Thesis(M Tech(Biomedical Technology))--Cape Peninsula University of Technology, 2009 / This thesis provides the first scientific evidence of the photoprotective properties of rooibos and honeybush herbal tea extracts and to some extent, two major honeybush polyphenols, hesperidin and mangiferin. These properties were demonstrated using in vivo models by: Providing evidence for the inhibition of tumour promotion by ultraviolet B (UVB) radiation in a two-stage skin carcinogenesis mouse model. Topical application of polyphenol-rich extracts of rooibos and honeybush prior to UVB tumour promotion of 7,12-dimethylbenz[a]anthracene (DMBA)-initiated mouse skin, inhibited the formation of tumours. The rooibos and honeybush extracts decreased the incidence and volume of the tumours. Topical application of hesperidin and mangiferin were less effective than the honeybush extracts as only the tumour volume was decreased, but not the incidence. Providing evidence for the inhibition of photodamage of the skin by UVB exposure in a mouse model. Topical application of polyphenolic rich extracts of honeybush prior to UVB irradiation of mouse skin reduced erythema, peeling, oedema and hyperplasia. The depletion of antioxidant enzymes catalase and superoxide dismutase (SOD) was prevented. The extracts protected the skin from oxidative and direct DNA damage, and reduced lipid peroxidation. The induction of cyclooxygenase-2 (COX-2) and ornithine decarboxylase (ODC) was also reduced. Topical application of the polyphenols hesperidin and mangiferin showed reduced protective effects compared to the extracts. Suggesting the possible mechanisms by which honeybush and the polyphenols protect against photocarcinogenesis such as reducing tumour promotion, inflammation and oxidative stress. Suggesting the benefits of including honeybush and rooibos as cosmeceuticals in skin care products and sunscreens as part of the strategy for preventing skin cancer. Discussing the recommendations for further study such as investigating more specific chemopreventive activities of these two South African herbal teas and their polyphenols, dose response studies and clinical evaluations.

Thesis (MSc Food Sc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Application of steam treatment to green Cyclopia maculata and C. longifolia to modulate their aroma and flavour profile, without detrimental effects on colour and individual phenolic content, was investigated. Steam treatment (96 °C, atmospheric pressure) of different time periods was applied to the shredded, fresh plant material before drying (STBD; 0, 30, 60, 90 and 120 s) or the herbal tea product after drying (STAD; 0, 1, 2, 3, and 4 min). Steam treatment of 60 s or longer resulted in a significant (p < 0.05) decrease in vegetative (‘green grass’ and ‘hay/dried grass’) and cereal (‘oats/porridge/grains’) aroma and flavour intensities, with an increase in some fruity (‘tropical fruit’ and ‘guava’) and ‘fruity-sweet' aroma intensities. These changes manifested to a greater degree for plant material subjected to STBD, while they were less prominent for C. longifolia compared to C. maculata. Additionally, 19 aroma compounds were instrumentally identified in the volatile fraction of C. maculata infusions prepared from STBD plant material. Of these, seven correlated well with green honeybush vegetative and cereal aroma attributes associated with unsteamed samples. Steaming thus resulted in the rapid loss of some of these highly volatile ‘green’-associated aroma compounds. Steam treatment of more than 60 s, regardless of STBD or STAD, was not detrimental to green colour, and individual phenolic content was not greatly affected. The 30 s STBD treatment of C. maculata, however, led to the rapid loss of green colour and oxidation of phenolic compounds, without considerable aroma improvement. This highlights the need for process control during green honeybush production. Storage stability of steamed (60 s, STBD) and unsteamed green C. maculata was investigated under low (0 °C; moisture impermeable packaging) and normal (25 °C at 60% relative humidity (RH); semi-moisture-permeable sachets) temperature storage conditions (LTS and NTS, respectively) over 6 months and high temperature storage (HTS) conditions (40 °C at 75% RH; semimoisture- permeable sachets) for 1 month. HTS conditions, after 1 month, seemed to emulate changes occurring over 6 months at NTS conditions, with little change detected at LTS conditions. Despite noticeable green colour loss over the respective storage periods at NTS and HTS conditions, individual phenolic compounds were not severely affected. NTS and HTS led to the progressive development of sought-after fermented honeybush sensory attributes, especially in unsteamed samples. These include prominent fruity (‘stewed fruit’, ‘apricot jam’ and ‘marmalade’) aromas and flavours, ‘general sweet’ and ‘fruity-sweet’ aromas and a sweeter taste. The collated descriptive sensory data were used to generate a preliminary sensory wheel for green honeybush aroma, and another for flavour, taste and mouthfeel. The sensory profile of green honeybush was finally described as a dominant vegetative aroma and flavour, prominent sweetassociated and slightly fruity aroma, with sweet and notably bitter tastes and an astringent mouthfeel. By combining the sensory data, it was clear that the major effects of storage outweighed those of steam treatments, suggesting that storage of 3 to 6 months may improve sensory quality, although colour may be slightly compromised. Immediate sensory manipulation and thus improvement, however, may be achieved to a lesser degree in the short term by STBD. / AFRIKAANSE OPSOMMING: Die toepassing van stoombehandeling van groen Cyclopia maculata en C. longifolia vir die modulasie van hul aroma en geur profiel, sonder gepaardgaande nadelige gevolge op kleur en individuele fenoliese inhoud, is ondersoek. Stoombehandeling (96 °C, atmosferiese druk) van verskeie tydstippe is toegepas op die gekerfde, vars plantmateriaal voor droging (SBVD; 0, 30, 60, 90 en 120 s) of die groen teeproduk ná droging (SBND; 0, 1, 2, 3 en 4 min). Stoombehandeling van 60 s of langer het tot 'n betekenisvolle (p < 0.05) afname in vegetatiewe (‘groen gras’ en ‘hooi/gedroogde gras’) en graan (‘hawer/pap/graan’) aroma en geur intensiteite gelei, met 'n toename in sommige vrugtige (‘tropiese vrugte’ en ‘koejawel’) en ‘vrugtige-soet’ aroma intensiteite. Hierdie veranderinge was meer merkbaar in die SBVD behandeling, terwyl C. longifolia minder vatbaar vir hierdie veranderinge was as C. maculata. Daarbenewens is 19 aromaverbindings in die vlugtige fraksie van C. maculata aftreksel voorberei van SBVD plantmateriaal instrumenteel geïdentifiseer. Sewe hiervan het goed gekorreleer met heuningbos vegetatiewe en graan aromas wat verband hou met ongestoomde monsters. Stoombehandeling het dus gelei tot die vinnige verlies van sommige van hierdie baie vlugtige ‘groen’-geassosieërde aromaverbindings. Stoombehandeling vir langer as 60 s, ongeag van SBVD of SBND, het nie die groen kleur en individuele fenoliese inhoud grootliks nadelig beïnvloed nie. ‘n SBVD behandeling van 30 s het egter tot die vinnige verlies van groen kleur en oksidasie van fenoliese verbindings, sonder aansienlike aroma verbetering van C. maculata gelei. Dít beklemtoon die noodsaaklikheid vir prosesbeheer tydens groen heuningbosproduksie. Stabiliteit van gestoomde (60 s, SBVD) en ongestoomde groen C. maculata tydens opberging onder lae (0 °C; vog-ondeurlaatbare verpakking) en normale (25 °C by 60% relatiewe humiditeit (RH); semi-vogdeurlaatbare sakkies) temperatuur opbergingtoestande (LTO en NTO, onderskeidelik) oor 6 maande en hoë temperatuur opbergingtoestande (40 °C by 75% RH; semivogdeurlaatbare sakkies; HTO) vir 1 maand, is ondersoek. HTO het na 1 maand soortgelyke veranderinge teweeggebring as NTO na 6 maande, terwyl min verandering by LTO bespeur is. Ten spyte van merkbare groen kleur verlies oor die onderskeie tydperke by NTO en HTO is individuele fenoliese verbindings min beinvloed. NTO en HTO het gelei tot die progresiewe ontwikkeling van gesogte sensoriese eienskappe van gefermenteerde heuningbos, veral in ongestoomde monsters. Dit sluit in prominente vrugtige (‘gestoofde vrugte’, ‘appelkooskonfyt’ en ‘marmelade’) aromas en geure , ‘algemene soet’ en ‘vrugtige-soet’ aromas en 'n soeter smaak. Al die beskrywende sensoriese data is saamgestel om 'n voorlopige sensoriese wiel vir groen heuningbos aroma op te stel, asook een vir geur, smaak en mondgevoel. Die sensoriese profiel van groen heuningbos kan beskryf word as 'n oorheersende vegetatiewe aroma en geur, prominente soet-verwante en effens vrugtige aroma, met soet en veral bitter smaak en ‘n vrank mondgevoel. Deur die sensoriese data te kombineer kon duidelik uitgewys word dat die belangrikste gevolge van opberging dié van stoombehandeling oortref. Dit dui daarop dat opberging van 3 tot 6 maande sensoriese kwaliteit kan verbeter, hoewel kleur effens benadeel kan word. In die kort termyn kan onmiddellike sensoriese manipulasie en dus verbetering egter in 'n mindere mate deur SBVD bereik word.

Thesis (MScAgric (Agronomy)--Stellenbosch University, 2008. / Honeybush (Cyclopia spp. Fabaceae) is indigenous to the fynbos botanical biome of the Eastern and Western Cape of South Africa. The increase in the international demand for honeybush tea for health benefits, concern over exploitation of wild populations and the lack of published agronomic information necessitated this study to evaluate different aspects of honeybush propagation. The main objectives of this study were to evaluate the effect of species and cutting position on rooting of cuttings of Cyclopia species using different rooting hormones, to evaluate the effect of an organic plant fertilizer and cutting position on growth and establishment of rooted cuttings and to study the influence of different seed pre-treatments on germination of Cyclopia species. Terminal and sub-terminal cuttings of C. intermedia and C. genistoides treated with different rooting hormones were rooted under day/night temperature controlled glasshouse conditions. Intermittent mist was used as means of moisture supply to the cuttings for 45-60 seconds daily every 30 minutes. C. genistoides rooted significantly better compared to C. intermedia as measured by rooting percentage, number of roots per cutting, length of longest root and mean root length during the summer season. The cutting position had a significant effect on rooting of the cuttings in summer compared to winter and spring season. The interactive effect of species, treatment and cutting position resulted into 86% of rooting in summer from the terminal cuttings of C. genistoides, while only 4% was recorded as the highest rooting percentage in both winter and spring seasons. The highest number of roots and the greatest root length per cutting were obtained with 2 and 4 g L-1 IBA from terminal cuttings of C. genistoides and these hormone concentrations were not significantly different to each other. To evaluate the effect of an organic plant fertilizer and cutting position on plant growth and establishment, rooted cuttings of two Cyclopia species (C. intermedia and C. genistoides) from two cutting positions (terminal and sub-terminal) were transferred to pots (576 cm3) and treated with Nitrosol fertilizer at application rates of 3.33 ml.L-1, 1.67 ml.L-1 and 0 ml.L-1 (control). Cyclopia plantlets were uniformly inoculated once with a symbiotic Rhizobium bacteria to improve the formation of nodules. Nitrosol® at 3.33 ml.L-1 significantly affected fresh and dry plant weight, fresh and dry root weight, number of shoots and nodules per plant compared to either 1.67 ml.L-1 or the control. Relative to species, C. genistoides performed better in terms of fresh and dry plant weight, fresh and dry root weight, and number of shoots and nodules per plant compared to C. intermedia. The origin of the cutting position did not significantly affect the above mentioned parameters. Plant mineral analysis revealed that most of the essential elements increased with increasing Nitrosol® application rates, with C. genistoides having higher levels of mineral elements than C. intermedia. This could be an indication of the differences between the two species in terms of nutrient uptake, utilization and distribution within the plant tissues. In the germination studies, seeds obtained from different seed sources of Cyclopia species were subjected to different pre-sowing treatments. Seed treatments were sulphuric acid (95%), hot water (100°C), water with smoke paper disk, and demineralised water (control). The study revealed that all the treatments had a significant effect on germination with the exception of eight year old seeds obtained from C. subternata (seed source two). Although hot water treatment improved germination compared to smoked paper disk and the control, seeds treated with hot water degenerated rapidly. The highest overall germination (77.33%) was found with one year old seeds compared to other seed sources older than one year. Although smoked paper disks generally did not improve germination compared to the control, in one year old seeds from seed source one, this treatment greatly influenced germination, suggesting that seed age might have influenced germination of these seeds. In terms of germination rate, germination generally started after four days in most treatments.

Thesis (MSc)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Phytoestrogens are perceived as a safer alternative to conventional hormone replacement therapy (HRT) for the alleviation of menopausal symptoms as they present a decreased side-effect profile. The Cyclopia subternata (honeybush) methanol extract, SM6Met, displays estrogenic attributes desirable for the development of an phytoestrogenic nutraceutical, namely, estrogen receptor (ER) α antagonism, ERβ agonism, and antagonism of 17β-estradiol (E2)-induced breast cancer cell proliferation. Activity-guided fractionation was employed in an attempt to isolate and identify the compounds inducing the specific estrogenic profile of SM6Met. Fractions were evaluated for estrogenic attributes and major polyphenols present. Initial liquid-liquid fractionation of SM6Met yielded a polar fraction (PF) and a non-polar fraction (NPF), with the estrogenic attributes of interest retained and concentrated in NPF. Subsequent high performance counter-current chromatography (HPCCC) fractionation of NPF yielded three fractions (F1-F3). Interestingly, the fractions revealed separation of the previously demonstrated positive estrogenic attributes of NPF into separate fractions, with F1 and F2 acting as ERα antagonists, only F2 inducing antagonism of E2-induced breast cancer cell proliferation and only F3 retaining ERβ agonist activity. Although ERβ agonism displayed by F3 was robust and significantly higher than that of 10-11 M E2, it also displayed weak ERα agonism. Fractionation also for the first time in the study revealed ERβ antagonism, as induced by F1. In terms of major polyphenols HPCCC fractionation resulted in a divergence with F1 emerging as the dihydrochalcone-rich fraction and F2 as the flavanone and benzophenone-rich fraction, while the xanthones, flavones and phenolic acids were retained in F3. In addition, a preliminary absorption study was conducted using the ex vivo flow-through diffusion assay whereby the permeability of porcine small and large intestine for polyphenols in SM6Met was evaluated. The major compounds present in SM6Met were not able to penetrate the large intestinal mucosa, but small intestinal permeation of all major compounds in SM6Met ensued, with apparent permeability coefficient (Papp) values ranging from 1.91-3.74 x 10-6 cm.s-1, indicative of good intestinal absorption. Open source programs used for theoretical prediction of absorption gave conflicting results, emphasising the need to confirm predictions experimentally. ACD/Labs predicted poor intestinal absorption of SM6Met compounds based on physicochemical profiling, while OSIRIS and ChemAxon anticipated good absorption. In conclusion, activity-guided fractionation results suggest that retention of all the positive estrogenic attributes of the original SM6Met in one fraction is not an attainable goal. This suggests that several of the polyphenols present in SM6Met or NPF, through antagonistic, synergistic, or additive effects, may together be conferring these desired estrogenic traits. Thus production or isolation of a mixture of compounds, i.e. an “intelligent” mixture, should serve as a superior strategy in designing a nutraceutical product tailored to user demand of estrogenic activity. / AFRIKAANSE OPSOMMING: Fitoestrogene word beskou as ‘n veiliger alternatief vir konvensionele hormoon-vervangingsterapie (HVT) vir die verligting van simptome geassosiaeer met menopause aangesien dit ‘n verminderde newe-effek profiel vertoon. Die metanol ekstrak van Cyclopia subternata (heuningbos), SM6Met, vertoon estrogeniese eienskappe wat wenslik is vir die ontwikkeling van ‘n fitoestrogeen nutraseutiese middel, naamlik, estrogeen reseptor (ER) α antagonisme en ERβ agonisme, asook antagonisme van 17β-estradiol (E2) geïnduseerde proliferasie van borskankerselle. Aktiwiteit-begeleide fraksionering (ABF) is gebruik om die verbindings wat die spesifieke estrogeniese profiel aan SM6Met verleen te probeer isoleer en identifiseer. Fraksies is ge-evalueer vir estrogeniese eienskappe, asook vir die hoof polifenole teenwoordig. Aanvankilike vloeistof-vloeistof fraksionering van SM6Met het ‘n polêre fraksie (PF) en ‘n nie-polêre fraksie (NPF) opgelewer met behoud en konsentrering van die wenslike estrogeniese eienskappe in NPF. Daaropeenvolgende hoë werkverrigting teen-vloei chromatografie (HPCCC) van NPF het drie fraksies (F1-F3) opgelewer. Interessant genoeg het hierdie fraksies ‘n verdeling van die wenslike estrogeniese eienskappe van NPF in die individuele fraksies teweeggebring, deurdat F1 en F2 ERα antagonisme getoon het, F2 E2-geïnduseerde proliferasie van borskankerselle antagoniseer het, en net F3 ERβ agonis-aktiwiteit behou het. Alhoewel die ERβ agonis-aktiwiteit van F3 betekenisvol hoër was in vergelyking met die aktiwiteit van 10-11 M E2, het dit ook swak ERα agonisme getoon. Verder het fraksionering, vir die eerste keer in hierdie studie, ERβ antagonisme meegebring soos getoon deur F1. HPCCC het ook ‘n skeiding van die hoof polifenole veroorsaak, waarvolgens F1 as die dihidrogalkoon-ryke fraksie, F2 as die bensofenoon-ryke en flavanoon-ryke fraksie, en F3 as die xantoon-, flavoon- en fenoliese suur-ryke fraksie tevoorskyn gekom het. ‘n Voorlopige absorpsie studie, wat gebruik gemaak het van die ex vivo deurvloei diffusie toetssisteem, is uitgevoer om die deurlaatbaarheid van vark dik- en dunderm vir SM6Met polifenole te evalueer. Die hoof verbindings van SM6Met kon nie die dikderm mukosa penetreer nie, maar die deurlaatbaarheid van die dunderm vir alle SM6Met hoof polifenole is aangetoon, met skynbare deurlaatbaarheidskoëffisiënt (Papp) waardes wat strek vanaf 1.91 tot 3.74 x 10-6 cm.s-1, ooreenstemmend met goeie intestinale absorpsie. Oopbron programme, wat gebruik is vir die teoretiese voorspelling van absorpsie deur gebruik te maak van fisiese-chemiese profilering van verbindings, het teenstellende resultate opgelewer wat daarop dui dat hierdie voorspellings eksperimenteel bevestig moet word. ACD/Labs, wat van die fisiese-chemiese eienskappe van die verbinding gebruik maak, het swak intestinale absorpsie van die polifenole voorspel, terwyl OSIRIS en ChemAxon goeie absorpsie voorspel het. Ten slotte, resultate van aktiwiteit-begeleide fraksionering het getoon dat behoud van al die gewensde estrogeniese eienskappe van SM6Met in een fraksie nie ‘n haalbare doelwit is nie. Hierdie bevinding dui daarop dat verskeie van die polifenole teenwoordig in SM6Met of NPF saam, deur middel van antagonistiese, sinergistiese of additiewe effekte, die wenslike estrogeniese eieskappe verleen. Die ontwikkeling of isolering van ‘n mengsel van verbindings, met ander woorde ‘n ―intelligente‖ mengsel is dus ‘n beter strategie vir die ontwerp van ‘n estrogeniese nutraseutiese produk om die verbruikers-aanvraag van estrogeniese aktiwiteit te voorsien.

Thesis (MTech (Consumer Science: Food and Nutrition))--Cape Peninsula University of Technology, 2011.Includes bibliographical references (leaves 139-156). / Oxidative stress has been implicated in the development of a number of chronic degenerative diseases. These diseases can often be prevented by dietary and lifestyle changes. One major dietary aspect is increasing the intake of plant foods, such as fruit and vegetables as they are rich in the major nutrient and nonnutrient antioxidants. The South African population does not have an adequate intake of fruit and vegetables and many barriers have been noted against increasing the intake of these foods. Other avenues should therefore be sought to increase the antioxidant intake. One such avenue is to use a basic food item consumed often as an antioxidant carrier to increase the antioxidant intake. Much interest has developed in functional foods and their therapeutic effects as well as the use of nutraceuticals in food product development and food fortification. Tea and its polyphenols, due to the antioxidant properties, is a potentially viable option for nutraceutical and functional food use. Rooibos and honeybush herbal teas, native to South Africa, can be potential nutraceutical sources to increase antioxidant levels of food due to their unique polyphenol content. The objective of the study was to increase the total antioxidant capacity (TAC) of a basic food item consumed regularly by most South Africans, such as brown bread through fluid manipulation using rooibos, honeybush and black teas respectively, as the replacement fluid for water in a standardised bread recipe formulation. The TAC of these experimental breads were compared to the TAC of the bread produced with no fluid manipulation (control bread) and a commercially processed bread (reference bread). The experimental breads were prepared using 2% weight per volume (w/v) and 5% (w/v) concentration levels for rooibos, honeybush and black tea respectively. To ascertain the TAC of the experimental breads, the fluorescein(FL)-based oxygen radical absorbance capacity (ORACFL) method was used. The hydrophilic (H)-ORACFL assay was used as the teas contain polyphenols which favour a water environment. The results indicated that the H-ORACFL of the control bread using water as the fluid and the commercial reference bread was significantly different (p < 0.05 for each) to all the experimental bread formulations besides for the 2% (w/v) honeybush experimental bread formulation. The 2% (w/v) honeybush experimental bread formulation was therefore not considered for the consumer acceptance testing. There was no significant difference (p > 0.05) in the H-ORACFL of the 2% (w/v) rooibos and black tea experimental bread formulations, but the H-ORACFL of the 5% (w/v) black tea experimental bread formulation was significantly (p < 0.05) higher than that of the 5% (w/v) rooibos experimental bread formulation. This particular experimental bread formulation had a “golden-orange” colour with a pungent smell due to the increase in rooibos concentration. The 5% (w/v) black tea experimental bread formulation, however, had noted negative sensory attributes of being densely textured with a very dark colour and a pungent and overpowering taste of black tea. These sensory attributes and TAC results lead to the rooibos experimental breads containing 5% (w/v) (as bread A) and 2% (w/v) (as bread B) chosen for the consumer acceptance testing and rooibos being a proudly South African product. The participants (n = 85) of the consumer acceptance testing participated voluntarily and were regular bread consumers. The sample was mostly female (57.6%), in the age group of 18 to 24 years (54.1%), with an education level of between three to four years post-matric/-grade 12 (71.7%) and single, living on their own without children (76.4%). They also represented a sample which had characteristics of a healthy lifestyle. Most participants perceived themselves as being of an optimal body weight (55.2%) and the majority was non-smokers (74.1%), physically active (63.5%), having no family history of chronic disease (69.4%) and not consuming dietary supplements (70.5%). Three sensory attributes of the experimental breads were rated for the acceptance testing and included appearance/colour, flavour/taste and texture/mouthfeel. Both the experimental bread formulations was accepted favourably regarding the above-mentioned sensory attributes. The three liking acceptance categories, ‘neither like nor dislike’ (neutral response), ‘like’ and ‘like very much’ were the most commonly indicated by the participants which also resulted in the experimental breads being rated as fair, good and very good according to the development scores. Although both breads were very similarly scored, there was a trend for bread A to be rated slightly higher regarding the above-mentioned sensory attributes. The acceptance category, ‘like’/good was scored the highest for bread A according to the sensory attributes of appearance/colour (56.5%), flavour/taste (55.3%) and texture/mouthfeel (50.6%). There was no significant association/difference (p > 0.05 for each) between the participants’ demographic characteristics and their overall acceptance of the experimental breads. Most of the participant biographic characteristics also had no significant (p > 0.05 for each) impact on the sensory attribute acceptance of the experimental breads. Although the majority (75%) of the participants consumed rooibos, there was no significant (p > 0.05) association/difference between the participants’ tea consumption and their acceptance of the experimental breads. Although most participants (64%) had no awareness of functional foods, most indicated willingness to purchase foods with added health benefits (93%) and to pay more for foods with added health benefits (73%). These results indicated a general acceptance of bread with functional food characteristics and the possibility of purchasing such products with added health benefits and even at a higher price premium.

Thesis (Msc Food Sc (Food Science))--Stellenbosch University, 2008. / Extraction efficiency of soluble solids (SS), total polyphenols (TP) and xanthones (AlCl3 assay) from dried, green Cyclopia subternata, as affected by mass-solvent ratio, extraction time and solvents, was investigated. In addition the effect of solvent composition on extraction of mangiferin and hesperidin was determined. Extraction of 5 g plant material as opposed to 0.5 and 1 g resulted in lower recoveries of SS, TP and xanthones (P<0.05). Extraction of SS and TP increased during the initial 20 min of contact time, where after it remained constant (P>0.05). Water, 33% acetonitrile, ethanol (50, 80 and 100%), methanol (50 and 100%) and 70% acetone were investigated as extraction solvents. Extraction for 30 min with 33% acetonitrile on a steam bath or 50% ethanol at 64°C on a water bath proved to be the most effective for extraction of SS, TP and xanthones, while 33% acetonitrile was most effective in extracting hesperidin from C. subternata. However, 70% acetone was most effective in extracting mangiferin. A poor correlation (r = 0.54) was observed for the total antioxidant activity (TAA) of C. subternata, as determined for water extracts and with the mangiferin content determined by HPLC. A moderate correlation (r = 0.85) was, however, obtained for TAA and TP content. The mangiferin content of green C. subternata can be determined using the aluminium chloride (AlCl3) colorimetric method. A moderate correlation (r = 0.87) was found for the xanthone content of the plant material determined using the AlCl3 colorimetric method and mangiferin content quantified by HPLC (y = 1.2x + 0.54) following extraction with hot water. For extraction using 33% acetonitrile a weaker correlation (r = 0.74; y = 1.3x + 0.87) was found between the xanthone and mangiferin contents. The xanthone content (determined by AlCl3) of the plant material as extracted by the two solvents, correlated well (r = 0.91). Good correlations were also obtained, when comparing extractions with water and 33% acetonitrile, for determination of the SS (0.94) and mangiferin contents (r = 0.97) of the plant material. Near infrared (NIR) spectroscopy was investigated as a rapid and more economical method for prediction of mangiferin and xanthone contents of dried, green C. subternata plant material. NIR spectroscopy calibration models can be used for screening purposes for the mangiferin and (SEP = 0.21 g.100 g-1; r = 0.82) and xanthone (SEP = 0.27 g.100 g-1; r = 0.81) contents. The effect of various pre-drying treatments and storage temperatures on the colour, soluble SS, TP, mangiferin and hesperidin contents of green C. subternata was investigated. By steaming green C. subternata directly after maceration, its colour retention can be improved. Good stability was shown for mangiferin and hesperidin during manufacture and storage of C. subternata.

Thesis (MSc (Chemistry and Polymer Science))--University of Stellenbosch, 2006. / Honeybush (Cyclopia spp.) is indigenous to South Africa and consists of more than 20 species of which only a few are used to make a herbal tea with a unique aroma and sweet taste. The aroma of unfermented and fermented honeybush tea differs considerably, indicating that changes take place in the chemical composition of the aroma during the fermentation process. Using a sample enrichment probe (SEP) and gas chromatography-mass spectrometry (GC-MS), the chemical composition of the aroma of unfermented and fermented C. genistoides, was analyzed and the resulting aroma profiles were compared. A total of 74 compounds were identified in the unfermented honeybush aroma, comprising, inter alia, a large number of saturated and unsaturated alcohols, aldehydes and methyl ketones that were found to be either absent, or present in lower relative concentrations, in the aroma of fermented honeybush. Most of these compounds, including 6-methyl-5-hepten-2-one, identified as one of the major constituents of unfermented honeybush, are probably responsible for its grassy, hay-like aroma. In the aroma of fermented honeybush 70 compounds were identified, of which 36 were found to be terpenoids. These compounds are probably responsible for the pleasant sweet aroma of the fermented honeybush. For instance, the major aroma constituent in the fermented honeybush, α-terpineol, is known to have a delicately floral and sweet odour. Other terpenoids occuring in significant quantities in the fermented honeybush, are linalool, cis and trans linalool oxide, nerol, geraniol, 2,6-dimethyl-1,7-octadien-3,6-diol, hexahydrofarnecyl acetone and phytol. These terpenoids are known to have sweet, sweet-woody en floral odours that probably contribute to the overall sweet aroma of the fermented honeybush. In the aroma of the unfermented honeybush a total of 25 terpenoids was identified, of which geranyl acetone, β-ionone, and dihydroactinidiolide are the most significant, since they are present in higher relative concentrations compared to the same terpenoids in the fermented honeybush. These three terpenoids have overall woody and even slightly green odours which contribute to the typical aroma of the unfermented honeybush. The terpenoids present in the aroma of unfermented and fermented honeybush belong to the following compound classes: • Terpenes • Terpene alcohols • Terpene aldehydes • Ketoterpenes • Terpene ethers • Terpene lactones Apart from the terpenoids present in the aroma of unfermented and fermented honeybush, the following classes of compounds are also present: • Aliphatic hydrocarbons (saturated) • Aliphatic alcohols (saturated and unsaturated) • Phenols • Aliphatic aldehydes (saturated and unsaturated) • Aliphatic ketones (saturated and unsaturated) • Aliphatic carboxylic acids (saturated) • Esters (methyl esters, ethyl and higher esters and aromatic esters) • Furane compounds • Lactones The present research was done to compare the chemical composition of the aroma of unfermented and fermented honeybush of one particular species, namely C. genistoides, to obtain an understanding of the evolution and/or disappearance of volatile compounds during fermentation that ultimately could help to identify compounds or compound types and their precursors responsible for its unique sweet aroma.

Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Wine knowledge has increased drastically in recent years with the aid of scientific experimentation. The producers of wine have increasingly incorporated new wine knowledge into the marketing and innovation of wine products. In parallel with the above, the consumer market has become more aware of winemaking practices and their influence on consumer health. The use of alternative methods and additives in wine are means by which producers have been able to create innovative wine products. This study focused on the addition of rooibos (Aspalathus linearis) and honeybush (Cyclopia spp.) plant material to red wine. Consumer liking, as well as descriptive sensory analysis (DSA) of different red wines made from a range of cultivars with the addition of rooibos and honeybush plant material, was investigated. Another aim of the study was to observe the effect of oxygen addition to red wines treated with rooibos and honeybush plant material. The consumer studies included a number of events where wines treated with rooibos and honeybush material were compared with other commercial wines. The results could not clearly indicate whether consumers liked or disliked the wine products with added rooibos and honeybush plant material. Further refinement of wines with the addition of rooibos and honeybush can serve as topics for future research. Focus on the potential health benefits which can be added to wine from rooibos and honeybush plant material, as a result of inherent antioxidants within both these species, may serve as a novel research topic. The DSA panels identified aroma descriptors that could be related to the addition of wood and leaf or only rooibos and honeybush wood to wine. The addition of oxygen to wines treated with rooibos and honeybush plant material indicated that aromas associated with these treatments are relatively stable in wine undergoing oxidation. This could possibly pave the way for future research on the topic of low sulphur dioxide containing wines. As a result of this study, a commercial wine product called “Rooibos wine” was developed and became available in retail. Further research on this topic and the impact of the addition of rooibos and honeybush material to wine may be of great benefit for the production of innovative wines with unique aroma profiles. / AFRIKAANSE OPSOMMING: Kennis oor wyn het oor die afgelope paar jaar vooruitgegaan met behulp van wetenskaplike eksperimentering. Wynprodusente sluit toenemend nuwe wynkennis in die bemarking en innovering van wynprodukte in. Parallel hiermee het die verbruikersmark baie meer bewus geword van wynbereidingspraktyke en die invloed daarvan op verbruikersgesondheid. Die gebruik van alternatiewe metodes en byvoegings in wyn is maniere waarop produsente innoverende wynprodukte kan skep. Hierdie studie het gefokus op die byvoeging van rooibos (Aspalathus linearis) en heuningbos (Cyclopia spp.) plantmateriaal by rooiwyn. Verbruikers se voorkeur, sowel as die beskrywende sensoriese analise (descriptive sensory analysis [DSA]) van diverse rooi wyn kultivars waarby rooibos en heuningbos plantmateriaal gevoeg is, is geanaliseer. Nóg ’n doelwit van die studie was om die effek van suurstofbyvoeging by rooiwyne wat met rooibos en heuningbos plantmateriaal behandel is waar te neem. Vir die verbruikerstudies is geleenthede aangebied waar wyne wat met rooibos en heuningbos materiaal behandel is met kommersiële wyne vergelyk is. Die resultate kon nie duidelik aandui of die verbruikers gehou het of nie van wynprodukte wat met rooibos en heuningbos behandel is. Verdere verfyning van wyne met bygevoegde rooibos en heuningbos kan as onderwerpe vir toekomstige navorsing dien. Die verhoogde gesondheidvoordele van wyn wat met rooibos en heuningbos plantmateriaal behandel is as gevolg van die inherente anti-oksidante eie aan die spesies, kan dien as fokus vir ʼn nuwe navorsingstema. Die DSA-panele het beskrywende terme vir aromas geïdentifiseer wat verband kan hou met die byvoeging van die hout/blaar of slegs die rooibos- en heuningboshout by die wyn. Die aromas in wyn wat met rooibos en heuningbos behandel is, is gevind om relatief stabiel te bly tydens oksidasie. Dit kan moontlik die weg baan vir toekomstige navorsing oor lae swaweldioksied-bevattende wyne. As gevolg van hierdie studie is ‘n kommersiële wynproduk, genaamd “Rooiboswyn”, ontwikkel en is dit in die handel beskikbaar. Verdere navorsing oor hierdie onderwerp en die impak van die byvoeging van rooibos en heuningbos materiaal by wyn sal moontlik van groot voordeel wees vir die produksie van innoverende wyne met unieke aroma profiele.

Thesis (MSc Food Sc)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: The sensory profiles, phenolic composition and colour of honeybush infusions, prepared from six Cyclopia species (C. sessiliflora, C. longifolia, C. genistoides, C. intermedia, C. subternata and C. maculata), were determined to establish the variation between species. The results of the sensory study were used to create a honeybush sensory wheel and lexicon. The “characteristic” sensory profile of honeybush tea can be described as a combination of floral, sweet, fruity and plantlike flavours with a sweet taste and a slightly astringent mouthfeel. Sensory results indicated that the species could be divided into three distinct groups; group A (C. sessiliflora, C. intermedia and C. genistoides), group B (C. longifolia and C. subternata) and group C (C. maculata). Group A was associated with fynbos floral, fynbos sweet and plantlike attributes, group B with rose geranium and fruity sweet attributes and group C with woody, boiled syrup and cassia/cinnamon attributes. Gas chromatography-olfactometry analysis of the C. maculata aroma fraction indicated that the spicy note of its aroma could possibly be explained by the high concentration of the volatile component eugenol. However, none of the aroma impact volatiles had a specific cassia/cinnamon note. Large variation in the composition of the honeybush infusions was revealed through the quantification of the soluble solids, total polyphenol and individual monomeric polyphenolic compounds, as well as the absorbance (“colour”). Infusions of C. genistoides, C. longifolia and C. sessiliflora had the highest soluble solids and total polyphenol content, as well as the highest absorbance values. Only mangiferin, isomangiferin, hesperidin and compound C (unidentified compound) were detected in all six Cyclopia species. Cyclopia genistoides, C. longifolia and C. sessiliflora, in order of prominence, contained the highest concentration of both mangiferin and isomangiferin whereas C. genistoides and C. maculata contained the highest hesperidin content. The bitter taste present in certain Cyclopia species appeared to be due to a high mangiferin content, however, compounds such as isomangiferin and compound C might also have played a role. The effect of fermentation (oxidation) temperature (80°C and 90°C) and time (8 h, 16 h, 24 h and 32 h) of C. genistoides, C. subternata and C. maculata on the sensory characteristics of their infusions was also investigated. Fermentation for longer than 8 h resulted in an increase in positive sensory attributes and a decrease in negative sensory attributes rather than the formation of new sensory attributes. A fermentation temperature/time combination of 80°C/24 hours or 90°C/16 h was required for C. genistoides, C. subternata and C. maculata. Fermenting C. genistoides at 90°C would result in a honeybush infusion with slightly less rose geranium notes whereas C. subternata can be fermented at either 80°C or 90°C, depending on whether floral or apricot jam notes are desired. Cyclopia maculata should preferably not be fermented at 90°C due to an increase in negative sensory attributes (hay/dried grass and green grass). Fermentation reduced the soluble solids content, total polyphenol content, colour and concentration of individual polyphenolic compounds. Changes in the taste and mouthfeel of honeybush tea could be attributed to changes in the polyphenolic composition caused by the high temperature oxidation. Mangiferin associated with the bitter taste of C. genistoides, while in C. subternata astringency may be partly attributed to the mangiferin and isomangiferin content. The study substantiated the need for further research on the contribution of the major phenolic compounds towards the taste and mouthfeel of Cyclopia species. / AFRIKAANSE OPSOMMING: Die sensoriese profiel, fenoliese samestelling en kleur van heuningbostee, berei van ses Cyclopia spesies (C. sessiliflora, C. longifolia, C. genistoides, C. intermedia, C. subternata en C. maculata), is bepaal ten einde die mate van variasie vas te stel. Die resultate van die sensoriese studie is gebruik om 'n sensoriese wiel en leksikon vir heuningbostee te ontwikkel. Die “karakteristieke” sensoriese profiel van heuningbostee kan beskryf word as 'n kombinasie van blomagtig, soet, vrugtig en plantagtige geure met 'n soet smaak en 'n effense frankheid. Sensoriese resultate het aangedui dat die spesies in drie groepe verdeel kon word; groep A (C. sessiliflora, C. intermedia and C. genistoides), groep B (C. longifolia and C. subternata) en groep C (C. maculata). Groep A is met fynbos blom, fynbos-soet en plantagtige geure geassosieer, groep B met roos geranium en vrugtige-soet geure en group C met houtagtige, gekookte stroop en kassia/kaneel geure. Gaschromatografie-olfaktometrie analises van C. maculata se aroma fraksie het getoon dat die speseryagtige aroma moontlik as gevolg van die hoë konsentrasie van die vlugtige komponent, eugenol, kon wees. Geen van die aroma-impak vlugtige verbindings het egter 'n spesifieke kassia/kaneelagtige noot gehad nie. Groot variasie in die samestelling van heuningbostee ten opsigte van die inhoud van oplosbare vastestowwe, totale polifenole en monomeriese fenoliese verbindings, asook die absorbansie (“kleur”) is aangetoon. Heuningbostee berei van C. genistoides, C. longifolia en C. sessiliflora het die hoogste oplosbare vastestowwe en totale polifenol inhoud, asook die hoogste absorbansie waardes gehad. Slegs mangiferien, isomangiferien, hesperidien en verbinding C (ongeïdentifiseerde verbinding) is in al ses Cyclopia spesies geïdentifiseer. Cyclopia genistoides, C. longifolia en C. sessiliflora, in volgorde van belangrikheid, het die hoogste konsentrasie van beide mangiferien en isomangiferin gehad teenoor C. genistoides en C. maculata wat die hoogste hesperidien konsentrasie gehad het. Die bitter smaak teenwoordig in sekere Cyclopia spesies blyk moontlik as gevolg van die hoë mangiferien inhoud te wees, hoewel komponente soos isomangiferien en komponent C dalk ook 'n rol mag speel. Die effek van die fermentasie temperatuur (80°C en 90°C) en tyd (8 h, 16 h, 24 h en 32 h) van C. genistoides, C. subternata en C. maculata op die sensoriese eienskappe van heuningbostee is ondersoek. Fermentasie vir langer as 8 h het tot 'n toename in positiewe sensoriese eienskappe en afname in negatiewe sensoriese eienskappe gelei eerder as die ontstaan van nuwe sensoriese eienskappe. Om heuningbostee met 'n optimum sensoriese profiel te verkry is 'n fermentasie temperatuur/tyd kombinaise van 80°C/24 h of 90°C/16 h nodig vir C. genistoides, C. subternata en C. maculata. Cyclopia genistoides wat by 90°C gefermenteer word sal minder van die roos geranium note bevat, terwyl C. subternata by 80°C of 90°C gefermenteer kan word, afhangende of 'n blomagtige of 'n appelkooskonfyt noot verlang word. Fermentasie by 90°C word nie aanbeveel C. maculata nie as gevolg van die toename van sekere negatiewe sensoriese eienskappe (hooi/droe gras aroma en -geur en groen gras aroma). Fermentasie het die inhoud van oplosbare vastestowwe, totale polifenole, individuele polifenoliese verbindings, asook kleur verminder. Veranderinge in die smaak en mondgevoel van heuningbostee kon toegeskryf word aan die veranderinge in die polifenoliese inhoud as gevolg van die hoë temperatuur oksidasie. Mangiferien is met die bitter smaak van C. genistoides geassosieer, terwyl mangiferien and isomangiferien moontlik deels frankheid in C. subternata veroorsaak. Die studie het die noodsaaklikheid vir verdere navorsing op die bydrae van die hoof fenoliese verbindings tot die smaak en mondgevoel van Cyclopia spesies gestaaf.

Thesis (MSc Food Sc )--Stellenbosch University, 2003. / ENGLISH ABSTRACT: Please refer to fulltext for abstract / AFRIKAANSE OPSOMMING: Sien volteks vir opsomming