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No description available.

No description available.

No description available.

The purpose of this research was to design a simple, novel interface for on-line coupling of Supercritical Fluid Extraction (SFE) with High Performance Reversed Phase Liquid Chromatography (HP-RPLC), and to explore its ability for quantitative analysis of analytes in different matrices. First, a simple interface was developed via a single one six-port injection valve to connect the SFE and LC systems. A water displacement method was utilized to eliminate decompressed CO2 gas in the solid phase SFE trap and connection tubes. To evalute this novel hyphenated system, spiked polynuclear aromatic hydrocarbons (PAHs) in a sand matrix were used as target analytes with the achievement of quantitative results. Also PAHs in naturally contaminated soil were successfully extracted and quantitatively determined by this hyphenated system. Compared to the EPA method (Soxhlet extraction followed by GC-MS), on-line SFE-LC gave precise (4-10% RSD) and accurate results in a much shorter time. Based on this hyphenated technique, a method for the extraction and analysis of hyperforin in St. John's Wort was developed under air/light free conditions. Hyperforin is a major active constituent in the antidepression herbal medicine-Hypericum Perforatum (St. John's Wort). Hyperforin is very sensitive to oxygen and light. There is no way to date to determine whether any degradation occurs during the sample-processing step in the analytical laboratory. On-line coupling of SFE-LC with UV absorbance/ electrospray ionization mass spectrometry (SFE-LC-UV/ESI-MS) provided an air/light free extraction-separation-detection system, which addressed this issue. Mass spectral data on the extract confirmed the presence of the major degradation compounds of hyperforin (i.e. furohyperforin and two of its analogues). Thus, the degradation process must have occurred during plant drying or storage. The feasibility of quantitative extraction and analysis of hyperforin by on-line SFE-LC was made possible by optimizing the extraction pressure, temperature, and modifier content. High SFE recovery (~90%) relative to liquid-solid extraction was achieved under optimized conditions. We then extended the interface's application to an aqueous sample by using a liquid-fluid extraction vessel. Quantitative extraction and transfer were achieved for the target analytes (progesterone, phenanthrene, and pyrene) spiked in water, as well as in real samples (urine and environmental water). During each extraction, no restrictor plugging was realized. Extraction temperature and pressure were optimized. Different amounts of salt were added to the aqueous matrix to enhance ionic strength and thus extraction efficiency. Methanol and 2-propanol were used as CO2 modifiers. Two modifier modes were compared, e.g. dynamically mixing modifier with the CO2 extraction fluid, and pre-spiking modifier in the extraction vessel. Surprisingly, we found pre-spiking the same amount of modifier in the vessel enhanced the recovery from ~70% to ~100% for progesterone, phenanthrene, and pyrene due to a "co-extraction effect". The last phase of our work explored the disadvantages/limitations of this hyphenated technique through the analysis of more highly polar phenolic compounds in grape seeds. Five types of SFE trapping adsorbent materials were evaluated in an effort to enhance the collection efficiency for the polar components. Pure supercritical CO2 was used first to remove the less polar oil in the seeds. Then methanol-modified CO2 was used to remove the polar components (e.g. phenolic compounds). Catechin and epicatechin (90%) were exhaustively extracted out of the de-oiled seed after 240 minutes with 40% methanol as modifier. Both singly linked (B-type) and doubly linked (A-type) procyanidins were identified by LC-ESI-MS, as well as their galloylated derivatives. Compared to the off-line SFE-LC approach, much less sample was required for extraction in the on-line method, since all the extracted components could be transferred to the LC column. Also, no extract processing/concentration step was needed in the on-line method. However, in the on-line mode, some polar compounds were lost (1) during the collection step (e.g. lower trapping efficiency on a single solid SFE trap when a high percentage modifier was used) and (2) during the water rinsing step (e.g. less retention of polar compounds on C18 trap). Therefore, this hyphenated technique is less desirable for the analysis of highly polar compounds. / Ph. D.

Semiochemicals released from logs infested by southern pine beetle (SPB), <i>Dendroctonus frontalis</i>, from a total of eight infestations located in Texas, South Carolina, and Virginia were collected four to eight days after initial attack. The quantities and chiralities of most semiochemicals, as analyzed by gas-liquid chromatography, showed geographic and temporal variations. Changes in the quantities of α-pinene (aP), frontalin (F), and <i>endo</i>-brevicomin (eB), are believed to result from responses of the host and the beetle to each other's activity at a given time and differences in their respective health. The chiralities of aP, F, and eB at all locations generally remained stable over time, yet variation across the insect's geographic range, particularly for aP and F, is believed to be due to genetic variation of individuals. Geographic and temporal variations in the quantities and chiralities of <i>cis</i>-verbenol (cV), <i>trans</i>-verbenol (tV), and verbenone (V) are presumed to be due to the multiple pathways of origin (SPB, autoxidation, and microorganisms). Analysis of the same semiochemicals isolated from hindguts of individual beetles from Texas, South Carolina, and North Carolina showed quantities of cV and tV to be substantially greater in females than in males; whereas, males contained much greater amounts of V. Geographic differences were found in quantities of tV and V in both sexes and in aP and F in males only. The chiralities of most semiochemicals present in SPB hindguts differed markedly from those released from infested logs. Males produced predominantly (+)-F and (-)-eB, (-)-cV, and (-)-V; however, the chirality of tV varied considerably among areas. In contrast, females produced predominantly (+)-cV and (+)-V and (-)-F, (-)-eB, and (-)-tV. The (+) enantiomer of aP predominated in both sexes, but the proportion of (+ )-aP was generally lower than that released from SPB-infested logs from the same areas. Geographic differences in chirality of tV and V were significant in males and for eB in females. In laboratory trials, aP autoxidized under ambient temperatures to form tV, myrtenol (M), V and to a lesser extent, cv. Both the quantities and chiralities of these compounds were dependent on the chirality of the aP precursor. Significantly greater amounts and proportions of the (+) enantiomer of each compound were produced when (+)-aP was predominant than when the antipode of the precursor was predominant. The extent to which autoxidation products playa role in bark beetle behavior is expected to be dependent on the proportion of aP in pine resin and its chirality present in a pine species. The information gained from this research was used to elaborate on previously proposed behavioral sequences occurring during the mass attack of host trees by SPB and to suggest new avenues to improve the use of semiochemicals in pest management efforts. / Ph. D.

The development of Infrared Spectrometry as a High Performance Liquid Chromatographic detector is presented. Early work with both a conventional dispersive instrument and a Fourier Transform Infrared (FTIR) Spectrometer is presented coupled via a flow cell to size exclusion chromatography. These were used for the analysis of the non-volatile components produced in the liquefaction of coal. Additional work is presented for the coupling of FTIR to analytical scale normal phase chromatography via a flow cell technique. Analysis of both model mixtures as well as a complex process solvent used in the liquefaction process is discussed. Use of deuterochloroform as an improved IR transparent solvent is demonstrated. Work with microbore (1 mm i.d.) columns coupled with on-line flow cell detection is presented. Modification of the flow cell design for microbore compatability is shown as well as the benefits of microbore columns for fiow cell FTIR. Detection limits as amount injected for both analytical and microbore scale HPLC-FTIR are shown. / Ph. D.

Thin film planar waveguides were originally introduced in microwave engineering. The spectroscopist began to use such waveguides as tools to solve chemical characterization problems since Harrick and Fahrenfort introduced the Attenuated Total'Reflection (ATR) in the early 1960's. Today, planar waveguides are playing an important and ever-increasing role in modem chemistry. In this thesis, a novel design for a refractometer involving the application of a thin film planar waveguide and coupling prism was demonstrated. This device shows the feasibility of refractive index measurements in a flowing stream. Therefore, an online detector for High Performance Liquid Chromatography (HPLC) was chosen as the vehicle to test out the concepts. The research works were devoted to studies of waveguide properties and flow dynamics in a chromatographic situation. It was found that microscale detection is possible. The sensitivity can be enhanced by using the highest propagation mode as the probe, and by selecting a proper refractive index liquid as the solvent carrier. A description of the investigation and the various factors involved in designing and optimizing a planar waveguide for refractive index detection is included. The results provide guidelines for the device as a realistic analytical detector. / Ph. D.

Atomic emission spectroscopy is a powerful method for chemical analysis because it offers convenient qualitative and quantitative determination of the elemental composition of an analytical sample. Although most frequently used to determine the total concentration of a given analyte, atomic spectroscopy may be coupled with a separation technique to aid in the elemental identification of individual species. Recently, supercritical fluid chromatography (SFC) has gained popularity among the research community partially because SFC is much more tolerant of polar functional groups than is gas chromatorgraphy (GC). SFC can be used in applications where derivatization is not possible due to sample complexity or steric hindrance. However, these polar compounds often require the addition of polar organic solvents to SFC to improve retention characteristics and peak shape. These polar modifiers reduce or eliminate the utility of flame-based detectors. Therefore, researchers have begun to look for alternative detection systems for SFC with polar modifiers. In particular, atomic emission based detectors using plasmas have become popular to fill this void. This research stems from several characteristics that the plasma based detectors possess which make them more appealing than flame-based photometric detectors. These characteristics include improved analytical sensitivity, fewer spectral interferences, a wide dynamic range and the ability to tolerate polar organic modifiers. Possibly the most important attraction of the plasma detector is, with respect to chromatographic analyses, that this detector allows analysis of compounds that co-elute from a column because the detector is element-specific. The evaluation of the high efficiency microwave induced plasma (HEMIP) as an elemental selective detector for packed column supercritical fluid chromatography has been explored in this dissertation. The effect of CO₂ introduction on the analytical parameters of the plasma, the feasibility of coupling packed column supercritical fluid chromatography to the argon HEMIP for the determination of metals, the use of a helium sustained plasma for nonmetal determinations, and the application of the He-HEMIP as a sensitive and selective detector for packed-column SFC. / Ph. D.

High performance liquid chromatographic separations employing on-line flow cell Fourier transform infrared spectroscopic detection were developed for polar compounds including phenols, alcohols, amines and azaarenes. Detection by FTIR gave information concerning hydrogen bonding and solvent effects occurring during the separations in addition to giving structural information about eluted species to aid in identification. Both analytical size and microbore normal phase columns were employed. Such experimental considerations as column overload, injected minimum detectable quantities, the use of analytical vs. microbore columns and flow cell pathlength were examined. The developed HPLC-FTIR systems were applied to the analysis of several coal liquefaction samples for heteroatom content. Confirmatory and additional information concerning the samples were provided by field ionization mass spectrometry, gas chromatography/ mass spectrometry and reversed phase liquid chromatography employing UV detection. An equation relating reversed phase retention times to structural parameters was developed and applied to the analysis of the coal-derived samples. Two process solvents were found to contain primarily alkyl-substituted phenols in addition to azaarenes such as pyridine and quinoline. Some non-distillable coal-derived samples were found to contain azaarenes such as alkyl quinolines. Evidence was also found concerning the presence of hydroxypyridine type compounds and the incorporation of process solvent molecules into the coal structure during liquefaction. / Ph. D.