Silva M et al. (2012), New gastropod vectors and tetrodotoxin potentia...

ECB-ART-50122

Mar Drugs 2012 Apr 01;104:712-726. doi: 10.3390/md10040712.

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New gastropod vectors and tetrodotoxin potential expansion in temperate waters of the Atlantic Ocean.

Silva M , Azevedo J , Rodriguez P , Alfonso A , Botana LM , Vasconcelos V .

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Tetrodotoxin is a potent low weight marine toxin found in warm waters, especially of the Indian and Pacific Oceans. Intoxications are usually linked to the consumption of the puffer fish, although TTX was already detected in several different edible taxa. Benthic organisms such as mollusks and echinoderms, with different feeding habits, were collected monthly along the Portuguese coast from the summer of 2009 until the end of 2010. The extraction and analysis techniques were optimized and TTX and some analogues were detected for the first time in two intertidal gastropod species-Gibbula umbilicalis and Monodonta lineata by LC-MS/MS and UPLC-MS/MS. Although the levels are low, these findings suggest that monitoring of TTX and analogues in North Atlantic species should be implemented so as to detect potentially new toxin vectors and seasonal and/or geographical patterns.

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	Figure 1. Location of the sampling points in the North Atlantic Portuguese coast: 1 Viana do Castelo; 2 Esposende; 3 Póvoa do Varzim; 4 Angeiras; 5 Memória; 6 Valadares; 7 Aguda; 8 São Martinho do Porto; 9 São Torpes; 10 Porto Côvo; 11 Monte Clérigos; 12 Vila Nova de Milfontes; 13 Almograve.
	Figure 2. Mass chromatograms of the LC-ESI-CID-MS/MS obtained under MRM operation of the TTX standard and naturally-contaminated samples of Charonia lampas and Lagocephalus sceleratus. (A) MRM of TTX standard (1500 ng/mL), m/z 320 > 302; (B) Extracted Ion Chromatogram (XIC) of a naturally-contaminated sample of C. lampas with TTX and the analogues 4-epiTTX; 5,6,11-trideoxyTTX; monodeoxyTTX; 4,9-anhydroTTX; (C) XIC of a naturally-contaminated sample of L. sceleratus with the analogues 11-norTTX-6-ol m/z 290 > 272.
	Figure 3. Mass chromatograms of the UPLC-MS/MS obtained under MRM operation of the TTX standard. TIC (total ion chromatogram). TTX standard 1000 ng/mL, m/z 320 > 302/162.
	Figure 4. Mass chromatograms of the LC-ESI-CID-MS/MS obtained under MRM operation of the positive sample of Gibbula umbilicalis for the analogue monodeoxyTTX (m/z 304 > 286/176). XIC extracted ions chromatogram.
	Figure 5. Mass chromatograms of the UPLC-MS/MS obtained under MRM operation of the positive samples of Monodonta lineata and C. lampas for (A): TTX (m/z 320 > 162/302), 4-epiTTX (m/z 320 > 162/302); and (B): 5,6,11-trideoxyTTX (m/z 272 > 254/162).