Biomedical

Automated Patch Clamp for the Detection of Tetrodotoxin in Pufferfish Samples

Institution: rnfinity

Email: info@rnfinity.com

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Mònica Campàs,

Mònica Campàs

Institution: IRTA, Marine and Continental Waters (AMiC) Programme, Ctra. Poble Nou del Delta, km. 5.5, 43540 La Ràpita, Spain

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Jaume Reverté,

Jaume Reverté

Institution: IRTA, Marine and Continental Waters (AMiC) Programme, Ctra. Poble Nou del Delta, km. 5.5, 43540 La Ràpita, Spain

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Àngels Tudó,

Àngels Tudó

Institution: Department of Basic Medical Sciences, Universitat Rovira i Virgili, C/Sant Llorenç 21, 43201 Reus, Spain

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Mounira Alkassar,

Mounira Alkassar

Institution: IRTA, Marine and Continental Waters (AMiC) Programme, Ctra. Poble Nou del Delta, km. 5.5, 43540 La Ràpita, Spain

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Jorge Diogène,

Jorge Diogène

Institution: IRTA, Marine and Continental Waters (AMiC) Programme, Ctra. Poble Nou del Delta, km. 5.5, 43540 La Ràpita, Spain

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Francesc X. Sureda

Francesc X. Sureda

Institution: Department of Basic Medical Sciences, Universitat Rovira i Virgili, C/Sant Llorenç 21, 43201 Reus, Spain

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Abstract

Tetrodotoxin (TTX) is a marine toxin responsible for many intoxications around the world. Its presence in some pufferfish species and, as recently reported, in shellfish, poses a serious health concern. Although TTX is not routinely monitored, there is a need for fast, sensitive, reliable, and simple methods for its detection and quantification. In this work, we describe the use of an automated patch clamp (APC) system with Neuro-2a cells for the determination of TTX contents in pufferfish samples. The cells showed an IC50 of 6.4 nM for TTX and were not affected by the presence of muscle, skin, liver, and gonad tissues of a Sphoeroides pachygaster specimen (TTX-free) when analysed at 10 mg/mL. The LOD achieved with this technique was 0.05 mg TTX equiv./kg, which is far below the Japanese regulatory limit of 2 mg TTX equiv./kg. The APC system was applied to the analysis of extracts of a Lagocephalus sceleratus specimen, showing TTX contents that followed the trend of gonads > liver > skin > muscle. The APC system, providing an in vitro toxicological approach, offers the advantages of being sensitive, rapid, and reliable for the detection of TTX-like compounds in seafood.

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2024-11-03

Doi: http://dx.doi.org/10.3390/md22040176
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