The thesis outlines an investigative study of the supercritical fluid extraction (SFE) of organic natural products from two Meliaceae species. Extractions from ground wood, bark and seeds of the selected species were studied. Qualitative and quantitative studies were performed using a number of techniques. In the qualitative study, both off-line and on-line combination of micro-SFE with capillary gas chromatography (SFE-GC) were performed on the wood and bark of Cedrela toona to show the applicability of the technique in the analysis of semi-volatile components from plant matrices. Results for the on-line and off-line SFE-GC were comparable and indicated that the on-line studies could be used to predict results obtainable from large scale SFE. Dynamic SFE of the limonoid (cedrelone) from the wood of Cedrela toona using pure and modified carbon dioxide was studied on the basis of a theoretically-derived dynamic extraction model. Extracts were collected for different lengths of time in order to investigate the extraction kinetics of this compound. The theoretically-derived model which involved extrapolating data obtained from shorter extraction times was used to estimate the amount of the compound present in the wood. The estimated quantitative results were found to be comparable to exhaustive extractions using liquid hexane as well as methanol-modified carbon dioxide. Further, data from the SFE extractions were fitted into the model to produce a characteristic kinetic curve from which factors controlling the extraction of the specific compound could be predicted. Following the dynamic extraction study, a statistical optimisation strategy was used to propose a model equation for the extraction of cedrelone from the wood of Cedrela toona. As the preliminary step, a screening design utilising a two level, three-factor full factorial analysis was used to study the effects of the variables pressure, temperature and the length of extraction time on the yields of cedrelone from the wood of Cedrela toona. By a statistical analysis of variance (ANOVA) at the 95% confidence level on the preliminary data collected, pressure and temperature were found to be the significant variables influencing the yields of the compound. Finally response surface methodology using the central composite design was used in an attempt to determine the maximum response space for the pressure and temperature effects by plotting a 3-dimensional response surface graph. An ANOVA study at the 99% confidence level was then employed to establish the true model equation for the extraction. / Thesis (M.Sc.)-University of Natal, 1995.

PART I: This work is part of an investigation to determine activity coefficients at infinite dilution (1'73) of hydrocarbons dissolved in the industrially important polar solvent tetrahydrothiophene-l, l-dioxide (sulfolane), by medium pressure gas liquid chromatography (g.l.c.). In this work the activity coefficients at infinite dilution for a series of l-alkenes (C6-CS) , l-alkynes (C6-CS) , and cycloalkanes (C7 and Cs) have been measured in the polar solvent, sulfolane, at 303.15 K and 313.15 K. The activity coefficients of some of the solutes discussed in this work would be difficult to determine by any other method because of their low solubility in sulfolane. The mixed second virial coefficients used in this work were determined assuming the principle of corresponding states, the Hudson and McCoubrey combining rules for T~2' the Lorentz rule for V~2' and the McGlashan-Potter equation. PART II: Unrefined sugar contains organic colour material originating In the sugarcane or formed during the extraction and purification processes. Sugar colour must be within the limits of acceptability for direct or indirect consumption. In this work, a cost effective technique to separate colourants from sugar through a sucrose packed medium pressure chromatographic column was investigated. Three dimensional perspective plots of wavelength/absorbance/time were developed to provide insight into the nature of the sugar colourants and to provide a means of investigating various decolourisation systems. In addition to the above experiment a procedure was developed to remove colourant species from unrefined sugar samples and from samples taken during the refining process for chemical analysis. In this work only one technique - gas chromatography-mass spectroscopy was used to identify the species. / Thesis (M.Sc.)-University of Natal, Durban, 1996.

Optimisation attempts within the petrochemical industry have led to interest in alternate solvents. The most widely used commercial solvents for the separation of hydrocarbons, by extractive distillation, are N-methylpyrrolidone and sulfolane. There has also been reference made to other solvents, such as N-formylmorpholine and the ethylene glycols [mono-, di-, tri- and tetra], being used. The alternate solvents proposed for this study were N-formylmorpholine, triethylene glycol and diethylene glycol. Infinite dilution activity coefficients, γ∞, provided a means of comparing the ease of separation of the different solutes using different solvents in extractive distillation. There is a substantial database of γ∞ measurements for systems involving N-methylpyrrolidone and hydrocarbons. A fairly large data set of γ∞ values of hydrocarbons in N-formylmorpholine has also been measured. Very little work has been conducted on the γ∞ values of hydrocarbons in either diethylene glycol or triethylene glycol. Gas liquid chromatography is one of the more common methods used to measure γ∞. To enable the measurement of γ∞ at higher temperatures, a pre-saturator was installed prior to the column. This ensured that the carrier gas entering the column was saturated with solvent and prevented the elution of solvent from the column. The γ∞ values of 25 solutes; including n-alkanes, alk-1-enes, alk-1-ynes, alcohols and aromatics; were measured at temperatures of 333.15, 348.15 and 363.15 K. The γ∞ measurements in N-formylmorpholine were used to verify this experimental set up and technique. Once the experimental set up had been proven, γ∞ in N-methylpyrrolidone, triethylene glycol and diethylene glycol were measured. Selectivities and capacities, at infinite dilution, of several solute combinations in the four solvents were then compared. In a few of these separation cases, the alternative solvents appeared to have better separation performance than N-methylpyrrolidone. The γ∞ values of three of the solutes in N-formylmorpholine and N-methylpyrrolidone were also measured using the novel cell design and measurement procedure suggested by Richon. It was found that this new technique required further development for the case of volatile solvents, since the results obtained using this technique did not compare favourably with the literature data. / Thesis (M.Sc.)-University of KwaZulu-Natal, Durban, 2012.

Supercritical fluid chromatography has recently been coupled to an ICPMS detector. The method has been shown to be suitable for the speciation and analysis of organometallic compounds at trace levels. This study has attempted to further the research initiated by other groups in this field by developing a new interface for coupling these two instruments. The new interface makes use of a modified join between the nebuliser and the torch in the ICP unit. The effect of the mobile phase on the plasma with time has been investigated and little spectral background interference has been observed. The chromatographic conditions were optimised using a flame ionisation detector and a series of tin, arsenic, iron, and mercury compounds were analysed using SFC-ICPMS. Mter focusing the ICP-MS on the element of interest, each compound was evaluated in terms of the change in peak intensity with change in concentration and the theoretical detection limits were compared to the practical detection limit. The restrictor temperature was determined using a rough calibration procedure with bench top experiments. The effect of the restrictor temperature on the peak intensity of each compound was then studied. All results were plotted and a theory for the observed trends and observations is proposed. The results obtained and the interface used have been compared to the results and interfaces of other groups and differences have been explained. Attempts to extract relevant compounds from topsoil using supercritical fluid extraction were made. Finally, sediment samples were collected from relevant points in Durban Bay and an attempt was made to extract these samples using supercritical fluid extraction. The extracted samples were analysed using SFC-ICPMS although little success was obtained. Reasons for the failure of this method on the real samples have been proposed. In the concluding section of this study SFC~ICPMS has been evaluated in terms of its future applicability and use as a viable analytical method. / Thesis (M.Sc.-)-University of Natal, Durban, 1994.

A mini-dissertation in partial compliance with the requirements for a Master's Degree in Technology: Homoeopathy, Durban Institute of Technology, 2002. / The purpose of this study was to compare the amount of caffeine extracted from triturated samples and ethanolic samples of Coffea tosta using high pressure liquid chromatography (HPLC) as a method of analysis. The study wanted to expand on homoeopathic pharmaceutical knowledge, specifically looking at the two methods of remedy preparation of plant materials. From the same batch of ground roasted coffee beans, using the decimal scale of dilution, the mother tincture (bill) and the first triturated (bill) samples were prepared. The subsequent 2xH and 3xH triturated and ethanolic potencies were then made in accordance with homoeopathic methodology. Each group contained three different dilution levels (bill, 2xH and 3xH), 18 samples per group giving a total of36 samples that were analysed using HPLC. Three samples were analysed from the three dilution levels in each Group, in total there were 18 samples from the triturated group and 18 from the ethanolic group. . The samples were analysed quantitatively using the highly accurate and advanced method of high pressure liquid chromatography. This method gives accurate readings of the caffeine concentrations of a sample compared to a caffeine standard. This allowed for quantification of the caffeine concentration of each sample. The percentage caffeine was calculated from each sample. The aim of the study was to evaluate the difference in each method of preparation by measuring the amount of caffeine extracted from the samples. The results obtained from the inter-Group Mann-Whitney and ANOVA tests showed that there was a significant difference between the ethanolic dilutions and triturated dilutions with regards to the 1xH and 2xH dilutions. In the 1xH dilution the ethanolic method retained / M

Many researchers have investigated ways to improve the separation power of conventional chromatography, most notable is the development of ultra-high performance liquid chromatography (UHPLC). However, only slight improvements in separation efficiency have been achieved up to this point, and unfortunately, modern reversed phase liquid chromatography (RPLC) methods do not have high enough resolving power to analyze complex proteomic mixtures.Uniformly sized silica particles from 10 nm to 1 micron are known to self-assemble into a highly ordered face centered cubic crystal. Silica colloidal crystals have shown recent promise in biological applications such as permselective nanoporous membranes, DNA sieving, reversed phase separation of small molecules on planar substrates, protein sieving, microarrays, total internal reflection fluorescence microscopy of live cells, and 3-D scaffolds for supported lipid films. In this work, silica colloidal crystals packed in capillaries are explored for their potential improvement in the efficiency of reversed phase chromatography.The silica colloidal crystal columns were chemically stabilized by with trichlorosilanes. The trichlorosilanes form chemical bonds between the particles and the particles and the substrate creating an increase in mechanical stability, and at the same time, providing an excellent chromatographic monolayer. After stabilization the fritless columns were able to withstand the pressure limit of the commercial UHPLC. Next, the columns were characterized using a small dye molecule, 1,1' - Didodecyl - 3,3,3',3' - tetramethylindocarbocyanine (DiIC12). The dye was run under capillary electrochromatography (CEC), and sub-micron plate heights were achieved. Further, a van Deemter plot of the dye molecule indicates that the plate height is largely due to the molecule's diffusion. This result suggests that the plate heights for proteins would be even smaller, since proteins have diffusion coefficients an order of magnitude smaller. The analysis of proteins by CEC yielded nanometer plate heights. Finally, pressure driven flow separations coupled with nano-electrospray ionization (n-ESI) MS have also been explored. The Poiseuille flow profile has been shown not to perturb the low plate heights. Gradient elution of peptides was also achieved, and the results demonstrate the highest chromatographic peak capacities for short analysis times to date.

Through chemical analysis and ion exchange chromatography of watersoluble antigens, this investigation supports the view that the majority of differences between the biotypes are quantitative. It was also found that strains demonstrate distinct, qualitative differences when compared to the attenuated strain 19 by immunodiffusion and thin-layer polyacrylamide gel, isoelectric focusing. These differences include the presence of antigens on virulent strains that are absent on strain 19. In addition, one antigen absent on strain 19, was found common to each virulent biotype. Finally, the results from immunodiffusion experiments, employing adsorbed and non-adsorbed immune globulins, indicate that at least some water-soluble antigens are exposed on the cell surface and that their distribution among the biotypes varies.

Continuous counter current extraction (CCCE) or dual flow counter current chromatography (DFCCC) is a promising technique where components can be separated continuously by two liquid phases that flow in opposite directions through a continuous length of coiled tubing. Specially designed end connectors and a coil planet centrifuge allow each respective phase to be pumped into each end of the tubing and the other phase to elute at each opposite end. In this thesis the feasibility and the mechanics of CCCE are investigated using stroboscopic photography on an experimental rig and a specially built pilot-scale CCCE centrifuge. The mechanics of the hydrodynamics in the coil was investigated systematically by comparing the measured volumes with photographic images of the process. This investigation revealed that the phases are not distributed evenly throughout the coil, which was previously assumed, but that there is a transition area where the phases switch from mainly upper phase at the head end of the tubing to mainly lower phase at the tail end. This means that the sample encounter three different phase distribution zones in the coil. At the head the upper phase is the dominant phase with a small volume of lower phase running through. At the tail the reverse situation is found and lower phase is dominant. The third zone is a short segment of the coil where there is a transition between the dominant phase conditions that exist at each end. The position of the transition zone and the volume of the other two zones are profoundly affected by the relative flow rates of the two phases. This work indicates that the volume distribution in the coil is affected most by the upper phase flow rate. The pilot-scale CCCE centrifuge was used to successfully separate industrially supplied samples. Crude reaction liquor was processed in both batch and continuous modes achieving the separation of the multi-component mixture into two groups. Changing the flow rate combinations changed the location of elution of some of the components in the mixture. Separation efficiency was maintained even when sample loading was increased. The separations were shown to be predictable with the dual flow theoretical model.

Heparin is a highly sulfated glycosaminoglycan with potent anticoagulant, antimetastatic, and anti-inflammatory effects. Polymeric and polyanionic nature of heparin makes dosing and side effects a nightmare for healthcare professionals. Our laboratory has proposed appropriately designed, small, highly sulfated aromatic molecules as potential mimetics of heparin. These easier-to-synthesize small molecules have been shown to possess interesting pharmacological and improved toxicological profiles. However, the detection and characterization of these highly sulfated molecules is challenging. A robust RP-IP UPLC-MS method was developed to successfully retain, resolve and quantify sulfated non-saccharide GAG mimetics without the requirement of pre- or post- column derivatization. Comparative analysis reveals intricate dependence of resolution and ionization on the structure of ion-pairing agents. This is the first report showing systematic use of MS cone voltage to fingerprint sulfated GAG mimetics, perhaps eliminating the need for tandem MS techniques.

New liquid phase microextraction (LPME) has been optimized. Plastic adapter with tapered tip was used as extraction device. Concentration of analytes was determined by GC-MS. Face centered design was used to optimize LPME and the results were evaluated by Minitab 16 programme. Volume of sample, volume of extraction solvent, stirring rate, extraction time, ionic strength and time after microextraction were optimized. Response of system was defined as the sum of the peak relative areas. Toluene, tetrachlorethylene, ethylbenzene, xylenes, mesitylene and naphthalene were the studied analytes. Methylhexadecanoate was used as an internal standard. Optimal system conditions were as follows: 20 mL sample volume, 300 μL extraction solvent volume, 20 minutes, stirring speed 700 rpm, no addition of salt, and time after microextraction 3.5 minutes. Determination coefficient was 0.9700 and the lack-of-fit was insignificant which indicated good agreement of the model with the experimental data. The preconcentration factor ranged from 26 (tetrachlorethylene) to 39 (xylenes). The microextraction yield was varied from 39.5% (tetrachlorethylene) to 59.1% (p-xylene and m-xylene). The new method was tested by analyzing real samples of tap water, river water and water from the sewage treatment plant. The samples were...