Thesis (MSc Food Sc)--Stellenbosch University, 2011. / ENGLISH ABSTRACT: Rooibos tea, produced from the endemic South African fynbos plant Aspalathus linearis, has various healthpromoting benefits which can largely be attributed to its phenolic composition. In this study, the reversedphase liquid chromatographic (RP-LC) separation of the principal phenolic constituents of aqueous rooibos infusions was optimised on conventional high-performance liquid chromatography (HPLC) instrumentation. The phenolic constituents comprised dihydrochalcones (aspalathin and nothofagin), flavones (orientin, isorientin, vitexin, isovitexin, luteolin, luteolin-7-O-glucoside and chrysoeriol), flavonols (quercetin, isoquercitrin, hyperoside and rutin), a hydroxycinnamic acid (ferulic acid) and a phenylpropanoid (phenylpyruvic acid glucoside, PPAG). A systematic approach towards method development was adopted: resolution was optimised by simultaneous optimisation of gradient conditions and temperature on a sub-2 6m phase to exploit the benefits of this phase for fast routine analyses. The developed HPLC method, utilising photodiode-array (PDA) detection, yielded complete separation of the 15 target analytes on the 1.8 µm C18 column, thermostatted to 37°C, within 37 min (total analysis time: 50 min). The method was successfully validated and demonstrated its suitability for the fast, quantitative analysis of aqueous infusions of unfermented and fermented rooibos. Mass spectrometric (MS) and tandem MS detection confirmed peak purity and permitted the tentative identification of 13 additional phenolic compounds, including a flavonol Odiglycoside (quercetin-3-O-robinobioside), a luteolin-6-C-pentoside-8-C-hexoside and a novel C-8-hexosyl derivative of aspalathin reported here for the first time. The HPLC-PDA method was subsequently applied to a large number of fermented rooibos samples representative of different production seasons (2009, 2010 and 2011) and quality grades (grades A, B, C and D) to capture as much potential variation in the phenolic composition as possible. Production season had no clear effect on the levels of the individual phenolic compounds in ‘cup-of-tea’ rooibos infusions, whilst high quality tea (grades A and B) was associated with higher levels of phenolic compounds and soluble solids than low quality tea (grades C and D). Steam-pasteurisation of the plant material, required to obtain a product of high microbiological quality, induced significant reductions in the mean values of most of the phenolic compounds in rooibos infusions. The major phenolic constituents of steam-pasteurised, fermented rooibos were isoorientin and orientin, whilst quercetin-3-O-robinobioside, PPAG and aspalathin were also present in high concentrations. Representative content values of the major phenolic compounds present in a typical ‘cup-of-tea’ rooibos infusion were thus obtained and the generated data are suitable for inclusion in food composition databases. The application of comprehensive two-dimensional liquid chromatography (LCxLC) was investigated as an alternative approach for the detailed investigation of rooibos phenolics. The combination of hydrophilic interaction chromatography (HILIC) in the first dimension and RP-LC in the second dimension offered different separation selectivities and hence a high degree of orthogonality. HILICxRP-LC provided a significant improvement in resolution, as is evident from practical peak capacities in excess of 2000 and 800 for the off-line and on-line methods, respectively. Further optimisation, particularly of the first dimension separation, is however required to improve the LCxLC separation of complex rooibos phenolic fractions. / AFRIKAANSE OPSOMMING: Gebruik van rooibostee, berei vanaf die eg Suid-Afrikaanse fynbosplant Aspalathus linearis, hou verskeie gesondheidsvoordele in wat grootliks toegeskryf kan word aan sy fenoliese samestelling. Die skeiding van die hoof fenoliese verbindings van ’n koppie rooibos is in hierdie studie deur middel van omgekeerde-fase vloeistof chromatografie (RP-LC) op konvensionele hoë-druk vloeistof chromatografiese (HPLC) toerusting geoptimiseer. Die fenoliese verbindings was verteenwoordigend van dihidrogalkone (aspalatien en notofagien), flavone (orientien, isoorientien, viteksien, isoviteksien, luteolien, luteolien-7-O-glukosied en krisoeriol), flavonole (kwersetien, isokwersetrien, hiperosied en rutien), ‘n hidroksiekaneulsuur (ferulasuur) en ‘n fenielpropanoied (fenielpirodruiwesuurglukosied, PPAG). Die ontwikkeling van die metode was sistematies benader: resolusie is op ‘n geselekteerde 1.8 6m stationêre fase met welombekende kinetiese voordele geoptimiseer deur die gradiëntkondisies en kolomtemperatuur gelyktydig te optimiseer. Die ontwikkelde HPLC metode, gekoppel aan ultraviolet-fotodiode deteksie (PDA), het binne 37 min (totale analiese tyd: 50 min) volledige skeiding van die 15 standaard verbindings op die 1.8 µm C18 kolom teen 37°C bewerkstellig. Die metode is suksesvol gevalideer en het sy toepaslikheid vir vinnige, kwantitatiewe analiese van ongefermenteerde en gefermenteerde rooibos gedemonstreer. Piek suiwerheid is deur middel van massa spektrometrie (MS) en tandem MS bevestig, wat ook die identifikasie van 13 addisionele verbindings toegelaat het, insluitende ‘n flavonol O-diglukosied (kwersetien-3-O-robinobiosied), ‘n luteolien-6-Cpentosied- 8-C-heksosied en ‘n unieke C-8-heksosiel afgeleide van aspalatien wat vir die eerste keer hier gemeld is. Die geoptimiseerde HPLC-PDA metode is gevolglik toegepas vir die analiese van ‘n groot aantal gefermenteerde rooibos monsters, verteenwoordigend van verskillende produksie seisoene (2009, 2010 en 2011) en kwaliteitsgrade (A, B, C en D). Hierdie lukraak-geselekteerde monsters het soveel as moontlik potensiële variasie in die fenoliese samestelling verseker. Produksie seisoen het geen definitiewe effek op die vlakke van die individuele fenoliese verbindings in ’n koppie rooibos gehad nie, terwyl hoë kwaliteit rooibos (grade A en B) geassosieër was met hoër vlakke van die individuele fenoliese verbindings en oplosbare vastestowwe in vergelyking met lae kwaliteit rooibos (grade C en D). Stoompasteurisasie van rooibos plantmateriaal, noodsaaklik om ‘n produk van hoë mikrobiologiese gehalte te verseker, het gelei tot ‘n betekenisvolle afname in meeste fenoliese verbindings in ’n koppie rooibos. Die hoof fenoliese verbindings van ‘n koppie stoom-gepasteuriseerde, gefermenteerde rooibos was orientien en isoorientien, terwyl kwersetien-3-O-robinobiosied, PPAG en aspalatien ook in noemenswaardige hoeveelhede aanwesig was. Verteenwoordigende waardes van die hoof fenoliese verbindings aanwesig in ‘n tipiese koppie rooibos is derhalwe verkry en die data is geskik vir insluiting in voedsel-samestelling databasisse. Die analiese van rooibos fenole met omvattende twee-dimensionele vloeistof chromatografie (LCxLC) is bestudeer as ‘n alternatiewe metode om verdere insig tot hierdie komplekse fenoliese fraksie te verkry. Die kombinasie van hidrofiliese interaksie chromatografie (HILIC) in die eerste dimensie en RP-LC in die tweede dimensie het ‘n uiters gesogte lae graad van korrelasie verskaf. HILICxRP-LC het ‘n besonderse toename in resolusie teweeg gebring, gekenmerk deur praktiese piek kapasiteite hoër as 2000 en 800 vir die af-lyn en aan-lyn metodes, onderskeidelik. Verdere optimisering, veral van die eerste dimensie skeiding, is egter nodig om die LCxLC skeiding van rooibos fenole te verbeter.

Thesis (MSc Food Sc)--University of Stellenbosch, 2011. / ENGLISH ABSTRACT: The sensory characteristics and chemical composition of rooibos infusions were analysed to establish the extent of variation in sensory attributes and compositional parameters, and investigate whether correlations exist between the sensory characteristics and the phenolic composition. To capture as much potential variation as possible, 69 rooibos samples were collected throughout the 2009 harvesting season from different production areas. The samples were also representative of the different quality grades (A, B, C or D) which are based on the appearance of the leaves, and the colour and flavour of the infusion. Quantitative descriptive analysis was used to develop sensory profiles for each of the rooibos infusions. Considerable variation in the sensory characteristics was observed, highlighting the need for standardised terminology that can be used to describe what is often referred to as “characteristic” rooibos flavour. Aroma, flavour, taste, and mouthfeel descriptors were generated and assembled into a rooibos sensory wheel. The most recurring attributes were selected to compile a rooibos sensory lexicon which provides a definition and a reference standard for clarification of each term. A combination of “honey”, “woody” and “fynbos-floral” notes with a slightly sweet taste and subtle astringency may be regarded as “characteristic” rooibos attributes indicative of good quality tea. Pleasant “caramel” and “fruity-sweet” notes were also observed in a number of infusions, whereas negative descriptors, associated with poor quality rooibos, included “green” and “hay” flavours. Large variation in the composition of rooibos infusions was revealed through the quantification of soluble solids (SS), total polyphenols (TP), tannins and 14 monomeric phenolic compounds, as well as spectrophotometric colour measurements. High quality rooibos was associated with higher levels of SS, TP, tannins and phenolic compounds than low quality rooibos. Correlations between the compositional parameters and sensory attributes of the infusions indicated that several non-volatile compounds, including enolphenylpyruvic acid-2-glucoside, quercetin-3-glucoside, and iso-orientin, were associated with the characteristic sweet taste of rooibos, while bitterness was related to certain flavonoids such as luteolin, quercetin and aspalathin. The only compound significantly correlated to astringency was rutin, although it is likely that aspalathin and several other monomeric flavonoids also contribute to astringency. The tannin content was not associated with astringency possibly because of the limitations associated with the tannin quantification method. To determine whether compositional changes resulting from steam pasteurisation of rooibos leaves influence the sensory quality of rooibos infusions, differences in the phenolic composition and sensory attributes of infusions, prepared from unpasteurised and pasteurised rooibos, were analysed. Steam pasteurisation significantly reduced the SS, TP and aspalathin content of rooibos infusions, as well as the absorbance, especially at a wavelength of 450 nm. It also resulted in significant reductions in the intensities of most of the aroma and flavour attributes, especially the “green” notes associated with low quality rooibos. After steam pasteurisation the prominent “green” flavour of certain samples was frequently replaced by a “hay” flavour. The taste attributes, sweetness and bitterness, remained unchanged, whereas the astringency of rooibos infusions decreased significantly. / AFRIKAANSE OPSOMMING: Die sensoriese eienskappe en chemiese samestelling van rooibostee is geanaliseer ten einde die mate van variasie van die sensoriese sowel as die chemiese profiel te bepaal, asook om vas te stel of korrelasies bestaan tussen die sensoriese eienskappe en die fenoliese samestelling van rooibostee. Om soveel as moontlik potensiële variasie in te sluit, is 69 rooibos monsters tydens die 2009 oesseisoen van verskillende produksiegebiede versamel. Die monsters is ook verteenwoordigend van die verskillende kwaliteitgrade (A, B, C en D), wat op grond van die voorkoms van die teeblare, en die kleur en geur van die tee toegeken is. Kwantitatiewe beskrywende analise is gebruik om ‘n sensoriese profiel vir elke rooibostee op te stel. Aansienlike variasie in die sensoriese eienskappe is waargeneem. Dit het die behoefte aan gestandaardiseerde terminologie onderskryf, wat gebruik kan word om die term “karakteristieke” rooibos geur in meer detail te omskryf. Spesifieke terme vir aroma, geur, smaak en mondgevoel is geformuleer, en gebruik om ‘n sensoriese wiel vir rooibostee saam te stel. Terme wat die meeste voorgekom het, is gekies om in ‘n sensoriese leksikon vervat te word wat ‘n definisie en ‘n verwysingstandaard vir elke term voorstel. Die “karakteristieke” sensoriese eienskappe van goeie kwaliteit rooibostee kan soos volg gedefinieer word: ’n kombinasie van “heuning”, “houtagtige” en “fynbos-blom” geure met ‘n effense soet smaak en ‘n sagte vrankheid. Aangename “karamel” en “vrugtige-soet” geure is ook waargeneem, terwyl terme soos “groen” en “hooi” met swak kwaliteit rooibostee geassosieer is. Groot variasie in die samestelling van rooibostee is verkry deur kwantifisering van die inhoud van oplosbare vastestowwe (SS), totale polifenole (TP), tanniene, en 14 monomeriese fenoliese verbindings, asook spektrofotometriese kleurmetings. Hoë kwaliteit rooibostee is geassosieer met hoër vlakke van SS, TP, tanniene en fenoliese verbindings in vergelyking met lae kwaliteit rooibos. Korrelasies tussen laasgenoemde parameters en die sensoriese eienskappe het aangedui dat sekere nie-vlugtige verbindings, bv. fenielpirodruiwesuurglukosied, kwersetien-3-glukosied en iso-orientien, geassosieer is met die kenmerkende soet smaak van rooibos, terwyl ‘n bitter smaak geassosieer is met spesifieke flavonoïede soos luteolien, kwersetien and aspalatien. Die enigste verbinding wat betekenisvol gekorreleer het met vrankheid was rutien, maar dit is waarskynlik dat aspalatien en ander monomeriese flavonoïede ook bygedra het tot vrankheid. Die tannieninhoud het nie verband gehou met vrankheid nie, vermoedelik as gevolg van die beperkinge van die chemiese metode vir tannienbepaling. Om te bepaal of stoompasteurisasie van rooibos die sensoriese kwaliteit en chemiese samestelling van rooibostee beïnvloed, is die verskille in fenoliese samestelling en sensoriese eienskappe van die tee, berei van ongepasteuriseerde en gepasteuriseerde rooibos, ondersoek. Stoompasteurisasie het ‘n betekenisvolle verlaging teweegebring van die SS, TP en aspalatien inhoud, sowel as van die absorbansie van die tee, veral by ‘n golflengte van 450 nm. Dit het ook gelei tot ‘n statisties betekenisvolle vermindering van die intensiteit van die meeste aroma en geur eienskappe, veral die sogenaamde “groen” eienskap wat met swak kwaliteit rooibos verbind word. As gevolg van stoompasteurisasie is die opmerklike “groen” geur van sekere ongepasteuriseerde monsters herhaaldelik vervang deur ‘n “hooi” geur. Die smaakeienskappe, soet en bitter, was onveranderd, terwyl vrankheid statisties betekenisvol afgeneem het as gevolg van pasteurisasie.

Philosophiae Doctor - PhD / Camellia sinensis (tea) and Aspalathus linearis (rooibos) may improve reproductive function owing to their antioxidant properties. To test this hypothesis, male and female rats were given 2% and 5% green tea (Gt), black tea (Bt), unfermented rooibos (Ur) or fermented rooibos (Fr) as sole source of drinking for 52 and 21 days respectively. Control rats received tap water. In addition, TM3 Leydig cells were exposed to 0.025, 0.05, 0.1 and 0.5 % aqueous extracts of green tea, black tea, unfermented and fermented rooibos for 24h. In vitro analysis of tea and the herbal infusion revealed the phenolic property and antioxidant capacity (FRAP) in the order Gt > Bt > Ur > Fr. Camellia sinensis and Aspalathus linearis revealed no significant effect on serum antioxidant capacity (p > 0.05) and lipid peroxidation (MDA) in the kidney or liver in both male and female rats and in the testes of the male rats (p > 0.05). In addition, the antioxidant levels were maintained in the testes, liver and kidneys in both the male and female rats. In the male rats, no significant alterations were observed in body weight gain, liver and reproductive organs weight, and serum testosterone (p > 0.05). Only, 5% green tea significantly increased testosterone level (p < 0.05). Seminiferous tubules displayed complete spermatogenesis with abundant sperm in the lumen in all treated groups. However, a significant decrease in diameter and germinal epithelial height of these tubules were observed (p < 0.05). In the epididymides, epithelial height of caput region showed a significant increase (p < 0.01), while the cauda region was increased by Camellia sinensis but decreased by Aspalathus linearis. Sperm concentration improved significantly by green tea and unfermented rooibos (p < 0.05), while black tea and fermented rooibos produced a non significant effect (p > 0.05). Sperm viability was enhanced in all treatment groups (p < 0.05). Furthermore, green tea, black tea and unfermented rooibos significantly improved the motility of rat sperm (p < 0.05); fermented rooibos tended to improve it (p > 0.05). In addition, green tea, black tea and fermented rooibos enhanced acrosome reaction (p < 0.05). Creatinine activity was significantly higher in rats treated with black tea, unfermented rooibos or fermented rooibos (p < 0.05), green tea tended to increase it (p > 0.05) reflecting the significant increased kidney weight in the treatment groups at high concentrations. Liver markers, ALT and AST, decreased significantly in all treated groups (p < 0.05), except in 5% fermented rooibos where a significant increase in AST level was observed (p < 0.01). In the female rats, the body weight gain, and reproductive organs weight was no affected (p > 0.05). However, 5% fermented rooibos reduced the ovarian weight (p < 0.05), while 5% unfermented rooibos significantly increased the uterine weight (p < 0.05). Liver weight increased significantly by black tea and unfermented rooibos (p < 0.05) while the kidney weight increased significantly by 5% black tea (p < 0.05). No significant effect was observed in the level of FSH produced, on the other hand, Camellia sinensis significantly lowered the level of LH (p < 0.05), while Aspalathus linearis had no effect (p > 0.05). Creatinine activity was enhanced significantly only by 5% fermented rooibos (p < 0.05). Liver markers, ALT and AST were reduced in most treated groups except in fermented rooibos where an increase was observed. In addition, histological sections revealed no obvious alteration in the ovaries, uteri, kidneys and liver of all treated female rats. Camellia sinensis and Aspalathus linearis significantly reduced the level of testosterone produced in TM3 Leydig cells under stimulated conditions in vitro (p< 0.05). Furthermore, both plants maintained the viability and morphology of the cells. However, at 0.5% of either plant extracts, a significant decrease in the viability (p < 0.05) and altered morphology of the TM3 Leydig cells was observed. In conclusion, Camellia sinensis and Aspalathus linearis significantly improved certain sperm function which might be attributed to their high level of antioxidant activity. However, the prolonged exposure of both plant extracts might result in subtle structural changes in the male reproductive system and impair kidney function. In addition, fermented rooibos at high concentration may also impair the functions of the liver. In vitro, both plants were shown to possess anti-androgenic property on TM3 Leydig cells. Furthermore, both Camellia sinensis and Aspalathus linearis may be classified as weak phytoestrogens due to the changes in the weight of the uterus and ovaries observed. / South Africa

Thesis (MSc Food Sc)--University of Stellenbosch, 2001. / ENGLISH ABSTRACT:Aspalafhus linearis is an indigenous fynbos plant cultivated in the Clanwilliam area of the Western Cape, South Africa. The rooibos tea that is prepared from this plant, has become popular worldwide mainly due to the alleged health properties. Studies on the anti-microbial properties of green, black and oolong teas have shown that these teas have strong anti-microbial activity against a wide range of microbes. No studies have been done on the anti-microbial activity of rooibos tea and the aim of this study was to determine what impact rooibos tea extracts would have on the growth of different food spoilage and potential pathogenic microbes. Water and ethyl acetate extracts of fermented and unfermented rooibos tea were used to determine the inhibitory effect on the growth of an Escherichia coli strain. The E. coli culture was grown in tea-MRS with either added fermented or unfermented rooibos tea extracts. Both the water and ethyl acetate extracts showed a strong inhibitory effect against the E. coli strain in that there was a decrease in the final bacterial cell density (Nmax)(from 0.59 00 to 0.25 00) and the maximum specific growth rate (~max)(from 1.12 h-1 to 0.20 h-1) and an increase in the doubling time (~) (from 0.59 h to 1.80 h) and lag time (tlag)(from 4.81 h to 6.60 h) as the concentration of the soluble solids of the tea extracts was increased from 0.5 to 5.0 g.r1 . Furthermore, it was found that the fermented rooibos tea had a much stronger inhibitory effect (69% decrease in growth at 5.0 g.r1 soluble solids) compared to the unfermented rooibos tea extracts (35.1% decrease in growth at 5.0 g.r1 soluble solids). The resulting data indicated that rooibos tea had a very strong inhibitory effect on the growth of the E. coli strain. It was also found that the water extracts of rooibos tea showed a stronger inhibitory effect on the growth of the E. coli than the ethyl acetate extracts, indicating that the antimicrobial activity of rooibos tea is not exclusively due to the polyphenolic content - individual compounds. It was also determined that the rooibos tea water extracts showed a bacteriostatic action against the E. coli strain in that as soon as the tea is no longer part of the growth medium, the E. coli resumed a normal growth pattern. The data obtained showed that the inhibitory effect of rooibos tea water extracts (69% decrease in growth) against the growth of E. coli was more pronounced than that found when black tea water extracts (25.7% decrease in growth) at the same concentrations were used.Rooibos tea water extracts (0.5 - 5.0 g.r1) of fermented and unfermented tea were also used to determine the inhibitory effect on other food spoilage microbes and potential pathogens. Strains of Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes, Streptococcus mutans, Saccharomyces cerevisiae and Zygosaccharomyces rouxii were grown in the presence of fermented and unfermented rooibos tea water extracts. The effect that fermented rooibos tea had on the growth of all the microbes tested was in the following order: Staph. aureus (90.8% decrease in growth) > L. monocytogenes (89.2% decrease in growth) > Strep. mutans (84.1% decrease in growth) > B. cereus (80.3% decrease in growth) > Sacch. cerevisiae (77.7% decrease in growth) > E. coli (69.0% decrease in growth). The rooibos tea clearly had an inhibitory effect on the growth of all the microbes, with the exception of the Z. rouxii strain where the presence of the tea water extracts was found to enhance the growth. The inhibitory effect of rooibos tea on the growth of these microbes was shown by changes in the growth parameters with Nmax and IJmaxshowing decreases, while the ld and tlagincreased as the concentration of the tea soluble solids was increased. As with E. coli, the fermented rooibos tea water extracts showed the stronger inhibitory effect on the growth of the various microbes. The data obtained in this study suggests that rooibos tea is not effective as an anti-microbial agent against all yeast species, but will strongly retard the growth of specific Gram-positive and Gram-negative bacteria. As long as rooibos tea is present, strong anti-microbial activity will be observed at a cup of tea concentration of 2.5 g.r1 soluble solids. These results may be of value to support the health claims associated with rooibos tea and may in the future lead to the use of rooibos tea as a "natural" food preservative. / AFRIKAANSE OPSOMMING:Aspalathus linearis is 'n inheemse fynbosplant wat gekultiveer word in die Clanwilliam area van die Wes Kaap, Suid-Afrika. Rooibostee, wat gemaak word van hierdie plante, het baie gewild geword wereldwyd a.g.v. die gesondheidsaspekte van hierdie tee. Studies toon dat groen, swart en oolong tee sterk anti-mikrobiese aktiwiteit het teen 'n wye reeks mikrobes. Aangesien daar voorheen geen studies gedoen is op die anti-mikrobiese aktiwiteit van rooibostee nie, was die doel van hierdie studie om die effek van rooibostee te bepaal op die groei van verskillende voedselbederwers en potensiele patogeniese mikrobes. Water- en etielasetaat-ekstrakte van gefermenteerde en ongefermenteerde rooibos tee is gebruik om die inhiberende effek op die groei van Escherichia coli te bepaal. Escherichia coli is gegroei in tee-MRS met bygevoegde gefermenteerde of ongefermenteerde rooibostee-ekstrakte. Seide die water- en etielasetaatekstrakte van rooibostee het 'n sterk inhiberende effek gewys teen E. coli en dit word gestaaf deur 'n afname in die finale bakteriese seldigtheid (Nmax)(vanaf 0.59 00 tot 0.25 00) en die maksimum spesifieke groeitempo (lJmax) (vanaf 1.12 h-1 tot 0.20 h-1) en 'n toename in die verdubbelingstyd (~) (vanaf 0.59 h tot 1.80 h) en die sloerfase (tlag)(vanaf 4.81 h tot 6.60 h) 5005 wat die konsentrasie van oplosbare vastestowwe van die tee toeneem van 0.5 tot 5.0 g.r1 . Verder is daar gevind dat die gefermenteerde rooibostee 'n baie sterker inhiberende effek het (69% afname in groei by 5.0 g.r1 oplosbare vastestowwe) in vergelyking met die ongefermenteerde rooibostee-ekstrakte (35.1% afname in groei by 5.0 g.r1 oplosbare vastestowwe). Die resultate van die data dui aan dat rooibos tee 'n baie sterk inhiberende effek het op die groei van die E. coli spesie. Die waterekstrakte van rooibostee het 'n sterker inhibisie getoon teen die groei van E. coli as die etielasetaat-ekstrakte, wat aandui dat die anti-mikrobiese aktiwiteit van rooibostee nie eksklusief toegeskryf kan word aan die polifenoliese samestelling nie. Daar is ook gevind dat rooibostee water-ekstrakte 'n bakteriostatiese effek het teen E. coli, want sodra die tee ekstrakte nie meer teenwoordig is in die groeimedium nie, hervat E. coli normale groei. Die data wys ook dat die inhiberende effek van rooibostee water-ekstrakte (69.0% afname in goei) teen E. coli baie sterker is as die van swart tee water-ekstrakte (25.7% afname in groei) by dieselfde konsentrasies.Rooibostee water-ekstrakte (0.5 - 5.0 g.r1) van gefermenteerde en ongefermenteerde rooibostee is ook gebruik om die inhiberende effek te bepaal teen ander voedselbederwers en potensiele patogene. Spesies van Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes, Streptococcus mutans, Saccharomyces cerevisiae en Zygosaccharomyces rouxii is gegroei in die teenwoordigheid van gefermenteerde en ongefermenteerde rooibostee waterekstrakte. Die effek wat gefermenteerde rooibostee het op die groei van die getoetste mikrobes is 5005 volg: Staph. aureus (90.8% afname in groei) > L. monocytogenes (89.2% afname in groei) > Strep. mutans (84.1% afname in groei) > B. cereus (80.3% afname in groei) > Sacch. cerevisiae (77.7% afname in groei) > E. coli (69.0% afname in groei). Rooibostee het 'n duidelike inhiberende effek gehad teen al die organismes, behalwe teen Z. rouxii spes ie, waar die teenwoordigheid van rooibostee die groei van die organisme bevorder het. Die inhiberende effek van rooibostee teen die groei van hierdie mikrobes word ondersteun deur die groei parameters waar die Nmaxen IJmaxafgeneem het terwyl die ~ en tlagtoegeneem het 5005 wat die konsentrasie van die oplosbare vastestowwe toeneem. Die gefermenteerde rooibostee water-ekstrakte het ook 'n sterker inhiberende effek op die groei van die verskillende mikrobes net 5005 met E. coli. Die data wat verkry is van hierdie studie dui aan dat rooibostee nie effektief sal wees as 'n anti-mikrobiese middel teen aile gis spesies nie, maar dit sal die groei van spesifieke Gram-positiewe en Gram-negatiewe bakterie sterk vertraag. So lank as wat rooibostee teenwoordig is, sal sterk anti-mikrobiese aktiwiteit waargeneem word by 'n koppie-tee konsentrasie van 2.5 g.r1 oplosbare vastestowwe. Hierdie resultate kan help om die gesondheidseienskappe geassosieer met rooibostee te ondersteun en help om die gebruik van rooibostee as 'n "natuurlike" preserveermiddel te bevorder. dedicated to my parents

Magister Scientiae - MSc (Physics) / Zinc oxide (ZnO) is a wide and direct semiconductor with a wurtzite crystal structure. Its multifunctionality as the ideal candidate in applications such as blue-UV light emitting diodes, transparent conducting oxide, selective gas sensor and efficient catalyst support among others, has attracted a significant interest worldwide. Nano-scaled ZnO has been synthesized in a plethora of shapes. A rich variety of physical and chemical methodologies have been used in the synthesis of undoped or doped ZnO. However, such methods either necessitate relatively high vacuum infrastructures, elevated temperatures, or the use of toxic reagents. The "green chemistry" synthesis of metal oxide nanoparticles which is based on using natural plant extract as an effective 'reducing agent' of metal precursor, has been reported to be a cleaner and environment-friendly alternative to the physical and chemical methods. The thesis is based on the synthesis and the main physical properties of pure ZnO nanoparticles synthesized by a completely green chemistry process using the natural extract of Aspalathus Linearis to bio-reduce the zinc acetate precursor. The obtained ZnO nanopowdered samples were annealed at different temperatures from 300 °C to 600 °C. The samples were characterized using Scanning Electron Microscopy, Energy Dispersive Spectroscopy, Transmission Electron Microscopy, X-ray Diffraction, Differential Scanning Calorimetry, Thermogravimetric Analysis and Fourier Transform Infrared. Highly pure quasi-spherical ZnO nanoparticles with an average crystallite size of 24.6 nm (at 300 °C), 27.2 nm (at 400 °C), 27.6 nm (at 500 °C), and 28.5 nm (at 600 °C) were found. The results also showed that the average crystallite size increased with an increase in annealing temperature. It was successfully demonstrated that the natural plant extract of Aspalathus Linearis can be used in the bio-reduction of zinc acetate dihydrate to prepare highly pure ZnO nanoparticles.

Magister Scientiae - MSc (Medical BioSciences) / In the past three decades there has been a huge increase in the use of herbal medicine globally. The active principles of these herbal medicines are mostly unknown with supportive evidence for safety and efficacy very rare. SeptilinTM is a phytopharmaceutical formulation which is recommended for the treatment and management of various infections. It has been claimed to have immunomodulatory actions that potentiates the body's immune response. The immunomodulatory activity of SeptilinTM has not been well investigated via appropriate in vitro models. Therefore this study was undertaken to investigate the in vitro effects of SeptilinTM on biomarkers of specific immune pathways by using WBC. Stimulated and unstimulated WBC were incubated with the product. Enzyme linked immunosorbent assays were used to screen for IL-6, IL-10, and IFN? as biomarkers for inflammation, humoral immunity, and cell mediated immunity, respectively. Results show that the presence of SeptilinTM in LPS stimulated WBC has no effect on the release of IL-6 and IFN? production but stimulated IL-10 production. SeptilinTM in unstimulated WBC has no effect on the release of IL-10 and IFN? production but stimulatory effects on IL-6 production.