Multistage Detection of Tetrodotoxin Traces in Diodon hystrix Collected in El Salvador

Toxins (Basel). 2023 Jun 25;15(7):409. doi: 10.3390/toxins15070409.

Abstract

This study describes a multistage methodology to detect minute amounts of tetrodotoxin in fishes, a plan that may be broadened to include other marine organisms. This methodology was applied to porcupinefish (Diodon hystrix) collected in Punta Chiquirín, El Salvador. A three-stage approach along with post-acquisition processing was employed, to wit: (a) Sample screening by selected reaction monitoring (HPLC-MS/MS-SRM) analyses to quickly identify possible toxin presence via a LC/MS/MS API 3200 system with a triple quadrupole; (b) HPLC-HRFTMS-full scan analyses using an ion trap-Orbitrap spectrometer combined with an MZmine 2-enhanced dereplication-like workflow to collect high-resolution mass spectra; and (c) HPLC-HRMS2 analyses. This is the first time tetrodotoxin has been reported in D. hystrix specimens collected in El Salvador.

Keywords: Diodon hystrix; HPLC-HRMS; HPLC-HRMS2; HPLC-MS/MS-SRM; MZmine; natural products detection; tetrodotoxin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, Liquid / methods
  • El Salvador
  • Tandem Mass Spectrometry* / methods
  • Tetraodontiformes*
  • Tetrodotoxin

Substances

  • Tetrodotoxin

Grants and funding

This work was supported by Grant PID2021-122732OB-C22 funded by Ministerio de Ciencia e Innovación (MCIN)/Agencia Estatal de Investigación (AEI)/10.13039/501100011033 and “ERDF A way of making Europe”, by the “European Union”. The study was also funded by Grant ED431C 2022/39 from Xunta de Galicia and Grant ELS7005 from the International Atomic Energy Agency (IAEA).