Thesis (PhD (Chemistry and Polymer Science))--University of Stellenbosch, 2008. / The main goal of this study was the development of new technologies based on modern analytical techniques for analysis of volatiles in wines. Due to the exponential growth of the wine industry and consumer demands for an enjoyable, safe-to-consume, and high quality product, the need for arose for methodologies aiding the understanding of wine better arose. Chemical analysis is a valuable way of studying the composition of wine in depth. Very sophisticated instrumentation is available nowadays but almost always the sample needs to be cleaned up or concentrated before such analysis. This study investigates the use of stir bar sorptive extraction (SBSE) as such a technique. It is shown that SBSE combined with gas chromatography-mass spectrometry (GC-MS) is extremely suited for a wide number of analyses and during the course of the study the technique was applied for troublesome analytical challenges in various beverages and foodstuffs. The study focuses on the development of a screening technique for volatiles in wine using SBSE and the application of the data to various chemometrical techniques for classification purposes. A second part of the study shows the applicability of SBSE for extraction of pesticides, contaminants and preservatives from wine, water, lemon flavoured beverages and yoghurt. The method is also elaborated upon by development of faster analysis methods for wine and beer and the investigation of using SBSE for headspace sampling of wine. In all the applications, SBSE technology was shown to be sensitive, repeatable, robust and very simple to use.

Thesis (PhD)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: In this work a robust reversed phase ion-pairing high performance liquid chromatographic (RP-HPLC) method has been developed for the separation, characterization and quantification of all possible [PtCl6-nBrn]2- (n = 0 – 6) and [PtCl4-nBrn]2- (n = 0 – 4) complex anions using UV-Vis detection. High resolution 195Pt NMR of more concentrated PtII/IV solutions served to validate the relevant species assignments, particularly those of the stereoisomer species, cis- and trans- [PtCl4Br2]2-, [PtCl2Br4]2- and mer- and fac-[PtCl3Br3]2-. Quantification of the PtII/IV species was achieved by means of IP-RP-HPLC coupled to either ICP-MS or ICP-OES, and together with the UV-Vis absorption spectra obtained by photodiode array (PDA) recording of all eluted species, allowed for the determination of the photometric characteristics (λmax and ε) of all the PtII/IV species. This data enables practical speciation studies of such PtII/IV complex anions using standard analytical equipment. The hyphenation of ion-pairing RP-HPLC to ICP-OES allows for the successful determination of the Pt to halide mole ratios of individually separated species in order to characterize these species in a novel manner. The Pt to chloride and/or Pt to bromide mole ratio of the [PtCl4]2- and the series of [PtCl6-nBrn]2- (n = 0 – 6) complexes were determined using HPLC-ICP-OES based on the 177.708 nm Pt, 134.724 nm Cl and 148.845 nm Br emission lines and served as a technique for the unambiguous chemical speciation of such complexes. An increase in sensitivity of the developed method was achieved by the use of an ion-pairing reversed phase ultra high performance liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-Q-TOF-MS) method. This method proved capable of separating and characterizing the homoleptic and heteroleptic [PtIVCl6-nBrn]2- (n = 0 – 6) and mono-aquated [PtIVCl5-nBrn(H2O)]- (n = 0 – 5) complex anions in well defined acidic aqueous solutions. Ion-pairing ultra high performance liquid chromatography separation based on the volatile ion-pairing reagent, tributylamine, provided adequate chromatographic resolution as well as sufficiently low background noise for high resolution ESI-Q-TOF-MS detection. The wealth of structural information contained in the mass spectra obtained for each PtIV species simplified the identification of individual species. Moreover, the general fragmentation trends encompassing a constant incremental change of 44 Da (79/81Br - 35/37Cl) resulting from the successive substitution of Cl- by Br-, in combination with the observed elution order, facilitated the relevant species assignments. The developed method enabled the relative rapid (<13 min) characterization of all 22 [PtCl6-nBrn]2- (n = 0 – 6) and mono-aquated [PtCl5-nBrn(H2O)]- (n = 0 – 5) species. Quantification of each individual [PtCl6-nBrn]2- (n = 0 – 6) species by means of ion-paring HPLC-UV-Vis allowed for the determination of all 17 stability constants for the PtIV chloridobromido halide exchange reaction network. Determination of the associated Gibbs free energies for each ligand exchange reaction step, o rxnK ΔG n (n = 1 - 17), together with energy conservation relationships, served to validate the accuracy of the experimentally calculated stability constants. The experimentally determined overall formation constant, or ΔGo rxn, and those calculated using the standard reaction half cell reduction potentials of [PtCl6]2- and [PtBr6]2- were in good agreement, further confirming the experimentally obtained thermodynamic parameters. The thermodynamic driving force for the PtIV chloride-bromido exchange reactions is attributed to the hydration of the halide ligands, which drives the reaction towards the bromido PtIV species in aqueous solutions, even though the chlorido PtIV complexes are energetically favoured in this reaction network. Evaluation of other metal cation halido exchange reactions shows that all metal halido complexes exhibit the F- >> Cl- > Br- > I- order of thermodynamic stability and is only inverted due to the solvation of the relevant halide ligands. Furthermore, density functional theory (DFT) was used to predict the thermodynamic stabilities with respect to the isodesmic reactions involving chlorido-bromido PtIV stereoisomer pairs and chlorido-bromido PtIV ligand exchange reactions of the [PtCl6-nBrn]2- (n = 0 – 6) species and confirm the F- >> Cl- > Br- > I- order of thermodynamic stability as well as determining the ΔΔGo rxn within the range of 8 - 20 kJ.mol-1 to the experimentally determined ΔΔGo rxn. / AFRIKAANSE OPSOMMING: Tydens hierdie studie is „n robuuste “reverse-phase” ioonparing hoë-verrigting vloeistof chromatografie, RP-IP-HPLC, metode ontwikkel vir die skeiding, karakterisering en kwantifisering van alle moontlike [PtCl6-nBrn]2- (n = 0 – 6) en [PtCl4-nBrn]2- (n = 0 – 4) kompleks anione waar UV-Vis as detektor gebruik word. Die relavante spesies toedelings wat gemaak is, veral ten opsigte van die cis- en trans-[PtCl4Br2]2-, [PtCl2Br4]2- en mer- en fac-[PtCl3Br3]2- stereo-isomeerpare, is deur middel van hoë-resolusie 195Pt KMR van meer gekonsentreerde PtII/IV oplossings bevestig. Die PtII/IV spesies was gekwantifiseer deur die IP-RP-HPLC aan of „n ICPMS of „n ICP-OES te koppel. Daarenbowe was dit moontlik om die fotometriese eienskappe (λmax en ε) van elke individuele PtII/IV komplex anion te bepaal deur die UV-Vis absorpsie spektrum van elke elueerende spesies met PDA op te neem. Die nuwe metode wat tydens hierdie studie ontwikkel is het dit dus moontlik gemaak om sulke PtII/IV komplek sanione met standaard analitiese toerusting prakties te skei. Verder is gevind dat deur IP-RP-HPLC aan ICP-OES te koppel dit moontlik is om die Pt tot halied mol verhoudings van elke individueel geskeide spesies te bepaal en dus hierdie spesies op „n oorspronklike, nuwe manier te karakteriseer. Die Pt tot chloried en/of Pt tot bromied mol verhoudings van die [PtCl4]2- en die reeks van [PtCl6-nBrn]2- (n = 0 – 6) kompleks anione, soos bepaal deur gebruik te maak van HPLC-ICP-OES, is gebasseer op die 177.708 nm Pt, 134.724 nm Cl en 148.845 nm Br emissie lyne. Hierdie metode kan gebruik word vir die eenduidige chemiese skeiding van hierdie komplekse. Die sensitiwiteit van hierdie metode was egter verder verbeter deur gebruik te maak van ioonparing “reverse-phase” ultra hoë-verrigting vloeistof chromatografie gekoppel met elektrosprei ionisasie quadropool “time-of-flight” massa spektrometrie (UHPLC-ESI-Q-TOFMS). Deur dit te doen is dit nou selfs moontlik om die homoleptiese en heteroleptiese [PtIVCl6-nBrn]2- (n = 0 – 6) spesies, asook die “mono-aqauted” [PtIVCl5-nBrnH2O]- (n = 0 – 5) spesies in „n goed gedefinieërde aangesuurde waterige oplossings te skei en te karakteriseer. Die vlugtige ioon-paringsreagent, tributielamien, is vir die skeidingsproses op die IP-UHPLC gebruik om te verseker dat voldoende chromatografiese resolusie, so wel as lae genoeg agtergrondgeraas, verkry word vir hoë-resolusie ESI-Q-TOF-MS deteksie. Die rykdom informasie vervat in die massaspektrum van elke PtIV spesies het die indentifikasie van elke spesies vergemaklik. Daarenbowe het die fragmentasie tendens, aanduidend van „n konstante inkrementele verandering van 44 amu (71/81Br – 35/37Cl) weens die opeenvolgende substitusie van Cl- met Br-, tesame met die elusie volgorde, die spesies-toedelings gefasiliteer. Met hierdie nuut ontwikkelde metode is dit nou moontlik om al 22 [PtCl6-nBrn]2- (n = 0 – 6) en “mono-aquated” [PtCl5-nBrnH2O]- (n = 0 – 5) spesies in „n relatiewe kort tydperk (< 13 min) te karakteriseer. Deurdat elke [PtCl6-nBrn]2- (n = 0 – 6) spesies nou individueel met IP-HPLC-UV-Vis gekwantifiseer kan word, is dit moontlik om al 17 stabiliteitskonstantes vir die PtIV chloridobromido halied uitruilingsreaksienetwerk te bepaal. Die geassosieerde Gibbs vrye energie, ΔG°rxnKn (n = 0 – 17), wat vir elke stap in die uitruilingsreaksienetwerk bepaal is, tesame met die energiebewaring verhoudings, was gebruik om die akkuraatheid van die eksperimenteel bepaalde stabiliteitskonstantes te bekragtig. Verdermeer was die waarde van die algehele formasie konstante wat eksperimenteel bepaal is, ΔG°rxn, in goeie ooreenstemming met dit wat bereken is deur die standaard reaksie halfsel reduksie potensiale van [PtCl6]2- en [PtBr6]2-. Dus is die eksperimenteel verkrygde termodinamiese parameters bevestig. Die termodinamiese dryfkrag vir die PtIV chloried-bromied uitruilingsreaksies is toegereken aan die hidrasie van die halied ligande, wat in waterige oplossings die reaksie na die bromied PtIV spesies dryf, al is die chloried PtIV spesies energeties bevoordeel in hierdie reaksienetwerk. Evaluering van ander metaalkatioon- halied-uitruilreaksies wys dat alle metaal-halied komplekse die F- >> Cl- > Br- > I- orde van termodinamiese stabiliteit volg en dat hierdie volgorde slegs omgekeer sal word weens solvasie van hierdie halied ligande. Darenbowe digtheids funksionele teorie (DFT) gebruik om die termodinamiese stabiliteit met betrekking tot isodesmiese reaksies wat chloried-bromied PtIV stereoisomeer pare behels te voorspel, sowel as van chloried-bromied PtIV liganduitruilingsreaksies van die [PtCl6-nBrn]2- (n = 0 – 6) spesies, en bevestig die F- >> Cl- > Br- > I- volgorde van termodinamiese stabiliteit. Verder was dit ook moontlik om met DFT die ΔΔG°rxn tot so naby as 8 – 20 kJ.mol-1 te bereken.

An assessment of heavy metal and pesticide pollution was conducted at a commercial airport in Pietermaritzburg, which is the base for a local crop spraying operation, as well as a small-scale aircraft maintenance centre. The study investigated the presence of heavy metals from the aircraft activities (tyre wear, emission and aircraft body attrition) along the edges of the runway. In addition, the study assessed the pollution of the soil in different parts of the airport (areas adjacent to the crop sprayer hangar and maintenance hangars drainage ditch) caused by organic chemicals (pesticides) originating from the washing and maintenance of the crop spraying aircrafts. A total of 37 samples were taken from the top soil (0-15 cm) adjacent to both sides of the runway, approximately 100 meters apart, for heavy metal assessment. Total metal concentrations were determined on a 1:1 HNO3 acid extract analysed using inductively coupled plasma spectroscopy and the results compared with the baseline concentrations of heavy metals in the soils of South Africa. The average levels of the heavy metals in the soils were: Cd - 4.4, Co - 76, Cr - 142, Mn - 6580, Pb - 64 and Zn - 70 mg/kg. The Cd and Co concentrations of all samples and the Pb and Zn concentrations of 86% of the samples were above the South African guidelines for the maximum permissible contents of these metals (2 mg/kg Cd, 20 mg/kg Co, 6.6 mg/kg Pb, 46.5 mg/kg Zn) in soils. In addition, the concentration of Cr was significantly above these guidelines (80 mg/kg Cr ) in 51% of the samples. All the metals, except zinc, were highly correlated with one another indicating the source for these heavy metals is the same. In addition the concentration of these metals was high at sampling points adjacent to the runway where pronounced aircraft tyre wear marks were observed, suggesting that the aircraft activities are the sources of these heavy metals. A total of 13 samples were collected from areas adjacent to the crop sprayer hangar and the drainage ditch (at the maintenance hangars) in order to determine the qualitative presence of organic pollutants. The samples were prepared using soxhlet extraction. Cypermethrin, deltamethrin, azoxystrobin, fusillade and fluazifop-p-butyl as well as fuel oils Jet Al and Avgas were qualitatively determined using GC/MS. Only fusillade and fluazifop-p-butyl were detected, in some of the samples. No quantitative analysis of the organic chemicals was attempted due to time constraints. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2004.

In this investigation the chemistry of three southern African plants used for medicinal purposes was investigated. The plants were Dioscorea dregeana, Avonia rhodesica and Equisetum ramossisimum. Extracts of all three of these plants have been found to be active on the central nervous system. The structures of the compounds isolated were determined by using 1H and 13C n.m.r., i.f., UV-VIS, - mass spectroscopy and chemical methods. D. dregeana yielded two alkaloids and three aromatic compounds. A. rhodesica contained a large amount of wax which was analysed by GC-MS and its distribution and physical nature on the surface of the leaves was determined by SEM (scanning electron microscopy). This plant also contained two sterols. Equisetum ramossisimum extracts contained a carotenoid and several porphyrins, as well as large amounts of silica. A DTSA X-ray microanalysis system (which was a component of the scanning electron microscope) was used to determine the distribution of silica in the stem. / Thesis (M.Sc.)-University of Natal, Durban, 1998.

Polycyclic (or polynuclear) aromatic hydrocarbons (PAHs) are environmental pollutants produced during the incomplete combustion of organic matter. Since many of these compounds have been shown to be mutagenic and/or carcinogenic, an investigation was initiated into determining the PAH content and mutagenicity of the ash that remains after sugar cane crop burning, and the soot deposited on toll booths by vehicle exhaust emissions. Due to the large amount of sugar cane farming in the Natal coastal region and that the favoured method of disposing unwanted leafy trash is crop burning, concern was expressed as to the nature of the residue that is formed. PAHs have been identified in the residues from combusted wood and straw and, due to their intrinsic similarity to sugar cane, it was considered that the burning of sugar cane could generate PAHs. It is well documented that vehicle exhaust emissions exhibit mutagenic properties and PAHs have been identified as the major contributors of this observed mutagenicity. Since a toll plaza is an area of high traffic density, it was considered to be an ideal location for an investigation into the build-up of particles emitted by the passing vehicles, and to study to what extent the operators are exposed to harmful compounds. In addition, this sample acted as a control, since the detection of PAHs and mutagenic activity in the soot would be an indication that the correct experimental techniques were being employed. Samples were collected on site. The sugar cane ash was collected off a field immediately after burning had taken place, and the soot was collected either by scraping the toll booth walls and surrounding areas or by wiping the surfaces with cotton wool swabs. The organic portion of the samples was separated from the inorganic and carbonaceous substances by extraction into a suitable solvent; the use of both acetone and dichloromethane was investigated. The extracts were divided into two portions - one was used for the analysis of PAHs and the other for determining mutagenic activity. Analysis for PAHs involved subjecting the extracts to a sample clean-up routine and the use of a number of analytical techniques to characterise the components. The mutagenic properties of the samples were investigated by means of two bacterial mutagenicity tests: the Salmonella typhimurium assay (the Ames test) and a new commercially available test kit, the SOS Chromotest. A number of PARs were identified in the extracts by means of reverse phase high performance liquid chromatography (HPLC) with both ultraviolet and fluorescence detection, the latter being the more sensitive method. Mutagenic activity was detected for both samples in the Ames test and for the toll booth soot in the SOS Chromotest, and this observed mutagenicity was attributed to the presence of the PAHs. / Thesis (M.Sc.)-University of Natal, Durban, 1992.

The goal of this project was the development of a computer-based system using artificial intelligence to classify the limonoids, protolimonoids and triterpenoids isolated from the family Meliaceae by the Natural Products Research Group of the University of Natal, Durban. A database of samples was obtained between 1991 and 1996, part of which time the author was a member of the group and isolated compounds from Turraea obtusifolia and Turraea floribunda. Over and above the problem of complexity and similarity in structures of the above mentioned natural products, are other difficulties. These include very small amounts of sample being isolated producing very weak peak signals in the C-13 NMR spectra, extraneous peaks in the NMR spectra due to different impurities and instrument noise, non-reproducible spectra due to the pulsed Fourier transform intervals and the nuclear Overhauser effect, impure samples often isolated as stereoisomeric mixtures or as mixed esters and superposition of peak signals in the NMR spectra due to carbons in the same environment within the same compound. These factors make identification by traditional computational and expert systems impossible. As a result of these shortcomings, the author has developed a novel approach using artificial neural network techniques. The artificial neural network system developed used real data from the 300 MHz NMR spectrometer in the Department of Chemistry, Durban. The system was trained to discriminate between limonoids, triterpenoids and flavonoids/coumarins from the C-13 NMR spectra of pure, impure and unseen compounds with an accuracy of better than 90%. Further differentiation of the glabretals from the rest of the protolimonoids as well as from the rest of the triterpenoids showed similarly significant results. Finally, individual limonoid discrimination within the limonoid dataset was extremely successful. Apart from its application to the extractives from Meliaceae, the methodology and techniques developed by the author can be applied to other sets of extractives to provide a robust method for the spectral classification of pre-identified natural products. / Thesis (Ph.D.)-University of Natal, Durban, 1998.