No description available.

\kur{Beauveria bassiana} is a entomopathogenic polyphagous fungus commonly found in soil and it is parasite of soil insects, mainly of the stages of insect that occur in soil. At the present time it is used in plant protection against more than 70 species of insects. In the Czech Republic \kur{Beauveria bassiana} has the greatest importance in the fight against bark beetle \kur{Ips typographus} in the NP Šumava in these days. This study was focused on the evaluation of genetic variability \kur{Beauveria bassiana} strains on the basis of microsatellite analysis and the comparison of four separation methods: electrophoresis in 2% agarose gel, electrophoresis in 3% synergel, chip electrophoresis and fluorescent capillary electrophoresis in term of the most precise separation of PCR products. We used 41 strains which were collected in the NP Šumava and 20 strains from long-term collection determined as an exotic in this study. This large geographical scale group contains the strains from whole world and in addition it was upgraded by the strains collected from the NP Krkonoše and South Moravia. For the microsatellite analysis there were used 11 pairs of primers but for inter-comparison of separative methods were chosen only 4 pairs of primers. The population of \kur{Beauveria bassiana} strains collected from the NP Šumava were evaluated by analysis of microsatellites as a conservative and fully closed regardless of the source and the location. The strains from the large geographical scale group showed the great genetic variability. In terms of separation, the best and most suitable separation method was proved, the fluorescent capillary electrophoresis. Despite of its difficult financial aspect, this method was evaluated as the most precise and the most sensitive. Its advantage is in possibility to detect the smallest differences in the length of single allele in the range 1-2 bp, which is for the gel electrophoresis impossible.

Doctor of Philosophy / Department of Chemistry / Christopher T. Culbertson / During the last five years, great advancements in microfluidics have been achieved with the development of “sample-in-answer-out” systems. Such systems have begun to realize the true potential of analytical miniaturization since the concept of the “micro-Total Analysis System” was first envisioned. These systems are characterized by the elegant integration of multiple fluid-handling channel architectures that enable serial execution of sample preparation, separation and detection techniques on a single device. While miniaturization and portability are often identified as key advantages for microfluidics, these highly integrated systems are heavily reliant upon large off-chip equipment, i.e. the microchip is often tethered to the laboratory via multiple syringe pumps, vacuum pumps, solenoid valves, gas cylinders and high voltage power supplies. In this dissertation, a procedure for the facile integration of dielectric elastomer (DE) actuators (called IDEAs) onto microfluidic devices is described. Poly(dimethylsiloxane) (PDMS) is commonly used as a microchip substrate because it is cheap and easy to fabricate, mechanically robust and optically transparent. The operation of an IDEA exploits the ability of PDMS to behave as a smart material and deform in the presence of an electric field. In Chapter 2, the fabrication of IDEA units on a standard microchip electrophoresis device is described. IDEA-derived injections were used to evaluate the physical performance of this novel actuator configuration. In Chapter 3, the analytical merits of IDEA-derived injections were evaluated. Sampling bias caused by electokinetic injection techniques has been problematic for conventional microchip electrophoresis systems due to the lack of fluid access. The hydrodynamic injections created by IDEA operation were found to be highly reproducible, efficient, and possess a negligible degree of sampling bias. In Chapter 4, the spatial characteristics of microchannel deformation due to IDEA actuation have been investigated using fluorescence microscopy. It was determined that the DE compresses more along the edge of the channel than in the middle of the channel. This information can be used to design a new generation of more efficient IDEAs.

Submitted in fulfilment of the requirements of the Degree of Master of Technology: Chemistry, Durban University of Technology, 2012. / The present work involves the interaction studies of chiral compounds with the Human Serum Albumin (HSA) protein using computational and experimental methods. The HSA protein has multiple binding sites that forms the basis for its exceptional ability to interact with many organic and inorganic molecules, which makes this protein an important regulator of intercellular fluxes and the pharmacokinetic behaviour of many drugs. This study was undertaken to evaluate the related pharmacokinetic and enantioselective binding parameters of the racemic catechin enantiomers with the HSA. Accordingly, this work involved a method development for the chiral separation of a racemic compound, by capillary electrophoresis-electrokinetic chromatography (CE-EKC) with a highly sulphated beta-cyclodextrin (HS--CD) as a chiral selector. The experimental work was supported by two molecular docking studies. The first included the mimicking of the host-guest interactions between a chiral selector and an enantiomeric compound. The second study included the estimation of the pseudo enantioselective (ES) binding of catechin to HSA. Overall, it was found that CE-EKC is the preferred method for the(±)-catechin binding to HSA protein evaluation. Moreover, the technique used in this work is not restricted to HSA or polyphenols, but can also be applied to other proteins and ligands that possess chirality. Furthermore, the molecular docking approaches also proved to be very useful for the evaluation of chiral recognition systems and for elucidation of the ligand-protein interactions.

Thesis (MSc)--Stellenbosch University, 2004. / ENGLISH ABSTRACT: Capillary electrophoresis (CE) has recently attracted considerable attention as a promising analytical technique for the separation of cations and anions in complex matrices. Determination of ions in aqueous samples using capillary electrophoresis can be accomplished with indirect UV detection. Most inorganic ions have weak absorption profiles in the UV-Vis wavelength range. These mostly non-absorbing species are commonly detected by indirect UV absorbance through addition of an absorbing co-ion (chromophore) into the electrolyte. Inorganic cations most often require an additional complexing agent to selectively alter their similar mobilities and proper separation. For optimal determination of alkali, alkaline, and transition metal ions, several electrolytes systems were studied. These include pyridine, imidazole and 4- aminopyridine as UV-absorbing species and glycolic acid, a-hydroxyisobutyric acid and their mixture were used as complexing reagents. A mixture of 10 metal ions (K+, Na+,Ca2+, Mg2+, Mn2+, Fe2+, Cd2+, Pb2+, Ni2+and Zn2+) was successfully separated. Detectionwas performed at 210,214 and 254 nm. In the anion determination chromate and 2, 6 pyridine dicarboxylic acids (PDC) were used as back ground electrolytes for inorganic ions (F-, CI- en SO₄² ̄ ) and organic acids (tartaric acid, malic acid, succinic acid and citric acid) separations respectively. Electroosmotic flow (EOF) was reversed in the direction of the anode by adding Cetyltrimethylammonium bromide (CTAB) to the electrolyte. Highly alkaline conditions were used to confer a negative charge on inorganic and organic anions to promote their migration towards the anode. The detection wavelength was 200 nm. All peaks were completely resolved and well separated. The limit of detection (LOD) of cations and anions were in the range of 0.5 - 3 ppm and 2 - 3.5 ppm respectively. The described methods were used successfully in routine analysis of real samples. This includes the qualitative and quantitative analysis of an environmental water samples from the areas surrounding Stellenbosch, beverages and orange juice. / AFRIKAANSE OPSOMMING: Kapillêre Elektroforese (CE) het in die onlangse verlede heelwat aandag getrek as "n belowende analitiese tegniek vir die skeiding van katione en anione in komplekse monsters. Die bepaling van ione in waterige medium met kapillêre elektroforese word gedoen deur indirekte Ultraviolet (UV) deteksie aangesien meeste anorganiese ione swak absorbsie in die die sigbare UV (UV-Vis) golflengtegebied toon. Deteksie van hierdie meestal nie-absorberende spesies word algemeen gedoen deur indirekte UV absorbansie deur die byvoeging van "n ko-ioon (chromofoor) tot die elektroliet. Anorganiese katione benodig dikwels "n addisionele komplekserings reagens om selektief hulle eenderse mobiliteite te verander en sodoende goeie skeiding te bewerkstellig. Vir die optimale bepaling van alkali-, alkali-aard- en oorgansmetaal ione is verskeie elektrolietsisteme bestudeer. Hierdie sluit in piridien, imidasool en 4-aminopiridien as UV absorberende spesies en glikoliensuur, a-hydroksie-isobottersuur asook "n mengsel van die twee as komplekserings reagense. "n Mengsel van 10 metaalione (K+, Na+, Ca2+, Mg2+, Mn2+, Fe2+, Cd2+, Pb2+, Ni2+ en Zn2+) is sukselvol op hierdie wyse geskei. Deteksie is gedoen by golflengtes van 210, 214 en 254 nm. Vir die anioon bepaling is chromaat en 2,6-piridiendikarboksielsuur gebruik as agtergrond elektroliete vir die skeiding van anorganiese anione (F-, CI- en SO₄² ̄ ) en organiese sure (tartaarsuur, malonsuur, suksiensuur en sitroensuur), onderskeidelik. Elektroosmotiese vloei (EOF) is omgekeer na die rigting van die anode deur byvoeging van setieltrimetielammoniumbromied (CTAB) by die elektroliet. Sterk alkaliese kondisies is gebruik om 'n negatiewe lading op die anorganiese en organiese anione te konsentreer en sodoende hul migrasie na die anode te bevorder. Die deteksiegolftengte hier gebruik was 200 nm. Volkome resolusie en goeie skeiding is gerealiseer vir al die pieke. Die deteksielimiete (LOD) vir die katione en die anione was 0.5 - 3 ppm en 2 - 3.5 ppm, onderskeidelik. Die metodes wat beskryf word is suksesvol aangewend vir roetiene analise van werklike monsters. Dit sluit in kwalitatiewe sowel as kwantitatiewe analise van omgewingswater monsters uit die Stellenbosch area, koeldranke en lemoensap.

Thesis(PhD)--Stellenbosch University, 2008. / ENGLISH ABSTRACT: Aminopolycarboxylic acids (APCAs) are chelating agents widely used to inactivate various metal ions by complex formation in industrial and household applications. Ethylenediaminetetraacetic acid (EDTA) and diethylenetriaminepentaacetic acid (DTPA) are the widely used agents. Their use is under scrutiny due to their persistence in the environment because they cannot readily biodegrade. This led to the introduction of readily biodegradable agents namely ethylenediaminedisuccinic acid (EDDS) and iminodisuccinic acid (IDS) as alternatives especially to EDTA. Therefore, there was an interest to study the separation of EDDS, IDS and other APCAs using a simple, quick and accurate method. Capillary electrophoresis was used to determine the separation and speciation of iminodisuccinic acid with various metal ions at various pH levels. Speciation modelling was also utilized to compare and validate the presence and distribution of metal-ligand species. The obtained CE results were compared with speciation profiles and a reasonable agreement was obtained. The degradation studies at various time intervals for the metal-ligand (ML) complexes of DTPA, S,S-EDDS, IDS and R,S-IDS with various metal ions (Cd2+, Cr3+, Cu2+, Fe3+, Mn2+, Pb2+ and Zn2+) at pH 7 and 9 was evaluated using CE. New peaks were observed in some ML complexes when the pH was changed from pH 9 to 7. Sharp peaks were seen for CuL (L = DTPA, S,S-EDDS, IDS and R,S-IDS), FeDTPA and FeEDDS at both pH s. While small broad peaks were observed for FeIDS, CrL and MnL complexes. CuDTPA and CuEDDS complexes showed a greater stability over some considerable time as compared to CuIDS, CuR,S-IDS and other metal complexes at pH 9. This work also investigated the effect of various cationic electroosmotic flow (EOF) modifiers and counter anions on the CE separation of EDTA, EDDS and IDS as Cu(II) complexes. The performance of the modifiers was evaluated in terms of migration times, resolution and plate numbers. The best results were observed when Tetradecyltrimethylammonium bromide (TTAB) and Cetyltrimethylammonium bromide (CTAB) was used as modifiers in order to reverse the EOF in the fused silica capillary. This resulted in short analysis time and better peak shapes. The effect of different counteranions attached to EOF modifiers on the separation was also shown. It was also found that the counter anions of EOF modifiers used influences the separation of the complexes. The EOF modifiers namely Cetyltrimethylammonium chloride (CTAC) and Tetradecyltrimethylammonium chloride (TTAC) were further utilized for the determination of EDTA in South African river waters and industrial effluents. A method for the simultaneous separation of Fe (III) complexed with EDTA, S,S -EDDS and IDS was developed by CE and high-performance liquid chromatography (HPLC). The recalcitrant EDTA is used in combination with readily biodegradable analogues like EDDS and IDS in many commercial products. The methodology performance was evaluated in terms of linearity, limit of detection (LOD), limit of quantitation (LOQ) and reproducibility for both CE and HPLC methods. The LOD values obtained from HPLC were low when compared with CE. The applicability of both methods was demonstrated for the analysis of cosmetic products such as foam bath and shower cream. The results obtained by both CE and HPLC were found to be comparable and are in good agreement. / AFRIKAANSE OPSOMMING: Aminopolikarboksiel sure (APCAs) is komplekseer middels wat algemeen gebruik word om verskillende metaal ione te deaktifeer deur kompleks vorming in industriële en huishoudige toepassings. Etileendiamientetraasynsuur (EDTA) en dietileentriamienpentaasyn suur (DTPA) is die mees algemene APCA s. Hul gebruik word deesdae noukerig ondersoek as gevolg van hul volharding in die omgewing. Daar is n intense soektog na bio-afbreekbare agente soos etieleendiamiendisuksien suur (EDDS) and iminodisuksien suur (IDS) wat as plaasvervangers kan dien vir nieafbreekbare EDTA. Daar is dus n behoefte om te kyk na eenvoudige, vinnige en noukeurige metodes vir die bepaling van EDTA, EDTA , IDS en ander APCA s. Kapillere elektroforese (CE) was gebruik vir die skeiding en spesiering van iminodisuksien suur met verskillende metale by verskillende pH s. Spesiasie modellering was ook gebruik om die teenwoordigheid en verspreiding van metaal- spesies te vergelyk. Die CE uitslae was met die spesiasie profiele vergelyk en n redelike ooreenkoms was gevind. Die degraderings studies as n funksie van tyd was met CE bestudeer vir verskillende metal-ligand (ML) samestellings van DTPA, S,S-EDDS, IDS en R,S-IDS met verskillende metal ione (Cd2+, Cr3+, Cu2+, Fe3+, Mn2+, Pb2+ en Zn2+) by pH 7 en 9. Nuwe pieke was opgemerk in sommige ML samestellings wanneer die pH van pH 9 na 7 verander. Hoë pieke was vir CuL (L = DTPA, S,S-EDDS, IDS and R,S-IDS), FeEDTA en FeEDDS by alle pH s gevind. Lae, breë pieke was vir FeIDS, CrL en MnL komplekse gevind. CuDTPA en CuEDDS komplekse het n goeie stabiliteit gewys oor ʼn redelike tyd as dit met CuIDS, CuR,S-IDS en ander metal komplekses by pH 9 vergelyk word. In hierdie werk was die effek van verskillende kationiese elektro-osmotiese stroom (EOF) modifiseerders ook ondersoek. Veral die effek wat teen ioone op die CE skeidings het van EDTA, EDDS en IDS asook Cu(II) komplekse was ondersoek. Die effek van die modifiseerders was ondersoek en ge-evalueer in terme van migrasie tye, resolusie en plaat getalle. Die beste skeidingskondisies was bereik wanneer tetradesieltrimetielammonium bromied (TTAB) and setieltrimetielammonium bromied (CTAB) as modifiseerders gebruik word. Hierdie kondisies het ook aanleiding gegee tot korter ondersoek tye en beter piekvorme. Die effek van verskillende teen ioone wat aan die EOF modifiseerders gekoppel was het ook interresante resultate opgelewer. Die EOF modifiseerders setieltrimetielammonium chloried (CTAC) en tetradesieltrimetielammonium chloried (TTAC) was ook gebruik vir die bepaling van EDTA in Suid Afrikaanse rivier waters en industrieel afloop. ʼn Metode vir die gelyktydige skeiding van Fe(III) met EDTA, S,S’-EDDS en IDS was met behulp van CE en hoedruk vloeistof chromatography (HPLC) ontwikkel. Die metodologie was ondersoek ingevolge lineariteit, limiet van deteksie (LOD), limiet van kwantifisering (LOQ) en die herhaalbaarheid van CE en die HPLC metodes. Die LOD waardes verkry vanaf HPLC was swakker vergeleke met die verkry deur CE. Die toepaslikheid van al die metodes was vir die ontleding van kosmetiese produkte soos bad skuim en stortbad room getoets. Die uitslae deur CE en HPLC was vergelykbaar en ʼn goeie ooreenkoms was gevind.

Thesis (PhD)--Stellenbosch University, 2004. / ENGLISH ABSTRACT: Separation of the metal ions Cu2+, Pb2+, Cd2+ Mn2+, Cr3+, Fe3+ and Zn2+ with poly ethylenediamine tetra-acetic acid (EDTA), ethylenediamine disuccinic acid ([S,S']- EDDS) and ethylenediamine dimalonic acid (EDDM) was performed by Capillary Electrophoresis. The electropherograms obtained by Capillary Electrophoresis were validated with a speciation model Joint Expert Speciation System (JESS). Excellent agreement was observed for the speciation diagrams for these species with the pH curves determined by capillary electrophoresis with those determined by speciation modelling using JESS. The ligands EDDS and EDDM are readily biodegradable. They have been proposed as substitute ligands for EDTA. Detection of the above species was performed using both an electrochemical and a UV detector. The obtained electropherograms were used for the determination of the detection limits of these species. The electrochemical detector has a lower detection limit for these species than the UV detector in conformity with reported literature for these detectors. / AFRIKAANSE OPSOMMING: Skeiding van die metaal ione Cu2+, Pb2+, Cd2+, Cr3+, Fe3+ en Zn2+ met behulp van etileendiamientetra-asyn suur (EDTA), etileendiamiendisuksien suur ([S,S']-EDDS) en etileendiamiendimaleen suur (EDDM) was gedoen deur kapilêre elektroferose. Die elektroferogramme, verkry deur kapilêre elektroferose was gebruik om die spesiasie model "Joint Expert Speciation System (JESS)" te valideer. Uitstekende ooreenstemming was waargeneem vir die verspreidings diagramme van hierdie spesies as 'n funksie van pH wat verkry was deur kapilêre elektroferose met die' wat vasgestel was deur die spesiasie modeleeringsproses van JESS. Die ligande EDDS en EDDM toon onmiddelike biodigradeerbaarheid. Hulle was voorgehou as alternatiewe ligande vir EDTA. Bepaling van die bogenoemde spesies was ook gedoen deur beide elektrochemiese en UV deteksie. Deur gebruik te maak van hierdie detektore was die deteksie limiete van hierdie spesies bepaal. Die elektroochemiese detektor het 'n laer deteksie limiet vir hierdie spesies in vergelyking met die UV detector. Dit stem ooreen met die literatuur vir hierdie detektore.

Thesis (PhD)--Stellenbosch University, 2004. / ENGLISH ABSTRACT: Separation of the metal ions Cu2+, Pb2+, Cd2+ Mn2+, Cr3+, Fe3+ and Zn2+ with poly ethylenediamine tetra-acetic acid (EDTA), ethylenediamine disuccinic acid ([S,S']- EDDS) and ethylenediamine dimalonic acid (EDDM) was performed by Capillary Electrophoresis. The electropherograms obtained by Capillary Electrophoresis were validated with a speciation model Joint Expert Speciation System (JESS). Excellent agreement was observed for the speciation diagrams for these species with the pH curves determined by capillary electrophoresis with those determined by speciation modelling using JESS. The ligands EDDS and EDDM are readily biodegradable. They have been proposed as substitute ligands for EDTA. Detection of the above species was performed using both an electrochemical and a UV detector. The obtained electropherograms were used for the determination of the detection limits of these species. The electrochemical detector has a lower detection limit for these species than the UV detector in conformity with reported literature for these detectors. / AFRIKAANSE OPSOMMING: Skeiding van die metaal ione Cu2+, Pb2+, Cd2+, Cr3+, Fe3+ en Zn2+ met behulp van etileendiamientetra-asyn suur (EDTA), etileendiamiendisuksien suur ([S,S']-EDDS) en etileendiamiendimaleen suur (EDDM) was gedoen deur kapilêre elektroferose. Die elektroferogramme, verkry deur kapilêre elektroferose was gebruik om die spesiasie model "Joint Expert Speciation System (JESS)" te valideer. Uitstekende ooreenstemming was waargeneem vir die verspreidings diagramme van hierdie spesies as 'n funksie van pH wat verkry was deur kapilêre elektroferose met die' wat vasgestel was deur die spesiasie modeleeringsproses van JESS. Die ligande EDDS en EDDM toon onmiddelike biodigradeerbaarheid. Hulle was voorgehou as alternatiewe ligande vir EDT A. Bepaling van die bogenoemde spesies was ook gedoen deur beide elektrochemiese en UV deteksie. Deur gebruik te maak van hierdie detektore was die deteksie limiete van hierdie spesies bepaal. Die elektroochemiese detektor het 'n laer deteksie limiet vir hierdie spesies in vergelyking met die UV detector. Dit stem ooreen met die literatuur vir hierdie detektore.

Thesis (MSc)--Stellenbosch University, 2000. / ENGLISH ABSTRACT: This study was initialized to introduce capillary electrophoresis (CE) as a useful technique in the analysis of fluid inclusions in quartz. lts advantages are low detection limits for the dissolved ionic content of the fluid, the small amount of sample (1 g or less) for a detailed qualitative and quantitative analysis, and the short time required to obtain results (one run for either cations or anions take approximately 10 minutes). The study area from which quartz veins were selected is situated within the Neoproterozoic Saldania belt. Syn- and post-tectonic S-, 1- and A-type granitoids from the Cape Granite Suite intruded the metamorphosed Malmesbury greywacke and pelites between 550 and 510 Ma. Additional periods of tectonism and metamorphism occurred during Cape Supergroup sedimentation (480 - 400 Ma) as well as Karoo sedimentation and the simultaneous Cape Orogeny (280 -215 Ma). The quartz-biotite±chlorite vems are hosted by Cape Granite as well as Malmesbury sediments. These barren quartz veins are part of two vein sets, one dipping at an angle between 15 and 500 to the S to SE and striking W, similar to Sn-mineralized quartz veins in the SW-cape, while the other is near-vertical and striking W to NW. Except for their orientation, no differences regarding associated minerals, inclusion characteristics or fluid chemistry indicated a difference in origin. Four fluid phases within a temperature range of 160 - 390 °C were identified as being largely late-magmatic and released from the underlying Cape Granite plutons, namely an early 370- 390 °C population, followed by the 310 - 360, 230 - 300 and lastly the 160 - 200 °C populations. Initiation of this fluid system occurred from at least SlOMa, after final granite intrusion, but the age of the final stage is unknown. Renewed fluid circulation occurred during a later period of metamorphism, possibly during the Cape Orogeny. These fluids had temperatures between 240 and 360°C and are of sedimentary origin, most likely released from the Malmesbury metamorphites. The technique of capillary electrophoresis has been evaluated for its application to bulk fluid inclusion analysis, and the crush-leach fluid extraction procedure of Bottrell, et al., (1988) optimized for CE analysis. Contamination factors were identified and minimized or eliminated, where possible. Bulk fluid inclusion chemistry obtained by CE was therefore proved to provide valuable information regarding the various fluid generations as long as inclusion populations are investigated individually to explain and correlate bulk data. / AFRIKAANSE OPSOMMING: Die doel van die studie was om te toon dat die tegniek van kapillêre elektroforese bruikbaar is in die analiese van vloeistofmsluitsels in kwarts. Die voordele van hierdie tegniek is lae deteksie limiete vir die opgeloste ioon inhoud van die vloeistof, die klein monstergrootte (< 1g) wat nodig is vir 'n omvattende kwalitatiewe en kwantitatiewe analise, en die kort tydsduur waarin resultate verkry word ('n katioon of anioon analise vir een monster duur lO minute). Die studie gebied waar kwarts are gemonster is, is binne die Neoproterosoïese Saldania Gordel geleë. Sin- en laat-tektoniese S-, I- en A-tipe graniete van die Kaapse Graniet Suite het die gemetamorfiseerde Malmesbury grouwakke en peliete tussen 550 en 510 Ma binnegedring. Latere periodes van tektonisme en metamorfose het tydens deponering van die Kaap Supergroep (480 - 400 Ma), en die gelyktydige episodes van Karoo sedimentasie en Kaapse Orogenese (280 - 215 Ma) plaasgevind. Die gasheer gesteentes vir die kwarts-biotiet±chloriet are is Kaapse Graniet sowel as Malmesbury sedimente. Hierdie ongemineraliseerde are is deel van twee aarstelsels, nl. een met 'n duik hoek tussen 15 en 50° S tot SO en 'n westelike strekking, soortgelyk aan die Sn- ,.gemineraliseerde are in die SW-Kaap, terwyl die ander stel are feitlik vertikaal is en W tot NW strek. Behalwe vir die verskil in oriëntasie was daar geen aanduiding, wat betref 'n verskil in geassosieerde minerale, vloeistofinsluitsel kenmerke of vloeistof chemie, dat hierdie twee aarstelsels van verskillende oorsprong is nie. Vier vloeistof fases binne 'n temperatuur gebied van 160 - 390 °C en 'n vloeistof saliniteit van 0 - 5.7 gewig % NaC1 ekw. is geïdentifiseer, met 'n laat-magmatiese assosiasie en vrygestel deur die onderliggende Kaapse Graniete. Dit behels 'n vroeë 370 - 390 °C populasie, gevolg deur die 310 - 360, 230 - 300 en laastens die 160 - 200 °C populasies. Inisiasie van hierdie sisteem kon moontlik rondom 510 Ma gelede plaasgevind het, maar die ouderdom van die finale fase is onbekend. Hernude vloeistof sirkulasie het tydens 'n later stadium van metamorfose onstaan, moontlik tydens die Kaapse Orogenese. Hierdie vloeistowwe het temperature tussen 240 en 360 °C en is van sedimentêre oorsprong waar dit moontlik deur metamorfose van die reeds gemetamorfiseerde Malmesbury gesteentes vrygestel is. Die tegniek van kapillêre elektroforese is vir die toepassing daarvan in die analise van vloeistof insluitsels in kwarts geëvalueer, terwyl die vloeistof vrystellingsmetode van Bottrell en Yardley (1988) vir hierdie tegniek geoptimaliseer is. Kontaminasie faktore is geïdentifiseer en verminder of uitgeskakel waar moontlik. Daar is getoon dat die vloeistof chemie, wat verteenwoordigend is van al die insluitsel populasies in 'n monster, wel bruikbaar is t.o.v. afsonderlike vloeistof generasies, solank elke populasie individueel bestudeer is om die omvattende chemiese data te verduidelik en met 'n enkele populasie te korrelleer.

Superoxide Dismutase (SOD) activity in cell-free extracts prepared from healthy mature flowers of Petunia hybrida (variety 'Hurrah') was studied. The SOD activity in the crude extracts was stable for more than one month when stored at -20 oC. It was found that pH 7.8 is optimal for SOD activity. Different flower tissues of petunia (stigma, style and ovary) at various stages of development were extracted and analysed for SOD activity. SOD activity was found to be significantly highest in the ovary tissue of dehiscent petunia flowers. Three SOD isozymes were detected after crude extracts of the different female reproductive tissues of petunia flowers were analysed on a non-denaturing polyacrylamide gel electrophoresis system. Based on a difference in the sensitivity of the SOD isoforms to H2O2 and KCN, it is suggested that Mn-SOD, Fe-SOD and Cu/Zn-SOD were present in the crude extracts of the female reproductive tissues of petunia flowers. The response of the female reproductive parts of petunia flowers was also tested under water deficiency and high temperature (35 oC) stress. The SOD activity seemed to increase more in response to the high temperature than the water deficiency stress. Intense blue staining was observed from developing younger buds, and much lower formazan deposition was detected at the later stage. This indicates the lower O2- produced during later stages mainly due to increasing SOD synthesis. DEAE cellulose chromatography was successfully used to partially purify SOD from the ovaries of petunia flowers. The characteristics of the partially purified enzyme fraction were found to be very similar to those of the crude extracts.