Determination of Lipophilic Marine Biotoxins in Mussels Harvested from the Adriatic Sea by LC-MS/MS.

Maria Schirone,Nicola Ferri,Francesco Chiumiento,Federica Di Giacinto,Giacomo Migliorati,Pierina Visciano,Rosanna Tofalo,Giovanna Suzzi,Miriam Berti

doi:10.3389/fmicb.2018.00152

Abstract

Lipophilic marine biotoxins include okadaic acid, pectenotoxin, yessotoxin and azaspiracid groups. The consumption of contaminated molluscs can lead to acute food poisoning syndromes depending on the exposure level. Regulatory limits have been set by Regulation (European Community, 2004a) No 853/2004 and LC-MS/MS is used as the official analytical method according to Regulation (European Community, 2011) No 15/2011. In this study specimens of mussels (Mytilus galloprovincialis) were collected along the coasts of the central Adriatic Sea during the years 2015–2017 and analyzed by the European harmonized Standard Operating Procedure. The method was validated for linearity, specificity, repeatability and reproducibility and it revealed able to be used for the detection of the lipophilic marine biotoxins. Levels of okadaic acid, pectenotoxin, yessotoxin and its analogs were detected at different concentrations in 148 (37%) out of a total of 400 samples, always below the maximum limits, except for 11 (4.3%) of them that were non-compliant because they exceeded the regulatory limit. Moreover, some samples were exposed to a multi-toxin mixture with regards to okadaic acid, yessotoxin and 1-Homo yessotoxin. Following these results, the aquaculture farms from which the non-compliant samples derived were closed until the analytical data of two consecutive samplings returned favorable. Besides the potential risk of consumption of mussels contaminated by lipophilic marine biotoxins, these marine organisms can be considered as bio-indicators of the contamination status of the marine ecosystem.

Highlights

Lipophilic marine biotoxins (LMB) are toxic metabolites produced by some species of unicellular algae developing during natural phenomena known as harmful algal blooms
The aim of this study was the application of the EU Harmonised Standard Operating Procedure for determination of Lipophilic marine biotoxins in molluscs by LC-MS/MS (2015) in specimens of Mytilus galloprovincialis coming from different aquaculture farms located along the central Adriatic coasts
The individual content of each detected biotoxin/analog was multiplied with the corresponding Toxicity Equivalent Factors (TEFs) before summarizing the total equivalents for the respective group toxins (European Food Safety Authority, 2009)

Summary

Introduction

Lipophilic marine biotoxins (LMB) are toxic metabolites produced by some species of unicellular algae developing during natural phenomena known as harmful algal blooms. They are grouped in different classes, i.e., okadaic acid (OA), azaspiracids (AZA), yessotoxins (YTX), pectenotoxins (PTX), and spirolides (Ferron et al, 2016). The OA group consists of OA and its isomers, the dinophysistoxins and 2 (DTX1, DTX2) and in addition the fatty acid ester derivatives of OA or DTX1 and DTX2 named DTX3 (Braga et al, 2017) These marine biotoxins are responsible of the human diarrhetic shellfish poisoning (DSP) characterized by gastrointestinal disorders such as nausea, vomiting, severe diarrhea and abdominal. The European Legislation set maximum levels in Regulations (European Community, 2004a) No 853/2004 and (European Community, 2013) No 786/2013, corresponding to 160 μg of OA equivalent kg−1 for OA, DTX and PTX together, 160 μg of AZA equivalent kg−1 for AZA and 3.75 mg YTX equivalent kg−1 for YTX-group

Objectives

Results

Conclusion