Presented diploma thesis is dedicated to the determination of selected chemical parameters of fruits of new currant varieties. The theoretical part is focused on the characteristics of currant, the biologically active substances of its fruits and also the utilization of current in the food industry. Further part of the theory deals with the main chemical parameters which were examined: polyphenols, anthocyanins, ascorbic acid and antioxidant activity. In the practical part there were determined selected chemical parameters in six white, twelve red and thirteen black varieties of currant. Anthocyanins and polyphenols have been determined spectrophotometrically. The content of ascorbic acid in selected varieties of currant was determined by HPLC. As a technique for the determinationof the antioxidant activity was selected the method of free radical DPPH.

Вещества, обладающие антирадикальной активностью (АРА) играют ведущую роль в системе защиты организма от свободных радикалов. Общие недостатки известных способов оценки АРА выражаются в том, что измерения проводятся непрямыми методами, что затрудняет количественно оценивать содержание веществ с антирадикальными свойствами. Результаты измерений представляются в относительных единицах, в качестве эталонных веществ чаще всего используются: тролокс, галловая и аскорбиновая кислоты, проявляющие разную АРА, что не позволяет сравнивать результаты между собой. Таким образом, существует необходимость в разработке прямого, безэталонного метода оценки антирадикальной активности. Одним из потенциально возможных решений данной проблемы является метод ЭПР-спектроскопии на модели 2,2-дифенил-1-пикрилгидразил. Цель и задачи исследования. Исследование безэталонным методом оценки антирадикальной активности природных и синтетических антиоксидантов на модели 2,2-дифенил-1-пикрилгидразил с использованием ЭПР-спектроскопии. Отработка методики определения антирадикальной активности на модели ДФПГ методом ЭПР-спектроскопии. Получение и анализ значений антирадикальной активности для ряда веществ с антирадикальными свойствами. Сравнение с результатами полученными методом потенциометрического определения антиоксидантной активности с использованием медиаторной системы. Полученные результаты. Установлены оптимальные условия для проведения анализа. Проведено исследование - новым, безэталонным методом оценки антирадикальной активности, ряда природных и синтетических антиоксидантов. Установлены значения антирадикальной активности ряда природных и синтетических антиоксидантов. Рассмотрены различные варианты механизмов ингибирования свободных радикалов и их зависимость от структуры антиоксиданта. / Substances with anti-radical activity (ARA) play a leading role in the body's defense system against free radicals. General disadvantages of the known methods of evaluating the APA expressed in the fact that measurements are carried out by indirect methods, making it difficult to quantify the content of substances with antiradical properties. The measurement results are presented in relative units, as reference substances are most often used: Trolox, ascorbic and gallic acid, exhibit different ARA, which does not allow to compare the results with each other. Thus, there is a need to develop direct, reference-free method of evaluating anti-radical activity. One possible solution to this potential problem is the method of EPR model 2,2-diphenyl-1-picrylhydrazyl. The purpose and objectives of the study. Study standardless method for assessing the antiradical activity of natural and synthetic antioxidants model 2,2-diphenyl-1-picrylhydrazyl using EPR spectroscopy. Testing methods for determining the anti-radical activity in the DPPH model EPR spectroscopy. Obtaining and analyzing anti-radical activity values for a number of substances with antiradical properties. Comparison with the results obtained by potentiometric determination of antioxidant activity using a mediator system Results. The optimum conditions for analysis. The research - the new reference-free method of evaluating anti-radical activity, a number of natural and synthetic antioxidants. Established values antiradical activity of natural and synthetic antioxidants. Various embodiments of the mechanisms of inhibition of free radicals and their antioxidant dependency structure.

The objects of research are low-molecular antioxidants, their mixtures and extracts of plant raw materials. Objective: To determine the antioxidant activity of tea and extracts of medicinal plant raw materials by potentiometric method using hexacyanoferrate (III) potassium as an oxidant model. The paper presents the causes of oxidative stress, exogenous antioxidants of natural origin, describes methods for determining the integral index of antioxidant activity. In the process of work, the antioxidant activity of low-molecular antioxidants was determined by potentiometric method using hexacyanoferrate (III) of potassium as an oxidizer model, potentiometric titration, photometric titration. As a result of the study, the antioxidant activity of extracts of various tea varieties and herbal plant raw materials was determined by the potentiometric method using potassium hexacyanoferrate (III) as an oxidant model. / Цель работы: определение антиоксидантной активности чая и экстрактов лекарственного растительного сырья потенциометрическим методом с использованием гексацианоферрата (III) калия в качестве модели окислителя. В работе представлены причины возникновения окислительного стресса, экзогенные антиоксиданты природного происхождения, описаны методы определения интегрального показателя антиоксидантной активности. В процессе работы определена антиоксидантная активность низкомолекулярных антиоксидантов потенциометрическим методом с использованием гексацианоферрата (III) калия в качестве модели окислителя, потенциометрическим титрованием, фотометрическим титрованием. В результате исследования определена антиоксидантная активность экстрактов различных сортов чая и лекарственного растительного сырья потенциометрическим методом с использованием гексацианоферрата (III) калия в качестве модели окислителя.

No / Flavonoids, of which flavones are a sub-group, are plant secondary metabolites found in a variety of natural food sources (e.g., vegetables) and wines. They elicit beneficial roles in health and disease through their antioxidant activity, but some of them have also now been found to exert specific effects on cell signalling. We recently showed that methoxylation of the flavone structure at the 4ʹ position, or additionally at the 3ʹ position, to produce 4ʹ-methoxyflavone (4MF) and 3ʹ,4ʹ-dimethoxyflavone (DMF), respectively, significantly enhanced activity against the cell death (“parthanatos”) mediated by poly (ADP-ribose) polymerase (PARP). We report here our attempt to correlate the antioxidant and parthanatos-inhibitory activities of these methoxylated flavones with those of the hydroxylated flavonoids. Cultures of HeLa and HaCaT cells were exposed to MNNG (50µM, up to 25min), which induces parthanatos, and the oxidant hydrogen peroxide (100µM – 2mM, up to 24h). The effects (up to 20µM) of the methoxylated flavones 4MF and DMF, the hydroxylated flavone luteolin (LN), and the non-flavone flavonoids quercetin (QE), naringin (NG) and epigallocatechin gallate (EGCG) on the reduction in viability (indicative of cell death) and morphological changes induced by MNNG or peroxide were then investigated. Both alamar blue and MTT assays were used to quantify viability. MNNG induced significant reduction in cell viability, which was not affected by the pan-caspase inhibitor Z-VAD-fmk but significantly blocked by DPQ, a PARP-1 inhibitor, consistent with the biochemical profile of parthanatos. Hydrogen peroxide also elicited a significant decrease in cell viability, with partial or no protection afforded by either Z-VAD-fmk or DPQ (dependent on peroxide concentration and treatment duration). 4MF and DMF demonstrated significant protection against MNNG-induced cell death but LN, QE, NG and EGCG showed little or no protection. On the other hand, 4MF and DMF elicited mostly negligible effects against hydrogen peroxide, whereas LN, QE, NG and EGCG elicited various levels of protection against it. We conclude that methoxylation at the 4ʹ or 3ʹ, 4ʹ positions of flavones favours anti-parthanatos but not antioxidant activity, whereas hydroxylation enhances antioxidant but not anti-parthanatos activity. / Abstract of conference paper.

<p>U okviru ove disertacije su ispitane mogućnosti primene savremenih postupaka za dobijanje ekstrakata rtanjskog čaja (Satureja montana L.).<br />U ekstraktima dobijenim primenom superkritičnog ugljendioksida (različitih gustina) utvrđeno je prisustvo karvakrola kao najzastupljenije komponente. Ekstrakt sa najvećim sadržajem karvakrola je dobijen pri uslovima 350 bar i 50 &deg;C (60,82%), dok je najveći prinos karvakrola ostvaren pri pritiscima 325 i 350 bar i temperaturi 60 &deg;C, i iznosio je 2,4 g/100 g droge. Primenom ugljendioksida na većim pritiscima se ostvaruje efikasnija ekstrakcija karvakrola iz droge, kao i proizvodnja ekstrakata sa većim sadržajima karvakrola. Druge komponente prisutne u ekstraktima u znatno nižim koncentracijama su: p-cimen, borneol, trans-kariofilen, kariofilen-oksid, &gamma;-terpinen i linalool.<br />Da bi se utvrdila mogućnost unapređenja prinosa ekstrakcije superkritičnim ugljendioksidom, kao i pobolj&scaron;anja kvaliteta ekstrakata, ispitan je uticaj različitih predtretmana biljnog materijala. Ustanovljeno je da predtretman vodom najznačajnije povećava prinos ekstrakcije (za 25%), dok je ultrazvučni predtretman najadekvatniji izbor za dobijanje ekstrakata sa najvećim sadržajem karvakrola (66,46%). Međutim, najveći prinos ekstrakcije karvakrola iz 100 g droge postignut je primenom etanola i vode kao predtretmana.<br />Ekstrakti dobijeni primenom subkritične vode (uz variranje temperature i vremena ekstrakcije) su hemijski okarakterisani i radi utvrđivanja uslova ekstrakcije pri kojima se postiže najveći kvalitet ekstrakata u pogledu sadržaja polifenolnih komponenti, kao i antioksidantne aktivnosti, primenjena je metoda odzivne povr&scaron;ine (RSM). Utvrđena je prednost primene subkritične vode u odnosu na klasičnu metodu ekstrakcije u pogledu sadržaja polifenolnih komponenti i antioksidantne aktivnosti.<br />Suvi ekstrakti S. montana dobijeni spray drying tehnologijom su&scaron;enja (sa različitim koncentracijama maltodekstrina) su ispitani u pogledu fizičkih i hemijskih osobina. Takođe, radi ispitivanja mogućnosti primene u vidu funkcionalnih ili prehrambenih proizvoda, izvr&scaron;ena je senzorna analiza dobijenih suvih ekstrakata, i analizirana je njihova farmakolo&scaron;ka aktivnost u smislu ACE inhibitornog i antioksidantnog delovanja.<br />Ustanovljeno je da je najniža koncentracija maltodekstrina (10%) najadekvatnija za dobijanje suvog ekstrakta S. montana sa najvećim sadržajem polifenolnih komponenti, sadržajem etarskog ulja i u pogledu antioksidantne i ACE inhibitorne aktivnosti. Takođe, suvi ekstrakt sa 10% maltodekstrina ispunjava i ostale fizičko-hemijske parametre, kojima se osigurava kako efikasno su&scaron;enje, tako i kvalitet ekstrakta.<br />In vivo aktivnost odabranih ekstrakata S. montana ispitana je na modelu hepatotoksičnosti indukovane primenom ugljentetrahlorida. Analiziran je uticaj ekstrakata na funkciju jetre, enzimske parametre oksidativnog stresa i serumske parametre oksidativnog o&scaron;tećenja izazvanog ugljentetrahloridom i ustanovljen je hepatoprotektivni učinak ekstrakata S. montana. Takođe, ispitano je dejstvo ekstrakata na rast Ehrlich-ovih ascitnih tumora implantiranih mi&scaron;evima, kao i na biohemijske parametre oksidativnog stresa u ćelijama tumora i utvrđeno je da je vreme aplikacije ekstrakata od izuzetnog značaja.</p> / <p>This dissertation investigates the possibilities of applying contemporary processes for obtaining extracts of winter savory (Satureja montana L.).<br />Carvacrol was determined to be the most abundant component in extracts acquired by using supercritical carbon dioxide (of various densities). Extract with the richest content of carvacrol was obtained under conditions of 350 bar and 50 &deg;C (60,82%), while the highest carvacrol yield was produced under 325 bar and 350 bar pressures and at a 60 &deg;C temperature reaching 2.4 g/100 g of dry weight. By applying carbon dioxide on higher pressures, a more efficient carvacrol extraction from herbal material is performed, alongside production of extracts with higher carvacrol content. Other components in the extracts present in significantly lower concentrations are the following: p-cymen, borneol, trans-caryophyllene, caryophyllene-oxide, &gamma;-terpinene and linalool.<br />In order to recognize the possibility of improving the extraction yield produced by employing supercritical carbon dioxide and enhancing the quality of extracts, the influence of several different pre-treatments was investigated. It was determined that the water pre-treatment raises the extraction yield the most (for 25%), while the ultrasonic pre-treatment was the most adequate choice for acquiring extracts with the highest content of carvacrol (66.46%). However, the highest carvacrol extraction yield from 100 g of dry weight was achieved by applying ethanol and water as pre-treatment.<br />Extracts which were attained with subcritical water (by varying the temperature and time of extraction) were chemically characterized and Response Surface Method (RSM) was applied for the purposes of determining conditions of extractions under which the highest quality of extracts in terms of content of polyphenolic compounds and antioxidant activity is reached. The advantage of employing subcritical water over the classical method of extraction in respect of content of polyphenolic components and antioxidant activity was demonstrated.<br />Dry extracts of S. montana obtained by using the spray drying technology (with different concentrations of maltodextrine) were examined in terms of their physical and chemical characteristics. Also, in order to investigate the possibility of their use as functional or food products, sensory analysis of attained extracts was performed, and furthermore, their pharmacological activity in terms of the ACE inhibitory and antioxidant effect was analysed as well.<br />It was determined that the lowest concentration of maltodextrin (10%) was the most adequate for obtaining dry extracts of S. montana with the richest content of polyphenolic components and essential oil, and in respect of antioxidant and ACE inhibitory activities. Also, dry extract with 10% of maltodextrin meets all other physical-chemical parameters, which provide efficient drying and quality of extracts.<br />In vivo activity of selected extracts of S. montana was examined on a model of hepatotoxicity induced by applying carbon tetrachloride. The influence of extracts on liver function, enzyme parameters of oxidative stress, and serum parameters of oxidative damage caused by carbon tetrachloride was analysed and hepatoprotective effect of S. montana extracts was determined. In addition, the influence of extracts on the growth of Ehrlich ascite tumors implanted in mice was investigated, and their impact on biochemical parameters of oxidative stress in tumor cells as well, and it was determined that the time of application of extracts is of great significance.</p>

<p>Cilj ispitivanja čiji su rezultati prikazani u ovoj doktorskoj disertaciji bio<br />je da se ispita mogućnost dobijanja kombuha napitka od/sa CoffeeBerry&reg;<br />ekstraktom i da se ispitaju njegove funkcionalne karakteristike.<br />CoffeeBerry&reg; ekstrakt kao bogat izvor biolo&scaron;ki aktivnih jedinjenja bi<br />obezbedio dodatne funkcionalne karakteristike kombuha napitku u<br />poređenju sa onim pripremljenim na tradicionalan način od zaslađenog<br />crnog čaja. U disertaciji je nakon optimizacije sastava podloge za<br />kultivaciju čajne gljive ispitana (in vitro) antioksidativna, antimikrobna i<br />citotoksična aktivnost kombuhe/fermentacione tečnosti obogaćene<br />CoffeeBerry&reg; ekstraktom. Antioksidativna aktivnost je ispitana ESR<br />(elektron-spin rezonantnom) spektroskopijom na reaktivne hidroksil i<br />stabilne DPPH (1,1-difenil-2-pikrilhidrazil) radikale, kao i kvalitativni i<br />kvantitativni sastav fenolnih jedinjenja, potencijalnih nosilaca biolo&scaron;ke<br />aktivnosti (HPLC metodom). Antimikrobna aktivnost je ispitana agardifuzionom<br />metodom na odabrane referentne i izolovane (&bdquo;divlje&ldquo;)<br />sojeve bakterija (Gram-pozitivnih i Gram-negativnih) i kvasaca.<br />Citotoksična aktivnost uzoraka na rast odabranih ćelijskih linija: Hep2c<br />(Human larynx carcinom), RD (Rhabdomyosarcoma) i L2OB (mi&scaron;ija<br />tumorska fibrolastna linija u koju su transfektovani neki humani geni)<br />ispitana je MTT testom. U navedenim ispitivanjima je tradicionalna<br />kombuha dobijena od zaslađenog crnog čaja kori&scaron;ćena kao kontrolni<br />uzorak. Rezultati su pokazali da CoffeeBerry&reg; ekstrakt značajno<br />doprinosi antioksidativnoj i citotoksičnoj aktivnosti napitka &scaron;to govori o<br />punoj opravdanosti upotrebe ovog ekstrakta kao funkcionalnog dodatka<br />podlozi za kultivaciju čajne gljive. Kombuha napici od crnog čaja i sa<br />dodatkom CoffeeBerry&reg; ekstrakta pokazali su približno isto delovanje na<br />bakterijske sojeve, dok je delovanje na kvasce izostalo u potpunosti.<br />Primarni nosilac antibakterijske aktivnosti napitaka je sirćetna kiselina.</p> / <p>The aim of this study was to investigate fermentation of sweetened<br />medium prepared with CoffeeBerry&reg; еxtract and functional<br />characteristics of a kombucha beverage enriched with CoffeeBerry&reg;<br />еxtract. Total phenol concentration in unfermented samples,<br />fermentation broths and kombucha beverages made of CoffeeBerry&reg;<br />еxtract and black tea was determined spectrophotometrically whereas<br />qualitative and quantitative concentration of polyphenolic compounds<br />was determined by HPLC method. Antioxidant activity on DPPH and<br />hydroxyl radicals in the same samples was determined on an ESR<br />spectrometer. Fermentation broth and kombucha beverage enriched with<br />CoffeeBerry&reg; еxtract had higher antioxidant activity against both<br />radicals than control samples. Antimicrobial activity of kombucha<br />beverages optimal acidity was tested against selected strains of bacteria<br />(Gram-positive and Gram-negative) and yeast. The main active<br />component of antibacterial activity was acetic acid, but samples did not<br />show any activity against yeast Candida albicans and Saccharomyces<br />cerevisiae. Antiproliferative activity of fermentation broth with and<br />without CoffeeBerry&reg; еxtract was measured by MTT test on following<br />cell lines: Hep2c (Human larynx carcinom), RD (Rhabdomyosarcoma) i<br />L2OB (murine tumor fibrolast lines trаnsfеcted by some human genes).<br />The highest antiproliferative activity shown fermentation broth enriched<br />with CoffeeBerry&reg; еxtract, wherein Hep2c cells was the most<br />susceptible.</p>

Dans ce travail nous avons réalisé une évaluation de la composition et des activités biologiques des HEs extraites de L. stoechas récoltées sur 11 sites dans le nord de l'Algérie. Les HEs ont été analysées par GC/FID et GC/MS, où un total de 121 composés ont été identifiés, représentant de 69.88 à 91.2% du contenu total de l'huile. Les principaux constituants étaient le fenchone (11.27-37.48%), le camphre (1.94-21.8%), le 1,8-cinéol (0.16-8.71%) et le viridiflorol (2.89-7.38%). Les activités biologiques in vitro ont démontré que les activités de piégeage du radical DPPH et l’oxydation des lipides du couple β- carotène/acide linoléique différaient d’un facteur 8 et étaient liées à différents ensembles de molécules. Nos 11 HEs ont présenté de bonnes activités antimicrobiennes envers la plupart des 16 souches pathogènes testées à des valeurs de concentration minimale inhibitrice (CMI) allant de 0.16 à 3.5 mg/ml. A l’aide d’amorces dégénérées nous avons isolé trois ADNc en pleine longueur, LpFENS, LpPINS et LpGEAS, de feuilles de L. pedunculata. Six ADNc homologues en pleine longueur, LSFENS, LsPINS, LsGEAS, LvFENS, LvPINS et LvGEAS, ont été également isolés des feuilles de L. stoechas et L. viridis en utilisant des amorces spécifiques aux trois premiers ADNc clonés. L’expression hétérologue dans E. coli et l’analyse de l’activité catalytique par GC/MS des enzymes natives recombinantes purifiées de ces TPSs ont permis leur caractérisation fonctionnelle en tant que, α-fenchol synthases (LpFENS, LsFENS et LvFENS), α-pinène synthases (LpPINS, LsPINS et LvPINS) et germacrène A synthase (LpGEAS, LsGEAS et LvGEAS) / In this work, we have assessed the composition and the biological activities of essential oils (EO) extracted from L. stoechas collected from 11 locations in northern Algeria. The oils were analyzed by GC/FID and GC/MS. A total of 121 compounds were identified that accounted for 69.88-91.2 % of the total oil contents. Major components were fenchone (11.27-37.48 %), camphor (1.94-21.8 %), 1,8-cineol (0.16-8.71 %) and viridiflorol (2.89-7.38 %). In vitro biological activities demonstrated that the DPPH-based radical scavenging and the β-carotene/linoleic acid-based lipid anti-oxidation activities differed by an 8 fold factor and were linked to different sets of molecules in different EOs. Our 11 EOs exhibited good antimicrobial activities against most of the 16 tested strains of pathogenic bacteria, filamentous fungi and yeast at minimum inhibition concentration (MIC) values of 0.16 to 3.5 mg/ml. The use of degenerate primers allowed us to isolate three full-length cDNAs, LpFENS, LpPINS and LpGEAS, from the leaves of L. pedunculata. Six full-length homologous cDNAs, LsFENS, LsFENS, LsPINS, LsGEAS, LvFENS, LvPINS and LvGEAS were also isolated from the leaves of L. stoechas and L. viridis with specific primers designed on the cDNAs previously cloned. Heterologous expression in E. coli and GC/MS analysis of the catalytic activity of native recombinant enzymes of these TPSs have led to their functional characterization as α-fenchol synthases (LpFENS, LsFENS and LvFENS), α-pinene synthases (LpPINS, LsPINS and LvPINS) and germacrene A synthases (LpGEAS, LsGEAS and LvGEAS)

Consommateurs, chercheurs et industriels cherchent de plus en plus à associer à la valeur nutritionnelle des aliments, un effet bénéfique pour la santé. Les aliments caractérisés par des pouvoirs antioxydants élevés semblent correspondre à cette demande. En effet, plusieurs travaux mentionnent les bénéfices santé de ces aliments. Pour répondre à cette demande les industriels mettent en avant ce critère. La fonctionnalité alimentaire, aux bénéfices prometteurs pour la santé publique, reste un concept néanmoins complexe dans la pratique. Ce travail met l’accent sur le fait que les propriétés fonctionnelles d’un aliment ne sont pas directement liées à la composition en ingrédients. L’évaluation du pouvoir antioxydant d'un aliment est dans la plupart des cas réalisée à partir des valeurs des différents ingrédients avant formulation et mise en œuvre des procédés. Pourtant, plusieurs phénomènes sont susceptibles de modifier le pouvoir antioxydant lors de la durée de vie d'un produit alimentaire. D’un point de vue industriel, il est nécessaire de développer une méthode de prédiction de cette activité. Les objectifs de ce travail étaient d’adapter une méthode reproductible et facile à réaliser de quantification du pouvoir antioxydant sur des aliments complexes et de comparer cet index d'un ensemble de produits contenant une large gamme de matières premières. Enfin, a été discuté l’intérêt que représente, dans le domaine des propriétés anti oxydantes, la quantification de l’index créé. Au moment où les industriels vont être amenés à proposer des allégations liées à l’effet bénéfique de l’aliment choisi parmi un échantillon de plus en plus ample / Consumers, researchers and industrialists try more and more to associate with the nutritional value of food, a beneficial effect for the health. Food characterized by high antioxidant powers seems to correspond to this demand. From an industrial point of view, it is necessary to develop methods of predicting this antioxidant capacity. The objectives of this work were to adapt a method reproducible and easy to realize of quantification of the antioxidant power on complex food and to compare this index of a set of products containing a wide range of raw materials. Until now, it has been considered that the use of raw materials characterized by a high antioxidant capacity also leads to a preparation with a high antioxidant activity. We evaluated many of the factors affecting the antioxidant activity of convenience foods (phenol content, effects of formulation, culinary reheating, and preservation) and to provide data on convenience foods consumed by the French population. The total antioxidant capacity of the ethanolic extracts was evaluated by the method of the equivalent Trolox (TEAC) using the radical cation ABTS•+. The concentration of the total phenolic compounds of the same extracts was determined by the Folin-Ciocalteu method. The results show that the food matrix is an important factor for the modulation of activities of antioxidants. A standardised testing protocol for evaluating antioxidative effects is necessary. Then, we discussed the interest of an index, as the industrialists are going to be brought to claim to the beneficial effect of the food chosen among a more and more ample sample

Dans cette étude nous avons évalué les activités biologiques de la mûre tropicale de montagne (Rubus adenotrichos) et les effets de la digestion ou des procédés de pasteurisation sur ces activités. L'extrait polyphénolique de mûre présente un pouvoir antioxydant déterminé par la méthode ORAC de 4339 ± 144 pM TE/g, plus élevé que celui de la quercétine et de l'acide ellagique. L'extrait de mûre présente des propriétés protectrices des liposomes et des homogénats de foie contre la peroxydation lipidique avec des IC50 de 7.0 ± 0.5 et 20.3 ± 4.2 mg/ml, respectivement. Les polyphénols de mûre inhibent la production de nitrite dans par les macrophages murins J774A.1 stimulées par le mélange LPS, IFN. Cette activité est confirmée par la diminution du taux de NO synthase inductible dans ces mêmes cellules. Les activités biologiques des fruits pourraient être affectés par les procédés industriels. Le processus de pasteurisation du jus de mûre a provoqué une diminution d'environ 7% de la concentration en anthocyanes pour la température la plus élevée évalué, 92°C. Pour le même échantillon, une réduction significative de 27% de l'activité anti-oxydante mesurée par la méthode DPPH et de 16% de la capacité de complexassions des radicaux NO. Toutefois, aucune différence significative n'a été mise en évidence autant pour la capacité d'inhibition de la peroxydation que pour l'activité anti-oxydante intracellulaire. Les effets in vivo sont largement dépendants de la biodisponibilité des composés phénoliques et de ce fait un jus de mûre a été digéré in vitro pour obtenir des échantillons qui représentent les grandes étapes du processus de digestion. La digestion gastrique a peu d'influence au contraire de la digestion intestinale sur la composition polyphénolique de l'extrait de mûre en particulier au niveau du rapport anthocyanines/elagitannins. La fraction dialysable présente une réduction de 33% et de 35% de l'activité contre les radicaux dérivés de l'azote DPPH et NO, respectivement. De même, la capacité d'inhiber au niveau intracellulaire les radicaux libres (ROS) a diminué d'environ 85%. Toutefois, aucune différence significative n'a été mise en évidence au niveau de la capacité d'inhiber la peroxydation lipidique. Finalement, cette étude a utilisé un modèle in vivo pour évaluer l'effet de la consommation d'une boisson de mûre chez des rats diabétiques. L'apport de mûre a provoqué une diminution du glucose (-48.6%), des triglycérides (-43.5%) et du taux de cholestérol (-28.6%) plasmatique des rats diabétiques par rapport aux témoins diabétiques qui n'ont pas consommé la boisson de ce fruit. De plus, les rats diabétiques qui consomment de la mûre ont vus améliorées les valeurs de la capacité antioxydante du plasma (+7%), tandis que les niveaux de peroxydation lipidique dans le plasma (-19%) et dans les tissus rénaux (-23%) ont été réduits. Cette étude suggère que l'incorporation de la mûre comme un complément alimentaire pourrait avoir des effets bénéfiques sur la santé, même si l'on considère les aspects de biodisponibilité et les effets du procédé de pasteurisation. / This study evaluated biological activities of blackberries (Rubus adenotrichos) and the effects of digestion or pasteurization processes on these activities. The blackberries phenolic extract showed an ORAC value of 4339 ± 144 μM TE/g, higher than those of quercetin and ellagic acid. It also protected liposomes and liver homogenates against lipid peroxidation; with IC50 of 7.0 ± 0.5 and 20.3 ± 4.2 μg/mL, respectively. The blackberry polyphenols inhibited nitrite production in J774A.1 cells stimulated with LPS+IFNγ due to down-regulation of iNOS protein expression, suggesting an anti-inflammatory potential. The biological activities of the fruits could be affected by the industrial processes. The pasteurization of a blackberry juice caused a 7% decrease in the anthocyanins concentration of the highest temperature tested, 92 °C. For the same sample, a significant reduction of 27% and 15% were showed in the DPPH and NO scavenging capacity, respectively. However, non-significant differences were evident neither for the peroxidation inhibitory capacity nor for the intracellular antioxidant activity after the pasteurization step. The antioxidant capacity is largely dependent of their bioavailability and because of this a blackberry juice was digested in vitro to obtain samples that represent the major stages of the digestion process. The gastric digestion had slight impact on polyphenolic composition while the pancreatin/bile digestion had marked effects on polyphenolic composition, particularly in the ratio anthocyanins-ellagitannins. These changes in phenolic composition caused in the dialyzed fraction a reduction of 33% and 35% in the activity against nitrogen derived radicals DPPH and NO respectively. As well the capacity to inhibit intracellular ROS was decrease by an 85% in the dialyzed fraction. Nevertheless, no significant differences were evident in the capacity to inhibit lipid peroxidation. Finally, this study used an in vivo model with diabetic rats to evaluate the effect of the consumption of a blackberry beverage. The blackberry intake provoked a decrease in plasmatic glucose (-48.6%), triglycerides (-43.5%) and cholesterol (-28.6%) levels in the diabetic rats with respect to the diabetic controls animals. The diabetic rats which consumed blackberry, improved (+7%) the plasma antioxidant capacity (PAC) values and reduced the levels of lipid peroxidation in plasma (-19%) and in kidney tissue (-23%). This study suggests that selecting blackberry as a dietary complement could have health beneficial effects even when considering the bioavailability aspects of the phenolic compounds and the pasteurization effects.

Les objectifs de cette thèse sont d’une part d'étudier les effets d’un traitement thermique et de l’environnement physico-chimique sur la stabilité de 6 flavonoïdes de structure différente et sur l'évolution de leur activité anti-oxydante. Les conditions du traitement thermique ont été les suivantes : (i) chauffage dans des conditions isothermes durant 2h pour des températures allant de 30 à 130°C et (ii) chauffage dans des conditions non isothermes par microcalorimétrie (de 30 à 130°C, 4°C/ heure). Les flavonoïdes ont été solubilisés dans de l’eau. Nous avons constaté que les flavonoïdes glycosylés sont plus résistants que les flavonoïdes aglycones. Les énergies d'activation de dégradation calculées dépendent aussi de la structure du flavonoïde. Pour se dégrader, les flavonoïdes glycosylés ont besoin d’une énergie élevée par rapport à la forme aglycone. L’exposition à la lumière a été réalisée durant 15 jours avec et sans oxygène, le témoin de l’expérience étant un stockage à l’obscurité avec et sans oxygène. La dégradation des flavonoïdes est influencée par la présence de lumière et par la quantité d'oxygène. Les molécules ont une sensibilité différente en fonction de leur structure, le classement suivant est obtenu d’après : naringine, ériodictyol puis rutine, lutéoline, lutéoline 7-O-glucoside et enfin le mesquitol. En effet, la présence d'un groupe hydroxyle en position 3 et une double liaison C2-C3 diminue la stabilité des flavonoïdes. En outre, il a été observé que, malgré la dégradation totale de certains flavonoïdes par le traitement thermique et l’environnement physico-chimique, les solutions traitées conservent une activité anti-oxydante / The objectives of this thesis are to study the effects of a heat treatment and the physicochemical environment on the stability of 6 flavonoids of different structure and on the evolution of their antioxidant activity. The heat treatment conditions were as follows: (i) heating under isothermal conditions for 2 h at temperatures ranging from 30 to 130 ° C and (ii) heating under non-isothermal conditions by microcalorimetry (30 to 130 ° C, 4 ° C / hour). The flavonoids were solubilized in water. We found that the glycosylated flavonoids are more resistant than the aglyconic flavonoids. The calculated degradation activation energies also depend on the structure of the flavonoid. To degrade, glycosylated flavonoids require high energy relative to the aglycone form. Exposure to light was carried out for 15 days with and without oxygen, the experimental control being a dark storage with and without oxygen. The degradation of flavonoids is influenced by the presence of light and by the amount of oxygen. The molecules have a different sensitivity according to their structure, the following classification is obtained according to: naringine, ériodictyol then rutin, luteolin, luteolin 7-O-glucoside and finally the mesquitol. Indeed, the presence of a hydroxyl group at position 3 and a C2-C3 double bond reduces the stability of the flavonoids. Furthermore, it has been observed that, despite the total degradation of certain flavonoids by the heat treatment and the physical-chemical environment, the treated solutions retain an antioxidant activity