Single-cell analysis of paralytic shellfish toxins in Alexandrium tamarense by HPLC with post-column fluorescent derivatization

Yuko Cho, Ryoko Ozeki, Mari Yotsu-Yamashita, Yasukatsu Oshima

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

Abstract

We developed a methodology for analyzing the C-toxin (C2) content in single Alexandrium tamarense cells; this method was based on high performance liquid chromatography (HPLC). C2 is the main paralytic shellfish toxin (PST) detected in a clonal culture of A. tamarense, which is a common causative organism in cases of paralytic shellfish poisoning in Japan. This HPLC method employs post-column fluorescent derivatization (FL). Mobile phase, column size, flow rate, reagent concentrations, and lamp type for the fluorescent detector were all optimized for the detection of C2. With this improved methodology, we could measure 1 fmol of C2 with a signal to noise ratio (S/N)=2. Clonal heterogeneity within the toxic strain, which was maintained for 13 years after re-isolation from the original clonal culture, ranged from <1fmol to 700fmolcell-1. This report is the first to demonstrate definitively that PST content varies on a cell-by-cell basis in a clonal culture of a dinoflagellate that causes paralytic shellfish poisoning.

Original languageEnglish
Pages (from-to)47-53
Number of pages7
JournalHarmful Algae
Volume25
DOIs
Publication statusPublished - 2013 May

Keywords

  • Alexandrium tamarense
  • Paralytic shellfish toxins
  • Single-cell analysis

ASJC Scopus subject areas

  • Aquatic Science
  • Plant Science

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