Validation d’un procédé automatisé d’extraction sur phase solide de glandes digestives de moules pour l’identification et la quantification des dinophysistoxines en LC/ESI/SM2 par piégeage d’ions quadripolaire

This report presents a new extraction method of the dinophysistoxins (DTXs), confirmed by quantification using high-performance liquid chromatography coupled to mass spectrometry with an ion trap and electro spray interface (HPLC/ESI/MS2). The method originality consists on the adaptation of DTXs basic extraction procedure (liquid/ liquid) to a solid phase extraction (SPE) via a robotic station: ASPEC XLi The parameters of the automatization procedure were optimized to obtain the best DTXs recovery rate. These improvements were loaded with digestive gland mussel homogenat realized on a silica cartridge SPE, activated in hexane/chloroform (50:50), washed with hexane/chloroform (50:50) and extracted by an elution gradient (chloroform methanol (65:35) and methanol (100%)). This method was validated according to two normative referentials (linearity, detection quantification limits and accuracy…) : - The Guide of the Pharmacy industry: Analytical Validation, report of the commission SFSTP 1992 (French Corporation of the Sciences and Technical Pharmaceutical), - - The Procedure of validation of an alternative method in compare to a reference method. (AFNOR, 1998. NF V 03-110). Comparison with the classical liquid/liquid extraction and the automated method present clear advantages. In an analytical method the extraction is generally considered to be the most labor-intensive and error-prone step. This new procedure allowed us to increase throughput, to improve the reproducibility and to reduce the error risks due to the individual manual treatments.

Les petites pêches côtières et estuariennes Françaises du sud du golfe de Gascogne. Typologie des flottilles et approche socio-économique et commerciale

The European contract PECOSUDE (Small inshore fisheries of the south of Europe) accepted in 2000 for 2 years concerns French, Spanish and Portuguese partners. The aim is to assess the inshore and estuarine fisheries from Loire estuary to Portugal. Three levels of study are considered : fisheries (data 1999), socioeconomy and marketing (data 1999 or 2000). Exploitation in the 12 miles area for France and on continental shelf for northern Spain and Portugal (within 1000 m depth), as well as the fishing period lower than 96 hours was determined as the definition of this fishery for the study. Socioeconomic analyses are made only on boats smaller than 15 m. The fisheries part analyses the activity of the fishing vessels taking into account gears used, landed species, areas and fishing seasons. Within the 39 harbours of the south Bay of Biscay 1 799 coastal or estuarine vessels with 3 580 fishermen are located. Statics gears are predominant, and more than half of the vessels are polyvalent (using several gears). In 1999, landings were 20 6441 weight for 84 M¿ value. The fishing activity of these boats was also analysed by typology of the fleet, based on fishing gears and/or species landed. These characterisations allowed the application of stratified sampling for the implementation of socioeconomic investigations. The analysis of the information received from the fishermen allow to describe the production factors, to estimate their costs, as well as the turnover, the richness creation and the efficiency of the production means. The capital value of the south Bay of Biscay " ship " is established about 94 k¿, the average turnover amount to 83 k¿ and the average rate of added value to 70%. Products commercialisation vary from component to component, the importance of the sells in auction places depends on the landed species. Some valuable species are directly sold to particular

Typologie de la flottille chalutière du port de Sète par une méthode de segmentation

This work aims to define a typology of trawler f1eet in Sète, the main fishing harbour along the French Mediterranean coast, using several multivariate analysis methods. The fishing ships taken to account are represented by annual profiles of landing specific compositions. Five fishing strategies have been identified. A segmentation method using symbolic objects allows a formaI characterisation of the different strategies. These strategies are studied according to several general characteristics usually used for management rules elaboration (power, length, ship age). The typological analysis allows to characterise two main exploitation ways, one directed to the catch of a few species (Engraulis encrasicolus, Sardina pilchardus), the other characterised by the exploitation of a great diversity of species. By this way, it is possible to estimate how the catch of low represented species can significantly contribute to the exploitation of a resource.

Silanisation de polysaccharides en milieu liquide ionique et réactivité d’un organoalkoxysilane vis-à-vis de nucléophiles simples : vers des hydrogels injectables pour l’ingénierie tissulaire du cartilage

Tissue engineering is a real challenge for the treatment of cartilage pathologies. In this field, biomimetic hydrogels based on natural polymers are among the most commonly used matrices. A hydrogel made of silanized hydroxypropylmethylcellulose (HPMC-Si) is especially promising because it can be injected in cartilaginous lesions by minimally invasive surgery. However, the current synthesis of HPMC-Si is limited by the insolubility of hydroxypropylmethylcellulose (HPMC). This thesis work was focused on finding new synthesis conditions for the design of HPMC-Si hydrogel. In order to obtain a complete solubilization of HPMC and to improve its functionalization by the (3-glycidyloxypropyl) trimethoxysilane (GPTMS), the use of ionic liquids (IL), which are excellent solvents for polysaccharides, was undertaken. The beginning of this study was first devoted to the selection of an IL and then to the development of new reaction conditions. With these new conditions, higher silicon rates were obtained for HPMC modified in ionic liquid medium, however no hydrogel could be formed. The second part was therefore devoted to the synthesis of GPTMS 13C. Indeed, thanks to this radiolabeling, a structural characterization by 13C NMR of the HPMC-Si could be achieved. Finally, the reactivity in organic solvents of three organosilanes, including the GPTMS, was investigated toward nucleophiles representing the common functions found in natural polymers (e.g. -NH2, -OH, -SH). The results of this thesis have provided insights into the GPTMS reactivity in organic medium and thus paves the way to new conditions for the silanization of polysaccharides.

Incertitudes des méthodes d’évaluation « eaux littorales » : utilisation de modèles linéaires dynamiques pour l’évaluation des incertitudes (chlorophylle a, phytoplancton)

L’incertitude associée à une mesure a pour origine d’une part la variabilité environnementale et d’autre part l’ensemble du processus d’acquisition depuis le prélèvement jusqu’à la saisie de la donnée dans une base. L’estimation de l'ensemble de cette variabilité est un exercice complexe à réaliser dans le cadre d’un plan d’expérience. En revanche, les séries temporelles présentent la caractéristique d’intégrer toutes les variabilités et ainsi l’analyse de ces séries en terme de signal et bruit doit permettre de quantifier l’amplitude des incertitudes. Toutefois, les séries temporelles d’observation présentent un ensemble de caractéristiques les rendant difficiles à analyser. Les modèles linaires dynamiques constituent une approche adaptée à ces données particulières en faisant l’hypothèse de paramètres variables dans le temps. Ainsi, l’objet du présent travail consiste à estimer les variances liées au processus d’observation à l’aide de modèles linéaires dynamiques. Plus particulièrement, les mesures considérées sont la chlorophylle a et l’abondance phytoplanctonique aux lieux de surveillance REPHY « Arcachon-Bouée- 7 » et « Teychan bis ». Les résultats montrent que pour la chlorophylle a, la variabilité d’observation est responsable de l’ordre de 80 % de la variabilité totale. Pour l’abondance phytoplanctonique, elle est également de 80 % à « Arcachon-Bouée 7 » mais de l’ordre de 70 % à « Teychan bis ». Ainsi la part de « bruit » est liée au lieu et au paramètre considéré. Exprimée en pourcentage de la médiane de la chlorophylle a, la variance d’observation place les bornes de l’intervalle de confiance à 95 % des observations à des valeurs de l’ordre de -40 % et +120 % de la médiane, l’intervalle étant sous estimé car ne prenant pas en compte la variabilité structurelle. Pour l’abondance phytoplanctonique en log10 cell./L, les ordres de grandeur correspondant en pourcentage de la moyenne sont de ± 13.5 %. Pour les deux paramètres, ces valeurs sont compatibles avec l’expérience des experts. Ainsi, l’approche mise en oeuvre s’est avérée riche d’enseignements en matière d’incertitude de mesure et les nombreuses améliorations méthodologiques envisagées ouvrent des perspectives fécondes à tout point de vue.