Marine Biotechnology

, Volume 12, Issue 5, pp 579–585

Quantitative Determination of Gymnodimine-A by High Performance Liquid Chromatography in Contaminated Clams from Tunisia Coastline

  • Riadh Marrouchi
  • Faten Dziri
  • Nawel Belayouni
  • Asma Hamza
  • Evelyne Benoit
  • Jordi Molgó
  • Riadh Kharrat
Original Article

Abstract

Quantitative determination by high performance liquid chromatography (HPLC) was performed for gymnodimine-A (GYM-A), a phycotoxin responsible for the contamination of Tunisian clams. This study demonstrates a rapid and reproducible HPLC-ultraviolet (UV) method for extraction, detection and quantification of GYM-A in toxic clams. The extraction of GYM-A from the digestive gland of clams in acetone, subsequent clean-up with diethyl ether and extraction with dichloromethane is the more valid protocol. Chromatography analyses were performed using a gradient of acetonitrile–water (10:90 to 90:10), containing trifluoroacetic acid (0.1%) for 20 min at 1 mL/min rate with a C18 column. Recovery rates exceeded 96%, and limits of detection and quantification were 5 ng/mL and 8 ng/g digestive gland, respectively. Repeatability and reproducibility were tested for various samples containing different levels of GYM-A. A significant correlation was observed between toxicity level of samples and the determined amount of GYM-A. Also, the persistence of GYM-A in contaminated clams from Boughrara lagoon was demonstrated. The kinetics discharge study of GYM-A in controlled medium, during 1 month, showed that the process of depuration was biphasic with an exponential discharge of 75% of the total amount of sequestered GYM-A during the first 12 days followed by a slow discharge (>10%) for the subsequent days up to the seventeenth day. This is the first time that a quantitative study of GYM-A in clams from Tunisian coasts is performed through the development of a new method for detection and quantify of this phycotoxin. We found HPLC-UV a reliable and suitable alternative to the mouse bioassay.

Keywords

Clams depuration Gymnodimine-A HPLC Mouse bioassay 

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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Riadh Marrouchi
    • 1
  • Faten Dziri
    • 1
  • Nawel Belayouni
    • 1
  • Asma Hamza
    • 3
  • Evelyne Benoit
    • 2
  • Jordi Molgó
    • 2
  • Riadh Kharrat
    • 1
  1. 1.Laboratoire des Toxines AlimentairesInstitut Pasteur de TunisTunis-BelvédèreTunisia
  2. 2.Laboratoire de Neurobiologie Cellulaire et Moléculaire–UPR9040CNRS, Institut de Neurobiologie Alfred Fessard–FRC2118Gif sur YvetteFrance
  3. 3.Institut National des Sciences et Technologie de la Mer, Centre de SfaxSfaxTunisia

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