Thesis (MScFoodSc)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: The flavanone aglycones, hesperetin and eriodictyol, have been identified as potential taste modulators with reported sweetness-enhancing and bitterness-masking properties, respectively. Reduction of the sugar content of food products has become important in view of the global obesity epidemic. Taste modulators have shown potential to enhance the sweet taste of reduced-sugar foods without unfavourably affecting their flavour profile. On the other hand, bitterness-masking taste modulators are useful to mask the bitter taste of functional phytochemical ingredients. In the current study, Cyclopia maculata (honeybush) was investigated as potential source of hesperetin- and eriodictyol-enriched extracts. Hesperetin and eriodictyol were present mainly below the quantification limit in C. maculata plant material, including unfermented leaf and stem material, unfermented and fermented tea, as well as the fermented by-product (< 40 mesh and > 12 mesh). Conversely, their rutinoside and modulatinginactive derivatives, hesperidin and eriocitrin were present at substantially higher concentrations in the plant material. The stems and by-product were shown to be good sources of hesperidin, but not eriocitrin. The qualitative and quantitative phenolic profile of the by-product was similar to that of the stems. The tea processing by-product was therefore selected to optimise extraction of flavanone glycosides for subsequent de-glycosylation of the flavanone glycosides to aglycones. The by-product was subjected to ultrasound-assisted extraction to investigate its potential as renewable source of the flavanone glycosides. Response surface methodology (RSM) was employed to optimise and study the individual and interactive effects of the process variables, namely ethanol concentration (% v/v), time (min), temperature (°C), and solvent:solid ratio (mL/g), on flavanone glycoside extraction. The hesperidin yield and content (of extract), as well as extract yield, increased with an increase in extraction time, temperature and solvent:solid ratio. Practical process restrictions limited global optimisation and only an optimum of 52.8% (v/v) ethanol for extract and hesperidin yield could be reached. Temperature was the parameter with the most significant effect (p < 0.05) on extraction efficiency among those studied. Practical process parameter values that were feasible for industrial application (52.8% (v/v) ethanol, 20 mL/g solvent:solid ratio, 60°C and 30 min) were selected for the preparation of a flavanone glycoside-enriched extract from the tea processing by-product. The flavanone glycoside-enriched extract was subjected to acid-catalysed hydrolysis to deglycosylate hesperidin and eriocitrin to hesperetin and eriodictyol, respectively. RSM was employed to optimise the acid hydrolysis process and to study the effect of the hydrolysis parameters (temperature (°C) and time (min)) on hydrolysis efficiency. At the maximum temperature (92.1°C) and corresponding optimum time (98.4 min) ca 80% conversion of hesperidin to hesperetin was achieved. Substantially more eriodictyol formed during acid hydrolysis than eriocitrin present in the initial extract owing to the deglycosylation of unidentified glycosides with the same aglycone. Unidentified breakdown products imparting a red colour to the acid-hydrolysed extract were also observed. The total phenolic content of the acid-hydrolysed extract was significantly higher (p < 0.05) than that of the unhydrolysed extract, indicating the formation of unidentified compounds with the ability to reduce the Folin-Ciocalteau reagent, although no significant difference (p ≥ 0.05) between the antioxidant activities of these extracts, as assessed with the DPPH radical scavenging and ORAC assays, was observed. The potential of enzymatic bioconversion as an alternative to acid-catalysed hydrolysis was investigated using commercial hesperidinase. Bioconversion resulted only in de-rhamnosylation with ca 100% conversion of hesperidin to hesperetin-7-O-glucoside in an aqueous C. maculata extract at pH 4.0 and 40°C. / AFRIKAANSE OPSOMMING: Die flavanoon aglikone, hesperetien and eriodiktiol, is geïdentifiseer as potensiële smaakmoduleerders met berigte soetheid-versterkende en bitter-maskerende eienskappe, onderskeidelik. Vermindering van die suikerinhoud van voedselprodukte het belangrik geword in die lig van die wêreldwye vetsugepidemie. Smaakmoduleerders het die potensiaal getoon om die soet smaak van voedsel met verlaagde suikerinhoud te versterk sonder om hul geurprofiel ongunstig te beïnvloed. Andersyds is bittermaskerende smaakmoduleerders nuttig om die bitter smaak van funksionele fitochemiese bestanddele te maskeer. In die huidige studie is Cyclopia maculata (heuningbos) ondersoek as ‘n potensiële bron van hesperetien- and eriodiktiol-verrykte ekstrakte. Hesperetien and eriodiktiol was hoofsaaklik teenwoordig onder die kwantifiseringsperk in C. maculata plantmateriaal, insluitend ongefermenteerde blaar- en stokmateriaal, ongefermenteerde en gefermenteerde tee, asook die gefermenteerde byproduk (< 40 maas en > 12 maas). Hierteenoor was hul rutinosiedes en modulerend-onaktiewe derivate, hesperidien and eriositrien, teenwoordig in aansienlik hoër konsentrasies in die plantmateriaal. Die stokmateriaal en byproduk is getoon om goeie bronne van hesperidien, maar nie eriositrien nie, te wees. Die kwalitatiewe en kwantitatiewe fenoliese profiel van die byproduk was soortgelyk aan dié van die stokke. Die teeprosesseringsbyproduk is dus geselekteer om die ekstraksie van flavanoonglikosiede, voorafgaande hul de-glikosilering na aglikone, te optimeer. Die byproduk is aan ekstraksie met behulp van ultrasoniese klank onderwerp om die potensiaal daarvan as hernubare bron van flavanoonglikosiede te ondersoek. Respons-oppervlak Metodologie (ROM) is gebruik om die individuele en wisselwerking effekte van die proses veranderlikes, naamlik etanolkonsentrasie (% v/v), tyd (min), temperatuur (°C), en oplosmiddel:vastestof verhouding (mL/g), op flavanoonglikosied ekstraksie te optimiseer en te bestudeer. Die hesperidienopbrengs en -inhoud (van ekstrak), sowel as die ekstrakopbrengs, het toegeneem met ‘n toename in die ekstraksietyd, - temperatuur en oplosmiddel:vastestof verhouding. Praktiese prosesbeperkings het die globale optimisering beperk en slegs ‘n optimum van 52.8% (v/v) etanol vir ekstrak- en hesperidienopbrengs kon bereik word. Temperatuur was die parameter met die mees beduidende effek (p < 0.05) op ekstraksie doeltreffendheid van dié wat bestudeer is. Praktiese prosesparameterwaardes wat haalbaar is vir industriële toepassing (52.8% (v/v) etanol, 20 mL/g oplosmiddel:vastestof verhouding, 60°C en 30 min) is geselekteer vir die voorbereiding van 'n flavanoonglikosied-verrykte ekstrak uit die teeprosesseringsbyproduk. Die flavanoonglikosied-verrykte ekstrak is aan suur-gekataliseerde hidrolise onderwerp om hesperidien en eriositrien na hesperetien en eriodiktiol, onderskeidelik, te de-glikosileer. ROM is gebruik om die suurhidrolise proses te optimeer en die effek van die hidrolise parameters (temperatuur (°C) en tyd (min)) op hidrolise doeltreffendheid te bestudeer. Ongeveer 80% omskakeling van hesperidien na hesperetien is behaal teen die maksimum temperatuur (92.1 °C) en ooreenstemmende optimum tyd (98.4 min). Aansienlik meer eriodiktiol is tydens suurhidrolise gevorm as eriositrien wat in die oorspronklike ekstrak teenwoordig was, as gevolg van de-glikosilering van ongeïdentifiseerde glikosiede met dieselfde aglikoon. Ongeïdentifiseerde afbreekprodukte, wat 'n rooi kleur aan die suurgehidroliseerde ekstrak gegee het, is ook waargeneem. Die totale fenoliese inhoud van die suurgehidroliseerde ekstrak was beduidend hoër (p < 0.05) as dié van die ongehidroliseerde ekstrak, wat die vorming van onbekende verbindings met die vermoeë om die Folin-Ciocalteau reagens te reduseer aandui, hoewel daar geen beduidende verskil (p ≥ 0.05) tussen die antioksidant-aktiwiteite van hierdie ekstrakte, soos bepaal met die DPPH radikaal blussings- en ORAC toetse, waargeneem is nie. Die potensiaal van ensiematiese bio-omskakeling as 'n alternatief vir suur-gekataliseerde hidrolise is ondersoek met behulp van kommersiële hesperidinase. Bio-omskakeling het slegs tot de-ramnosilering gelei met ca 100% omskakeling van hesperidien na hesperetien-7-O-glukosied in 'n C. maculata waterekstrak by pH 4.0 en 40°C.

Les agrumes très consommés au niveau mondial, représente des sources de caroténoïdes tels que la β-cryptoxanthine, qui contribuent avec les flavonoïdes (hespéridine) aux effets santé de ces fruits et en particulier dans la prévention du diabète de type 2 (DT2). Le DT2 est une pathologie mondiale en croissance exponentielle. L’objectif général de ce travail est par conséquent d’étudier les propriétés nutrition-santé de concentrés de jus de clémentine, obtenus par une technologie membranaire innovante et spécifiquement enrichis en β-cryptoxanthine (βCX) et hespéridine (HES) mais également en pectines. Le devenir digestif de ces phytomicronutriments, garants de la qualité nutritionnelle des concentrés a évalué dans un premier temps en couplant le modèle de digestion in vitro aux cellules intestinales de type Caco-2. Pour une meilleure vision physiologique des mécanismes associés aux premières étapes de la digestion, des modèles de digestion, statique et dynamique, sont comparés. La bioaccessibilité et l’absorption intestinale des caroténoïdes sont fortement influencées par la teneur et structure des pectines. Dans un second temps, les activités biologiques (disglycémie, dislipidémie, inflammation) impliquées dans la prévention du DT2 sont évaluées in vivo chez le rat après administration du concentré d’agrume. L’étude a démontré qu'un aliment à base d'agrume spécifiquement enrichi en βCX et HES est efficace dans la prévention du syndrome métabolique/DT2, soulignant le rôle possible de la βCX et de sa bioconversion en rétinoïdes. Une étude in vitro complémentaire sur macrophages murins révèle des effets anti-inflammatoires synergiques des 2 phytomicronutriments. / Citrus fruits, which are widely consumed worldwide, represent sources of carotenoids such as βCX, which contribute with flavonoids (HES) to the health effects of these fruits, particularly in the prevention of type 2 diabetes (T2D). T2D is a worldwide pathology that is growing exponentially. The general objective of this work is therefore to study the nutritional and health properties of clementine juice concentrates obtained by innovative membrane technology and specifically enriched in both βCX and HES but also in pectins. The digestive behaviour of these phytomicronutrients, guarantors of the nutritional quality of the concentrates, was first assessed by coupling the in vitro digestion model with the Caco-2 intestinal cells. For a better physiological vision of the mechanisms associated with the first stages of digestion, models of static and dynamic digestion are compared. The bioavailability and intestinal absorption of carotenoids are strongly influenced by the content and structure of pectins. In a second step, the biological activities (disglycemia, dislipidemia, inflammation) involved in the prevention of T2D are evaluated in vivo in rat after administration of the citrus concentrate. The study demonstrated that a citrus food specifically enriched in βCX and HES is effective in preventing metabolic syndrome/T2D, highlighting the possible role of βCX and its retinoid bioconversion. A complementary in vitro study on murine macrophages reveals synergistic anti-inflammatory effects of the two phytomicronutrients.