Detection of yessotoxin by three different methods in Mytilus galloprovincialis of Adriatic Sea , Italy

Samples of Mytilus galloprovincialis collected from shellfish aquaculture plans located in the Adriatic Sea were analysed for yessotoxin (YTX) by three methods, in vivo (Mouse Bioassay, MBA), in vitro (functional assay) and chemical test (Liquid Chromatography–Mass Spectrometry, LC–MS/MS). As YTX coexists with other phycotoxins in shellfish, namely the diarrhoetic shellfish poisoning, okadaic acid, dinophysistoxins, pectenotoxins, and azaspirazids, the MBA is not completely satisfactory because it is difficult to identify which toxin causes the death of the mice. So, the two other techniques were proposed to detect and quantify YTX and its analogues in order to avoid this problem. The global results showed no difference among the three methods and the correlation between the functional assay and LC–MS/MS was positive (Spearman r = 0.72). Both analytical methods demonstrated advantages; the functional assay is specific, very sensitive and correlates well with real toxicity, whereas LC–MS/MS is convenient because it allows the detection of YTX and some analogues which are currently included in the EU regulation. For this reason LC–MS/MS will become the official method starting 1st January 2015 (Regulation 15/2011/EU). Only four samples exceeded the current EU regulation limit of 1 mg of YTX equivalent kg1. However, all samples belonged to a monitoring program and they were not suitable for consumers.