La pollution atmosphérique liée aux aérosols organiques secondaire (SOA) représente un des enjeux majeurs du XXIème siècle. La photochimie multiphasique des SOA constitue le coeur et l'originalité de cette thèse.Le réacteur photochimique permet de simuler en laboratoire, l'oxydation en phase aqueuse atmosphérique des composés organiques volatils biogéniques (BVOC), et notamment, la méthyl vinyl cétone (MVK), afin d'étudier la formation ces SOA.Nous étudions la réactivité de la MVK en présence de ●OH et sa capacité à oligomériser en fonction des concentrations initiales de MVK, d'oxygène, et de ●OH. Une large stratégie analytique basée sur la chromatographie liquide couplée à la spectrométrie de masse (MS) permet d'identifier des produits de réaction, et d'établir un mécanisme réactionnel, expliquant la formation des oligomères, leurs rendements et leur vieillissement.Les données colligées servent d'entrées à un modèle de boîte multiphasique, afin d'explorer la sensibilité de l'oligomérisation aux conditions atmosphériques.Ensuite, nous comparons la réactivité de la MVK en présence de ●OH à celle induite par la photolyse de l'acide pyruvique; puis nous mesurons la tension de surface engendrée par ces deux systèmes d'oligomères. Enfin, la mobilité ionique couplée à la MS permet d'observer la co-oligomérisation d'une gamme étendue de BVOC en présence de ●OH.L'oligomérisation atmosphérique implique (i) une concentration minimale de précurseurs pouvant être atteinte dans les aérosols humides via la co-oligomérisation; (ii) une réactivité en compétition avec l'oxygène dissous dans la phase aqueuse, et dont la pertinence atmosphérique reste à explorer. / Air pollution caused by secondary organic aerosol (SOA) is one of the major challenges of this century. We focus this thesis on SOA , through an innovative approach, i.e. multiphase photochemistry.The photochemical reactor allows to simulate in laboratory, the atmospheric aqueous phase oxidation of biogenic volatile organic compounds (BVOC) and in particular, methyl vinyl ketone (MVK), and thus, to study SOA.We study the reactivity of MVK in the presence of ●OH and its ability to oligomerize under various initial concentrations of oxygen, MVK and ●OH. A wide analytical strategy based on liquid chromatography-mass spectrometry is used to identify the reaction products, and establish a chemical mechanism. We focus on these oligomers systems, formation, yield and aging. Collected data are used as inputs to a multiphase box model to explore the sensitivity of oligomerization to the variations of physical and chemical atmospheric parameters. The photochemistry of pyruvic acid generates radical chemistry and initiates MVK oligomerization. We closely compare this reaction to MVK ●OH oxidation. Then, we measure the surface activity of both systems. The ability of oligomers to partition to the interface could affect the climate. Finally, we used ion mobility - mass spectrometry to observe ●OH co-oligomerization of a mixture of organic compounds most representative of the atmosphere.Atmospheric oligomerization implies (i) a minimal concentration of precursors that could be reached in wet aerosol via the co-oligomerization; (ii) a reactivity in competition with the addition of the dissolved oxygen, whose the atmospheric relevance remains to be explored.

Present organic carbon fluxes from an upland peat catchment were quantified through measurement of in-situ direct and indirect greenhouse gas fluxes. To predict future greenhouse gas (GHG) fluxes, peat from eroded (E) and uneroded (U) site of an upland peat catchment was characterized.Composition of peat from E and U sites at the Crowden Great Brook catchment, Peak District Nation Park, UK that was characterized by Pyrolysis-Gas Chromatography-Mass Spectrometry (Py-GC-MS) at 700 oC. Pyrolysis products of the peat were then classified using the Vancampenhout classification into 6 compound classes - viz. aromatic and polyaromatic (Ar), phenols (Ph), lignin compounds (Lg), soil lipids (Lp), polysaccharide compounds (Ps) and N-compounds (N). There was no significant difference in the composition between the eroded and uneroded sites within the study area or between peats from different depths within each site. Nevertheless, there was a significant difference between sites in the proportions of Sphagnum that had contributed to the peat. Pyrolysis products of the peat were also classified into pedogenic (Pd) and aquagenic (Aq) OC – the mean percentage of Pd in both eroded and uneroded peats was 43.93 ± 4.30 % with the balance of the OC classified as Aq.Greenhouse gas (GHG) fluxes were quantified directly by in-situ continuous measurement of GHG was carried out at the E and U sites of the catchment using a GasClam: mean in-situ gas concentrations of CH4 (1.30 ± 0.04 % v/v (E), 0.59 ± 0.05 % v/v (U) and CO2 (8.83 ± 0.22 % v/v (E), 1.77 ± 0.03 % v/v (U)) were observed, with both the CH4 and CO2 concentrations apparently unrelated to atmospheric pressure and temperature changes. Laboratory measurements of ex-situ gas production - for both CH4 and CO2 this was higher for U site soils than for E site soils. At the U site, maximum production rates of both CH4 (46.11±1.47 mMol t-1 day-1) and CO2 (45.56 ± 10.19 mMol t-1 day-1) were observed for 0-50 cm depth in soils. Increased temperature did not affect gas production, whilst increased oxygen increased gas production. The CH4/CO2 ratios observed in-situ are not similar to those observed in the ex-situ laboratory experiments; suggest that some caution is advised in interpreting the latter. However, the maximum OC loss of 2.3 wt. % observed after 20 weeks of ex-situ incubation is nevertheless consistent with the long-term degradation noted by Bellamy et al (1985) from organic-rich UK soils. Indirect greenhouse gas (GHG) fluxes were quantified through the mass flux of suspended organic carbon (SsOC) drained from studied catchments. The SsOC was quantified by interpolating and rating methods. Unfiltered (UF) organic carbon (OC) fluxes in 2010 were calculated to be 8.86 t/km2/yr for the eroded sub-catchment and 6.74 t/km2/yr for the uneroded sub-catchment. All the rating relationships have a large amount of scatter. Both UF OC and <0.2 µm fraction OC are positively correlated with discharge at the eroded site, whilst there is no discernable relationship with discharge at the uneroded site. SsOC is dominated by Pd type OC (95.23 ± 10.20 % from E; 92.84 ± 5.38 % from U) far more so than in sources of the peats, suggesting slower oxidation of Pd (cf. Aq) OC.

<p>Volatile organic compounds (VOCs) are endogenous byproducts of metabolic pathways that can be altered by a disease or condition, leading to an associated and unique VOC profile or signature. Current methodologies for VOC detection include canines, gas chromatography-mass spectrometry (GC-MS), and electronic nose (eNose). Some of the challenges for canines and GC-MS are cost-effectiveness, extensive training, expensive instrumentation. On the other hand, a significant downfall of the eNose is low selectivity. This thesis proposes to design a breathalyzer using chemiresistive gas sensors that detects VOCs from human breath, and subsequently create an interface to process and deliver the results via Bluetooth Low Energy (BLE). Breath samples were collected from patients with hypoglycemia, COVID-19, and healthy controls for both. Samples were processed, analyzed using GC-MS and probed through statistical analysis. A panel of 6 VOC biomarkers distinguished between hypoglycemia (HYPO) and Normal samples with a training AUC of 0.98 and a testing AUC of 0.93. For COVID-19, a panel of 3 VOC biomarkers distinguished between COVID-19 positive symptomatic (COVID-19) and healthy Control samples with a training area under the curve (AUC) of receiver operating characteristic (ROC) of 1.0 and cross-validation (CV) AUC of 0.99. The model was validated with COVID-19 Recovery samples. The discovery of these biomarkers enables the development of selective gas sensors to detect the VOCs. </p><p><br></p><p>Polyethylenimine-ether functionalized gold nanoparticle (PEI-EGNP) gas sensors were designed and fabricated in the lab and metal oxide (MOX) semiconductor gas sensors were obtained from Nanoz (Chip 1: SnO₂ and Chip 2: WO₃). These sensors were tested at different relative humidity (RH) levels, and VOC concentrations. Contact angle which measures hydrophobicity, was 84° and the thickness of the PEI-EGNP coating was 11 µ m. The PEI-EGNP sensor response at RH 85% had a signal 10x higher than at RH 0%. Optimization of the MOX sensor was performed by changing the heater voltage and concentration of VOCs. At RH 85% and heater voltage of 2500 mV, the performance of the sensors increased. Chip 2 had higher sensitivity towards VOCs especially for one of the VOC biomarkers identified for COVID-19. PCA distinguished VOC biomarkers of HYPO, COVID-19, and healthy human breath using the Nanoz. A sensor interface was created to integrate the PEI-EGNP sensors with the printed circuit board (PCB) and Bluno Nano to perform machine learning. The sensor interface can currently process and make decisions from the data whether the breath is HYPO (-) or Normal (+). This data is then sent via BLE to the Hypo Alert app to display the decision.</p>

<p>U okviru ove doktorske disertacije ispitane su i<br />poređene različite ekstrakcione tehnike korena<br />gaveza, lista crnog duda i peteljki tre&scaron;nje. Poređene<br />su konvencionalne i savremene ekstrakcione<br />tehnike i izvedena je optimizacija ekstrakcionih<br />postupaka s ciljem dobijanja ekstrakata ispitivanih<br />biljnih vrsta sa najvećom bioaktivno&scaron;ću. Za svaku<br />ekstrakcionu tehniku ispitan je uticaj najznačajnijih<br />parametara: maceracija (rastvarač i vreme<br />ekstrakcije), ubrzana ekstrakcija (temperatura i<br />rastvarač), supekritična ekstrakcija (pritisak i<br />koncentracija ko-rastvarača) i ekstrakcija<br />subkritičnom vodom (temperatura, pritisak, vreme<br />ekstrakcije i brzina me&scaron;anja). Ekstrakcija polifenola<br />korena gaveza, lista crnog duda i peteljki tre&scaron;nje je<br />bila najefikasnija ekstrakcijom subkritičnom<br />vodom. Primenjene su tehnika visokopritisne tečne<br />hromatografije spregnute sa masenom<br />spektrometrijom uz primenu različitih masenih<br />analizatora, masenog analizatora na bazi vremena<br />preleta jona i tandema kvadrupolnog i analizatora<br />na bazi vremena preleta jona. Za karakterizaciju<br />dobijenih ekstrakata instrumentalnom analizom su<br />određene kvalitativne i kvantitativne karakteristike dobijenih ekstrakata korena gaveza, lista crnog duda i peteljki tre&scaron;nje. HPLC-ESI-QTOF-MS/MS analizom ekstrakata identifikovano je vi&scaron;e od 27 novih jedinjenja različitih hemijskih klasa u svakom od ispitivanih biljnih matriksa.</p> / <p>Within the scope of this doctoral dissertation,<br />different extraction techniques of comfrey root,<br />black mulberry leaves and sweet cherry stems were<br />applied and compared. Conventional and novel<br />extraction techniques were optimized and<br />compared in order to obtain plant extracts with the<br />highest content of bioactive compounds. The<br />influence of the most important parameters was<br />investigated for each extraction technique:<br />maceration (solvent and extraction time),<br />accelerated solvent extraction (temperature and<br />solvent), supercritical fluid extraction (co-solvent<br />percentage and pressure) and subcritical water<br />extraction (temperature, pressure, extraction time<br />and agitation rate). Extraction of polyphenolic<br />compounds from comfrey root, black mulberry<br />leaves and sweet cherry stems was the most<br />effective using subcritical water. High-pressure<br />liquid chromatography coupled to mass<br />spectrometry techniques using time-of-flight and<br />quadrupole-time-of-flight mass analyzers have<br />been used to characterize bioactive compounds.<br />Qualitative and quantitative characteristics of the<br />obtained comfrey root, black mulberry leaves and sweet cherry stem extracts were determined. HPLC-ESI-QTOF-MS/MS revealed the presence of more than 27 compounds of different chemical classes that have been identified for the first time in&nbsp; the plant matrix.</p>

Natural resins have been in use for a long time and for manifold purposes resulting in a long and complex terminological history. The investigation of this history has so far been based on the connection between nomenclature and chemical composition. Because resin chemistry and the botanical classification of source plants are connected as well, the investigation of natural resins can be enhanced by adding taxonomy as an additional dimension, providing a more complex and complete picture of resin chemistry and resin use. The Vigani Cabinet, a collection of 300-year-old pharmaceutical and chemical materials owned by Queens’ College, Cambridge (UK), allows doing just that. A wide range of historical literature provides information about contemporary terminology, botanical and geographical origin, manufacture, trade and properties of resinous materials from the 18th century. This contemporary context is a particular feature of the Cabinet, which allows adding a historical dimension to the correlations between terminology, chemical composition and taxonomy. The dissertation thesis presented here provides an investigation of 17 botanical, 80 reference materials and samples from 24 natural resins from the Vigani Cabinet, studying these complex correlations and changes over time. The analytical method employed in this study was gas chromatography-mass spectrometry (GC-MS) with and without methylation with trimethylsulfoniumhydroxide. This technique provided detailed molecular compositions of the studied materials. Analysed botanical samples are taken from Pinaceae, Cupressaceae and Pistacia resins, commerical references from Araucariaceae, Copaifera, Fabaceae, Myroxylon and Burseraceae. Additionally, the soluble fraction of Baltic amber was analysed. Materials from the Vigani Cabinet analysed in this work were labelled as "turpentines", "pix burgundica", "sandaracha", "copaiba", "balsamum peruvianum and tolutanum", "mastiche", "anime", "copal", "elemi", "tacamahaca" and "succinum". Historical nomenclature of natural resins has not always been unequivocally associated with a botanical origin. The availability of natural resins changed throughout the centuries. Lack of knowledge, in particular about resins from over-seas, or adulterations resulting from changing harvesting methods, led to changes in trade names or variations in the composition of products traded under the same name. Generic names were used for resins with similar properties but different botanical (and geographical) origin. The thesis shows that a chemotaxonomic reference system is suitable for the identification of unknown resinous materials, and a number of new insights into the nomenclature of natural resins from the 17th and 18th century is obtained. The study of historical literature contributed in a significant way to the historico-cultural and archeometric research of the samples from the Vigani Cabinet and of natural resins in general and provided a basis for the interpretation of the chemical data from the Vigani samples.:CONTENTS 1 INTRODUCTION 1 1.1 Natural resins in a historical and modern context 1 1.2 The Vigani Cabinet and its historical background 3 1.3 Aim of the thesis - outline 6 2 LITERATURE REVIEW 8 2.1 Gymnosperm resins – conifer resins and products 9 2.1.1 Pinaceae 9 2.1.2 Cupressaceae 17 2.1.3 Araucariaceae 20 2.2 Angiosperm resins I – Fabales 21 2.3 Angiosperm resins II – Sapindales 30 2.3.1 Anacardiaceae 30 2.3.2 Burseraceae 35 2.3.3 Rutaceae 43 2.4 Fossil resins 45 2.5 Summary and research deficits 49 3 EXPERIMENTAL 53 3.1 Coupled gas chromatography and mass spectrometry 53 3.1.1 Materials 53 3.1.2 Sample preparation 54 3.1.3 Instrumentation 54 3.1.4 Data-Evaluation 58 3.2 Fourier transformation infrared spectroscopy 60 3.2.1 Sample preparation 61 3.2.2 Instrumentation 61 3.2.3 Data evaluation 61 4 RESULTS – REFERENCE MATERIALS 62 4.1 Gymnosperm resins – conifer resins and products 62 4.1.1 Pinaceae – Coniferous turpentines 62 4.1.1.1 Phytochemical markers – detection of adulterations 62 4.1.1.2 Aging by heat and light 73 4.1.2 Cupressaceae – Sandarac 80 4.1.3 Araucariaceae – Coniferous copals 88 4.1.4 Discussion 91 4.2 Angiosperm Resins I - Fabales 94 4.2.1 Copaifera – Copaiba balsam 94 4.2.2 Legume copals 102 4.2.3 Myroxylon – Balsam of Tolu and Peru 108 4.2.4 Discussion 117 4.3 Angiosperm resins II - Sapindales 120 4.3.1 Anacardiaceae – Pistacia resins 120 4.3.2 Burseraceae – Elemi, copal and others 127 4.3.3 Discussion 142 4.4 Fossil resins 144 4.4.1 Baltic amber 144 4.4.2 Discussion 153 4.5 Summary and research deficits 155 5 RESULTS – RESINOUS MATERIALS FROM THE VIGANI CABINET 160 5.1 Gymnosperm resins – conifer resins and products 162 5.1.1 1/8 Terebin. Strasb. 163 5.1.2 1/9 Tereb Com 170 5.1.3 1/10 Venice Turpentine 176 5.1.4 1/11 Venic. Turpent. 183 5.1.5 1/13 Tereb E Chio 188 5.1.6 A/23 Pix Burgundica 194 5.1.7 A/26 Sandaracha 203 5.2 Angiosperm resins I - Fabales 210 5.2.1 1/4 Balsam Cipivi 211 5.2.2 A/5 Gum Animi 218 5.2.3 La2/7 Unknown resin 228 5.2.4 1/31 Bals Peruv 230 5.2.5 2/1 Bals Peru 237 5.2.6 Z/17 Balsam Tolutanum 240 5. 3 Angiosperm resins II – Sapindales 245 5.3.1 A/11 Mastiche 246 5.3.2 1/14 Tereb i E Cypri 252 5.3.3 A/21 Gum Copal 258 5.3.4 A/24 [.] Elemi 268 5.3.5 A/22 Tacamahaca 276 5.3.6 Z/1 Tacamahaca 283 5.4 Fossil Resins 287 5.4.1 E/13 Succinum Citrinum 288 5.4.2 E/14 Succinum flavan 295 5.4.3 E/15 Succinum albam 302 5.4.4 E/16 Succinum nigram 307 5.4.5 F/13 L. Gagatis 313 6 CONCLUSIONS 316 7 REFERENCES 324 APPENDIX 365 Investigated materials from the Vigani Cabinet 366 Annotated list of historical literature 367 List of figures 374 List of tables 379 Compound lists 381 Atlas of mass spectra 422 / Naturharze werden schon lange für sehr unterschiedliche Zwecke verwendet. Dies hat zu einer oft komplizierten Terminologie geführt, deren Untersuchung sich bisher auf den Zusammenhang zwischen dem Namen des Harzes und seiner chemischer Zusammensetzung stützte. Letztere ist aber auch mit der botanischer Herkunft und damit der Biochemie der Stammpflanze verknüpft, weshalb man chemotaxonomische Aspekte für die systematische Untersuchung von Naturharzen als zusätzliche Variablen nutzen kann. Dadurch erhält man, wie die gezeigt werden soll, ein vollständigeres und komplexeres Bild der Chemie und Nutzung von Naturharzen. Die hier präsentierte Untersuchung beschäftigt sich mit dem Vigani-Kabinett, einer 300 Jahre alten pharmazeutischen Materialiensammlung, die sich im Queens‘ College, Cambridge (UK), befindet. In der Literatur des ausgehenden 17. und des 18. Jahrhunderts finden sich zahlreiche Informationen zu Terminologie, botanischer und geographischer Herkunft, Verarbeitung, Handel und Eigenschaften von Naturharzen. Dadurch wird die historische Dimension des oben beschriebenen Zusammenhangs zwischen Terminologie, chemischer Zusammensetzung und Taxonomie erfahrbar. In der Arbeit werden 17 botanische Proben, 80 moderne Referenzmaterialien und 24 Proben aus dem Vigani-Kabinett im Hinblick auf diese Zusammenhänge und Veränderungen untersucht.Die chemischen Analysen wurden mit gekoppelter Gaschromatografie-Massenspektrometrie mit und ohne Methylierung mit Trimethylsulfoniumhydroxid durchgeführt. Damit konnte die molekulare Zusammensetzung der Proben detailliert untersucht werden. Die untersuchten botanischen Proben stammten von Pinaceae, Cupressaceae und Pistaciaharzen, kommerzielle Referenzen von Araucariaceae, Copaifera, Fabaceae, Myroxylon und Burseraceaeharzen. Zusätzlich wurde noch die lösliche Fraktion von Baltischem Bernstein untersucht. Die untersuchten Proben aus dem Vigani-Kabinett waren sowohl englisch als auch Latein mit "turpentines", "pix burgundica", "sandaracha", "copaiba", "mastiche", "anime", "copal", "elemi", "tacamahaca", "balsamum peruvianum and tolutanum" und "succinum" beschriftet. Zusammenfassend lässt sich sagen, dass die historische Nomenklatur von Naturharzen nicht immer eindeutig mit ihrem botanischen Ursprung verknüpft war. Zusätzlich veränderte sich die Erhältlichkeit der Harze im Laufe der Jahrhunderte. Durch fehlendes Wissen, insbesondere für Materialien und Pflanzen aus Übersee, oder Verfälschungen aufgrund von veränderten Fördermethoden veränderten sich die Handelsnamen dieser Materialien oder die Zusammensetzung von Materialien, die unter demselben Namen gehandelt wurden. Harze mit ähnlichen Eigenschaften aber unterschiedlichen botanischen (und geographischen) Ursprungs trugen generische Namen. Die Arbeit zeigt jedoch, dass ein chemotaxonomisches Bezugssystem die Identifizierung von unbekannten Harzen ermöglicht, und zeigt eine Reihe neuer Erkenntnisse über die Nomenklatur von Naturharzen des 17. und 18. Jahrhunderts. Die Untersuchung historischer Quellen trug dabei sehr zur Erhellung des historisch-kulturellen und archeometrischen Hintergrundes und zur Interpretation der chemischen Daten der Vigani-Proben bei.:CONTENTS 1 INTRODUCTION 1 1.1 Natural resins in a historical and modern context 1 1.2 The Vigani Cabinet and its historical background 3 1.3 Aim of the thesis - outline 6 2 LITERATURE REVIEW 8 2.1 Gymnosperm resins – conifer resins and products 9 2.1.1 Pinaceae 9 2.1.2 Cupressaceae 17 2.1.3 Araucariaceae 20 2.2 Angiosperm resins I – Fabales 21 2.3 Angiosperm resins II – Sapindales 30 2.3.1 Anacardiaceae 30 2.3.2 Burseraceae 35 2.3.3 Rutaceae 43 2.4 Fossil resins 45 2.5 Summary and research deficits 49 3 EXPERIMENTAL 53 3.1 Coupled gas chromatography and mass spectrometry 53 3.1.1 Materials 53 3.1.2 Sample preparation 54 3.1.3 Instrumentation 54 3.1.4 Data-Evaluation 58 3.2 Fourier transformation infrared spectroscopy 60 3.2.1 Sample preparation 61 3.2.2 Instrumentation 61 3.2.3 Data evaluation 61 4 RESULTS – REFERENCE MATERIALS 62 4.1 Gymnosperm resins – conifer resins and products 62 4.1.1 Pinaceae – Coniferous turpentines 62 4.1.1.1 Phytochemical markers – detection of adulterations 62 4.1.1.2 Aging by heat and light 73 4.1.2 Cupressaceae – Sandarac 80 4.1.3 Araucariaceae – Coniferous copals 88 4.1.4 Discussion 91 4.2 Angiosperm Resins I - Fabales 94 4.2.1 Copaifera – Copaiba balsam 94 4.2.2 Legume copals 102 4.2.3 Myroxylon – Balsam of Tolu and Peru 108 4.2.4 Discussion 117 4.3 Angiosperm resins II - Sapindales 120 4.3.1 Anacardiaceae – Pistacia resins 120 4.3.2 Burseraceae – Elemi, copal and others 127 4.3.3 Discussion 142 4.4 Fossil resins 144 4.4.1 Baltic amber 144 4.4.2 Discussion 153 4.5 Summary and research deficits 155 5 RESULTS – RESINOUS MATERIALS FROM THE VIGANI CABINET 160 5.1 Gymnosperm resins – conifer resins and products 162 5.1.1 1/8 Terebin. Strasb. 163 5.1.2 1/9 Tereb Com 170 5.1.3 1/10 Venice Turpentine 176 5.1.4 1/11 Venic. Turpent. 183 5.1.5 1/13 Tereb E Chio 188 5.1.6 A/23 Pix Burgundica 194 5.1.7 A/26 Sandaracha 203 5.2 Angiosperm resins I - Fabales 210 5.2.1 1/4 Balsam Cipivi 211 5.2.2 A/5 Gum Animi 218 5.2.3 La2/7 Unknown resin 228 5.2.4 1/31 Bals Peruv 230 5.2.5 2/1 Bals Peru 237 5.2.6 Z/17 Balsam Tolutanum 240 5. 3 Angiosperm resins II – Sapindales 245 5.3.1 A/11 Mastiche 246 5.3.2 1/14 Tereb i E Cypri 252 5.3.3 A/21 Gum Copal 258 5.3.4 A/24 [.] Elemi 268 5.3.5 A/22 Tacamahaca 276 5.3.6 Z/1 Tacamahaca 283 5.4 Fossil Resins 287 5.4.1 E/13 Succinum Citrinum 288 5.4.2 E/14 Succinum flavan 295 5.4.3 E/15 Succinum albam 302 5.4.4 E/16 Succinum nigram 307 5.4.5 F/13 L. Gagatis 313 6 CONCLUSIONS 316 7 REFERENCES 324 APPENDIX 365 Investigated materials from the Vigani Cabinet 366 Annotated list of historical literature 367 List of figures 374 List of tables 379 Compound lists 381 Atlas of mass spectra 422

[ES] Debido a la naturaleza (poli)iónica de los conjugados polipéptido-fármaco (PDC), su traslado a clínica puede ser una tarea complicada y costosa, que requiere técnicas de polimerización reproducibles y escalables, la implementación de herramientas analíticas sofisticadas, pasos de caracterización exhaustivos y la recopilación de datos detallados de seguridad y eficacia. Algunas técnicas clásicas implementadas en el control de calidad de los PDC no se pueden utilizar para el análisis cualitativo y cuantitativo de productos de degradación y metabolitos. Como alternativa, la espectrometría de masas (MS) y el análisis de fraccionamiento de flujo de campo de flujo asimétrico (AF4) se han posicionado de forma relevante en dicha caracterización de polipéptidos y PDC. Mediante el uso de métodos de LC-MS, se puede realizar el análisis tanto de fármacos como de productos o metabolitos de degradación. Además, dado que la separación en AF4 no se basa en la interacción del analito con la columna, como ocurre en SEC, sino aplicando el flujo cruzado en un canal vacío; en AF4 es posible una separación de moléculas "a medida" según su tamaño y peso molecular dando lugar a la mejora selectiva de la separación. Teniendo en cuenta lo descrito anteriormente con respecto al diseño de PDC y su caracterización, la investigación incluida en esta tesis se centra en el desarrollo de nuevos procedimientos analíticos que ayuden a la selección de candidatos PDC con las características adecuadas que les permitirán avanzar a su evaluación preclínica. Implementaremos una herramienta de inteligencia artificial, diseño de experimentos, para desarrollar métodos analíticos adecuados y determinar las condiciones óptimas en la síntesis de nanoconjugados polipeptídicos. Además, exploraremos técnicas relativamente nuevas, como AF4 para desarrollar nuevos nanoconjugados polipeptídicos simples y de combinación y además, generaremos imágenes de espectrometría de masas, para entender su comportamiento en modelos preclínicos relevantes, lo que nos permitirá identificar terapias eficaces para una gran variedad de patologías (tumores sólidos avanzados incluyendo cáncer de mama triple negativo, cáncer de próstata y cáncer de páncreas, así como, lesión medular). / [CAT] A causa de la naturalesa (poli)iònica i, de vegades, proteica dels conjugats polipèptid-fàrmac (PDC), el seu trasllat a clínica pot ser una tasca complicada i costosa, que requereix tècniques de polimerització reproduïbles i escalables, la implementació d'eines analítiques sofisticades, passos de caracterització exhaustius i la recopilació de dades detallades de seguretat i eficàcia. Algunes tècniques clàssiques implementades en el control de qualitat dels PDC no es poden utilitzar per l'anàlisi qualitativa i quantitativa de productes de degradació i metabòlits. Com a alternativa, l'espectrometria de masses (MS) i l'anàlisi del fraccionament de flux de camp de flux asimètric (AF4) han trobat el seu camí en la caracterització de PDC. Mitjançant l'ús de mètodes LC-MS, es pot realitzar l'anàlisi tant de fàrmacs com de productes/metabòlits de degradació. A més, atès que la separació en AF4 no es basa en la interacció de l'analit amb la columna com en SEC sinó aplicant el flux creuat en un canal buit, una separació de molècules "a mesura" segons la seua grandària i pes molecular i la millora selectiva de la separació és possible en l'instrument AF4. Tenint en compte tot el descrit anteriorment respecte al disseny de PDC i la seua caracterització, la investigació inclosa en aquest projecte es centra en el desenvolupament de nous procediments analítics que ajuden a la selecció de candidats PDC per als seu estudi preclínic posterior. Implementarem una eina d'intel·ligència artificial, disseny d'experiments, per a desenvolupar mètodes analítics i la síntesi de nanoconjugats basats en polipèptids de manera adequada. A més, explorarem tècniques relativament noves, com AF4, i generarem imatges d'espectrometria de masses, per a desenvolupar nous conjugats a la recerca de teràpies eficaces per a tractar una varietat de malalties (càncer de mama triple negatiu, càncer de pròstata, càncer de pàncrees, així com en la lesió de medul·la espinal). / [EN] Due to the (poly)ionic and proteinic nature of polypeptide-drug conjugates (PDCs), their translation "from bench to bedside" represents a complex and expensive undertaking, requiring reproducible and scalable polymerization techniques, the implementation of sophisticated analytical tools, exhaustive characterization steps, and the collection of detailed safety and efficacy data. Classical techniques, such as liquid chromatography (LC) - UV/Vis and size exclusion chromatography (SEC) implemented in the quality control of PDCs during and after synthesis, cannot always support a qualitative and quantitative analysis of degradation products and metabolites. As an alternative, mass spectrometry (MS) and asymmetric flow field flow fractionation (AF4) have grown in influence on polypeptide and PDC characterization. The analysis of drug and degradation products/metabolites can take advantage of LC when coupled to MS. Meanwhile, AF4-mediated separation does not suffer from problems related to the interaction of the analyte with the column like in SEC; instead, AF4 applies a cross flow in an empty channel, which supports the "tailor-made" separation of molecules according to size and molecular weight. The research included in this Ph.D. thesis focuses on developing new analytical procedures that will aid the selection of PDC candidates for further preclinical studies. We implemented an artificial intelligence tool (design of experiments) to develop analytical methods and optimize the synthesis of genipin-crosslinked PDCs. Moreover, we explored relatively new techniques, such as AF4 and mass spectrometry imaging, in developing novel single and combination PDCs and studying their biological fate in the search for efficient therapies for a range of diseases (advanced solid tumors, including triple negative breast, prostate, and pancreatic cancer, as well as spinal cord injury). / Dordevic, S. (2023). Development of Analytical Procedures for the Characterization of Polypeptide-Based Nanoconjugates [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/194554

The distribution and potential sources of 15 polycyclic aromatic hydrocarbons (PAHs) in soils and Digitaria eriantha in the vicinity of three South African coal-fired power plants, Matla, Lethabo and Rooiwal were determined by gas chromatography–mass spectrometry. An ultrasonic assisted dispersive liquid-liquid microextraction (UA-DLLME) method was developed for the extraction of polycyclic aromatic hydrocarbon in soil, followed by determination using gas chromatography mass spectrometry. The study showed that an extraction protocol based on acetonitrile as dispersive solvent and C2H2Cl2 as extracting solvent, gave extraction efficiencies comparable to conventional soxhlet extraction for soil samples. The extraction time using ultrasonication and the volume of the extraction solvent was also investigated. Using a certified reference material soil (CRM), the extraction efficiency of UA-DLLME ranged from 64 to 86% in comparison with the Soxhlet result of 73 to 95%. In comparison with the real sample, the CRM result did not show a significant difference at 95% C.I. The UA-DLLME proved to be a convenient, rapid, cost-effective and greener sample preparation approach for the determination of PAHs in soil samples. PAH compound ratios such as phenanthrene/phenanthrene + anthracene (Phen/ Phen + Anth) were used to provide a reliable estimation of emission sources. The total PAH concentration in the soils around three power plants ranged from 9.73 to 61.24 μg g−1, a range above the Agency for Toxic Substances and Disease Registry levels of 1.0 μg g−1 for a significantly contaminated site. Calculated values of the Phen/Phen + Anth ratio were 0.48±0.08, 0.44±0.05, and 0.38+0.04 for Matla, Lethabo and Rooiwal, respectively. The flouranthene/fluoranthene + pyrene (Flan/ Flan + Pyr) levels were found to be 0.49±0.03 for Matla, 0.44±0.05 for Lethabo, and 0.53±0.08 for Rooiwal. Such values indicate a xx pyrolytic source of PAHs. Higher molecular weight PAHs (five to six rings) were predominant, suggesting coal combustion sources. The carcinogenic potency B[a]P equivalent concentration (B[a] Peq) at the three power plants ranged from 3.61 to 25.25, indicating a high carcinogenic burden. The highest (B[a] Peq) was found in samples collected around Matla power station. It can, therefore, be concluded that the soils were contaminated with PAHs originating from coal-fired power stations. Nine metals (Fe, Cu, Mn, Ni, Cd, Pb, Hg, Cr and Zn) were analysed in soil and the Digitaria eriantha plant around three coal power plants (Matla, Lethabo and Rooiwal), using ICP-OES and GFAAS. The total metal concentration in soil ranged from 0.05 ± 0.02 to 1835.70 ± 70 μg g-1, 0.08 ± 0.05 to 1743.90 ± 29 μg g-1 and 0.07 ± 0.04 to 1735.20 ± 91 μg g-1 at Matla, Lethabo and Rooiwal respectively. The total metal concentration in the plant (Digitaria eriantha) ranged from 0.005 ± 0.003 to 534.87 ± 43 μg g-1 at Matla, 0.002 ± 0.001 to 400.49 ± 269 μg g-1 at Lethabo and 0.002 ± 0.001 to 426.91 ± 201 μg g-1 at Rooiwal. The accumulation factor (A) of less than 1 (i.e. 0.003 to 0.37) at power plants indicates a low transfer of metal from soil to plant (excluder). The enrichment factor values obtained (2.4 – 5) indicate that the soils are moderately enriched, with the exception of Pb that had significant enrichment of 20. The Geo-accumulation Index values of metals indicate that the soils are moderately polluted (0.005 – 0.65), except for Pb that showed moderate to strong pollution (1.74 – 2.53). / Chemistry / D. Phil. (Chemistry)

Soil organic matter (OM) is a complex mixture of compounds, mainly derived from plants and microbes at various states of decay. It is part of the global carbon cycle and is important for maintaining soil quality. OM protection is mainly attributed to its association with minerals. However, clay minerals preferentially sorb specific OM structures, and clay sorption sites become saturated as OM concentrations increase. Therefore, it is important to examine how OM structures influence their association with soil minerals, and to characterize other protection mechanisms. Several techniques, which provide complementary information, were combined to investigate OM composition: Biomarker (lignin phenol, cutin-OH acid, and lipid) analysis, using gas chromatography/mass spectrometry; solid-state 13C nuclear magnetic resonance (NMR) spectroscopy; and an emerging method, solution-state 1H NMR spectroscopy. OM composition of sand-, silt-, clay-size, and light fractions of Canadian soils were compared. It was found that microbial-derived and aliphatic structures accumulated in clay-size fractions, and lignin phenols in silt-size fractions may be protected from further oxidation. Therefore, OM protection through association with minerals may be structure-specific. OM in soils amended with maize leaves, stems, and roots from a biodegradation study were also examined. Over time, lignin phenol composition, and oxidation; and aliphatic structure contribution changed less in soils amended with leaves compared to soils amended with stems and roots. Compared to soils amended with leaves and stems, amendment with roots may have promoted the more efficient formation of microbial-derived OM. Therefore, plant chemistry influenced soil OM turnover. Synthetic OM-clay complexes and soil mineral fractions were used to investigate lignin protection from chemical oxidation. Coating with dodecanoic acid protected lignin from chemical oxidation, and overlying vegetation determined the relative resistance of lignin phenols in clay-size fractions from chemical oxidation. Therefore, additional protection from chemical oxidation may be attributed to OM composition and interactions between OM structures sorbed to clay minerals. Overall, these studies suggest that while association with minerals is important, OM turnover is also influenced by vegetation, and protection through association with clay minerals was modified by OM structure composition. As well, OM-OM interaction is a potential mechanism that protects soil OM from degradation.

Soil organic matter (OM) is a complex mixture of compounds, mainly derived from plants and microbes at various states of decay. It is part of the global carbon cycle and is important for maintaining soil quality. OM protection is mainly attributed to its association with minerals. However, clay minerals preferentially sorb specific OM structures, and clay sorption sites become saturated as OM concentrations increase. Therefore, it is important to examine how OM structures influence their association with soil minerals, and to characterize other protection mechanisms. Several techniques, which provide complementary information, were combined to investigate OM composition: Biomarker (lignin phenol, cutin-OH acid, and lipid) analysis, using gas chromatography/mass spectrometry; solid-state 13C nuclear magnetic resonance (NMR) spectroscopy; and an emerging method, solution-state 1H NMR spectroscopy. OM composition of sand-, silt-, clay-size, and light fractions of Canadian soils were compared. It was found that microbial-derived and aliphatic structures accumulated in clay-size fractions, and lignin phenols in silt-size fractions may be protected from further oxidation. Therefore, OM protection through association with minerals may be structure-specific. OM in soils amended with maize leaves, stems, and roots from a biodegradation study were also examined. Over time, lignin phenol composition, and oxidation; and aliphatic structure contribution changed less in soils amended with leaves compared to soils amended with stems and roots. Compared to soils amended with leaves and stems, amendment with roots may have promoted the more efficient formation of microbial-derived OM. Therefore, plant chemistry influenced soil OM turnover. Synthetic OM-clay complexes and soil mineral fractions were used to investigate lignin protection from chemical oxidation. Coating with dodecanoic acid protected lignin from chemical oxidation, and overlying vegetation determined the relative resistance of lignin phenols in clay-size fractions from chemical oxidation. Therefore, additional protection from chemical oxidation may be attributed to OM composition and interactions between OM structures sorbed to clay minerals. Overall, these studies suggest that while association with minerals is important, OM turnover is also influenced by vegetation, and protection through association with clay minerals was modified by OM structure composition. As well, OM-OM interaction is a potential mechanism that protects soil OM from degradation.

The distribution and potential sources of 15 polycyclic aromatic hydrocarbons (PAHs) in soils and Digitaria eriantha in the vicinity of three South African coal-fired power plants, Matla, Lethabo and Rooiwal were determined by gas chromatography–mass spectrometry. An ultrasonic assisted dispersive liquid-liquid microextraction (UA-DLLME) method was developed for the extraction of polycyclic aromatic hydrocarbon in soil, followed by determination using gas chromatography mass spectrometry. The study showed that an extraction protocol based on acetonitrile as dispersive solvent and C2H2Cl2 as extracting solvent, gave extraction efficiencies comparable to conventional soxhlet extraction for soil samples. The extraction time using ultrasonication and the volume of the extraction solvent was also investigated. Using a certified reference material soil (CRM), the extraction efficiency of UA-DLLME ranged from 64 to 86% in comparison with the Soxhlet result of 73 to 95%. In comparison with the real sample, the CRM result did not show a significant difference at 95% C.I. The UA-DLLME proved to be a convenient, rapid, cost-effective and greener sample preparation approach for the determination of PAHs in soil samples. PAH compound ratios such as phenanthrene/phenanthrene + anthracene (Phen/ Phen + Anth) were used to provide a reliable estimation of emission sources. The total PAH concentration in the soils around three power plants ranged from 9.73 to 61.24 μg g−1, a range above the Agency for Toxic Substances and Disease Registry levels of 1.0 μg g−1 for a significantly contaminated site. Calculated values of the Phen/Phen + Anth ratio were 0.48±0.08, 0.44±0.05, and 0.38+0.04 for Matla, Lethabo and Rooiwal, respectively. The flouranthene/fluoranthene + pyrene (Flan/ Flan + Pyr) levels were found to be 0.49±0.03 for Matla, 0.44±0.05 for Lethabo, and 0.53±0.08 for Rooiwal. Such values indicate a xx pyrolytic source of PAHs. Higher molecular weight PAHs (five to six rings) were predominant, suggesting coal combustion sources. The carcinogenic potency B[a]P equivalent concentration (B[a] Peq) at the three power plants ranged from 3.61 to 25.25, indicating a high carcinogenic burden. The highest (B[a] Peq) was found in samples collected around Matla power station. It can, therefore, be concluded that the soils were contaminated with PAHs originating from coal-fired power stations. Nine metals (Fe, Cu, Mn, Ni, Cd, Pb, Hg, Cr and Zn) were analysed in soil and the Digitaria eriantha plant around three coal power plants (Matla, Lethabo and Rooiwal), using ICP-OES and GFAAS. The total metal concentration in soil ranged from 0.05 ± 0.02 to 1835.70 ± 70 μg g-1, 0.08 ± 0.05 to 1743.90 ± 29 μg g-1 and 0.07 ± 0.04 to 1735.20 ± 91 μg g-1 at Matla, Lethabo and Rooiwal respectively. The total metal concentration in the plant (Digitaria eriantha) ranged from 0.005 ± 0.003 to 534.87 ± 43 μg g-1 at Matla, 0.002 ± 0.001 to 400.49 ± 269 μg g-1 at Lethabo and 0.002 ± 0.001 to 426.91 ± 201 μg g-1 at Rooiwal. The accumulation factor (A) of less than 1 (i.e. 0.003 to 0.37) at power plants indicates a low transfer of metal from soil to plant (excluder). The enrichment factor values obtained (2.4 – 5) indicate that the soils are moderately enriched, with the exception of Pb that had significant enrichment of 20. The Geo-accumulation Index values of metals indicate that the soils are moderately polluted (0.005 – 0.65), except for Pb that showed moderate to strong pollution (1.74 – 2.53). / Chemistry / D. Phil. (Chemistry)