by Kwok Chiu Nga. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 1-3 (4th gp.)). / Abstract also in Chinese. / TABLE OF CONTENTS --- p.i / ABSTRACT --- p.iv / 摘要 --- p.vi / LIST OF FIGURES --- p.vii / LIST OF TABLES --- p.x / ABBREVIATION --- p.xi / Chapter CHAPTER ONE --- RESEARCH BACKGROUND / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.1.1 --- Alkaloids --- p.1 / Chapter 1.1.2 --- Diterpenoid alkaloids --- p.2 / Chapter 1.1.3 --- Aconitine-type alkaloids --- p.2 / Chapter 1.1.4 --- Toxicity --- p.4 / Chapter 1.1.5 --- Safety concerns --- p.4 / Chapter 1.2 --- Summary of the Previous Work --- p.8 / Chapter 1.3 --- Objectives and Outline of the Present Work --- p.13 / Chapter CHAPTER TWO --- INSTRUMENTATION AND EXPERIMENTAL / Chapter 2.1 --- Instrumentation --- p.15 / Chapter 2.1.1 --- High performance liquid chromatography (HPLC) --- p.15 / Chapter 2.1.2 --- Triple-stage quadrupole (TSQ) mass spectrometer --- p.17 / Chapter 2.1.2.1 --- Atmospheric pressure chemical ionization (APCI) --- p.17 / Chapter 2.1.2.2 --- Electrospray ionization (ESI) --- p.20 / Chapter 2.1.2.3 --- Quadrupole system --- p.20 / Chapter 2.1.2.4 --- Ion detection system --- p.22 / Chapter 2.1.2.5 --- Data system --- p.22 / Chapter 2.2 --- Experimental --- p.22 / Chapter 2.2.1 --- Sample and reagents --- p.22 / Chapter 2.2.2 --- Sample preparation --- p.23 / Chapter 2.2.3 --- High performance liquid chromatography conditions --- p.23 / Chapter 2.2.4 --- Mass spectrometry conditions --- p.25 / Chapter 2.2.4.1 --- Atmospheric pressure chemical ionization conditions --- p.25 / Chapter 2.2.4.2 --- Electrospray ionization conditions --- p.25 / Chapter CHAPTER THREE --- SELECTION AND OPTIMIZATION OF HPLC/MS METHOD / Chapter 3.1 --- Introduction --- p.26 / Chapter 3.2 --- Experimental --- p.29 / Chapter 3.3 --- Results and Discussion --- p.29 / Chapter 3.3.1 --- Triethylamine concentration --- p.31 / Chapter 3.3.2 --- Ammonium acetate concentration --- p.34 / Chapter 3.3.3 --- Acetic acid concentration --- p.37 / Chapter 3.3.4 --- HPLC/MS interface --- p.40 / Chapter 3.3.5 --- MS/MS conditions --- p.40 / Chapter 3.4 --- Conclusions --- p.43 / Chapter CHAPTER FOUR --- DETERMINATION OF ACONITINE-TYPE ALKALOIDS IN ACONITE ROOTS / Chapter 4.1 --- Introduction --- p.48 / Chapter 4.2 --- Experimental --- p.48 / Chapter 4.3 --- Results and Discussion --- p.50 / Chapter 4.3.1 --- Selection of internal standard --- p.50 / Chapter 4.3.2 --- Method validation --- p.50 / Chapter 4.3.2.1 --- Precision of measurement --- p.50 / Chapter 4.3.2.2 --- Accuracy of measurement --- p.50 / Chapter 4.3.2.3 --- Limits of detection and quantitation --- p.58 / Chapter 4.3.3 --- Determination of aconitine-type alkaloids in aconite roots --- p.58 / Chapter 4.4 --- Conclusions --- p.60 / Chapter CHAPTER FIVE --- CONCLUSIONS AND FUTURE WORK / Chapter 5.1 --- Conclusions --- p.67 / Chapter 5.2 --- Future Work --- p.68 / ACKNOWLEDGMENT --- p.A1 / APPENDIX --- p.A2 / REFERENCES --- p.R1

Mass spectrometry imaging (MSI) is a powerful tool that provides mass-specific surface images with micron or sub-micron spatial resolutions. In a microscope MSI experiment, large sample surfaces are illuminated with a defocused laser or primary ion beam, enabling all surface molecules to be desorbed and ionised simultaneously before being electrostatically projected onto a position-sensitive imaging detector at the end of a time-of-flight mass analyser. Traditionally only the image of one mass-to-charge ratio can be obtained in a single acquisition, which limits its applicability. However, the development of event-triggered sensors, such as CMOS-based cameras, revives the microscope MSI method by allowing multi-mass imaging. Therefore, the challenges facing microscope have MSI shifted to improving its mass resolution, effective mass range, and mass accuracy. This thesis proposes effective solutions to each of them, and thus significantly improves the performance and applicability of microscope MSI. To increase the mass range, two modified post-extraction differential acceleration (PEDA) techniques, double-field PEDA and time-variable PEDA, were used to demonstrate mass-resolved stigmatic imaging over a broad m/z range. In double-field PEDA, a potential energy cusp was introduced into the ion acceleration region of an imaging mass spectrometer, creating two m/z foci that were tuned to overlap at the detector plane. This resulted in two focused m/z distributions that stretched the mass-resolved window with m/Δm >= 1000 to 165 Da without any loss in image quality; a range that doubled the 65 Da achieved under similar conditions using the original PEDA technique. In time-variable PEDA, a dynamic pulsed electric field was used to maximize the effective mass range of PEDA. By simultaneously focusing ions between 300 to 700 m/z using an exponentially rising voltage pulse, time-variable PEDA provides an effective mass range more than six times wider than the original PEDA method. Although reflectrons are widely used to improve the mass resolving power of ToF-MS, incorporating them in a microscope MSI instrument is novel. A reflectron MSI instrument was designed and implemented. Simulations demonstrated that one-stage gridless reflectrons were more compatible with the spatial imaging goal of the microscope MSI instrument than the gridded reflectrons. Preliminary experimental results showed that coupling the gridless reflectron with single-field PEDA achieved a mass resolution above 8,000 m/Δm while keeping a spatial resolution of 20 um. In conclusion, the gridless reflectron was able to triple the mass resolving power without losing any spatial imaging power. The poor mass accuracy hurdle was overcome by machine learning algorithms, which can construct clinical diagnostic models that recognise the peak pattern of biological mass spectra and classify them accurately without knowing the actual mass of each peak. After a proof of concept "experiment", where the mass spectra of dye molecules were classified by various learning algorithms, three pairs of datasets (ovarian cancer, prostate cancer, chronic fatigue and their respective controls) were used to build classifiers that accurately distinguish blood samples from controls. Possible biomarkers were also discovered by evaluating the importance of each m/z feature, which may assist further studies.

Com a demanda cada vez maior por corantes alimentícios não tóxicos e sustentáveis, fontes naturais têm emergido como uma potencial alternativa aos corantes sintéticos artificiais. O urucum, planta tropical, tem sido cada vez mais explorado para suprir as demandas por corantes naturais, visto que sua semente apresenta uma alta concentração de carotenoides com poder colorífico. O Brasil que já era grande produtor de urucum tem sido destaque na produção de suas sementes para extração de pigmentos, devido à alta na demanda por corantes de origem natural. Das sementes do urucum é possível a extração de dois dos principais compostos para fabricação de corantes - bixina e norbixina. Visando contribuir para o melhor entendimento das potencialidades das sementes de urucum, além de fornecer evidência experimental sobre as propriedades antioxidantes e conteúdo de compostos fenólicos, flavonoides e carotenoides, foram determinados o perfil químico, o potencial antioxidante a partir de extratos de sementes de urucum, bem como a caracterização do carotenoide bixina por HPLC-PDA e ESI-Orbitrap. Sendo assim possível observar que o perfil químico e os teores de compostos bioativos em sementes de urucum não apresentam diferença independente da região produtora. Por fim, o projeto de pesquisa focou na conversão de bixina (carotenoide lipossolúvel que representa 80% do total de carotenoides presentes na semente) em norbixina (carotenoide solúvel em água) para suprir o aumento de demanda por corantes hidrossolúveis, visto que a indústria de alimentos tem visado a redução de gorduras nos alimentos. / With the increasing demand for non-toxic and sustainable food coloring, natural sources have appeared as a potential alternative to artificial synthetic dyes. The tropical plant known as annatto has been increasingly exploited to attend the demands for natural colorings as its seed are rich in carotenoids with high coloring potential. Brazil already considered a big producer of annatto seeds has been prominent in the production of its seed for extraction of its pigments, due to increasingly demand for natural-source food coloring. From annatto seeds it is possible to extract two of the main compounds for fabrication of coloring - bixin and norbixin. In order to a better understanding of the potential of annatto seeds as well as providing experimental evidence on the antioxidant properties and content of phenolic, flavonoids and carotenoids, the chemical profile, antioxidant potential as well as the characterization of bixin carotenoid by HPLC-PDA and ESI-Orbitrap, were determined from annatto seeds extract. It was found that the chemical profile and the content of bioactive compounds in annatto seeds did not differ independently of its producing site. In addition, the research foccused on the conversion of bixin (liposoluble carotenoid, responsible for 80% of the seeds\' carotenoids) into norbixin (water soluble carotenoid) to attend the demand for water-soluble coloring, as the food industry has been aiming a reduction in food fat content.

Chondroitin sulfate proteoglycans (CSPGs) are found in specialized brain extracellular matrix structures termed perineuronal nets (PNNs). The chondroitin sulfate chains of these CSPGs are thought to have a strong effect on neuroplasticity, along with development, injury, and diseased states of the brain. Wisteria floribunda agglutinin (WFA) is a plant lectin used to identify PNN via staining; the pattern of this staining is changed upon schizophrenia. As such, one powerful method of probing the identity of the CS chains of PNNs and addressing what changes in CS identity occur during schizophrenia is to characterize the features of the CS which bind to the lectin. Methods for characterization of WFA-CS binding and their biological relevance were developed and evaluated. Commercially available CS was used to probe the binding affinity of the agglutinin to various regions of CS via hemagglutination inhibition assays and affinity gradient elution of CS bound to WFA. The size, sulfation extent, and fragment location in the CS chain from these eluates were determined using HILIC-LC-MS. As commercial sources can be used to elucidate the binding specificity of WFA, but not the actual relevant binding partner of WFA within the brain, PNN CS extractions were performed with a modified method aimed at reducing the timescale at which PNN CS can be obtained so as to allow similar experimentation on CS directly from PNN. The results pave the way for further determination of WFA-CS binding.

Spurious, falsely-labeled, falsified or counterfeit (SFFC) pharmaceuticals are a health concern that claims hundreds of thousands of lives annually1, a violation of intellectual property rights which cost legitimate companies billions2, and a low-risk high yield revenue stream for organized crime2. While ports of entry and border control points are the primary access control points for SFFC3,4, advances in field portable detection and equipment offers an increasingly effective method for the assessment of pharmaceuticals at regional centers and points of distribution. This is particularly important for less developed countries (LDC) who do not maintain satellite or regional testing facilities. As part of a proposed protocol to assess field portable detection equipment, an ultrafast liquid chromatography, tandem mass spectrometry (UFLC-MS/MS) method for the quantification of liquid formulation Oxytocin was developed. The six minute method was found to have a within run %bias of +/-16%, a linear dynamic range of 150-1000 nanograms/milliliter (ng/ml), and an accuracy within acceptability criteria for all tested concentrations. The effectiveness of three identified transition ions, 723.1, 86.2 and 70.1 Daltons, for the analysis of oxytocin by mass spectrometry was assessed across several figures of merit to include signal to noise ratio, %CV, calibration sensitivity, and analytical sensitivity. The 723.1 ion fragment was recommended for quantification, while the 70.1 dalton ion was recommended as a qualifier ion, although 86.2 also performed within acceptability criteria. A method for the UFLC-MS/MS assessment of degradation products for oxytocin was proposed for specificity testing. Degradation of oxytocin by exposure to highly acidic, basic, and thermal conditions for one hour was attempted. Formation of degraded products was not observed. Additionally, existing High Performance Liquid Chromatography (HPLC) methods for the simultaneous assessment of Artesunate and Amodiaquine HCl were modified to assess compatibility with UFLC. No method assessed produced sufficient quality signal to continue with method development.

The Centers for Disease Control and Prevention deemed the increase in overdose fatalities, due to the use of opioids, an “opioid epidemic” in the United States. Heroin, fentanyl, and other synthetic opioids are commonly abused and are contributing to the opioid epidemic. In 2016, the Drug Enforcement Administration temporarily placed three fentanyl analogs (beta-hydroxythiofentanyl, butyryl fentanyl, and furanyl fentanyl) under Schedule I due to their imminent threat to public health. These drugs elicit analgesic effects similar to heroin making them desirable drugs to abuse. Novel fentanyl analogs and designer opioids are expected to become more prominent in forensic casework in the near future as the opioid epidemic continues. These drugs can be seen in forensic seized drug and urine casework samples either alone or mixed with other drugs of abuse. It is therefore necessary to have an efficient methodology to identify these new compounds. Currently, gas chromatography-mass spectrometry (GC/MS) is used to identify drugs of abuse and is considered the “gold standard” in forensic casework. However, analysis times can often range from 15 to 60 minutes in length. Another drawback is the need for spectral library matching, which requires analytical reference materials for identification. Therefore, the identification of novel fentanyl analogs and designer drugs is limited until a reference material becomes available. In this study, direct sample analysis time-of-flight mass spectrometry (DSA-TOFMS) was evaluated to provide rapid identification of fentanyl and other synthetic opioids in seized drug and urine casework samples. DSA is a direct ambient ionization source, which requires no chromatography and minimal sample preparation. TOFMS is a high resolution mass spectrometer that uses collision-induced dissociation (CID) to produce precursor ion and characteristic fragmentation ions, which provide additional structural and molecular formula information, allowing for the identification of compounds without a reference material. The analytes explored in this study include: heroin, 6-monoacetylmorphine (6-MAM), morphine, fentanyl, norfentanyl, 4-anilino-N-phenethylpiperidine (4-ANPP), acetyl fentanyl, beta-hydroxythiofentanyl, butyryl fentanyl, furanyl fentanyl, valeryl fentanyl, AH-7921, U-47700, buprenorphine, norbuprenorphine, desomorphine, MT-45, W-15, and W-18. Direct sample analysis time-of flight mass spectrometry (DSA-TOFMS) is a novel instrumentation that could be utilized in the forensic sciences field to qualitatively identify illicit substances in casework samples. In this study, 19 compounds of interest containing heroin, fentanyl, fentanyl analogs, and other synthetic opioids were evaluated using DSA-TOFMS. DSA-TOFMS abbreviated the workload of the analysis and was utilized to provide precursor ion and characteristic fragmentation ions within an analysis time of 20 seconds. Certified reference standards were used to optimize instrumentation settings, to determine precursor ions and characteristic fragmentation ions, and to determine the limit of detection of the instrument. A carryover study determined there were no persisting ions present when entering the capillary inlet between runs. A repeatability study revealed the DSA-TOFMS repeated results within the acceptable criteria range of above 500 counts and within 10ppm error 93% (10ppm) and 83% (1ppm). Forensic seized drug casework samples were evaluated with DSA-TOFMS and qualitatively identified. Out of the 64 samples, 89% were qualitatively identified as heroin, 4% were qualitatively identified as fentanyl, 1% was qualitatively identified as heroin and fentanyl, 3% were qualitatively identified as acetyl fentanyl, and 3% were qualitatively identified as furanyl fentanyl. The casework samples containing furanyl fentanyl were considered “true unknown unknown samples,” as the Maine Health and Environmental Testing Laboratory gas chromatography-mass spectrometry library did not have a spectrum to use for the identification of these samples. Forensic urine casework samples were evaluated with DSA-TOFMS. Samples previously confirmed to contain compounds of interest were prepared using minimal sample preparation technique (filtered using 0.45 microns syringe filters and diluted (1:10) with LC/MS grade water). Analysis displayed the limitations of DSA-TOFMS as only twelve of the forty compounds of interest were present and only three of the twelve were within the acceptable criteria range. DSA-TOFMS is a fast and reliable technique with minimal sample preparation for forensic seized drug samples. However, the concentration in complex matrixes, such as urine and blood, were unable to be qualitatively identified using this sample preparation method by DSA-TOFMS.

No description available.

Thesis (Sc.D.)--Massachusetts Institute of Technology, Dept. of Chemical Engineering, 1981. / MICROFICHE COPY AVAILABLE IN ARCHIVES AND SCIENCE. / Bibliography: leaves 936-960. / by James D. Bittner. / Sc.D.

Dentre os herbicidas atualmente comercializados, o glifosato é um dos mais utilizados no Brasil. A eficácia do glifosato, bem como de outros herbicidas, no combate às ervas daninhas, esbarra nos problemas que esses compostos acabam proporcionando ao meio ambiente. Embora estudos venham sendo realizados para elucidar melhor o comportamento de herbicidas no meio ambiente, a complexidade do comportamento desses compostos abre caminho para realização de um número significativo de trabalhos de pesquisa. Nesses estudos, é rotineiro o uso de compostos radiomarcados (traçadores radioativos) para avaliar a biodisponibilidade no solo. Todavia, o emprego, manipulação e estocagem de compostos radiomarcados exigem cuidados sob o ponto de vista da segurança, razão pelo qual o uso de isótopos não radioativos é uma tendência internacional, especialmente em pesquisas de campo. Dentre desse contexto, o trabalho descreve um método para a síntese do glifosato enriquecido no isótopo estável do nitrogênio (15N). A sintese do herbicida-15N foi realizada utilizando-se da reação de fosfometilação com diaquil fosfito e glicina-15N. Os testes foram realizados em microescala e em quantidades equimolares. Nas condições estabelecidas, foi possível alcançar um rendimento de reação de aproximadamente 25%. A otimização das condições de sintese, empregando a técnica isotópica, deverá disponibilizar uma importante ferramenta a ser utilizado em estudos referentes ao comportamento do glifosato no sistema solo-planta / Among the actual commercialized herbicides, the glyphosate is one of the most used in Brazil. Its efficiency, as well as the others herbicides, on weeds control is attached to problems that this composts cause to the environment. Although studies about the herbicides behavior in the environment have been done, the complexity of this composts behavior leads to the accomplishment of an expressive number of research works. In these works, it?s common the use of radio labeled compounds (radioactive tracers) to evaluate its bio-availability in the soil. However, the use of this radio labeled composts, its manipulation and storage demands a careful handling in the chemical security point of views. This is the reason the use of non radiactive isotopes is an international tendency, especially for field researches. According to this context, the present work describes a method for the synthesis of enriched glyphosate for the nitrogen stable isotope (15N). The tests were undertaken in micro-scale and in equimolar quantities. Under these established conditions, the reaction reached the yield of approximately 25%. The optimization of the synthesis conditions with the use of isotopic techniques shall offter an important tool to be used in studies that are referred to glyphosate behavior in soil- plant system

No description available.