IMPROVED METHOD FOR PREPARATION AND USE OF 9-ANTHRYLDIAZOMETHANE FOR DERIVATIZATION OF HYDROXYCARBOXYLIC ACIDS APPLICATION TO DIARRHETIC SHELLFISH POISONING TOXINS

Application of a method for the ''in situ'' generation of 9-anthryldia zomethane (ADAM) to the derivatization of the carboxyl function in dia rrhetic shellfish poisoning (DSP) toxins revealed the formation of art ifact products. Using liquid chromatography-mass spectrometry, it was determined that these artifacts were due to base-catalyzed reactions b etween the solvent, ethyl acetate, and the hydroxyl groups of the anal yte to produce O-acetylated ADAM derivatives. Using a new formulation, with tetrahydrofuran as solvent, it was possible to eliminate these a rtifact reactions. Various reaction parameters have also been re-optim ized to ensure quantitative derivatizations. An assessment method was developed that was useful not only for optimizing reaction parameters, but also for evaluating the reagent potency before use on important s amples. Finally, application of the method to the determination of DSP toxins in plankton and mussel tissue was demonstrated. Published by E lsevier Science B.V.