Les stilbénoïdes, sont des composés phénoliques majoritairement issus du règne végétal. La Vigne par l’intermédiaire du vin et du raisin est la principale source alimentaire de stilbènes. La mise en évidence de leur rôle dans les mécanismes de défense des plantes et leurs activités biologiques, y compris sur l’Homme, en font un sujet d’étude en plein essor. L’un des objectifs de cette thèse a été de développer un ensemble de stratégies à la fois analytiques et préparatives utilisant la Chromatographie de Partage Centrifuge (CPC) pour l’étude et l’obtention de ces molécules. Dans un premier temps, nous avons développé une approche de couplage entre cette technique et un spectromètre à Résonance Magnétique Nucléaire (RMN) par l’intermédiaire d’un système d’Extraction sur Phase Solide automatisé (EPS). Dans un second temps, nous avons mis au point un ensemble de méthodes et d’approches séparatives permettant d’obtenir ces composés. Nous avons, grâce au développement d’une méthodologie de dosage de solvants par spectrométrie RMN, étudié une gamme dérivée d’une gamme très utilisée en CPC, l’ARIZONA. Nous avons montré que ces systèmes dérivés, peuvent être utilisés au travers de stratégies d’élution telles que des pas et des gradients afin d’optimiser les séparations. L’une des finalités de notre travail est d’offrir des méthodes permettant d’obtenir ces composés dans des quantités et des qualités suffisantes pour pouvoir constituer une chimiothèque interne au laboratoire pouvant prétendre à s’intégrer dans la Chimiothèque Nationale. Une dernière partie fait état du développement d’outils informatiques, dont la création d’une base de donnée Libre pour les chercheurs en Substances Naturelles. / Stilbenoids are phenolic compounds mostly found in the vegetable kingdom. Vine through wine and grape is the main source of stilbenes in the human diet. The involvement of these compounds in plants resistance mechanisms and their diverse biological activities, including on the human health are continuously highlighted. Making this topic a fast-growing one. One of the objectives of this thesis has been to develop a whole set of analytical and preparative strategies using Centrifugal Partition Chromatography (CPC) in order to study and obtain these molecules. Over a first phase, we developed a hyphenated approach between this technique and Nuclear Magnetic Resonance (NMR) through an automated Solid Phase Extraction (SPE) system. Then, we developed a set of separative methods and approaches with the aim of obtaining these compounds. Then, we carried out the development of a solvent quantifications methodology using NMR spectrometry. This allowed us to study a spin-off scale of a widely-used solvent systems collection called ARIZONA. We showed that these systems are good candidates for different elution strategies using steps and gradients with the aim to optimize separations. One of our purpose was to provide methods for effective and efficient purification of these compounds. This would allow, together with a lab-scale compounds library, their integration into the Chimiothèque Nationale, a nationwide chemical library. Last part accounts for computer tools development, including the creation of a Free database system for Natural Substances researcher.

<p>Infrared absorption spectra can provide analytically useful information on a large variety of compounds, ranging from small ions to large biological molecules. In fact, all analytes that possess a dipole moment that changes during vibration are infrared-active. The infrared (IR) spectrum can be subdivided into far-, mid- and near- regions. The focus of attention in this thesis is the mid-IR region, in which the fundamental vibrations of most organic compounds are located, thus providing scope for positive structural identification. However, while such near-ubiquitous signals can be very useful for monitoring simple molecules in simple systems, they can be increasingly disadvantageous as the number of analytes and/or the complexity of the sample matrix increases. Thus, hyphenation to a separation system prior to detection is desirable. Paper I appended to this thesis presents (for the first time) the on-line hyphenation between Fourier transform infrared spectroscopy, FTIR, and capillary zone electrophoresis, CZE. CZE is a highly efficient separation technique that separates ionic analytes with respect to their charge-to-size ratio. It is most commonly performed in aqueous buffers in fused silica capillaries. Since these capillaries absorb virtually all infrared light an IR-transparent flow cell had to be developed. In further studies (Paper II) the applicability of CZE is expanded to include neutral analytes by the addition of micelles to the buffer, and micellar electrokinetic chromatography, MEKC, was successfully hyphenated to FTIR for the first time. Paper III describes an application of the on-line CZE-FTIR technique in which non-UV-absorbing analytes in a complex matrix were separated, identified and quantified in one run.</p><p>Measuring aqueous solutions in the mid-IR region is not straightforward since water absorbs intensely in this region, sometimes completely, leaving no transmitted, detectable light. For this reason, quantum cascade lasers are interesting. These lasers represent a new type of mid-IR semiconducting lasers with high output power due to their ingenious design. The laser action lies within one conduction band (intersubband) and can be tailored to emit light in the entire mid-IR region using the same semiconducting material. To investigate their potential to increase the optical path length in aqueous solutions, these lasers were used with an aqueous flow system (Paper IV), and the experience gained in these experiments enabled hyphenation of such lasers to a CZE system (Paper V).</p>

Infrared absorption spectra can provide analytically useful information on a large variety of compounds, ranging from small ions to large biological molecules. In fact, all analytes that possess a dipole moment that changes during vibration are infrared-active. The infrared (IR) spectrum can be subdivided into far-, mid- and near- regions. The focus of attention in this thesis is the mid-IR region, in which the fundamental vibrations of most organic compounds are located, thus providing scope for positive structural identification. However, while such near-ubiquitous signals can be very useful for monitoring simple molecules in simple systems, they can be increasingly disadvantageous as the number of analytes and/or the complexity of the sample matrix increases. Thus, hyphenation to a separation system prior to detection is desirable. Paper I appended to this thesis presents (for the first time) the on-line hyphenation between Fourier transform infrared spectroscopy, FTIR, and capillary zone electrophoresis, CZE. CZE is a highly efficient separation technique that separates ionic analytes with respect to their charge-to-size ratio. It is most commonly performed in aqueous buffers in fused silica capillaries. Since these capillaries absorb virtually all infrared light an IR-transparent flow cell had to be developed. In further studies (Paper II) the applicability of CZE is expanded to include neutral analytes by the addition of micelles to the buffer, and micellar electrokinetic chromatography, MEKC, was successfully hyphenated to FTIR for the first time. Paper III describes an application of the on-line CZE-FTIR technique in which non-UV-absorbing analytes in a complex matrix were separated, identified and quantified in one run. Measuring aqueous solutions in the mid-IR region is not straightforward since water absorbs intensely in this region, sometimes completely, leaving no transmitted, detectable light. For this reason, quantum cascade lasers are interesting. These lasers represent a new type of mid-IR semiconducting lasers with high output power due to their ingenious design. The laser action lies within one conduction band (intersubband) and can be tailored to emit light in the entire mid-IR region using the same semiconducting material. To investigate their potential to increase the optical path length in aqueous solutions, these lasers were used with an aqueous flow system (Paper IV), and the experience gained in these experiments enabled hyphenation of such lasers to a CZE system (Paper V).

The scientific progress about the important chemotherapeutic drug substance cisplatin (CDDP) and its function has often been rendered by data difficult to interpret, and still many questions about its mode of action remains to be clarified by the scientific community. However, studies of CDDP possess a high complexity due to; i) low intracellular concentration, ii) many potential biomolecule targets, iii) poor or unknown stability of the intact drug and its biomolecule adducts and iv) complex and varying sample matrices. Metallomic studies, using advanced analytical techniques may contribute to clarify the interactions between CDDP and intracellular biomolecules. For a successful outcome sample preparation conditions as well as separation and detection techniques must be carefully selected and optimized to achieve accurate results and correct interpretation of data.         This thesis describes some new and improved analytical methodologies for characterizing the Pt metallome in CDDP-exposed malignant cells. The developed methods are based on powerful liquid chromatography (LC) methods hyphenated to sensitive detection by inductively coupled plasma- (ICP) and electrospray ionization mass spectrometry (ESIMS). Consideration has also been taken about sample preparation conditions.         By selecting “chemically inert” sample preparation (cell lysis by osmosis) and separation (using only nonreactive or no additatives) conditions we could avoid the formation of platinum artifact compounds previously described in the literature (Paper I and II). Using oxygen containing organic solvents with high boiling points (dimethylformamide; DMF, 1,4-dioxane, n-propanol and ethanol) as alternatives to acetonitrile in the LC separations, significant improvements were achieved in ICPMS sensitivity and robustness. When evaluated in combination with chromatographic performance and ESIMS detection the overall best performance was achieved with n-propanol (Paper II, III and IV). From the studies in Paper II we could show that free intact CDDP can be found in malignant cells, as supporting evidence for passive or endocytotic uptake of the drug and further estimate a half-life for intracellular CDDP to about 15 minutes. Such data has not been shown before. In Paper V, the above improved LC methods were used to demonstrate differences in the platinum and cupper metallome from sensitive and resistant T289 melanoma cells exposed to CDDP at near clinical levels.         In a wider perspective we have shown the potential of using hydrophilic liquid interaction chromatography (HILIC) hyphenated to ICPMS detection as a general approach for analysis of hydrophilic metallo-compounds (Paper II). Taking advantage of the superior ICPMS performance using n-propanol gradients for reversed phase liquid chromatography (RPLC) possess a true alternative and /or complimentary technique to size exclusion chromatography (SEC) commonly applied within metallomic studies of biomolecules (Paper V). Using n-propanol in HILIC as well as in RPLC enables parallel detection by ICP- and ESIMS using only one set of chromatographic parameters (Paper III and IV), something commonly called for by scientists in the field.

This thesis investigates the changes in whether compound nouns were closed (written as one word), open (written as separate words) or hyphenated in Early New High German between 1550 and 1710. Due to the fact that there were no orthographic norms in the German of this time, graphematic phenomena in this period of the German language are very fruitful to examine. The study is based on a corpus of 249 sermons in 90 different postils. Since this thesis aims to show a diachronic development, the corpus texts originate from six time windows centred around the years 1550, 1570, 1600, 1620, 1660 and 1710. The results of the study show a general development from 1550, when around 80% of the occurrences of compound nouns were written as one word, to 1620, when this way of writing dominated almost entirely. In the texts from the last two time windows, the hyphenation spreads, and by 1710, nearly two thirds of the instances of compound nouns were written with a hyphen. The present study also shows that the geographical origin of a text is of lesser importance for the writing of compound nouns as one word, separate words or with a hyphen. However, the distinction between genuine compound nouns (a compound noun with the modifier in an unmarked case) and artificial ones (a compound noun with the modifier in an oblique case) seems to be of greater relevance. The artificial compound nouns are closed to a lesser extent in the period between 1550 and 1620 and hyphenated to a higher extent from 1660 onwards than the genuine compound nouns. In a second part of the study, the compound nouns of the different time windows are examined from a lexical point of view, showing that many compound noun lexemes were almost consistently written in the same way (either as one word, as separate words, or with a hyphen) in all occurrences within each time window.

<p>The present paper examines the N₁ by N₁ construction using corpus linguistic methodology.The distribution of types of the construction that occur more than once either unhyphenated or hyphenated in any subcorpus of the British National Corpus accessed through the BrighamYoung University interface is examined. Written and spoken language as well as variousgenres are compared. Hyphenation is also investigated. A collocation analysis of some typesof the construction is further carried out and it is concluded that the N₁ by N₁ construction canbe part of the on a N₁ by N₁ basis construction. Results from the quantitative analysis as wellas the qualitative discussion suggest that the N P N construction may be undergoinglexicalisation starting as an adverbial and moving to functioning as a premodifier. Thissuggestion is indicated through complementary diachronic searches in the Oxford EnglishDictionary. It is also indicated that the construction may follow a development pattern similarto that of N₁to N₁. The notion of construction is discussed in relation to the N₁ by N₁ construction, and a hierarchical view of constructions is proposed as a solution to some of theproblems with the term.</p>

Text in Afrikaans / Summaries in Afrikaans and English / In Afrikaans, soos in NederJands en Duits, word saamgestelde woorde aanmekaar geskryf. Nuwe woorde word dus voortdurend geskep deur woorde aanmekaar te haak Dit bemoeilik die proses van woordafkapping tydens teksprosessering, wat deesdae deur rekenaars gedoen word, aangesien die verwysingsbron gedurig verander. Daar bestaan verskeie afkappingsalgoritmes en tegnieke, maar die resultate is onbevredigend. Afrikaanse woorde met korrekte lettergreepverdeling is net die elektroniese weergawe van die handwoordeboek van die Afrikaanse Taal (HAT) onttrek. 'n Neutrale netwerk ( vorentoevoer-terugpropagering) is met sowat. 5 000 van hierdie woorde afgerig. Die neurale netwerk is verfyn deur 'n gcskikte afrigtingsalgoritme en oorfragfunksie vir die probleem asook die optimale aantal verborge lae en aantal neurone in elke laag te bepaal. Die neurale netwerk is met 5 000 nuwe woorde getoets en dit het 97,56% van moontlike posisies korrek as of geldige of ongeldige afkappingsposisies geklassifiseer. Verder is 510 woorde uit tydskrifartikels met die neurale netwerk getoets en 98,75% van moontlike posisies is korrek geklassifiseer. / In Afrikaans, like in Dutch and German, compound words are written as one word. New words are therefore created by simply joining words. Word hyphenation during typesetting by computer is a problem, because the source of reference changes all the time. Several algorithms and techniques for hyphenation exist, but results are not satisfactory. Afrikaans words with correct syllabification were extracted from the electronic version of the Handwoordeboek van die Afrikaans Taal (HAT). A neural network (feedforward backpropagation) was trained with about 5 000 of these words. The neural network was refined by heuristically finding a suitable training algorithm and transfer function for the problem as well as determining the optimal number of layers and number of neurons in each layer. The neural network was tested with 5 000 words not the training data. It classified 97,56% of possible points in these words correctly as either valid or invalid hyphenation points. Furthermore, 510 words from articles in a magazine were tested with the neural network and 98,75% of possible positions were classified correctly. / Computing / M.Sc. (Operasionele Navorsing)

Text in Afrikaans / Summaries in Afrikaans and English / In Afrikaans, soos in NederJands en Duits, word saamgestelde woorde aanmekaar geskryf. Nuwe woorde word dus voortdurend geskep deur woorde aanmekaar te haak Dit bemoeilik die proses van woordafkapping tydens teksprosessering, wat deesdae deur rekenaars gedoen word, aangesien die verwysingsbron gedurig verander. Daar bestaan verskeie afkappingsalgoritmes en tegnieke, maar die resultate is onbevredigend. Afrikaanse woorde met korrekte lettergreepverdeling is net die elektroniese weergawe van die handwoordeboek van die Afrikaanse Taal (HAT) onttrek. 'n Neutrale netwerk ( vorentoevoer-terugpropagering) is met sowat. 5 000 van hierdie woorde afgerig. Die neurale netwerk is verfyn deur 'n gcskikte afrigtingsalgoritme en oorfragfunksie vir die probleem asook die optimale aantal verborge lae en aantal neurone in elke laag te bepaal. Die neurale netwerk is met 5 000 nuwe woorde getoets en dit het 97,56% van moontlike posisies korrek as of geldige of ongeldige afkappingsposisies geklassifiseer. Verder is 510 woorde uit tydskrifartikels met die neurale netwerk getoets en 98,75% van moontlike posisies is korrek geklassifiseer. / In Afrikaans, like in Dutch and German, compound words are written as one word. New words are therefore created by simply joining words. Word hyphenation during typesetting by computer is a problem, because the source of reference changes all the time. Several algorithms and techniques for hyphenation exist, but results are not satisfactory. Afrikaans words with correct syllabification were extracted from the electronic version of the Handwoordeboek van die Afrikaans Taal (HAT). A neural network (feedforward backpropagation) was trained with about 5 000 of these words. The neural network was refined by heuristically finding a suitable training algorithm and transfer function for the problem as well as determining the optimal number of layers and number of neurons in each layer. The neural network was tested with 5 000 words not the training data. It classified 97,56% of possible points in these words correctly as either valid or invalid hyphenation points. Furthermore, 510 words from articles in a magazine were tested with the neural network and 98,75% of possible positions were classified correctly. / Computing / M.Sc. (Operasionele Navorsing)

The present paper examines the N1 by N1 construction using corpus linguistic methodology.The distribution of types of the construction that occur more than once either unhyphenated or hyphenated in any subcorpus of the British National Corpus accessed through the BrighamYoung University interface is examined. Written and spoken language as well as variousgenres are compared. Hyphenation is also investigated. A collocation analysis of some typesof the construction is further carried out and it is concluded that the N1 by N1 construction canbe part of the on a N1 by N1 basis construction. Results from the quantitative analysis as wellas the qualitative discussion suggest that the N P N construction may be undergoinglexicalisation starting as an adverbial and moving to functioning as a premodifier. Thissuggestion is indicated through complementary diachronic searches in the Oxford EnglishDictionary. It is also indicated that the construction may follow a development pattern similarto that of N1 to N1. The notion of construction is discussed in relation to the N1 by N1 construction, and a hierarchical view of constructions is proposed as a solution to some of theproblems with the term.

Text in Afrikaans / Die doel van hierdie studie was om te bepaal tot watter mate ’n suiwer patroongebaseerde benadering tot woordafbreking bevredigende resultate lewer. Die masjienleertegnieke kunsmatige neurale netwerke, beslissingsbome en die TEX-algoritme is ondersoek aangesien dit met letterpatrone uit woordelyste afgerig kan word om lettergreep- en saamgesteldewoordverdeling te doen. ’n Leksikon van Afrikaanse woorde is uit ’n korpus van elektroniese teks genereer. Om lyste vir lettergreep- en saamgesteldewoordverdeling te kry, is woorde in die leksikon in lettergrepe verdeel en saamgestelde woorde is in hul samestellende dele verdeel. Uit elkeen van hierdie lyste van ±183 000 woorde is ±10 000 woorde as toetsdata gereserveer terwyl die res as afrigtingsdata gebruik is. ’n Rekursiewe algoritme is vir saamgesteldewoordverdeling ontwikkel. In hierdie algoritme word alle ooreenstemmende woorde uit ’n verwysingslys (die leksikon) onttrek deur stringpassing van die begin en einde van woorde af. Verdelingspunte word dan op grond van woordlengte uit die samestelling van begin- en eindwoorde bepaal. Die algoritme is uitgebrei deur die tekortkominge van hierdie basiese prosedure aan te spreek. Neurale netwerke en beslissingsbome is afgerig en variasies van beide tegnieke is ondersoek om die optimale modelle te kry. Patrone vir die TEX-algoritme is met die OPatGen-program gegenereer. Tydens toetsing het die TEX-algoritme die beste op beide lettergreep- en saamgesteldewoordverdeling presteer met 99,56% en 99,12% akkuraatheid, respektiewelik. Dit kan dus vir woordafbreking gebruik word met min risiko vir afbrekingsfoute in gedrukte teks. Die neurale netwerk met 98,82% en 98,42% akkuraatheid op lettergreep- en saamgesteldewoordverdeling, respektiewelik, is ook bruikbaar vir lettergreepverdeling, maar dis meer riskant. Ons het bevind dat beslissingsbome te riskant is om vir lettergreepverdeling en veral vir woordverdeling te gebruik, met 97,91% en 90,71% akkuraatheid, respektiewelik. ’n Gekombineerde algoritme is ontwerp waarin saamgesteldewoordverdeling eers met die TEXalgoritme gedoen word, waarna die resultate van lettergreepverdeling deur beide die TEXalgoritme en die neurale netwerk gekombineer word. Die algoritme het 1,3% minder foute as die TEX-algoritme gemaak. ’n Toets op gepubliseerde Afrikaanse teks het getoon dat die risiko vir woordafbrekingsfoute in teks met gemiddeld tien woorde per re¨el ±0,02% is. / The aim of this study was to determine the level of success achievable with a purely pattern based approach to hyphenation in Afrikaans. The machine learning techniques artificial neural networks, decision trees and the TEX algorithm were investigated since they can be trained with patterns of letters from word lists for syllabification and decompounding. A lexicon of Afrikaans words was extracted from a corpus of electronic text. To obtain lists for syllabification and decompounding, words in the lexicon were respectively syllabified and compound words were decomposed. From each list of ±183 000 words, ±10 000 words were reserved as testing data and the rest was used as training data. A recursive algorithm for decompounding was developed. In this algorithm all words corresponding with a reference list (the lexicon) are extracted by string fitting from beginning and end of words. Splitting points are then determined based on the length of reassembled words. The algorithm was expanded by addressing shortcomings of this basic procedure. Artificial neural networks and decision trees were trained and variations of both were examined to find optimal syllabification and decompounding models. Patterns for the TEX algorithm were generated by using the program OPatGen. Testing showed that the TEX algorithm performed best on both syllabification and decompounding tasks with 99,56% and 99,12% accuracy, respectively. It can therefore be used for hyphenation in Afrikaans with little risk of hyphenation errors in printed text. The performance of the artificial neural network was lower, but still acceptable, with 98,82% and 98,42% accuracy for syllabification and decompounding, respectively. The decision tree with accuracy of 97,91% on syllabification and 90,71% on decompounding was found to be too risky to use for either of the tasks A combined algorithm was developed where words are first decompounded by using the TEX algorithm before syllabifying them with both the TEX algoritm and the neural network and combining the results. This algoritm reduced the number of errors made by the TEX algorithm by 1,3% but missed more hyphens. Testing the algorithm on Afrikaans publications showed the risk for hyphenation errors to be ±0,02% for text assumed to have an average of ten words per line. / Decision Sciences / D. Phil. (Operational Research)