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      Liquid Chromatography with a Fluorimetric Detection Method for Analysis of Paralytic Shellfish Toxins and Tetrodotoxin Based on a Porous Graphitic Carbon Column

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          Abstract

          Paralytic shellfish toxins (PST) traditionally have been analyzed by liquid chromatography with either pre- or post-column derivatization and always with a silica-based stationary phase. This technique resulted in different methods that need more than one run to analyze the toxins. Furthermore, tetrodotoxin (TTX) was recently found in bivalves of northward locations in Europe due to climate change, so it is important to analyze it along with PST because their signs of toxicity are similar in the bioassay. The methods described here detail a new approach to eliminate different runs, by using a new porous graphitic carbon stationary phase. Firstly we describe the separation of 13 PST that belong to different groups, taking into account the side-chains of substituents, in one single run of less than 30 min with good reproducibility. The method was assayed in four shellfish matrices: mussel ( Mytillus galloprovincialis), clam ( Pecten maximus), scallop ( Ruditapes decussatus) and oyster ( Ostrea edulis). The results for all of the parameters studied are provided, and the detection limits for the majority of toxins were improved with regard to previous liquid chromatography methods: the lowest values were those for decarbamoyl-gonyautoxin 2 (dcGTX2) and gonyautoxin 2 (GTX2) in mussel (0.0001 mg saxitoxin (STX)·diHCl kg −1 for each toxin), decarbamoyl-saxitoxin (dcSTX) in clam (0.0003 mg STX·diHCl kg −1), N-sulfocarbamoyl-gonyautoxins 2 and 3 (C1 and C2) in scallop (0.0001 mg STX·diHCl kg −1 for each toxin) and dcSTX (0.0003 mg STX·diHCl kg −1 ) in oyster; gonyautoxin 2 (GTX2) showed the highest limit of detection in oyster (0.0366 mg STX·diHCl kg −1). Secondly, we propose a modification of the method for the simultaneous analysis of PST and TTX, with some minor changes in the solvent gradient, although the detection limit for TTX does not allow its use nowadays for regulatory purposes.

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          Most cited references 45

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          Validation of new methods.

          Reliable analytical data are a prerequisite for correct interpretation of toxicological findings in the evaluation of scientific studies, as well as in daily routine work. Unreliable analytical data might not only be contested in court, but could also lead to unjustified legal consequences for the defendant or to wrong treatment of the patient. Therefore, new analytical methods to be used in forensic and/or clinical toxicology require careful method development and thorough validation of the final method. This is especially true in the context of quality management and accreditation, which have become matters of increasing relevance in analytical toxicology in recent years. In this paper, important considerations in analytical method validation will be discussed which may be used as guidance by scientists wishing to develop and validate analytical methods.
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            Marine algal toxins: origins, health effects, and their increased occurrence.

             F Van Dolah (2000)
            Certain marine algae produce potent toxins that impact human health through the consumption of contaminated shellfish and finfish and through water or aerosol exposure. Over the past three decades, the frequency and global distribution of toxic algal incidents appear to have increased, and human intoxications from novel algal sources have occurred. This increase is of particular concern, since it parallels recent evidence of large-scale ecologic disturbances that coincide with trends in global warming. The extent to which human activities have contributed to their increase therefore comes into question. This review summarizes the origins and health effects of marine algal toxins, as well as changes in their current global distribution, and examines possible causes for the recent increase in their occurrence. Images Figure 2 Figure 3
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              Paralytic Shellfish Toxins in Bivalve Molluscs: Occurrence, Transfer Kinetics, and Biotransformation

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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Toxins (Basel)
                Toxins (Basel)
                toxins
                Toxins
                MDPI
                2072-6651
                28 June 2016
                July 2016
                : 8
                : 7
                Affiliations
                [1 ]Department Analytical Chemistry, Faculty of Sciences, University of Santiago de Compostela, Lugo 27002, Spain; veronica.rey@ 123456rai.usc.es
                [2 ]CIFGA S.A., Plaza Santo Domingo 20-5ª, Lugo 27001, Spain; mercedes@ 123456cifga.es (M.A.); alvaro@ 123456cifga.es (A.A.)
                [3 ]Department Pharmacology, Veterinary Faculty, University of Santiago de Compostela, Lugo 27002, Spain
                Author notes
                [* ]Correspondence: anamaria.botana@ 123456usc.es (A.M.B.); luis.botana@ 123456usc.es (L.M.B.); Tel.: +34-982-824-071 (A.M.B.); +34-982-822-233 (L.M.B.)
                Article
                toxins-08-00196
                10.3390/toxins8070196
                4963829
                27367728
                © 2016 by the authors; licensee MDPI, Basel, Switzerland.

                This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license ( http://creativecommons.org/licenses/by/4.0/).

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