Single-cell analysis of paralytic shellfish toxins in Alexandrium tamarense by HPLC with post-column fluorescent derivatization |
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| Affiliation: | 1. Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiya, Aoba-ku, Sendai 981-8555, Japan;2. Faculty of Agriculture, Tohoku University, 1-1 Tsutsumidori-Amamiya, Aoba-ku, Sendai 981-8555, Japan;3. Graduate School of Life Sciences, Tohoku University, 1-1 Tsutsumidori-Amamiya, Aoba-ku, Sendai 981-8555, Japan;1. Centro de Investigação em Química and Faculdade de Ciências, University of Porto, 4169 007 Porto, Portugal;2. INFU, Faculty of Chemistry, Technical University of Dortmund, Otto-Hahn-Str. 6, D-44221 Dortmund, Germany;3. Department of Biomedical Sciences, Faculty of Health and Society, Malmö University, Malmö högskola, 205 06 Malmö, Sweden;1. UMR 5119, UM2-CNRS-IRD, Écologie des systèmes marins côtiers, université Montpellier 2, 34095 Montpellier cedex 5, France;2. Laboratoire « bioressources marines », université Badji-Mokhtar-Annaba, BP 12, El-Hadjar, 23000 Annaba, Algeria;3. Ifremer, laboratoire Environnement Ressources Languedoc-Roussillon, BP 171, 34203 Sète, France;1. Normandie Université, COBRA, UMR 6014 et FR 3038, Université de Rouen, INSA de Rouen, CNRS, IRCOF, 1 rue Tesnière, 76821 Mont Saint Aignan Cedex, France;2. IFREMER, Laboratoire Phycotoxines, F-44311 Nantes 03, France;3. DGA Maîtrise NRBC, département Analyse Chimique, F-91710 Vert Le Petit, France;1. Freshwater Fisheries Research Institute of Jiangsu Province, Nanjing 210017, PR China;2. Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, PR China;1. Department of Physical Chemistry, Chair of Chemistry, Medical University, Lublin, Poland;2. Department of Cosmetology, Faculty of Physical Education and Sport in Biala Podlaska, Jozef Pilsudski University of Physical Education, Warsaw, Poland;3. Chair of Environmental Chemistry and Ecotoxicology, University of Bayreuth, Germany |
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| Abstract: | We developed a methodology for analyzing the C-toxin (C2) content in single Alexandrium tamarense cells; this method was based on high performance liquid chromatography (HPLC). C2 is the main paralytic shellfish toxin (PST) detected in a clonal culture of A. tamarense, which is a common causative organism in cases of paralytic shellfish poisoning in Japan. This HPLC method employs post-column fluorescent derivatization (FL). Mobile phase, column size, flow rate, reagent concentrations, and lamp type for the fluorescent detector were all optimized for the detection of C2. With this improved methodology, we could measure 1 fmol of C2 with a signal to noise ratio (S/N) = 2. Clonal heterogeneity within the toxic strain, which was maintained for 13 years after re-isolation from the original clonal culture, ranged from <1 fmol to 700 fmol cell−1. This report is the first to demonstrate definitively that PST content varies on a cell-by-cell basis in a clonal culture of a dinoflagellate that causes paralytic shellfish poisoning. |
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| Keywords: | Single-cell analysis Paralytic shellfish toxins |
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