Thesis (MScEng) -- University of Stellenbosch, 2004. / ENGLISH ABSTRACT: The grape juice concentrate (GJC) plant of the KWV at Robertson spent significant amounts of money on the decolourisation of grape juice concentrate. A chemically activated powdered activated carbon (PAC) purchased from Norit, namely CA1, was used as decolourisation product. Apart from the expenses involved, it contributed largely to the solid waste produced at this plant. A way was sought to minimise or prevent GJC discolourisation (and possibly solid waste) without increasing operating expenses. Browning reactions in GJC are as old as the product itself. Numerous researchers have studied the origins of these reactions, the reactants and products involved, as well as the reaction kinetics of these reactions. From the work of these researchers four possible browning reaction pathways were identified, namely: • enzymatic oxidative browning, • non-enzymatic oxidative browning, • non-enzymatic browning (the Maillard reaction), and • caramelisation. It was also identified that 5-hydroxymethylfurfural (HMF) are indicative of the browning potential of GJC. A method to analyse for HMF (quantitative and qualitative) was develop for the purposes of this study, namely positive electron-spray ionisation preceded by high-pressure liquid chromatography (HPLC) and followed by dual mass spectrometry. This method showed good repeatability and was used to analyse all samples generated during this study. It was confirmed that the manufacturing process at this plant favours nonenzymatic browning reactions, since mild heat treatment deactivates enzymes. Further investigation indicated that the overruling browning reaction on this plant was non-enzymatic oxidative browning. It was shown that neither the presence, nor the absence of protein had any effect on the rate of formation of HMF. It was, however, confirmed that HMF formation could be attributed to high temperatures and prolonged exposure to these temperatures. Other adsorption products were evaluated against the then current PAC (CA1), namely a steam activated PAC supplied by Norit, SA4, and a polymeric adsorbent, Polyclar V (polyvinylpolypyrrolidone/PVPP). Both SA4 and PVPP indicated superior HMF adsorption capacities. Replacing CA1 with SA4 could result in operating expenses savings and possible solid waste reduction. However, PVPP were too expensive to be considered an economically viable replacement for CA1. Improved concentration technologies such as reverse osmosis (RO) membrane concentration followed by centrifugal evaporation (CE) or twostage CE should be considered as possible replacement for the existing concentration technology (multi-stage falling film evaporator). This should decrease heat treatment/exposure by more than 90% and thus reduce browning significantly. An added advantage could be the reduction of solid waste, since less (if not no) decolourisation will be required. Alternatively, juice should be stored with added sulphur dioxide (SO2), since it was shown that this juice contained much lower HMF concentrations than diluted concentrate (stored for the same time). This should reduce heat exposure by up to 50% and thus minimise browning reactions. / AFRIKAANSE OPSOMMING: Die druiwesapkonsentraat (DSK) aanleg van die KWV in Robertson het jaarliks aansienlike bedrae geld spandeer tydens die ontkleuringsproses van DSK. ‘n Chemies geaktiveerde verpoeierde koolstof (GVK) verkrygbaar van Norit, naamlik CA1, is gebruik as ontkleuringsproduk. Buiten die kostes verbonde aan hierdie produk het dit ook grootliks bygedra tot soliede afval by hierdie aanleg. Oplossings is gesoek om die verbruining/ontkleuring van DSK (en dalk ook soliede afval) te verminder (of selfs te voorkom) sonder om bedryfskostes te verhoog. Verbruiningsreaksies in DSK bestaan al so lank soos DSK self. Verskeie navorsers het die oorsake, reaktante, produkte en reaksiekinetika van hierdie reaksies oor die jare heen bestudeer. Uit die werk van sommige van hierdie navorsers kon vier moontlike verbruiningsreaksieroetes geïdentifiseer word, naamlik: • ensiematiese oksidatiewe verbruining, • nie-ensiematiese oksidatiewe verbruining, • nie-ensiematiese verbruining (die Maillard-reaksie), en • karamelisering. Daar was verder geïdentifiseer dat 5-hidroksiemetielfurfuraal (HMF) aanduidend is van die verbruiningspotensiaal van DSK. ‘n Analitiese metode (kwalitatief en kwantitatief) om vir HMF te analiseer is vir die doel van hierdie studie ontwikkel, naamlik positiewe elektronsproei ionisasie, voorafgegaan deur hoëdruk vloeistof chromatografie en gevolg deur dubbele massa spektrometrie. Hierdie analitiese metode het goeie herhaalbaarheid getoon en was deurgaans gebruik om monsters te analiseer gedurende hierdie studie. Dit was bevestig dat die vervaardigingsproses by hierdie aanleg nieensiematiese verbruiningsreaksies begunstig, aangesien geredelike hittebehandeling ensieme deaktiveer. Verdere navorsing het getoon dat die oorheersende verbruiningsreaksies by hierdie aanleg nie-ensiematiese oksidatief van aard is. Resultate het getoon dat proteinstabiliteit geen invloed op die vormingstempo van HMF het nie. Dit was egter bevestig dat vorming van HMF direk verband hou met hoë temperature en lang blootstellingsperiodes aan hierdie temperature. Ander adsorpsieprodukte was vergelyk met die huidige GVK (CA1), naamlik ‘n stoom geaktiveerde verpoeierde koolstof (Norit se SA4) en ‘n polimeriese adsorbant, Polyclar V (polivinielpolipirrolidoon/PVPP). Beide SA4 en PVPP het CA1 oortref wat betref HMF adsorpsie. Moontlike bedryfskostebesparings (en soliede afval verminderings) potensiaal bestaan indien CA1 vervang word met SA4. Die teenoorgestelde is egter waar vir PVPP wat bedryfskoste aangaan. Instede van die huidige verdampinstegnologie, naamlik vallendefilmverdamping, hoort verbeterde konsentrasietegnologieë soos tru-osmose membraankonsentrasie gevolg deur sentrifugale verdamping, of, alternatiewelik, twee-stadium sentrifugale verdamping, orrweeg te word. Op hierdie wyse behoort hittebehandeling (en dus verbruining) met sowat 90% verminder te word. ‘n Moonlike addisionele voordeel is die vermindering van soliede afval aangesien minder ontkleuring nodig sal wees. Indien die verbeterde tegnologieë te duur is moet daar gekyk word daarna om die ongekonsentreerde sap met addisionele swaweldioksied (SO2) te stoor, aangesien veel laer HMF konsentrasies in sulke sap waargeneem is as in verdunde direkte konsentraat wat vir dieselfde typerk gestoor is. Hittebehandeling sal op hierdie wyse met tot 50% verminder word (en dus verbruining ook).

Thesis (Masters of Environmental Health)--Cape Peninsula University of Technology, 2018. / In the Western Cape, South Africa and other regions globally, grape pomace (GP) is one of the abundant agro-waste from the winery industry. This study reports on the hyper-extraction of fermentable sugars from GP treated with white rot fungi (WRF) Phanerochaete chrysosporium BKMF 1767 to facilitate improved biovalorisation for total reducing sugars (TRS) extraction in conjunction with Nepenthes mirabilis digestive fluids. TRS were quantified using the 3,5-dinitrosalicylic acid (DNS) reagent method. The free readily dissolvable sugars from the GP recorded for the bio-treated (BT) samples was 206.39 ± 0.06 mg/L and for the untreated (UT) samples was 271.05 ± 0.02 mg/L. Overall, the TRS yield for the Bio-treated (BT) and untreated (UT) samples was recorded as 205.68 ± 0.09 and 380.93 ± 0.14 mg/L, respectively, using hot water pretreatment (HWP) with 2266.00 ± 0.73 (BT) and 2850.68 ± 0.31 mg/L (UT), respectively, for dilute acid pretreatment (DAP); with 2068.49 ± 6.02 (BT) and 2969.61 ± 8.054 mg/L (UT) respectively, using the cellulase pretreatment (CP) method. Using the HWP as a reference, the relative increases imparted by the biotreatment was higher (51%) for DAP and low (33%) for CP. The combination of conventional used pre-treatment methods (hot water pretreatment, dilute acid pre-treatment, and cellulase pre-treatment) in a single pot system was also done while monitoring the total residual phenolics (TRPCs) in the samples. Furthermore, powder X-ray diffraction (pXRD) were used to measure the crystallinity index (CrI) and functional groups of pre- and post-pretreated GP to ascertain the efficiency of the pre-treatment methods, with quantification of lignin, holocellulose, and ash. Overall, the TRS yield for N. mirabilis pre-treated agro-waste was 951 mg/L ± 4.666 mg/L, with biomass having a lower CrI of 33%, and 62% residual lignin content. Furthermore, reduced TRPCs were observed in hydrolysate, suggesting limited inhibitory by-product formation during N. mirabilis pre-treatment

CAPES / A busca por produtos naturais e os riscos para a saúde decorrentes do consumo de antioxidantes sintéticos incentivam a investigação para a descoberta de novas fontes potenciais de compostos bioativos. A produção de vinho e suco de uva geram grandes quantidades de coprodutos ricos em compostos fenólicos que podem ser utilizados como antioxidantes naturais. Este estudo teve como objetivo avaliar as melhores condições de extração de compostos bioativos, através de um planejamento fatorial fracionário 24-1, de um coproduto oriundo do processamento da indústria de sucos de uvas (BUS – Bagaço de Uvas Suco) e de um coproduto oriundo do processamento da indústria de vinhos (BUV – Bagaço de Uvas Vinho). E, após a determinação da melhor condição para ambos os coprodutos, avaliar a atividade antioxidante utilizando diferentes metodologias de quantificação, sendo que dentre as duas amostras (BUS e BUV), a que obtivesse maior potencial antioxidante, seria testada no processamento de linguiça frescal de frango com intuito de verificar a inibição da formação de malonaldeído em comparação ao antioxidante sintético ES (eritorbato de sódio). Para a extração dos compostos bioativos as variáveis independentes estudadas foram: tipo de coproduto (BUS ou BUV), temperatura do banho-maria (40 ou 60 °C), tempo de extração dos compostos bioativos (15 ou 45 minutos) e o tipo de solvente utilizado (acetona 80% ou etanol 80%). Os extratos dos coprodutos foram analisados em termos do conteúdo total de compostos fenólicos por Folin-Ciocalteu e atividade sequestradora do radical DPPH• (variáveis dependentes). A atividade sequestradora de radicais livres nos 16 testes gerados variaram 65-149 μmol Trolox.g-¹ e compostos fenólicos totais entre 18-35 mg GAE.g-¹ (equivalente ácido gálico). A análise de regressão linear mostrou que mais de 97% da variabilidade dos dados foi explicada pelo modelo gerado para a variável dependente atividade antioxidante e 93% para compostos fenólicos totais. Levando-se em conta os pontos ótimos de extração para BUS e BUV, fixaram-se extrações com acetona 80%, 60°C, durante 15 minutos para BUS e 45 minutos para BUV. Sob essas condições de extração, BUS apresentou: atividade antioxidante pelo método DPPH (IC50), ABTS•+, FRAP, e percentual de inibição do sistema de β- caroteno/ácido linoleico em valores de 349 μg.mL-1, 334 mmol TEAC.g-1, 151 mmol Fe²+.g-1 e 73%, respectivamente. Enquanto BUV resultaram em 489 μg.mL-1, 249 umol TEAC.g-1, 111 umol Fe²+.g-1 e 68%, respectivamente. As respostas do planejamento fatorial mostraram que BUS teve o maior teor de compostos fenólicos e atividade antioxidante quando comparados ao BUV, em suas melhores condições. Foram elaboradas quatro formulações de linguiça frescal de carne de frango: amostra controle (sem antioxidantes – T1), amostra contendo antioxidante sintético a 0,01% (T2), amostra contendo 2 mg.g-1 de extrato de BUS (T3) e uma amostra contendo 4 mg.g-1 de BUS (T4). Para a elaboração das linguiças com extrato de BUS, utilizou-se o extrato da melhor condição, determinada pelo planejamento fatorial (acetona 80%, 60°C, 15 minutos), sendo que os extratos foram evaporados e liofilizados. Os parâmetros físico-químicos e microbiológicos das formulações T1, T2, T3 e T4, resultaram em valores que estão em concordância com as legislações vigentes. A cor sofreu variações durante o tempo de armazenamento das linguiças frescais de frango, sendo que T3 e T4 foram os tratamento que tiveram maior ΔE e menores valores de L*. Os tratamentos T1 e T2 obtiveram maiores valores de a*, b* e C*. O pH decaiu, para as quatro formulações de linguiças, variando de 6,71 a 5,99, entre os tratamentos. Para medir a oxidação lipídica foi utilizada a metodologia TBARS. As amostras contendo BUS foram capazes de inibir a oxidação lipídica. No tempo de 8 dias de armazenamento a 4°C, em relação ao controle, T2 conseguiu inibir a formação de malonaldeído em 0,31%, enquanto que os tratamentos contendo 2 e 4 mg.g-1 de extrato liofilizado de BUS (T3 e T4) inibiram 55% e 73% respectivamente. / The search for natural products and the risk to human health due to consumption of synthetic antioxidants encourage research for the discovery of new potential sources of bioactive compounds. The production of wine and grape juice generates large amounts of pomace rich in phenolic compounds that can be used as natural antioxidants. The objective of this work was to evaluate the extraction of bioactive compounds from pomace of grape juice (BUS) and wine (BUV) industry using a 24-1 fractional factorial design. After the determination of the better extraction condition for both co-products, was determinate with the total antioxidant activity using different determination methodology, and among the two samples (BUS and BUV), which obtained the largest quantifications by these methodologies, would be tested in the processing of chicken sausage in order to verify the inhibition of formation of malondialdehyde compared to synthetic antioxidant ES (sodium erythorbate)The independent variables studied were grape fruit pomace (GJP) and grape wine pomace (GWP), both temperature (40 and 60°C), extraction time of compounds (15- 45 min), and solvent type (acetone and ethanol, 80%). Co-products extracts were analyzed in terms of the contents of TPC by Folin Ciocalteu and scavenging activity of the DPPH• radical. After determining the best extraction method of bioactive compounds in each pomace (GJP and GWP), we determined the amount of antioxidants required to decrease the initial concentration of DPPH at 50% (IC50), ABTS• radical scavenging activity, Ferric Reducing Antioxidant Power (FRAP), and inhibition percentage by autoxidation of the system of β-carotene/linoleic acid. The DPPH radical scavenging activity in the 16 tests ranged from 65-149 μmol Trolox.g-1, and the TPC from 18-35 mg GAE.g-1 (gallic acid equivalent). The statistical analysis explains the significance of the effects and coefficients of the model generated in terms of their antioxidant activities. The linear regression analysis showed that the model generated explain more than 93% of the data variability. Thus, for both pomaces, acetone 80% was the best solvent for the extraction of antioxidant compounds and the ideal extraction temperature was 60°C. However, the best extraction time for GJP was 15 min and for GWP, 45 min. Under these extraction conditions, GJP showed activity in the DPPH method (EC50), ABTS•+, FRAP, and inhibition percentage at 339 μg.mL-1, 334 μmol TEAC.g-1, 151 μmol Fe2+.g-1and 73%, respectively. The values for GWP were 489 μg.mL-1, 249 μmol TEAC.g-1, 111 μmol Fe2+.g-1 and 68%, respectively. The responses of the factorial design showed that GJP had higher contents of phenolic compounds and antioxidant activity than GWP in its better extractions conditions. Was formulated four different chicken meat sausages: control sample (without antioxidant – T1), sample containing synthetic antioxidant 0.01% (T2), sample containing 2mg.g-1 of GJP extract (T3) and a sample containing 4 mg.g-1 of GJP extract (T4), the GJP extract was utilized lyophilized and in your optimized condition according to the factorial design (acetone 80%, 60ºC, 15 minutos), and the extracts were route-evaporated and lyophilized. The physicochemical and microbiological parameters of all formulations resulted in values that are consistent with current laws. Was observed color variations during the storage period of fresh chicken sausages, treatments T3 and T4 had the highest ΔE and the smaller L * values. T1 and T2 treatments had higher values of a *, b * and C *. The pH showed a reduction for the four formulations of sausages, ranging from 6.71 to 5.99, between treatments. To measure lipid oxidation TBARS methodology was used. Samples containing BUS were able to inhibit lipid oxidation. At time 8 days of storage at 4 ° C, compared to the control, T2 could inhibit the formation of malonaldehyde 0.31%, while the treatments with 2 ad 04 mg.g-1 of lyophilized BUS extract (T3 and T4) inhibited 55% and 73% respectively.