Cost-Effective and Rapid Detection of Tetrodotoxin Using Indium Tin Oxide Electrodes via In Vitro Electrophysiology and Electrochemistry.

IF 4 3区医学 Q2 FOOD SCIENCE & TECHNOLOGY

Toxins Pub Date : 2025-09-13 DOI:10.3390/toxins17090462

Naga Adithya Chandra Pandurangi, Manel M Santafe, Angels Tudo, Nagihan Ozsoy, Fransesc X Sureda, Mark L Dallas, Ioanis Katakis

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引用次数: 0

Abstract

The real-time, cost-effective detection of marine toxins like tetrodotoxin (TTX) remains a significant challenge for the scientific community. Traditional methods, including cell-based assays (CBAs), high-performance liquid chromatography (HPLC), and automated patch clamp (APC), are time-consuming, requiring expensive lab-based equipment and highly trained personnel. Enzyme-linked immunosorbent assays (ELISAs), lateral flow assays (LFAs), and immunosensors may not be suitable for toxin analogues. Thus, a simplified approach has been developed in this study, which involves the electrophysiological and electrochemical interrogation of N2a cells grown on ITO-coated glass electrodes by measuring extracellular field potentials (EFP) in conjunction with whole-cell patch clamp recordings and electrochemical impedance spectroscopy (EIS) measurements both before and after incubation with TTX. The ITO substrate proved biocompatible and non-toxic for N2a cells. TTX exposure caused 102% inhibition in EFP values at 300 nM, confirmed by sodium current inhibition of 93% at 300 nM and 22% at 1 nM in patch clamp studies (IC₅₀ = 6.7 nM). EIS measurements indicated concentration-dependent impedance changes in the range of 6-300 nM. This research aims to provide a proof-of-concept for integration of electrophysiological and electrochemical approaches to simplify toxin detection systems.

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基于体外电生理和电化学的氧化铟锡电极高效快速检测河豚毒素。

对河豚毒素（TTX）等海洋毒素进行实时、经济有效的检测仍然是科学界面临的重大挑战。传统的方法，包括基于细胞的测定（cba）、高效液相色谱（HPLC）和自动膜片钳（APC），耗时长，需要昂贵的实验室设备和训练有素的人员。酶联免疫吸附测定法（elisa）、侧流测定法（LFAs）和免疫传感器可能不适用于毒素类似物。因此，本研究开发了一种简化的方法，通过测量细胞外场电位（EFP），结合全细胞膜片钳记录和电化学阻抗谱（EIS）测量，在TTX孵育前后，对生长在ito涂层玻璃电极上的N2a细胞进行电生理和电化学询问。ITO底物对N2a细胞具有生物相容性和无毒性。在膜片钳研究中，钠电流在300 nM处抑制93%，在1 nM处抑制22% (IC50 = 6.7 nM)，证实了TTX暴露对300 nM处EFP值的抑制作用为102%。EIS测量表明，阻抗随浓度变化在6-300 nM范围内。本研究旨在为电生理和电化学方法的集成提供概念验证，以简化毒素检测系统。

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来源期刊

Toxins TOXICOLOGY-

CiteScore

7.50

自引率

16.70%

发文量

765

审稿时长

16.24 days

期刊介绍： Toxins (ISSN 2072-6651) is an international, peer-reviewed open access journal which provides an advanced forum for studies related to toxins and toxinology. It publishes reviews, regular research papers and short communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced.