the use of solid phase extraction (SPE) columns were investigated as a means of dealing with high levels of dissolved solids. OVB based SPE columns were found to give high retentions of Au and Pt when chelated with ammonium pyrrolidinediethylcarbamate (APOC). A second alternative that avoided digestion of the aluminosilicate matrices, was to carry out the analysis using laser ablation (LA). LA-ICP-MS is becoming increasingly used for trace elemental analysis but as yet no universal calibration method is available. The general problems associated with matrix matched standards are inherent as the ablation mechanism and plasma conditions can differ dramatically with very small changes in matrix composition. Hence the addition of chromophores was employed to increase the absorption of the laser energy. The use of vanillic, nicotinic and pyrazinoic acid were used to improve the ablation of pressed powder discs at the laser wavelength of 213 nm. Synthetic aluminosilicate discs and standard additions were both employed for the calibration and determination of Ti.

A significant issue affecting the aquaculture and water industries is the presence of off-flavour compounds in water, which cause problems by imparting an undesirable earthy/musty flavour and smell to water and fish. Two predominant off-flavour compounds are geosmin (GSM) and 2-methylisoborneol (MIB). These compounds are produced by several varieties of cyanobacteria and actinomycetes as metabolic products and can be detected by humans at concentrations as low as 0.015 mg L-1. Removal of GSM and MIB from potable waters has proven to be inefficient using standard water treatment such as filtration, coagulation, flocculation, sedimentation and chlorination. Activated carbon and membrane processes can physically remove GSM and MIB, but do not destroy them, and ozone treatment can be expensive. Titanium dioxide (TiO2) photocatalysis has recently been demonstrated to rapidly degrade GSM and MIB. When the semiconductor catalyst is illuminated with ultraviolet light simultaneous oxidation and reduction reactions occur. Pollutants are broken down into mineral acids, carbon dioxide and water. This study was conducted to determine if TiO2 photocatalysis, using a pelleted form of TiO2 called Hombikat K01/C, was a suitable method for the treatment of potable water. Additionally an analytical method was developed to rapidily analyse the large number of samples generated. Two reactors, a bench scale batch reactor and pilot scale flow reactor, were developed and used to evaluate the efficacy of Hombikat K01/C TiO2 photocatalysis in degrading GSM. The batch reactor, containing Hombikat K01/C, was used to investigate the effect of numerous experimental variables on the photocatalysis of GSM, including initial substrate concentration, pH, light intensity, aeration rate, the presence of additional reactants, and catalysis conducted in deuterated water. GSM was rapidly degraded using the TiO2 batch reactor, with the rate of GSM degradation most affected by light intensity and additional reactants, though pH also had a notable effect. A kinetic isotope effect of 1.61 was observed for the destruction of GSM using Hombikat K01/C TiO2. The flow reactor was also found to efficiently degrade GSM in raw waters. The rate of GSM destruction was found to be significantly lowered by UV shielding of the catalyst, caused by constituents of raw the water used, and the presence of additional reactants. The pilot scale flow reactor was also successfully evaluated in Denmark using gesomin contaminated water from an eel farm

The use of controlled substances and pharmaceuticals is a wide-spread problem that does not currently have a quick and effective way to gauge the abuse of drugs in the community. The analysis of wastewater allows for a timely method in observing the use of illicit drugs and non-controlled substances in the surrounding communities. Removal of chemical contaminants from wastewater is crucial to prevent further introduction into the surrounding ecosystems. All water samples utilized for optimization and the wastewater samples that were tested to identify controlled substances and pharmaceuticals underwent two forms of extraction and were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The Anton-Paar synthesis reactor was used to extract the drug components from the wastewater samples for analysis, followed by solid phase extraction, before analysis on the LC-MS/MS. The drug panel that was used for this research consisted of cocaine, methamphetamine, fentanyl, tetrahydrocannabinol (THC), 6-monoacetylmorphine (6-MAM), alprazolam, benzoylecgonine, buprenorphine, caffeine, clonazepam, codeine, diphenhydramine, hydrocodone, 3,4-methylenedioxymethamphetamine (MDMA), morphine, norfentanyl, norhydrocodone, noroxycodone, norsertraline, oxycodone, and sertraline. The results showed that there was no statistically significant difference between the method optimization run parameters; therefore, the lowest temperature, 90 ℃ for 5 minutes, was selected for wastewater analysis to prevent any further degradation. Alprazolam, benzoylecgonine, caffeine, diphenhydramine, methamphetamine, norfentanyl, oxycodone, sertraline, and THC were detected in the influent wastewater samples. The effluent samples contained all of the compounds listed above, with the exception of methamphetamine and THC. The detection of these compounds shows prevalence in the surrounding community of the testing site. With the knowledge of prevalent drugs of abuse and pharmaceuticals, law enforcement, social workers, and public health officials can take preventative measures such as educational and awareness programs.

With the increased practice of testing oral fluid for the detection of illicit drug use, the available active and passive collection methods have also increased. Active collection methods stimulate oral fluid production of the salivary gland through acidic or non-acidic methods. In contrast, passive collection methods simply use a collection device that is held in the oral cavity to accumulate oral fluid. This study focused on the recovery of drug analytes from solutions used in active and passive collection methods after experiencing the same solid-phase extraction method. A total of eleven drug analytes were examined including benzodiazepines, cocaine, methamphetamines, opioids, and delta-9-tetrahydrocannabinol (THC). Recovery samples were analyzed via Gas Chromatography–Mass Spectrometry (GC/MS). Of the eleven compounds tested during the recovery study, five were found to have statistically significant differences between the percent recoveries in active and passive oral fluid solutions. The active collection solution yielded higher percent recoveries than the passive collection solution. These findings suggest that active solution would be a more viable collection method to recover a greater majority of the drug analyte present in oral fluid.

The work contained within this dissertation focuses on the use of supercritical carbon dioxide (CO2) in the field of separation science, particularly with regards to the preparation of materials for sample preparation and greenness of supercritical fluid chromatography (SFC) as a separation technique. Supercritical CO2 was utilized in supercritical antisolvent (SAS) precipitation to fabricate a carbon sorbent from micronized polyacrylonitrile (PAN) for use in solid phase extraction (SPE). Additionally, improvements were suggested for the Analytical Method Greenness Score (AMGS) calculator so that the greenness of CO2-containing mobile phases could be assessed more accurately. Furthermore, an additional safety metric was developed to account for solvent release in the event of a leak during SFC analysis. Finally, recommendations for improving the greenness of SFC methods are outlined.The potential of using supercritical antisolvent precipitation for the fabrication of a carbon sample preparation sorbent was explored. Operating conditions, such as pressure, temperature, solution concentration, and CO2 mole ratio, for SAS precipitation were varied to study their effect on the morphology of micronized PAN. Stabilization and carbonization methods were examined to ensure that the morphology of the product was maintained, while maximizing the surface area. The performance of the PAN-based carbon material as an SPE sorbent was studied using a mixture of atrazine, an herbicide, and its degradation products. The solvents needed to effectively condition the sorbent and elute the desired analytes were studied and compared to a commercial carbon sorbent, allowing insight into the interactions available on the PAN-based sorbent. In addition, the greenness of CO2 in separation processes was studied. The AMGS calculator attempts to account for CO2-containing mobile phases, however, several improvements are needed to assess greenness more accurately. Here, EHS scores are altered to reflect operating conditions conventionally used in chromatography, as well as updating the density and cumulative energy demand (CED) value of CO2 used in greenness assessments. These changes allow for a more accurate representation of the hazards of solvents used in chromatography, as well as the mass of CO2 consumed. A comparison of SFC and reversed-phase liquid chromatography (RPLC) separations of pharmaceutical compounds are used to illustrate the impact of the changes suggested for the AMGS calculator. Most greenness metrics consider the hazards associated with handling and disposing of solvents, however, none account for additional hazards that arise from instrument failure, such as solvent leaks. Here, a method was developed to determine the composition of organic-CO2-air mixtures contained in the column compartment. The composition at various leak times was then compared to the flammability limits of the mixture to determine an overall hazard time. The effect of pressure, temperature, and mobile phase composition was assessed to determine methods to reduce additional safety risks. Increasing pressure was shown to most effectively reduce the hazard time. In addition, hazard times were assigned a score so that this metric could be readily incorporated into greenness assessments, such as the AMGS calculator. Finally, recommendations for improving the greenness of SFC methods are outlined. Pressure, temperature, and solvent choice were assessed to determine how to most effectively improve the greenness of SFC methods using the AMGS calculator. It was shown that increasing the pressure of the separation significantly reduces the analysis time of the separation, leading to a lower instrument energy consumption. In addition, the choice of organic co-solvent greatly affects the greenness of the SFC method. Simply choosing the most green solvent based on individual metrics may inhibit the chromatographic performance, yielding longer separation times, increased solvent and energy usage. Therefore, a greenness assessment that accounts for multiple components of the separation method, such as the AMGS calculator, should be used to screen methods and determine the most green option for the desired application.

With the increase of driving while under the influence of drugs and the legalization of marijuana there is a need to develop and validate solid phase extraction coupled with liquid chromatography tandem mass spectrometry methods using blood. The protocol that was followed was in the Clinical Lab Consulting Method Validation manual. The parameters that were tested were linearity/reportable range, limit of detection, and precision. The linearity/reportable range showed that 80/81 of the analytes had an average R^2 value of 0.950 or higher. The exception was cannabidiol with an average R^2 value of 0.932. The limit of detection showed that a cutoff of 1 ng/mL was sufficient for 80/81 analytes. THC requires a cutoff of 2 ng/mL. The precision showed issues with negative mode analytes such as cannabidiol and barbiturates.

In forensic toxicology, opioids are frequently associated with drug abuse or drug-related death cases. An optimal method for use in the identification and quantification of opioids in a complex blood matrix is of paramount importance. Along with the ability to identify and quantitate opioids, this method should be accurate, sensitive, and selective. The application of sample pre-treatment and solid-phase extraction are common to purify and concentrate the target analytes before analyzing with liquid chromatography-tandem mass spectrometry. The purpose of this study was to compare the performance of two standard operating procedures, adopted by the Massachusetts State Police Crime Laboratory Toxicology and the Biomedical Forensic Sciences– Toxicology Laboratory at Boston University School of Medicine, for detecting opioids in blood. A total of eight drugs were analyzed: 6-monoacetylmorphine, codeine, fentanyl, hydrocodone, morphine, norhydrocodone, oxycodone, and oxymorphone. Comparison was performed using the parameters studied as part of method validation, including calibration model, bias, precision, carryover, interferences, ionization suppression/enhancement, and recovery. The results indicated that the method from Massachusetts State Police provided a better performance with between-run precision, interferences from matrix and other commonly encountered drugs, matrix effect at high concentration (250 ng/mL) and matrix recovery. Meanwhile, the method from Biomedical Forensic Sciences showed less bias, within-run precision, and matrix effect at low concentrations. Carryover and internal standard interference were comparable in both standard operating procedures. The calibration models were adjusted by altering the selection of regression model for improved quantification method performance. The volume of solvents, sample matrix, as well as time, were taken into consideration in accessing the overall performance of identification and quantitation. Both procedures were comparable yet the one from Massachusetts State Police was more beneficial in identifying the target analytes with greater sensitivity and selectivity and the one from Biomedical Forensic Sciences was more economical and efficient.

Methyl anthranilate (MA), ortho-aminoacetophenone (o-AAP), and furaneol contribute to the characteristic “native grape” and “cotton candy” aromas of Vitis labruscana grapes and their hybrids, e.g. ‘Concord’ and ‘Niagara’. Convenient measurement of these compounds is of interest to grape breeders who wish to either increase or decrease their concentrations in grape varieties. Previous approaches for measurement of these odorants either cannot measure all compounds in a single analysis, or else have detection limits above the sensory thresholds for the odorants (low µg/L). We evaluated the use of an optimized reverse phase solid phase extraction (SPE) approach prior to selected ion monitoring (SIM) GC-MS to achieve low µg/L detection limits for MA, o-AAP, and furaneol in a single analysis.

In forensic toxicology, analysis of drugs and metabolites in biological fluids is performed to determine cause of death, suspected drug use, drug facilitated sexual assaults, or whether someone was driving under the influence. Analyte identification and concentration determination can be determined in a variety of matrices (e.g., blood, urine, or oral fluid) and can be complex. It is therefore necessary to have optimal sample preparation and instrumental conditions that work for all matrices of interests. Determining the best approach can be challenging due to the amount of time and resources to perform expansive evaluations of sample preparation, stationary/mobile phases, liquid chromatography (LC) conditions and mass spectrometry (MS) operating parameters. In this study three different sample preparation methods were validated for blood and urine. The three sample preparation methods were solid-phase extraction (SPE), supported liquid extraction (SLE), and liquid-liquid extraction (LLE). Six different drug groups were used as the analytes being tested by the methods. These drug groups were amphetamines, local anesthetics, opioids, hallucinogens, antidepressants, and novel psychoactive substances (NPS). A total of twenty-three drugs were used: amphetamine, methamphetamine, (3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy-N-ethylamphetamine (MDEA), and 3,4-methylenedioxymethamphetamine (MDMA), benzoylecgonine (BZE), cocaine, lidocaine, codeine, methadone, morphine, 6-monoacetylmorphine (6-MAM), fentanyl, oxycodone, lysergic acid diethylamide (LSD), phencyclidine (PCP), amitriptyline, citalopram, fluoxetine, trazodone, ethylone, α-pyrrolidinopentiophenone (α-PVP), and 25I-NBOMe. The methods were validated according to guidelines set forth by the Scientific Working Group for Forensic Toxicology (SWGTOX) Standard Practices for Method Validation in Forensic Toxicology and the American Academy of Forensic Science (AAFS) Standards Board (ASB) draft of Standard Practices for Method Validation in Forensic Toxicology. Parameters of calibration model, bias, precision, limit of detection (LOD), limit of quantitation (LOQ), dilution integrity, ion suppression/enhancement, interference studies, and stability were evaluated. Recovery was also assessed to determine the efficiency of the extraction. Calibration models met the 0.98 R2 minimum requirement. For all sample preparations the compounds evaluated in each were found to be stable for at least 72 hours. Interferences were found to be similar across all three sample preparation methods. Parameters of bias, precision, and dilution integrity were largely comparable between all three methods. Overall for LOD, SLE resulted in lower values for blood and urine ranging for 0.1 to 5 ng/mL. Overall for LOQ, SLE resulted in lower values for blood and LLE resulted in lower values for urine in the range of 0.5-10 ng/mL. SLE resulted in the highest recovery for all twenty-three analytes, due to LLE failing to extract consistently or completely for benzoylecgonine, morphine, and 6-monoacetylmorphine. Overall, SLE resulted in the lowest percent values for ion suppression and enhancement for both blood and urine. Overall, blood resulted in high ion suppression (exceeding -20%) for SPE and LLE. Final determination overall was that SLE was the best sample preparation method for all twenty-three analytes. This was determined based on the evaluation of recovery, ion suppression/enhancement, and LOD, as well as sample preparation time. Sample preparation time for SLE was approximately 1 hour, while SPE took 2.5 hours and LLE 2 hours.

A cannabinoid refers to any natural or synthetic compound that interacts with the CB1 and CB2 receptors. There are currently three different groups of cannabinoids: endogenous cannabinoids, phytocannabinoids and synthetic cannabinoids. The most common phytocannabinoid is delta-9-tetrahydrocannabinol (THC), which is the active component in the Cannabis sativa or marihuana plant1–3. Two examples of synthetic cannabinoids that are present in case reports from 2012 to 2018 are AB-FUBINACA and AB-PINACA4–7. THC and synthetic cannabinoids are commonly encountered drugs in forensic toxicology cases, therefore, being able to extract these compounds and their metabolites is imperative for toxicological interpretation. There are a variety of commercially available sample preparation techniques for these analytes. Companies such as UCT, Biotage, Millipore-Sigma, Tecan, and Thermo Fisher Scientific manufacture these products. The focus of this research was to evaluate these techniques for their cleanliness, efficiency and cost effectiveness. Sample preparation techniques are designed to remove the different components of the matrix and other prescription or illicit substances present in the sample that could interfere with the assay, increase the analyte recovery, extraction efficiency, decrease variability, and clean-up the sample to allow for less instrument downtime and longer column life8. This study focused on comparing a liquid-liquid extraction (LLE), solid phase extraction (SPE), and supported liquid extraction (SLE). The primary purpose of this study was to develop and validate the three above mentioned sample preparation techniques for the analysis of THC, 11-hydroxy-THC, 11-nor-9-carboxy-THC (THCCOOH), AB-FUBINACA, AB-FUBINACA metabolite 3, and AB-PINACA in blood and urine. Parameters assessed followed Academy Standards Board (ASB) Standard 036, Standard Practices for Method Validation in Forensic Toxicology, including recovery, suppression, and matrix effects. For urine and blood analysis, the calibration range was determined to be 1 ng/mL to 50 ng/mL for all three techniques. Urine recovery was highest for the LLE method, with all compounds having a recovery greater than 50%. The SLE method had the lowest LOQ results for urine, with 0.5 ng/mL for 11-hydroxy-THC and THCCOOH, 0.75 ng/mL for THC, AB-FUBINCA and AB-FUBINACA metabolite 3, and 1 ng/mL for AB-PINACA. Ion suppression was reduced using the SLE method for urine along with having the shortest sample preparation time of 1 hr for up to 48 samples. For blood analysis, the LLE method had the greatest recovery of all analytes. The LLE method also had reduced suppression and matrix effects compared to the SPE method. Sample preparation was shorter for the SPE method, consuming 2 hrs for an average sample batch, compared to 4 hrs for the LLE method, which included a 2 hr freezing step. In conclusion, for urine analysis, all three sample preparation techniques were acceptable for the analysis of THC, synthetic cannabinoids, and their metabolites, with the SLE method being the preferred method. For blood analysis a LLE and SPE method were developed and are adequate for the analysis of THC, synthetic cannabinoids, and their metabolites, with the LLE method being the preferred method.

The issue with maintaining stability of compounds during storage and transportation has been universal across many fields such as clinical, pharmaceutical, food industry and especially in forensic laboratories where case backlogs hinder immediate analysis of samples upon collection. This study compares various storage conditions and analyzes changes in compound stability over 28 days. The various storage conditions compared in this study are temperature (+4 °C and -20 °C); water sample and urine sample and solid state (loading onto Solid Phase Extraction cartridges) and storing in its liquid state (present in the urine or water sample). 51 compounds were analyzed in this study belonging to pharmaceutical and hormone classes. Two Dimensional Liquid Chromatography was used for separation allowing the analysis of varied compounds in terms of chemistry. Mass Spectrometry was used for detection of these compounds. This study conclusively helped determine that there is no significant difference in stability of 42 out of 51 of these compounds on comparing them in solid versus liquid state over a period of 28 days. This helps determine that Solid Phase Extraction (SPE) cartridges can be used as an alternative for storage and transportation of these compounds. 46 compounds showed no significant difference in +4 and -20 °C temperature storage conditions as well. Despite not including a wash step during sample preparation between loading and elution, 11 out of 51 compounds did not show suppression or enhancement due to matrix effect. This study not only highlights the importance of sample preparation prior to analysis but also shows how SPE technique could help maintain stability of compounds during storage. In future studies, stability changes using SPE for long term storage could provide beneficial results to conclude if SPE techniques can provide an advantage over liquid state for a period longer than a month.

In the field of forensic toxicology, several challenges exist with quantification analysis of cocaine and metabolites in post mortem samples. Cocaine can prove difficult to detect and quantify in blood, urine, and soft tissues following extensive decomposition. Alternative matrices, such as hair, nails, and bone could prove useful in detecting chronic drug use in post-mortem toxicology cases. Detection and quantification of drugs in complex matrices is difficult to accomplish due to time-consuming extraction processes, and inability to detect an analyte at trace levels. Further, analysis of drugs in hard tissues, such as hair and bone, has only been attempted in recent years. Even fewer studies have investigated detection of drugs following decomposition of remains, specifically outdoor decomposition. The objective of this study was to develop a robust extraction and clean up methodology, in which a homogenization step precedes, to efficiently extract drugs from complex matrices, reach a target limit of detection (LOD) and to maintain instrument performance using multidimensional chromatography. Multi-dimension chromatography platform such as two dimensional liquid chromatography tandem mass spectrometry ( 2D-LC/MS/MS,) offers options not compatible with single dimension v units. With large volume injection capabilities of aqueous and organic extracts, the analytical process be reduced from multiple hours to minutes. All rat specimens used for this study fell under an Institutional Animal Care and Use Committee (IACUC) protocol. The rodents underwent a 10-12 weeks chronic intravenous self-administration of cocaine. This was followed by a six-week period of abstinence, followed again by a three-week period of cocaine self-administration before being euthanized. Average daily dosages for each rat fell within a range of 13-19 mg/kg. A total of 14 cocaine positive rats were placed outside and above ground in the Boston University Forensic Anthropology Outdoor Research Facility (Holliston, MA, U.S.A) for a period of 12 months. All recoverable skeletal samples were collected for testing. Drug free control rat bones were also acquired by placing drug-free rats outdoors, above ground, until full decomposition occurred. In this study, a method analyzing cocaine and its major metabolites benzoylecgonine and ecgonine methyl ester was developed. After homogenization of whole bones, the extraction process was performed using a mixed mode reversed-phase/ion exchange sorbent. The use of a 2D LC/MS/MS technology eliminates the need for a lengthy evaporation step in the extraction method. The chosen 2D LC/MS/MS used in this application was identified using a 6x6 automated method development protocol. The manual extraction of the bone samples was completed in less than an hour. The analysis was performed using 100μL of the final organic solvent (MeOH) extracts. vi The limit of quantitation (LOQ) for cocaine and benzoylecgonine was measured at 0.05ng/g (0.05ng/mL or 50pg/g) of sample material and the LOQ for ecgonine methyl ester was measured at 0.1ng/g (0.1 ng/mL or 100pg/g). The extraction method for cocaine proved to give a linear dynamic range of 2.5 orders of magnitude (0.05 ng/g to 10ng/g with an R2 = 0.998. The micro extraction protocol combined with a multi-dimension chromatography used in this study decreased sample preparation time without sacrificing the quality seen with current single dimension chromatography techniques. The procedure developed in this study can be utilized on bone and completed in less than an hour before injection into the 2D-LC/MS/MS system.

The rare earth elements (REE), composed primarily of the fifteen lanthanides, find broad application in many modern devices making them indispensable and important to the sustainable, green world approach prevalent these days. Liquid-liquid extraction for the separation of lanthanides using a calix[4]arene scaffold with coordinating ligands on its lower rim has been studied for many years. New solid phase materials capable of coordinating metals are important to the field of rare earth metal separation and isolation as they will not only minimize solvent wastes but have the potential to be reused, thereby making the separation and recovery of rare earth metals economical. In the first part, the synthesis and characterization of new chlorine terminated 2-alkyl-p-tert-butylcalix[4]arenes with different coordinating ligands such as phosphine oxides, ketones, CMPO, and esters on the lower rim were investigated. Here, the extraction efficiencies of these new calix[4]arenes for rare earth metals in liquid-liquid extraction was investigated in order to confirm that functionalization at the 2-position of the calix[4]arenes does not interfere with the extraction properties of the coordinating ligands on the lower rim.In the second part of this project, further modification of the 2-position was carried out by converting the chlorine terminated end to the highly desirable amino group, which was then utilized in coupling reactions to immobilize these 2-position/tail modified calix[4]arenes bearing lower rim coordinating ligands on solid supports covalently via the attached linker group. These newly synthesized solid phase extractants were characterized using solid state NMR spectroscopy, FT-IR spectroscopy, elemental analysis and EDS and their thermal stability was further determined using TGA. This represents the first report of attachment of calix[4]arene ligands to solid supports via their 2-positions to the best of our knowledge. These new materials were further investigated in the separation of lanthanides.

O. horridus (a member of the family Araliaceae) is a plant traditionally used by the tribe of Pacific Northwest America for medicinal as well as spiritual purposes. O. horridus is found in the temperate forests of northwestern North America as an understory shrub capable of growing in well-drained to poorly drained, shady sites. Traditional medical preparations of O. horridus include decoction or infusion, preparation as a tea, preparation as a poultice, or preparation as a tonic. O. horridus is used as a form of treatment for cancer, hypoglycemia, diabetes, tough pneumonia, and for colds. A practical and reliable extraction method using Soxhlet extraction, Liquid-Liquid Extraction (LLE), and Solid Phase Extraction (SPE) was designed to extract, purify, and identify chemical compounds from the complex plant matrix of O. horridus. Soxhlet extraction, LLE, and SPE were used successively to obtain the desired sample for HPLC and LC-MS analysis. Soxhlet extraction was performed using methanol to extract the sample from the root bark of O. horridus. This sample was then subjected to LLE using A.C.S. grade solvents, i.e., hexane, chloroform, ethyl acetate, n-butanol, and water successively to extract compounds of different polarities. These fractions were dried down separately using a rotary evaporator, and the fraction of interest (chloroform) was subjected to SPE using a Silica, HyperSep Diol, and Sep-Pak C-18 SPE cartridge successively. The desired extract was separted by C-18 and analyzed using high performance liquid chromatography-UV (HPLC-UV). Extraction and separation by using Soxhlet, LLE, and SPE allowed for sample clean-up and concentration of the analytes of interest. HPLC-UV was used to separate compounds of interest from the post SPE extracted sample of the chloroform fraction. Fraction collection of major peaks allowed for further HPLC analysis of peaks of interest and LC-MS analysis provided rapid analysis of compounds in the collected peaks. Ferulic acid was identified in the chloroform fraction of O. horridus using LC-MS.

An increased interest has been placed on the miniaturization of chemical processes. Focus is placed on rapid chemical analysis, decreased operation cost, minimal generation of chemical waste, and reduced sample size. Presented here is an approached the goal of 'miniaturization' through two distinct areas of analytical chemistry: 1) employing capillary-channeled polymer (C-CP) fibers as a stationary phase in micro-SPE (solid phase extraction) tips for the extraction of proteins from buffered solutions (i.e. biofluids) with analysis by mass spectrometry (MS) and 2) the development of the liquid sampling-atmospheric pressure glow discharge (LS-APGD) as an excitation source for elemental analysis. Determination of proteins in biological matrices is vital for the identification of disease-specific biomarkers. With this in mind, research efforts have grown tremendously for protein analysis in biofluids. More importantly, research has focused on development of extraction/separation techniques necessary for performance on miniaturized levels (i.e. low volume aliquots/concentrations) prior to analysis with mass spectrometry (MS). MS is commonly used for detection due to its sensitivity and the abundant chemical information it provides on analysis. However, buffered species (i.e. salts and biofluids) can be detrimental to MS analysis due to mechanisms such as co-ionization. Presented here, is the use of C-CP fibers as a stationary phase for the extraction of proteins from these matrices. These C-CP mirco-SPE tips have shown to be effective for extraction of nanogram quantities of proteins from buffered species. More importantly, extractions have been optimized to be performed with samples sizes of ~1 µL compared to commercially available tips which require up to 100 µL of sample. Investigations of fiber packing density, tip length, loading capacities, and load/elution volumes were performed to improve the efficiency of C-CP tips for the extraction of proteins. After optimization, C-CP tips were utilized for extracting nanogram quantities of proteins (µL aliquots) from urine matrices. Additionally, there is a need to design spectrochemical instruments with lower power consumption, reduced sample sizes, compact footprint, and the ability to be operated under ambient conditions. In this dissertation the liquid sampling-atmospheric pressure glow discharge (LS-APGD) is employed as a miniaturized approach towards elemental analysis focusing on the determination of plasma characteristics (i.e. plasma temperatures) under parametric evaluations for elemental analysis of solutions and the use of the LS-APGD source as a secondary excitation/ionization source following laser ablation of solid materials. Excitation, rotation, and ionization temperatures along with electron number density and robustness characteristics of the LS-APGD were studied under parametric evaluation. It was determined that the LS-APGD show great promise in terms of handling complex samples (liquid and laser ablated particles) without presenting matrix effects. Both directions demonstrate promising results with regards to miniaturization of chemical analysis techniques in hopes of developing rapid procedures that require small quantities of sample while operating at low costs and producing little to zero chemical waste.

Synthetic cannabinoids were first created in a pharmaceutical setting where scientists were studying marijuana. Researchers were trying to develop medically beneficial marijuana analogs. The compounds, however, were found to give physiological effects that were more potent than marijuana. Presently, synthetic cannabinoids have become a psychoactive drug of abuse, sold in head shops and over the Internet. New compounds are constantly being synthesized, which makes analysis of the drugs difficult. Solid phase extraction (SPE) is a well-studied method used in toxicological analysis to extract drugs and their metabolites from biological fluids. This sample preparation method is necessary to isolate the desired components of a sample for analysis by gas chromatography and mass spectrometry (GC/MS). This study sought to compare four brands of commercially available SPE cartridges using a procedure from United Chemical Technologies (UCT) for the simultaneous extraction of the three synthetic cannabinoid metabolites, JWH-018 N-(4-hydroxypentyl), JWH-122 N-(5-hydroxypentyl), and JWH-250 N-(5-hydroxypentyl), from urine. The cartridges from UCT, Thermo Scientific, Agilent Technologies, and SiliCycle were evaluated to determine how they performed throughout the SPE procedure. A recovery efficiency study was conducted to measure the amount of extracted metabolites from the urine. The responses of the quantification ion of the metabolites from an extracted urine sample were compared to a neat sample and the percent recovery was calculated. A within-run precision study was also utilized to measure the reproducibility of the cartridges, which was determined by the coefficient of variation (CV) of the different brands. The outcome of this research led to a development of a GC/MS method for detection of the three metabolites, creation of calibration curves for quantification, use of SPE for the extraction of the metabolites from urine, and the quantification of the extracted compounds to determine the efficacy and consistency of four brands of SPE cartridges. Method optimization was able to minimize the interday variations seen in the results of aliquots of the same samples. Optimal parameters include initial validation of the GC/MS method, a clean liner for the analysis of synthetic cannabinoid metabolites, using a GC column with a high temperature limit, and derivatization of the extracts before injection into the GC. While this study shows it is possible to use GC/MS for the analysis of these metabolites, LC/MS does not have the same restrictions because a liner, temperature elution, and derivatization of the analytes are not utilized. It was determined from the results of these studies that SiliCycle had the most reproducible and efficient cartridges. SiliCycle cartridges had a consistent and fast flow rate with a percent recovery efficiency within ±20% of the actual value. The results from SiliCycle were followed by cartridges from UCT, Thermo Scientific, and Agilent brands, respectively.

ABSTRACT Vitamins are essential to human beings and support long-term healthy lifestyle. They exist widely in food, but in small amount. Hence, dietary supplements and fortified food products are more and more popular in the nutritional markets. According to the Infant Formula Act of 1980, the range of each of the nutrients should fall within a specific range or above the required minimum, so there is an increasing interest of accurate measurement of vitamins, particularly the B-group vitamins. However, traditional methods for vitamin B analyses are time-consuming and often in poor accuracy. As a result, rapid and reliable analytical methods for simultaneous determination of water-soluble vitamins in multivitamin and supplemented foods are important, indispensible and needed by food and nutraceutical industries for quality control during production, and for accurate evaluation of data and label clams. Due to low concentrations of B8 and B9 in nature, there is a critical need for preconcentration to facilitate their isolation and purification from a complex matrix like food. In order to minimize the consumption of solvents, solid phase extraction (SPE) has been introduced for sample preparation. In this study, two types of SPE, i.e., reversed-phase C18 and strong anion-exchange phase, were used for sample clean-up and pre-concentration. The average recoveries for vitamin B3, B5, B7 and B9 were 101.3%, 102.9%, 92.8% and 102.6%, respectively, by using C18 SPE. In contrast, the strong anion-exchange SPE provided the average recoveries of B3, B5, B7 and B9 in 91.5%, 93.0%, 109.1% and 106.7%, respectively. Furthermore, simultaneous determination of water-soluble vitamins was developed by a HPLC system installed with a ZORBAX Eclipse XDB-C18 (250mm x 4.6mm, 5 μm particle size, Aglient Technologies, Inc., Loveland, CO, USA) with a guard column (12.5mm x 4.6mm, 5 μm particle size), and compared by three detectors, including DAD, ELSD and MS. The B-complex supplement, multivitamin/multimineral tablets and powder and vitamin drink were tested to verify HPLC method. According to the results, LC-MS is the best to do the simultaneous determination of the B-group vitamins in lights of its analytical accuracy, precision, sensitivity and versatility.

The task of sample preparation encompasses many challenges and demands for analytical techniques. Appropriate preparative steps can be the determining factor between successful measurements and failures. Too little preparation can lead to the inability to detect analytes due to low concentrations, insufficient extraction of analytes, and/or strong matrix effects; exhaustive preparation steps increases analysis times, the potential for analyte losses, and the difficulty to obtain consistent reproducible results. Discussed here are several examples of analytical techniques where sample preparation is critical and can be used to improve existing techniques. Capillary-channeled polymer (C-CP) fibers used for solid phase extraction (SPE) protein analysis techniques is described first. The C-CP fibers have several advantageous attributes stemming from the unique shape, wide chemical and pH stability ranges, predictable surface interactions, and overall low material costs. The fibers have been employed in micropipette tip and microcolumn SPE formats for the desalting of proteins from high salt content buffer, synthetic urine and synthetic saliva matrices. Micropipette tips were prepared and used for an off-line method prior to UV-VIS detection, and a microcolumn format was developed as an on-line SPE technique for ESI-MS detection. The characterization of isoflavones present in soy, kudzu, and red clover products by liquid chromatography-particle beam electron ionization mass spectrometry (LC-PB/EIMS) is also described. Here, NIST Standard Reference Materials (SRMs) were extracted using 80% methanol without a hydrolysis step. Samples were then analyzed using a chromatographic separation and analytes quantified by an internal standard (IS) method using 7-hydroxy-4-chromone as the IS compound. The particle beam allows for simple interfacing between LC separations and EIMS detection, giving ability to attain qualitative and quantitative information simultaneously. Isoflavones were quantified and compared to NIST certified values.

Solid phase extraction (SPE) has become a popular sample preparation method for the extraction of analytes from complex matrices. Solid phase extraction overcomes such disadvantages as large solvent usage and waste, large amounts of time spent in sample preparation and labor intensive methods with little hope of automation. In this report, SPEC®⋅ Microcolumns are evaluated for the extraction of Zantac® (ranitidine hydrochloride) from urine for high performance liquid chromatography (HPLC) analysis. SPEC®⋅Microcolumns are rigid disc structures with glass fiber impregnated with silica gel. These microcolumns look like packed-bed SPE cartridges but have the advantages of low void volumes and low bed mass that give clean extracts with the use of small amounts of solvents. An SPE method was developed and evaluated for the recovery of Zantac® from water and spiked urine. Percent recoveries and matrix interferences are reported. The limit of detection and analytical range tor the HPLC method of analysis with ultraviolet detection will be presented.