Purification and characterization of saxitoxin from Mytilus chilensis of southern Chile

Toxicon. 2015 Dec 15:108:147-53. doi: 10.1016/j.toxicon.2015.09.045. Epub 2015 Oct 3.

Abstract

In the current communication we describe an innovative method to purify saxitoxin (STX), a toxin presents in contaminated muscle of Mylitus chilensis extracted in the southern part of Chile, using a liquid chromatographic methodology based on ionic pairs. The STX was extracted using HCl and treated with ammonium sulfate following a treatment with trichloroacetic acid and hexane/diethyl ether (97/3). The samples were analyzed by a semi-preparative HPLC in order to collect pure fractions of STX and these fractions were eluted in solid-phase cationic interchange SCX extraction columns. The purified STX was stable and homogeneous and its identity was confirmed by LC-MS-MS, which demonstrated a high quality purification of STX, without presence of analogs such as neosaxitoxin (Neo) and decarbamoyl saxitoxin (dcSTX). The STX biological activity was analyzed in a bioassay in mice model and compared to the standard STX produced by the FDA and no significant differences were observed.

Keywords: HPLC; Harmful algal blooms; Mylitus chilensis; Paralytic shellfish poison; Saxitoxin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ammonium Sulfate / chemistry
  • Animals
  • Chile
  • Chromatography, High Pressure Liquid / methods*
  • Chromatography, Liquid
  • Hydrochloric Acid / chemistry
  • Mice
  • Mytilus / chemistry*
  • Saxitoxin / chemistry
  • Saxitoxin / isolation & purification*
  • Solid Phase Extraction / methods
  • Tandem Mass Spectrometry

Substances

  • Saxitoxin
  • Hydrochloric Acid
  • Ammonium Sulfate