Phytochemical composition, antioxidant activity, pigment stability, bioactive properties, and in-vitro absorption of polyphenolics in acai fruit (Euterpe oleracea Mart.) were investigated. Detailed characterization of phenolic compounds present in acai fruit, acai fruit pulp, and a polyphenolic-enriched acai oil were conducted by HPLCESI- MSn analyses and their stability and influence on antioxidant capacity determined. Anthocyanins were predominant in acai fruits, which also contained several flavone and flavonol glycosides, flavanol derivatives, and phenolic acids. In-vitro absorption and antiproliferative effects of phytochemical extracts from acai pulp and acai oil were determined as a function of chemical composition. Polyphenolic mixtures from both acai pulp and acai oil extracts significantly inhibited HT-29 colon cancer cell proliferation, also inducing the generation of reactive oxygen species. In-vitro intestinal absorption using Caco-2 cell models demonstrated that phenolic acids and monomeric flavanol derivatives are readily transported through cell monolayers in-vitro. The influence of polyphenolic cofactors on the stability of anthocyanins in acai fruit under varying conditions of temperature and pH was evaluated. Significant time, temperature, and pH-dependent anthocyanin losses were observed in all models, yet the presence of phenolic acids, procyanidins, and flavone-C-glycosides had a positive influence on anthocyanin stability. External addition of flavone-C-glycosides significantly enhanced visual color, increased anthocyanin stability during exposures to high pH or storage temperatures, and had comparable effects to those of a commercial anthocyanin enhancer. Anthocyanin polymerization reactions occurring during storage of acai fruit juice models were investigated and potential mechanisms and reaction products identified. Polymeric anthocyanin fractions contained several anthocyanin-flavanol adducts based on cyanidin or pelargonidin aglycones and their presence was related to increased anthocyanin sulfite bleaching resistance and to the appearance of large, unresolved peaks in HPLC chromatograms. A reaction mechanism involving the nucleophilic addition of anthocyanins in their hydrated form to flavanol carbocations resulting from cleavage of interflavanic bonds was proposed for the formation of flavanol-anthocyanin adducts in acai fruit juices. Antiproliferative activity and in-vitro absorption of monomeric and polymeric anthocyanin fractions were also evaluated. Both fractions inhibited HT-29 colon cancer cell growth in a similar, concentration-dependent manner, yet in-vitro absorption trials using Caco-2 intestinal cell monolayers indicated the presence of anthocyanin polymers may influence anthocyanin absorption in acai fruit products.

Phenolic antioxidants reduce the effect of oxidative stress within cells. They are found in a various fruits, vegetables and as food additives to reduce spoilage. Consumption of antioxidants by humans has been linked with increased lifespan and reduced incidence of cancer and cardiovascular disorders (Cabrera et al. 2006; Kuriyama 2008). In cultured mammalian cells however, some of these phenolic antioxidants have been reported to generate reactive oxygen species (ROS), leading to chromosomal breakage (Long et al. 2007; Long & Halliwell 2001). It is clear then, that amongst this group of compounds, in vitro toxicological study is not a reliable prediction of human hazard. It is for this reason that the work described in this thesis was undertaken: the principal aim was to gain a better understanding of the reasons underlying this contradiction. It has been suggested that excessive ROS generated in vitro might be a result of the higher levels of oxygen (~20%) compared to (1-7%) in vivo: (Yusa et al. 1984; Turrens et al. 1982). With clearer understanding, new experimental approaches might be taken to highlight or reduce positive in vitro genotoxicity test results that might be considered misleading. A diverse set of test compounds was first chosen. It included polyphenolic (PPA), monophenolic (MPA) and non-phenolic antioxidants (NPA), in addition to mechanistically characterised oxidants, genotoxins and cytotoxic, non-genotoxins as controls. Genotoxicity was assessed in vitro using the GADD45a, GFP reporter assay and in silico using Derek Nexus™. Amongst the 19 antioxidants assessed, the 11 of 12 of PPAs, 0 of 4 MPAs and 1 of 3 NPAs (ethoxyquin) produced positive results in vitro and 8 of 12 PPAs generated alerts of at least plausible genotoxicity in silico. To discover whether these results were the result of cellular hyperoxia-promoted generation of physiologically irrelevant ROS in cells, genotoxicity was reassessed in the presence of 1 and 5% oxygen. This reduced oxygen exposure had no effect upon the qualitative result for any of the assessed compounds and a negligible effect upon the dose at which any positive result was produced. An assessment of the ability of antioxidants to generate potentially genotoxic ROS within cells was carried out using the intracellular fluorescent dye, dichlorofluorescin diacetate (DCFH-DA). 10 of 12 PPAs, 0 of 4 MPAs and 1 of 3 NPAs (ethoxyquin) were shown to increase the level of ROS within TK6 human lymphoblastoid cells within 4 hours of compound exposure. Within this same timeframe, the mitochondrial membranes in cells treated with 10 of 12 PPAs, 2 of 4 MPAs and 1 of 3 NPAs (ethoxyquin) were shown to become depolarised using JC-1 dye. It was unclear however, whether mitochondrial membrane depolarisation was a cause or a consequence of ROS generation within the cells. In order to assess whether the increase in intracellular ROS led to an increase in oxidised DNA within treated cells, 8-oxoguanine (8-OG) was quantified using a FITC conjugated anti8-OG antibody. This assessment revealed that levels of the oxidised base were only increased in cells exposed to two of the 12 PPAs (quercetin and resorcinol). The level of 8OG detected was lower than the vehicle control for cells treated with 10 of the 15 antioxidants. One interpretation of this is that these agents induce the repair pathway for oxidative damage, which leads to a lower level of oxidised DNA bases in the genome. The results showed that while a large proportion of PPAs produce genotoxic results in vitro and lead to increased levels of ROS, the amount of oxidised DNA is not higher in treated cells. This would suggest the presence of a different mechanism for the observed genotoxicity.

O mercúrio é um dos metais mais nocivos presentes no ambiente, advindo tanto de fontes naturais quanto antropogênicas. Os indivíduos estão expostos a diferentes formas do mercúrio através de diversas vias, como a alimentação, principalmente no caso do metilmercúrio presente em peixes. Suas formas orgânicas são muito significativas do ponto de vista toxicológico, considerando-se a exposição da população e seus efeitos pró-oxidantes e genotóxicos envolvidos na origem de inúmeras doenças. Por outro lado, é suposto que compostos polifenólicos e outros antioxidantes da dieta podem exercer atividade protetora contra os efeitos deletérios do mercúrio. A curcumina é um pigmento amarelo polifenólico extraído do rizoma da planta Curcuma longa Linn (Zingiberaceae). Diversas evidências apontam para suas propriedades antioxidantes, de modulação da sinalização celular e alteração da expressão gênica, além da possibilidade de sua utilização na prevenção dos efeitos deletérios dos metais no organismo. Assim sendo, este trabalho teve por objetivo avaliar os efeitos da associação entre o mercúrio e a curcumina, investigando a citotoxicidade de dois compostos orgânicos de mercúrio, metilmercúrio e etilmercúrio, e da curcumina, de forma isolada e combinada, em células HepG2, utilizando o ensaio do MTT. A genotoxicidade desses mesmos compostos também foi avaliada por meio do ensaio do cometa. Além disso, a alteração no estado oxidativo das células pela razão de glutationa (GSH/GSSG) e a alteração na expressão de 84 genes relacionados com vias de dano e reparo no DNA, utilizando-se de PCR-Array, também foram avaliados. Os compostos orgânicos de mercúrio apresentaram citotoxicidade em concentrações iguais ou maiores que 16 ?M. A curcumina apresentou citotoxicidade apenas na concentração de 128 ?M. Nos experimentos de associação entre o mercúrio e a curcumina foi observado um aumento da citotoxicidade, entre a concentração de 8 ?M das duas formas de mercúrio e a concentração de 64 ?M de curcumina. Na avaliação da genotoxicidade foi observado um efeito genotóxico significativo do metilmercúrio nas concentrações de 8, 16 e 32 ?M, enquanto o etilmercúrio apresentou genotoxicidade significativa apenas na concentração de 32 ?M. A curcumina não apresentou genotoxicidade. Na associação dos compostos não foi detectada ação antigenotóxica da curcumina e houve um aumento na genotoxicidade do metilmercúrio em associação com a concentração de 32 ?M de curcumina. A razão de glutationa mostrou um aumento significativo nas células tratadas com metilmercúrio, entretanto isso não foi observado quando o metilmercúrio foi associado com a curcumina. A análise da expressão de 84 genes relacionados com danos no DNA por PCR-Array mostrou alteração na expressão de 26 genes, sendo que 3 deles tiveram aumento na expressão, DDIT3, GADD45A e PPP1R15A, relacionados principalmente com o bloqueio do ciclo celular e o processo de apoptose, e 23 genes tiveram sua expressão reduzida, relacionados com diversas vias de reparo do DNA, checkpoints do ciclo celular e apoptose. Os resultados obtidos indicam que, nas condições avaliadas, a associação da curcumina com o mercúrio aumentou os efeitos deletérios do metal, causando um aumento na citotoxicidade e na genotoxicidade do mercúrio, não se caracterizando como uma possível estratégia de prevenção dos efeitos deletérios do mercúrio / Mercury is one of the most harmful metals present in the environment arising from both natural and anthropogenic sources. The individuals are exposed to different forms of mercury through various sources, such as food, especially in the case of methylmercury in fish, with this organic forms being very significant from a toxicological point of view, considering the exposure of the population and its pro-oxidant and genotoxic effects, involved in the origin of many diseases. On the other hand it is assumed that polyphenolic compounds and other dietary antioxidants can have protective activity against the harmful effects of mercury. Curcumin is a polyphenol extracted from the rhizome of Curcuma longa Linn. (Zingiberaceae). Several evidences show its ability to act as an antioxidant, modulate cell signaling and gene expression, and the possibility of its use in chemoprevention of the deleterious effects of metals. Therefore, this study aimed to evaluate the effects of the association between mercury and curcumin, investigating the cytotoxicity of two organic compounds of mercury, methylmercury and ethylmercury, and curcumin, alone and in combination, in HepG2 cells using the the MTT assay, the genotoxicity of these same compounds through the comet assay, the changes in oxidative state of the cells by the concentration of glutathione and GSH/GSSG ratio and the changes in the expression of 84 genes related to DNA damage anda repair pathways by PCR-Array. Organic mercury compounds showed cytotoxicity at concentrations equal to or greater than 16 uM. Curcumin only showed cytotoxicity at the concentration of 128 ?M. There was an increase in cytotoxicity when mercury (8 ?M) was associated with curcumin (64 ?M). In the genotoxicity assay there was a significant genotoxic of methyl mercury at concentrations of 8, 16 and 32 ?M while ethyl mercury whas genotoxic only at 32 ?M. Curcumin was not genotoxic. There was no anti-genotoxic activity in the association of compounds and there was an increase in genotoxicity of MeHg in association with 32 ?M of curcumin. The quantification of glutathione (GSH/GSSG) showed a significant increase in the GSH/GSSG ratio in cells treated with MeHg, however this was not observed when MeHg was associated with curcumin. Gene expression analysis showed changes in the expression of 26 genes, 3 of them were upregulated, DDIT3, GADD45A and PPP1R15A, mainly related to the cell cycle blockage and apoptosis, and 23 genes were down-regulated, related with DNA repair, cell cycle checkpoints and apoptosis. These results indicate that the combination of curcumin with mercury increased the deleterious effects of the metal, causing an increase in cytotoxicity and genotoxicity of mercury, and do not represent a possible strategy to prevent the harmful effects of mercury.

Rykštenė yra daugiametis augalas, priklausantis astrinių šeimai. Ji pasižymi keliais farmakologiniais veikimais, tokiais kaip skatinančiu diurezę, priešuždegiminiu, analgetiniu, antispazmolitiniu, antibakteriniu ir antigrybeliniu, taip pat antioksidaciniu ir priešvėžiniu aktyvumu. Iš rykštenės žaliavos gaminami vaistai skatinantys diurezę. Ji taip pat įeina į Lietuvoje gaminamos arbatos „Diuretiko“ (UAB „Acorus Calamus”) sudėtį. Šio darbo tikslas - atlikti polifenolinių rūgščių kokybinę ir kiekybinę analizę rykštenės ekstraktuose taikant efektyviąją skysčių chromatografiją. Buvo paruošti Solidago virgaurea ir Solidago gigantea lapų ir žiedų ekstraktai naudojant vandeninį metanolio tirpalą (70%). Jie buvo analizuojami ESC metodu. Detekcijai naudojamas fotodiodų matricos detektorius. Gauti rezultatai buvo lyginami su standartinių junginių (chlorogeno ir neochlorogeno rūgščių) duomenimis. Polifenolinės rūgštys nustatomos bangos ilgiui esant 324nm. Buvo identifikuotos dvi polifenolinės rūgštys: chlorogeno ir neochlorogeno. Solidago virgaurea chlorageno rūgšties yra 1.31%, o neochlorogeno rūgšties – 0.06%. Solidago gigantea chlorogeno rūgšties procentinis kiekis yra 2.35%, o neochlorageno rūgšties – 0.02%. / Goldenrod is a perennial plant belonging to the Asteraceae family. It has several pharmacological functions, such as promoting diuresis, anti-inflammatory, analgesic, antispazmolitiniu, antibacterial and antifungal as well as anti-oxidative and anti-tumor activity. Of goldenrod made a medicaments stimulating diuresis. It also includes in production of tea „Diuretiko“ (UAB „Acorus Calamus”) composition in Lithuanian. The aim of this research was to make polyphenolic acids qualitative and quantitative analysis of the goldenrod extracts using high-performance liquid chromatography. Was prepared Solidago virgaurea and Solidago gigantea leaves and flowers extracts of aqueous methanol (70%). They were analyzed in HPLC method. For detection was used photodiode array detector. The results were compared with standard compounds (chlorogenic and neochlorogenic acids) data. Polyphenolic acids determined at the wave length of 324nm. Has been identified two polyphenolic acids: chlorogenic and neochlorogenic. In Solidago virgaurea chlorogenic acid is 1.31% and neochlorogenic acid - 0.06%. In Solidago gigantea chlorogenic acid percentage is 2.35%, and neochlorogenic acid - 0.02%.

O mercúrio é um dos metais mais nocivos presentes no ambiente, advindo tanto de fontes naturais quanto antropogênicas. Os indivíduos estão expostos a diferentes formas do mercúrio através de diversas vias, como a alimentação, principalmente no caso do metilmercúrio presente em peixes. Suas formas orgânicas são muito significativas do ponto de vista toxicológico, considerando-se a exposição da população e seus efeitos pró-oxidantes e genotóxicos envolvidos na origem de inúmeras doenças. Por outro lado, é suposto que compostos polifenólicos e outros antioxidantes da dieta podem exercer atividade protetora contra os efeitos deletérios do mercúrio. A curcumina é um pigmento amarelo polifenólico extraído do rizoma da planta Curcuma longa Linn (Zingiberaceae). Diversas evidências apontam para suas propriedades antioxidantes, de modulação da sinalização celular e alteração da expressão gênica, além da possibilidade de sua utilização na prevenção dos efeitos deletérios dos metais no organismo. Assim sendo, este trabalho teve por objetivo avaliar os efeitos da associação entre o mercúrio e a curcumina, investigando a citotoxicidade de dois compostos orgânicos de mercúrio, metilmercúrio e etilmercúrio, e da curcumina, de forma isolada e combinada, em células HepG2, utilizando o ensaio do MTT. A genotoxicidade desses mesmos compostos também foi avaliada por meio do ensaio do cometa. Além disso, a alteração no estado oxidativo das células pela razão de glutationa (GSH/GSSG) e a alteração na expressão de 84 genes relacionados com vias de dano e reparo no DNA, utilizando-se de PCR-Array, também foram avaliados. Os compostos orgânicos de mercúrio apresentaram citotoxicidade em concentrações iguais ou maiores que 16 ?M. A curcumina apresentou citotoxicidade apenas na concentração de 128 ?M. Nos experimentos de associação entre o mercúrio e a curcumina foi observado um aumento da citotoxicidade, entre a concentração de 8 ?M das duas formas de mercúrio e a concentração de 64 ?M de curcumina. Na avaliação da genotoxicidade foi observado um efeito genotóxico significativo do metilmercúrio nas concentrações de 8, 16 e 32 ?M, enquanto o etilmercúrio apresentou genotoxicidade significativa apenas na concentração de 32 ?M. A curcumina não apresentou genotoxicidade. Na associação dos compostos não foi detectada ação antigenotóxica da curcumina e houve um aumento na genotoxicidade do metilmercúrio em associação com a concentração de 32 ?M de curcumina. A razão de glutationa mostrou um aumento significativo nas células tratadas com metilmercúrio, entretanto isso não foi observado quando o metilmercúrio foi associado com a curcumina. A análise da expressão de 84 genes relacionados com danos no DNA por PCR-Array mostrou alteração na expressão de 26 genes, sendo que 3 deles tiveram aumento na expressão, DDIT3, GADD45A e PPP1R15A, relacionados principalmente com o bloqueio do ciclo celular e o processo de apoptose, e 23 genes tiveram sua expressão reduzida, relacionados com diversas vias de reparo do DNA, checkpoints do ciclo celular e apoptose. Os resultados obtidos indicam que, nas condições avaliadas, a associação da curcumina com o mercúrio aumentou os efeitos deletérios do metal, causando um aumento na citotoxicidade e na genotoxicidade do mercúrio, não se caracterizando como uma possível estratégia de prevenção dos efeitos deletérios do mercúrio / Mercury is one of the most harmful metals present in the environment arising from both natural and anthropogenic sources. The individuals are exposed to different forms of mercury through various sources, such as food, especially in the case of methylmercury in fish, with this organic forms being very significant from a toxicological point of view, considering the exposure of the population and its pro-oxidant and genotoxic effects, involved in the origin of many diseases. On the other hand it is assumed that polyphenolic compounds and other dietary antioxidants can have protective activity against the harmful effects of mercury. Curcumin is a polyphenol extracted from the rhizome of Curcuma longa Linn. (Zingiberaceae). Several evidences show its ability to act as an antioxidant, modulate cell signaling and gene expression, and the possibility of its use in chemoprevention of the deleterious effects of metals. Therefore, this study aimed to evaluate the effects of the association between mercury and curcumin, investigating the cytotoxicity of two organic compounds of mercury, methylmercury and ethylmercury, and curcumin, alone and in combination, in HepG2 cells using the the MTT assay, the genotoxicity of these same compounds through the comet assay, the changes in oxidative state of the cells by the concentration of glutathione and GSH/GSSG ratio and the changes in the expression of 84 genes related to DNA damage anda repair pathways by PCR-Array. Organic mercury compounds showed cytotoxicity at concentrations equal to or greater than 16 uM. Curcumin only showed cytotoxicity at the concentration of 128 ?M. There was an increase in cytotoxicity when mercury (8 ?M) was associated with curcumin (64 ?M). In the genotoxicity assay there was a significant genotoxic of methyl mercury at concentrations of 8, 16 and 32 ?M while ethyl mercury whas genotoxic only at 32 ?M. Curcumin was not genotoxic. There was no anti-genotoxic activity in the association of compounds and there was an increase in genotoxicity of MeHg in association with 32 ?M of curcumin. The quantification of glutathione (GSH/GSSG) showed a significant increase in the GSH/GSSG ratio in cells treated with MeHg, however this was not observed when MeHg was associated with curcumin. Gene expression analysis showed changes in the expression of 26 genes, 3 of them were upregulated, DDIT3, GADD45A and PPP1R15A, mainly related to the cell cycle blockage and apoptosis, and 23 genes were down-regulated, related with DNA repair, cell cycle checkpoints and apoptosis. These results indicate that the combination of curcumin with mercury increased the deleterious effects of the metal, causing an increase in cytotoxicity and genotoxicity of mercury, and do not represent a possible strategy to prevent the harmful effects of mercury.

The thesis has focused on the separation of proanthocyanidins (Pas) in beer, raw beer material and beer intermediates (wort, hopped wort, green beer) by using high performance liquid chromatography coupled to high resolution mass spectrometric detection. A chromatographic column XSELECT HSS T3 was used for separation of individual proanthocyanidins. Mobile phase was formed by a mixture of 0,1% formic acid in water (A) and 0,1% formic acid in acetonitril (B), separation was conducted in gradient mode. Calibration range for catechin and procyanidin B1 a B2 was linear in the range from 10 µg.l-1 to 200 µg.l-1 and for epicatechin from 5 µg.l-1 to 200 µg.l-1 . Limit of quantitation (LOQ) was set to the lowest point of calibration curve, hence for catechin, procyanidin B1 and B2 was LOQ = 10 µg.l-1 and for epicatechin was LOQ = µg.l-1 . Yield of method of the observed Pas was very good, where yield oscillated around 100%. The method also had excellent repeatability, where the relative standard deviation (RSD) was lower then 20%. After optimization of this method it was used for observation of profile changes during brewery process in catechin, epicatechin, procyanidin B1, B2 and B3. It was discovered, that catechin, epicatechin, procyanidin B1 and B2 originate mainly from hops and procyaniding B3 is...

A incidência de doenças crônicas não transmissíveis como a diabetes tipo 2 e complicações cardiovasculares tem aumentado significativamente, e tem-se associado principalmente às mudanças nos hábitos alimentares tradicionais. O objetivo do presente estudo foi caracterizar diferentes cultivares de feijão, lupino e grãos da região dos Andes quanto a seus compostos fenólicos antioxidantes, capacidade antioxidante e potencial anti-hiperglicêmico e anti-hipertensivo in vitro. Dependendo do tipo de cultivar, o feijão é uma fonte promissora de taninos condensados, antocianinas, e flavonóis; enquanto que o lupino andino destacou-se pela presença de isoflavonas. Após o tratamento térmico, o feijão e lupino andino inibiram significativamente a enzima conversora da angiotensina I, relevante na prevenção da hipertensão, enquanto o milho roxo andino inibiu a α-glicosidase, relevante na prevenção da hiperglicemia. Uma combinação apropriada de grãos tradicionais como parte da dieta poderia contribuir na modulação dos níveis de glicose e na prevenção das complicações relacionadas ao desequilíbrio óxido-redução. / Incidence of chronic diseases such as diabetes type 2 and related cardiovascular complications has increased significantly due mainly to current changes in traditional food dietary habits. The objective of this study was to characterize several bean and lupin cultivars along with grains from the Andean region in relation to their phenolic compounds, antioxidant capacity and anti-diabetes and anti-hypertension potential using in vitro assays. Depending on the cultivar, beans are interesting sources of condensed tannins, anthocyanins and flavonols, whereas major phenolic compounds in Andean lupins were isoflavones. Following thermal treatment, selected beans and Andean lupins inhibited significantly the hypertension relevant angiotensin I-converting enzyme and among Andean grains, the purple corn inhibited the hyperglycemia relevant α-glucosidase. A good combination of traditional grains as a part of the overall diet can contribute to effective dietary strategies for managing Type 2 diabetes and associated complications linked to unbalanced cellular redox status.

Η ρύπανση από τα χημικά λιπάσματα είναι έντονη στα ελληνικά εδάφη (ιδιαίτερα από τα νιτρικά άλατα) και επομένως είναι κρίσιμη η μείωση των εισροών αγροχημικών στο περιβάλλον. Η μείωση της λίπανσης αυξάνει την συγκέντρωση αντιοξειδωτικών ουσιών στους φυτικούς ιστούς. Πολλές επιστημονικές μελέτες μέχρι σήμερα συσχετίζουν την διατροφή με φυτικά προϊόντα υψηλής διατροφικής αξίας που περιέχουν φαινολικές-αντιοξειδωτικές ουσίες με την πρόληψη καρδιαγγειακών παθήσεων, πολλών μορφών καρκίνου αλλά και την γήρανση. Έτσι η έρευνα αυτή είχε διπλό στόχο: τη μείωση της ρύπανσης του περιβάλλοντος μέσω της μείωσης των λιπασμάτων που χορηγούνται στις καλλιέργειες, και την παραγωγή φυτικών προϊόντων υψηλής βιολογικής και διατροφικής αξίας, αφού θα παρέχουν μεγαλύτερες ποσότητες αντιοξειδωτικών ουσιών στον ανθρώπινο οργανισμό. Επίσης θα πρέπει να αναφερθεί ότι η μείωση των ποσοτήτων των λιπασμάτων που θα απαιτούνται για την καλλιέργεια των φυτών θα επιφέρει και οικονομικό όφελος προς τους καλλιεργητές και τους καταναλωτές, αφού θα μειωθεί το κόστος παραγωγής των φυτικών προϊόντων. Για το σκοπό αυτό μελετήθηκε για πρώτη φορά η επίδραση της μεταβολής της συγκέντρωσης των χορηγούμενων νιτρικών ιόντων μέσω του θρεπτικού διαλύματος υδροπονικής καλλιέργειας στο ρυθμό παραγωγής πολυφαινολικών ενώσεων σε λαχανικά ευρείας κατανάλωσης (παντζάρι και μαρούλι) και διαπιστώθηκε ότι η μεγιστοποίηση της παραγωγής των φυσικών αντιοξειδωτικών παραγόντων είναι δυνατή μέσω του ελέγχου της αζωτούχου θρέψης. Παράλληλα, διαπιστώθηκε και ποσοτικοποιήθηκε η ύπαρξη ενός κρίσιμου σημείου στη συγκέντρωση του χορηγούμενου αζώτου που ενεργοποιεί τον δευτερογενή μεταβολισμό των καλλιεργούμενων φυτών παντζαριού και μαρουλιού, αυξάνοντας σημαντικά το ρυθμό παραγωγής των ιδιαίτερα ευεργετικών για την υγεία φυτοχημικών ενώσεων όπως φαινολικών και μπετακυανινών. Η αύξηση όχι μόνο της περιεκτικότητας αλλά και της ενεργότητας των αντιοξειδωτικών παραγόντων επιβεβαιώθηκε με τη χρήση προηγμένων μεθόδων προσδιορισμού όπως το EPR. Τέλος προσδιορίστηκαν και ποσοτικοποιήθηκαν με σύγχρονη μέθοδο φασματομετρίας μαζών (LC-MS/MS) στα διαφορετικά μέρη των φυτών παντζαριού και στα φύλλα του μαρουλιού συνολικά επτά διαφορετικές πολυφαινολικές ενώσεις. / The pollution from chemical fertilizers is pronounced in Greek soils (particularly nitrates) and is therefore critical to reduce inputs of agrochemicals in the environment. The decrease of fertilization increases the concentration of antioxidants in plant tissues. Many scientific studies to date relate the diet with plant products of high nutritional value containing phenolic-antioxidants with the prevention of cardiovascular disease, number of cancers and aging. So the aims of the present research are: to reduce environmental pollution by reducing fertilizer applied to crops, and the production of crops with high biological and nutritional value, providing greater amounts of antioxidants in the human body. It should also be noted that the decrease in the quantities of fertilizer that will be required for the cultivation of plants will also lead to economic benefits to farmers and consumers, as it will reduce the cost of production of plant products. For this purpose the effect of varying concentration of nitrate granted via hydroponic nutrient solution, in the production rate of polyphenolic compounds in vegetables (lettuce and beetroot) was studied. It was found that maximizing the production of natural antioxidants is possible through the control of nitrogen nutrition. Also, it was ascertained that a critical point in the concentration of the administered nitrogen exists below which the secondary metabolism of the studied crops i.e. beetroot and lettuce is activated, significantly increasing the rate of production of the highly beneficial for the health compounds such as phenolic phytochemicals and betacyanins. The augmentation of the activity of antioxidants was confirmed by using advanced methods such as EPR. Finally seven different polyphenolic compounds were identified and quantified by the use of liquid chromatography-tandem mass spectrometry (LC-MS/MS), in different plant parts of beetroot and lettuce leaves.

Darbo tikslas: Pagaminti vaistinių melisų ištraukas ir įvertinti jų kokybę. Darbo uždaviniai: 1. Remiantis mokslinės literatūros duomenimis parinkti tinkamą vaistinių melisų skystą vaisto formą, atsižvelgiant į veikliosios medžiagos farmakologines savybes bei parinkti analizės metodus preparatų kokybei įvertinti; 2. Nustatyti ekstrahento (etanolio ir vandens) įtaką melisų ištraukų kokybei; 3. Palyginti ekstrahavimo metodo daromą įtaką vaistinių melisų ištraukų kokybei; 4. Ištirti žaliavos ir ekstrahento santykio daromą įtaką vaistinių melisų ištraukų kokybei. Metodika: 1. Tyrimo objektas – vaistinių melisų ištraukos; 2. Vaistinių melisų vandeninių ir etanolinių ištraukų technologija: a) Ištraukos gamintos klasikinės maceracijos ir perkoliacijos metodais; b) Etanolinės ištraukos gamintos ekstrahuojant ultragarsu ir kokybė vertinama po 20 min., 30 min. ir 40 min. ekstrahavimo; 3. Taikytas spektrofotometrinis metodas pagal Folin – Ciocalteau metodiką, norint nustatyti suminį polifenolinių junginių kiekį pagal KRE; 4. Sauso likučio tyrimas atliktas gravimetriniu metodu; 5. Antimikrobinis aktyvumas nustatytas pagal Ph. Eu. 01/2002, 2.6.12 metodiką. Rezultatai: Spektrofotometriniu metodu nustatyta, kad ekstrahentas turi įtakos veikliųjų junginių išsiskyrimui: didesnis suminis polifenolinių junginių bei sauso likučio kiekis nustatytas, kai ekstrahentu pasirenkamas etanolis, o ne vanduo. Vertinant etanolio koncentraciją nustatyta, kad mažiausiai veikliųjų junginių... [toliau žr. visą tekstą] / Purpose of work: To make and evaluate the quality of Melissa officinalis extracts. Tasks of work: 1. According to the scientific literature to identify an appropriate Balm liquid dosage form according to the pharmacological properties of the active material and the selection of analytical methods for evaluating the quality of products; 2. To set an extraction solvent (ethanol and water) influence on the quality of extracts of Melissa officinalis; 3. To compare extraction method influence on the quality of extracts of Melissa officinalis; 4. To investigate the relationship between the raw material and extraction solvent influence on the quality of extracts of Melissa officinalis. Methods: 1. Object of analysis – extracts of Melissa officinalis; 2. Preparation of an aquous and ethanolic extracts: a) Extracts have been produced by tradicional maceration and percolation methods; b) Ethanol extracts have been produced by extraction by ultrasound. The quality of these extracts has been evaluated after 20, 30, 40 min. 3. Spectrophotometric method (Folin – Ciocalteua method) – common amount of total polyphenolic compounds by p-cumaric acid equivalent has been evaluated; 4. The evaluation of dry residual according to gravimetric analysis has been calculated; 5. Antimicrobial activity by Ph. Eu. 01/2002, 2.6.12 method has been evaluated. Results: By spectrophotometric method it has been evaluated that extrahent has the influence on active compounds emission: higher amount of... [to full text]

Thesis (MSc Food Sc)--University of Stellenbosch, 2011. / ENGLISH ABSTRACT: Nectar formulations containing red-fleshed plum pulp and varying amounts of red-fleshed plum skin extract were developed. Red-fleshed plum nectar formulations containing 0, 8, 16, 24, and 32% skin extract were benchmarked against twenty-two commercial beverages containing red, violet and blue fruits. The total soluble solid content, pH, titratable acidity, colour, total polyphenolic, individual polyphenolic, total anthocyanin, and ascorbic acid contents, as well as antioxidant activity (oxygen radical antioxidant capacity (ORAC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and ferric reducing/antioxidant power (FRAP)) of the commercial beverages and plum formulations were determined. The plum nectar formulations had similar or higher total polyphenolic content, antioxidant activity, and colour values than the average for the commercial beverages. The individual polyphenolic compounds analysed in the nectar formulations were cyanidin-3-glucoside, cyanidin-3- rutinoside, quercetin-3-glucoside, quercetin-3-rutinoside, quercetin-3-xyloside, and neochlorogenic acid. Increasing polyphenolic content and antioxidant activity was observed with an increase in skin extract content of the formulations. The sensory attributes of the formulations were plum and plantlike aroma, plum and plantlike flavour, sweetness, acidity, and astringency. Increases in plantlike aroma and flavour, acidity, and astringency in conjunction with decreases in plum aroma, plum flavour, and sweetness extract were observed with an increase in skin extract. Consumer analysis indicated that all formulations were acceptable. The 0 and 16% skin extract formulations were most preferred, while the 32% skin extract formulation was least preferred. A shelf-life study was conducted in two phases. In Phase 1, the 0, 16 and 24% skin extract formulations were stored at 0 and 5°C (analysis time points: before pasteurisation, after pasteurisation (week 0), and after 1, 2, 4, 6, 8, 12, 16, 18, 20, and 24 weeks of storage). Chemical analyses conducted included colour, total polyphenolic, individual polyphenolic compound, and total anthocyanin contents, and antioxidant activity (DPPH• scavenging activity). Results from Phase 1 indicated close associations between the 16 and 24% skin extract formulations, and between these formulations and all chemical attributes. Regression analysis of results indicated significant (P 0.05) decreases in red colour, total anthocyanins, cyanidin-3-glucoside, cyanidin-3- rutinoside, DPPH• scavenging activity, total polyphenolic content, quercetin-3-rutinoside, and quercetin- 3-xyloside for formulations stored at 0°C. The total and red colour, total anthocyanins, cyanidin-3- glucoside, and cyanidin-3-rutinoside in formulations stored at 5°C showed similar results. During Phase 2 of the shelf life study, sensory analysis was conducted on the 0 and 24% skin extract formulations stored at 5°C (preparation time points: 0, 1, 2, and 3 months). Sensory attributes, including plum, plantlike, and raisin flavour, sweetness, acidity, and astringency, were stable during storage. Chemical results from Phase 2 were similar to those of Phase 1. The chemical and sensory stability of formulations after 24 weeks of storage in Phase 1 and Phase 2 indicated that, with the exception of the anthocyanin degradation, the formulations could be beneficial to juice industries. Thus, red-fleshed plum nectars have the potential to compete with high-antioxidant fruit beverages. / AFRIKAANSE OPSOMMING: Nektarformulasies wat rooivleis pruimpulp en varieërende hoeveelhede rooivleis pruimskilekstrak bevat, is ontwikkel en ondersoek. Rooivleis pruimnektarformulasies wat 0, 8, 16, 24, en 32% skilekstrak bevat, is vergelyk met twee-en-twintig kommersiële drankies wat rooi, violet en blou vrugte bevat. Die totale oplosbare vastestof, totale polifenoliese, individuele polifenoliese, totale antosianien- en askorbiensuur inhoude, sowel as die pH, titreerbare suurheid, kleur, antioksidant aktiwiteit (suurstofradikale antioksidantkapasiteit (ORAC), 2,2-difeniel-1-pikrielhidrasiel (DPPH) radikaal blussingsaktiwiteit, en ysterreduserende/antikoksidantkrag (FRAP)) van die kommersiële drankies en pruimnektarformulasies, is bepaal. Die pruimnektarformulasies het soortgelyke of hoër totale polifenoliese inhoud, antioksidantaktiwiteit, en kleurwaardes gehad in vergelyking met die gemiddelde vir die kommersiële drankies. Die individuele polifenoliese verbindings wat in die nektarformulasies geanaliseer is, was sianidien-3-glukosied, sianidien-3-rutinosied, kwersetien-3-glukosied, kwersetien-3-rutinosied, kwersetien-3-xylosied, en neochlorogeniese suur. ‘n Toenames in die kleurwaardes, polifenoliese inhoud, en antioksidantaktiwiteit is waargeneem met ‘n toename in skilekstrak is in die formulasies. Die sensoriese eienskappe van die formulasies was pruim- en plantagtige aroma, pruim- en plantagtige geur, soetheid, suurheid, en frankheid. Toenames in plantagtige aroma en geur, suurheid en frankheid, sowel as ‘n afname in pruimaroma, pruimgeur, en -soetheid, is met ‘n toename in skilekstrak waargeneem. Verbruikersanalise het aangedui dat al die formulasies aanvaarbaar was. Die 0 en 16% skilekstrakformulasies was die mees aanvaarbaarste, terwyl die 32% skilekstrakformulasie die minste aanvaarbaar geag is. ‘n Rakleeftydstudie is in twee fases gedoen. In Fase 1 is die 0, 16, en 24% skilekstrakformulasies by 0 en 5°C gestoor (analiseringstydpunte: voor pasturisasie, na pasturisasie (week 0), en na 1, 2, 4, 6, 8, 12, 16, 18, 20, en 24 weke van berging). Chemiese analise wat gedoen is, sluit totale polifenoliese, individuele polifenoliese verbinding, en totale antosianien inhoude, sowel as kleur en antioksidantaktiwiteit (DPPH• blussingsaktiwiteit) in. Resultate van Fase 1 het ‘n nou verband tussen die 16 en 24% skilekstrakformulasies, sowel as tussen hierdie formulasies en hul chemiese kenmerke aangedui. Regressie-analise van resultate het betekenisvolle (P 0.05) afnames geïllustreer in rooi kleurwaardes, DPPH• blussingsaktiwiteit, sowel as totale antosianiene, sianidien-3-glukosied, sianidien-3-rutinosied, totale polifenoliese, kwersetien-3-rutinosied, en kwersetien-3-xylosied inhoude van die formulasies wat by 0°C gestoor is. Die totale kleur-, rooi kleurwaardes, sowel as totale antosianien, sianidien-3-glukosied, en sianidien-3-rutinosied in die formulasies wat by 5°C gestoor is, het soortgelyke resultate gegee. Gedurend Fase 2 van die rakleeftydstudie is sensoriese analise op die 0 en 24% skilekstrakformulasies wat by 5°C gestoor is, gedoen (voorbereidingstydpunte: 0, 1, 2, en 3 maande). Sensoriese eienskappe, insluitend pruim-, plantagtige-, en rosyntjiesmake, soetheid, suurheid, en frankheid, was stabiel gedurende berging. Chemiese resultate van Fase 2 was soortgelyk aan dié van Fase 1. Die chemiese en sensoriese stabiliteit van die formulasies na 24 weke van opberging in Fase 1 en Fase 2 nieteenstaande antosianienafname, het aangedui dat die formulasies voordelig kan wees vir die vrugtedrankiebedryf. Dus het rooivleis pruimnektars die vermoeë om met hoë antioksidant vrugtedrankies mee te ding.