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.

Supercritical fluids were introduced as mobile phases for chromatography and extractions in the early sixties. Over the past decade the technique has received increased attention, largely owing to the introduction of several commercial instruments and to the ease of adapting available commercial equipment for use in sfe and sfc. This dissertation examines the use of supercritical CO₂ as a mobile phase for chromatography and extraction. The first chapter explores the problems and provides background for the research. The physical properties of supercritical fluids are described and pertinent recent research is identified and summarized. The following chapter presents the design of three interrelated studies that investigate in-situ concentration, quantitative aspects of sfc, and quantitative aspects of sfe. The first of the three studies examined the exploitation of the innate properties of a supercritical fluid. Because the mechanism of elution with supercritical fluid mobile phases is based on solvation, it was possible in this study to concentrate analytes at the head of the supercritical zone. Concentration was followed by supercritical fluid chromatography. This allowed the analysis of sample components at part per billion levels. The use of supercritical fluids as mobile phases for chromatography and the quantitative nature of sfc with flame ionization detection was examined in the second study. Under numerous detector and injector configurations, a distinct decrease in response factors with an increase in carbon number was evident. Results showed that the decrease in response factors was related to an increase in CO₂ flow through the detector. The magnitude of the decrease was multivariably dependent. The third study dealt with the quantitative aspects of using a supercritical Oobile phase for the extraction of PNAs and pesticides from several matrices including contaminated soil. Results showed that sfe is a reliable, easy, and efficient (> 85% recovery) method of removing trace materials from contaminated soil. Implementation of a simple resistively heated collector was used to circumvent the problems of extracting damp matrices such as soil. The results and conclusions are presented in the final two chapters. / Ph. D.

Wood pulp mill effluents continue to attract much attention due to environmental consequences. However, in comparison, very little work has been published on wood pulp extracts themselves. In this investigation, chemithermomechanical (CTMP) pulps as well as Kraft (BKP) pulps were Soxhlet extracted with solvents of different polarity. These two types of pulp extracts were then compared qualitatively using GC-FID and GC-MSD as well as quantitatively based on the percent of extractives obtained. For all the pulps studied, the percent extractives of water > ethyl acetate > cyclohexane. The CTMP extracts exhibited many more components as compared to BKP extracts for all the extractions solvents. The presence of trace chlorinated phenolics in the above wood pulp extracts was also addressed utilizing GC-ECD, GC-EIMS and GC-NCIMS. 4-MCG, 4,5-DCG, 4,5,6-TCG, 3,4,5-TCG, 2,4,6- TCP, 2,3,4,6-TeCP, PCP and 6-MCVN were discovered. Due to a lack of knowledge of the complete history of the wood pulps studied, the exact causes for their discoveries are unknown. Attempts were also made to study the feasibility of Supercritical Fluid Extraction of the above mentioned wood pulps due to the difficulties faced with Soxhlet extractions. The percent extractives obtained using SF-CO₂ and cyclohexane were found to be comparable. / Ph. D.

An explosive vapor generator which produces transient "clouds" of explosive vapors for use in the evaluation of commercial explosive vapor detection systems has been developed based on a capillary gas chromatograph. The design of the vapor generator replaces a flame ionization detector on the gas chromatograph with an effluent heater system to provide a cloud of explosive vapor while preventing condensation onto the walls of the capillary column. This vapor source has been successfully used to produce vapors of dinitrotoluene (DNT), trinitrotoluene (TNT), and 1,3,5-trinitro-1,3,5- triazacyclohexane (RDX). The chemical composition of the output was confirmed by both ion mobility spectrometry, and quadrupole mass spectrometry. Adsorptive effects of TNT and RDX were studied in order to optimize the injection parameters, achieving detection limits three orders of magnitude lower than previously reported. The quantitative output of the system was verified using thermal tube desorption gas chromatography. / 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.

A rapid scanning square wave voltammetric detector has been developed for use with high performance liquid chromatography.The electrochemical cell used in the detector was designed so that the HPLC effluent flows through the center of a large diameter platinum disk electrode and is then forced to flow radially across the electrode surface. The arrangement of the electrodes in the cell was intended to result in large analytical currents while minimizing electrical resistance and analyte band spreading in the detection zone. The detector was evaluated in terms of its minimum detectable quantity, linear dynamic range, electrochemical efficiency, and analyte band spreading. The MDQ was found to be in the low parts per billion range for hydroquinone. The detector was shown to provide data that is qualitatively superior to data obtained by amperometric detection and was shown to be compatible with gradient elution HPLC over a broad range of solvent compositions. A sensor based on the quartz crystal microbalance was also developed for use in flowing solvent streams. Quartz crystals were treated with various compounds to produce close - packed monolayer coatings which could interact with solutes entering the flow cell. The solute capacity was determined for one of the monolayer coatings and various factors that influence the magnitude of the OCM signal were investigated. These factors include the solvent flow rate, the solvent strength, solute molecular structure, and bonded phase molecular structure. The QCM sensor was found to be a convenient probe for conducting surface adsorption studies and the molar free energy of adsorption was determined for some chemically related solutes on an amine modified crystal. / Ph. D.

In our previous studies, uncross-linked large diameter cellulose beads were optimized for solids content, bead size, pressure-flow limits, molecular accessibility and performance as an immunosorbent. Here, anion exchange (DEAE) cellulose beads were derivatized by two different procedures (defined as A and B) and the changes in bead morphology were correlated with transport and sorption kinetics. The kinetic characteristics clearly defined a minimum of two different types of protein binding site architecture. DEAE cellulose beads exhibited molecular exclusion of BSA near the edge of the bead in contrast to greater permeability seen in underivatized beads. Thus, accessible BSA binding sites are present only on the surface of the derivatized beads. DEAE cellulose beads derivatized by procedure B gave higher density of DEAE ligand as compared to beads derivatized by procedure A, as well as higher static and dynamic capacity for BSA. Even though DEAE cellulose beads (DP 2070, 450 μm diameter derivatized by procedure B) have lower small ion capacity than DEAE cross-linked agarose beads, as well as 1/4 the surface area, they exhibit equivalent binding capacity for BSA per volume of support. Thus, DEAE cellulose beads possess more sites per surface area as well as have lower ligand density per BSA site. Furthermore, BSA adsorption sites on DEAE cellulose beads derivatized by procedure B exhibit slow binding kinetics as compared to those derivatized by procedure A and also compared to DEAE crosslinked agarose beads. Thus, the rate limiting step for the adsorption of BSA on DEAE cellulose beads was not diffusion as suggested by the large diameter of the bead. Feasibility studies were performed for process scale applications to fixed and expanded bed anion exchange purification. The large diameter DEAE cellulose beads of this study maybe useful for process scale anion exchange as evident from purification of immunoglobulins from hybridoma cell culture in fixed bed. The balance of large diameter and density of these DEAE cellulose beads enable stable expanded bed purification of proteins such as recombinant human protein C from transgenic porcine whey. / Ph. D.