Objetivo The periodic development of toxicity in bivalve molluscs, referred to as paralytic shellfish poisoning (PSP) or diarrhetic shellfish poisoning (DSP), is becoming a worldwide problem endangering most European seafood productions. To ensure a safe supply of shellfish, it is necessary to have a reliable analytical method to detect the presence of these toxins. Recently, an alternative assay for PSP toxins using high performance liquid chromatography (HPLC) has been developed in Japan and North America that offers several advantages over the mouse bioassay (greater precision, less analytical time, ability to determine individually any of the different PSP toxins). The aim of the proposal is: to develop in Europe this HPLC analytical procedure; to quantify the PSP toxins in consumable shellfish as well as in contaminated waters; and to extend the method to the DSP toxins. This would provide a fast and reliable analytical service to PSP and DSP European monitoring programs and give a tool for research activities in this field. High performance liquid chromatography (HPLC) methods have been developed in order to quantify the paralytic shellfish poisoning (PSP) and diarrhetic shellfish poisoning (DSP) toxins in consumable shellfish as well as in contaminated waters. Progress has been made in the following areas: Improvement to the HPLC/fluorescence detection (FD) analysis method for the determination of acid DSP toxins in mussel hepatopancreas: One of the most promising approaches for determination of acidic polyether toxins is a method based on HPLC using FD after esterification of the carboxylic acid with 9-anthryldiazomethane (ADAM) to produce a highly fluorescent 9-anthrylmethyl derivative. In order to improve the reliability of the method, the influence of several parameters (extraction and sample clean up procedure, nature of the solvent, reagent product ratio, reaction time, reaction mixture clean up procedure, etc) has been investigated and a new more reliable analytical protocol has been determined. The chromatograms are devoid of interfering peaks in the area critical for the evaluation of the acid DSP toxins and a standardized synthetic method to prepare the ADAM reagent has been developed. Toxic microalgae cultures: Improved procedures of production of Prorocentrum lima, Alexandrium tamarense and Gymnodium catenatum at a large scale in batch cultures to obtain reference compounds (okadaic acid, Dinophysistoxin 1, PSP toxins) for HPLC analysis have been developed. Different culture media have been studied and data collected on the nutritional factors that influence algae growth and toxin production. It has been found that media containing organic compounds are particularly effective. They induce high total cell numbers. The chelating action of organic acids on trace metals is certainly an important factor explaining this promoting action. Preparation of DSP and PSP toxin standard: An effective method of isolation of okadaic acid and Dinophysistoxin 1 from mass culture of P lima and P concavum has been developed. Moreover, standard mixture or identified PSP toxins have been isolated from A tamarense and G catenatum. These samples can be used as standards for the determination of toxins in contaminated shellfish as well as a source of pure samples of individual toxins.The project is made up of 2 aspects: the study of the PSP toxins and the study of the DSP toxins. These 2 types of toxins differ from each other in their biological origin, their structures, their physiological activities and the isolation methods of their constituents. Ámbito científico natural scienceschemical sciencesorganic chemistryorganic acidsnatural sciencesbiological sciencesmicrobiologyphycology Programa(s) FP2-FAR - Community research and coordination programmes (EEC) in the fisheries sector, 1988-1992 Tema(s) Data not available Convocatoria de propuestas Data not available Régimen de financiación Data not available Coordinador Universite Libre de Bruxelles (ULB) Aportación de la UE Sin datos Dirección CP 160 Av. Fr. Roosevelt 50 1050 Bruxelles Bélgica Ver en el mapa Coste total Sin datos Participantes (2) Ordenar alfabéticamente Ordenar por aportación de la UE Ampliar todo Contraer todo RIJKSINSTITUUT VOOR VISSERIJONDERZOEK (RIVO) Países Bajos Aportación de la UE Sin datos Dirección Ijmuiden Ver en el mapa Coste total Sin datos UNIV BRUXELLES-ULB-LAB OCEANOGRAPHIE BIOLOGIQUE Bélgica Aportación de la UE Sin datos Dirección Bruxelles Ver en el mapa Coste total Sin datos
