<p>Prisustvo organskih zagađujućih supstanci (farmaceutski aktivnih komponenata i prirodnih toksina‐mikotoksina) u uzorcima životne sredine i namirnicama je u porastu kao posledica novih industrijskih procesa i ostalih antropogenih aktivnosti, kao i klimatskih promena. Takođe veliku pažnju javnosti privlače i neorganske zagađujuće supstance kao &scaron;to su te&scaron;ki elementi. S obzirom da zagađujuće supstance imaju negativan uticaj na životnu sredinu i zdravlje ljudi, u svetu se preduzimaju mere u cilju smanjenja stepena izloženosti toksičnim jedinjenjma i posledicama izlaganja. Trenutno, jedan od najvećih izazova, jeste procena rizika povezana sa velikim brojem zagađujućih supstanci u tragovima ili u tzv. ultratragovima, uključujući &ldquo;novo&rdquo; otkrivena zagađujuća jedinjenja, a jedan od osnovnih trendova je razvoj i primena brzih i efikasnih metoda za njihovu analizu u ispitivanim uzorcima na bazi naprednih hromatografskih i spektrometrijskih tehnika.<br />Tehnike bazirane na tečnoj hromatografiji sa različitim masenim analizatorima za<br />kvantifikaciju organskih zagađujućih supstanci kao i metode zasnovane na atomskoj<br />apsorpcijonoj spektrometriji za određivanje ultratragova neorganskih zagađujućih supstanci postale su referentne na međunarodnom nivou. Ovakve napredne spregnute tehnike postale su važne za identifikaciju, kvantifikaciju i praćenje različitih zagađujućih supstanci u uzorcima životne sredine i namirnicama i proceni njihovog &scaron;tetenog uticaja na zdravlje čoveka. S obzirom da su u literaturi retka istraživanja koja se bave razvojem i primenom metoda zasnovanih na naprednim hromatografskim i spektrometrijskim tehnikama i određivanju organskih i neorganskih zagađujućih supstanci u matriksima životne sredine i namirnicama sa prostora zapadnog Balkana, a uzimajući u obzir njihovu važnost, specifični ciljevi disertacije su:<br />&bull; unutra&scaron;nja (&bdquo;inhouse&ldquo;)<br />provera kvaliteta i pouzdanosti postojeće &bdquo;multi‐rezidualne&ldquo;<br />metode zasnovane na UHPLC‐QqLIT‐MS/MS za analizu 81‐e farmaceutski aktivne<br />komponente (PhAC) u otpadnoj, povr&scaron;inskoj, podzemnoj i pijaćoj vodi i po prvi put<br />dobijanje sveobuhvatnih rezultata njihovog prisustva u različitim tipovima vode sa<br />područja Srbije;<br />&bull; unutra&scaron;nja (&bdquo;inhouse&ldquo;) provera kvaliteta i pouzdanosti postojeće &bdquo;multi‐toksin&ldquo;<br />metode za analizu 8 Fusarium mikotoksina u uzorcima ozime p&scaron;enice različitih sorti<br />zasnovane na HPLC‐QqQ‐MS/MS radi određivanja regionalnih razlika između žitnih<br />regiona kao i otpornosti ispitivanih sorti p&scaron;enice na kontaminaciju Fusarium<br />toksinima;<br />&bull; modifikacija postojeće &bdquo;multi‐toksin&ldquo; metode zasnovane na UHPLC‐QqQ‐MS/MS za<br />analizu 11 osnovnih mikotoksina u uzorcima bra&scaron;na i njena unutra&scaron;nja (&bdquo;inhouse&ldquo;)<br />provera kvaliteta i pouzdanosti, kao i provera kroz interlaboratorijsko poređenje,<br />radi dobijanja podataka za procenu &scaron;tetnog uticaja ispitivanih mikotoksina na<br />zdravlje populacije;<br />&bull; razvoj &bdquo;multi‐toksin&ldquo; (vi&scaron;ekomponentne) i &bdquo;multi‐matriks&ldquo; (za vi&scaron;e matriksa) metode<br />bazirane na UHPLC‐QqQ‐MS/MS za analizu 10 mikotoksina u različitim vrstama<br />ko&scaron;tuničavog voća čija provera kvaliteta je zasnovana na intralaboratorijskoj proveri<br />tačnosti i preciznosti dobijenih rezultata;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &bull; primena postojeće analitičke procedure zasnovane na naprednoj tehnici pripreme<br />(mikrotalasnoj digestiji) različitih uzoraka biljnog i životinjskog porekla i provera<br />kvaliteta metode identifikacije i kvantifikacije zasnovane na atomskom apsorpcionom spektrometru sa grafitnom kivetom (GFAAS) radi dobijanja sveobuhavatnih rezultata o prisustvu te&scaron;kih elemenata (arsena, olova i kadmijuma) radi procene izloženosti stanovni&scaron;tva Srbije toksičnim neorganskim elementima.<br />Postignuti rezultati predstavljaju jedinstvene rezultate za područje Srbije dobijene<br />primenom naprednih spregnutih tehnika koje imaju značajnu ulogu u praćenju prisustva većeg broja organskih i neorganskih zagađujućih supstanci u izabranim uzorcima životne sredine i namirnica, (regulisanih postojećim zakonodavstvom) radi procene stepena zagađenosti ili u slučaju jedinjenja koja nisu regulisana zakonodavstvom radi sticanja novih saznanja o njihovom prisustvu i proceni mogućeg negativnog uticaja na životnu sredinu i zdravlje populacije.</p> / <p>The presence of organic pollutants in environmental samples and food (pharmaceutically active components and natural toxins‐mycotoxins) is increased as a result of new industrial processes and other anthropogenic activities, as well as climate change. Similarly heavy elements as inorganic pollutants have attracted worldwide attention. Since, these pollutants have negative impact on environment and human health, extremely efforts are undertaken in the world to reduce the level of exposure to these pollutants and consequences of the exposure. Currently, one of the highest challenges is to assess the risk associated with a large number of pollutants in trace or ultra trace levels, including &quot;new&quot; (emerging) discovered pollutants, and one of the main trends is development and implementation of fast and efficient methods for their analysis on the basis of advanced chromatographic and spectrometric techniques. Therefore, coupled techniques have become important for the identification, quantification and monitoring of various pollutants in environmental samples and food and assessment of their hazard impact on human health. Since, there are scarce data about the development and application of advanced methods based on chromatographic and spectrometric techniques for determination of organic and inorganic pollutants in environmental matrices and food from the Western Balkan, and taking into account their importance, specific objectives of<br />the dissertation were: &bull; internal (&quot;in‐house&quot;) quality control of the existing &quot;multi‐residual&quot; method based on UHPLC‐QqLIT‐MS/MS for analysis of 81 pharmaceutically active components (PhAC) in wastewater, surface, underground and drinking water due to obtained for the first time comprehensive results of their presence in different types of water from Serbia; &bull; internal (&quot;in‐house&quot;) quality control of the existing &quot;multi‐toxin&quot; method for the analysis of 8 Fusarium mycotoxins in samples of different winter wheat cultivars based on HPLC‐QqQ‐MS/MS to determine the differences among wheat-growing regions as well as the resistance of the analysed wheat cultivars towards Fusarium toxins; &bull; modification of existing &quot;multi‐toxin&quot; method based on UHPLC‐QqQ‐MS/MS for analysis of 11 principal mycotoxins in samples of flour and its internal (&quot;in‐house&quot;) quality control as well as verification through the interlaboratory comparison, in order to obtain data for assessing the hazard effect of these mycotoxins on the health of the population; &bull; the development of &quot;multi‐toxin&quot; and &quot;multi‐matrix&quot; method based on UHPLC‐QqQMS/ MS for the analysis of 10 mycotoxins in various types of nuts based on intralaboratory verification of the accuracy and precision of the obtained results; &bull; application of analytical procedure based on advanced preparation technique (microwave digestion) and atomic absorption spectrometer with a graphite furnace (GFAAS) and its verification in order to obtain comprehensive results on the presence of heavy elements (arsenic, lead and cadmium) in different samples of plant and animal origin to assess the exposure of the Serbian population to toxic inorganic elements. The obtained results are unique for the Serbia. They are obtained by applying advanced coupled techniques that have a significant role in monitoring the presence of a numerous organic and inorganic pollutants in analyzed samples of the environment and food. The presented results contribute to the assessment of pollution degree and in the case of new (emerging) not regulated polutant they might give new information about the possible negative impact on the environment and health of the population.</p>

L’un des objectifs du programme de recherche Opticook est d’équiper les fours avec des capteurs spectroscopiques qui permettront de piloter de façon non intrusive la cuisson de viande et de poisson. Le développement de ces capteurs est basé sur l’hypothèse que les protéines de ces aliments sont de bons marqueurs du degré de cuisson. Dans ce contexte, dans un premier temps, plusieurs outils ont été mis en œuvre pour caractériser l’effet de différents barèmes de cuisson sur les propriétés macroscopiques (texture, couleur) d’échantillons de bœuf, de poulet et de dos de cabillaud. Puis, pour étudier les transformations que subissent ces échantillons à l’échelle moléculaire, nous avons utilisé la calorimétrie et différentes spectroscopies. La spectroscopie dans le domaine du visible nous a permis d’observer la dénaturation des hémoprotéines grâce à l’évolution de l’état (oxy à met ). Ces protéines sont responsables du changement de couleur observé entre 50 et 75 °C. Les spectroscopies dans le moyen infrarouge et de fluorescence ont été utilisées pour suivre les changements de structures associées à la dénaturation des protéines fibrillaires. La myosine, purifiée à partir du blanc de poulet, a été étudiée en fonction de la température par fluorescence. Les résultats nous ont montré que des évolutions de la structure protéique se produisent dès 50 °C. À l’échelle microscopique, nous avons réalisé des expériences sur des échantillons de bœuf par RMN bas champ (séquences d’impulsion et gradient de champ), et par tomographie de neutrons pour rendre compte de la contraction des fibres et de la libération du jus selon le degré de cuisson. L’analyse des images 3D enregistrées par tomographie de neutrons et le traitement des données de RMN montrent des changements importants de la morphologie et de la densité de fibres des échantillons après la cuisson à haute température. Pour associer les deux approches (moléculaire et microscopique) des mesures couplées de spectroscopie et d’imagerie de neutrons ont été menées dans le four Opticook pendant la cuisson de tranches de bœuf. Ce couplage nous a permis de suivre l’évolution de la morphologie due à la contraction des fibres, la migration du jus, ainsi que le changement de couleur associé à la dénaturation des hémoprotéines. Nous en concluons qu’il est en effet possible de suivre le degré de cuisson de viande et de poisson à l’aide de différentes spectroscopies (la fluorescence et le visible) grâce aux signatures spectroscopiques de plusieurs marqueurs internes à ces matrices : la myosine, le collagène et les hémoprotéines. / One of the purposes of the Opticook project is to install spectroscopic sensors in ovens, so equipping them with non intrusive tools allowing following cooking process of meat and fish. The hypothesis on which sensors were developed was: are muscular proteins proper probes to discriminate among several cooking degrees? Thus, at the beginning of the project several tools were used to characterise effect of several cooking degrees on macroscopic properties (texture and colour) of beef, chicken and cod fillet samples. Following, calorimetry and spectroscopic techniques were used to study transformations at molecular scale. In particular, visible spectroscopy followed denaturation of haemproteins observing their evolution from oxy to met state. Colour change was observed between 50 and 75 °C. Middle infrared and fluorescence spectroscopies were used to observe evolution of fibrillar proteins which are associated with meat texture changes. One of myofibrils, myosin, was purified from chicken breast and was studied depending on temperature by fluorescence spectroscopy. Observations showed evolution of protein structure occurred around 50 °C. At microscopic scale, low field NMR (spin echo and gradient field configurations) and neutron tomography were used to observe morphology changes and juice release on beef samples according to cooking degree. 3D image analysis and NMR data showed morphology and fibre density changes at high temperature.We associated the two approaches (molecular and microscopic) in the last part of the research. So coupled measures were realised by combining neutron imaging and spectroscopy techniques during cooking process on beef slices by Opticook oven. Coupled measures allowed following morphology evolution because of fibre contractions, juice migration and colour modification as well associated to haemprotein denaturation. In conclusion, it is possible to follow cooking process of meat and fish by following several spectroscopic signatures (fluorescence and visible ones) of matrix own probes: myosin, collage and haemproteins.