Occurrence Of Tetrodotoxin Research Articles

Occurrence of tetrodotoxin in the gastropod mollusk Tutufa lissostoma (frog shell)

Paralytic toxicity was detected in gastropod mollusk Tutufa lissostoma (frog shell) specimens collected from Suruga Bay, Shizuoka Prefecture, Japan. Seventeen of the 22 digestive glands removed were toxic; the highest toxicity, expressed as tetrodotoxin (TTX), being as high as 700 mouse units (MU) per gram. Attempts were made to isolate the toxin from pooled digestive glands by activated charcoal treatment and column chromatography on Amberlite IRC-50, CM-Sephadex C-25 and Bio-Rex 70. The toxin showed a specific toxicity (as TTX) of 4200 MU/mg. It exhibited the same thin-layer chromatographic and electrophoretic behaviors and 1H-NMR spectrum as TTX. The toxin gave the same pattern as the TTX standard when alkali-hydrolyzed and analyzed by GC - MS, indicating that it contains the quinazoline skeleton specific to TTX. From these results the frog shell toxin was identified as TTX.

Another Occurrence of Tetrodotoxin in a Trumpet Shell Charonia sauliae

In December 1982, an outbreak of paralytic poisoning occurred due to ingestion of the viscera of a trumpet shell (“boshubora”, Charonia sauliae) in Wakayama Prefecture. Many specimens of this gastropod were collected from adjacent waters and analyzed for toxicity by the routine assay method for tetrodotoxin (TTX). Most of the specimens were toxic: the highest toxicity was 480MU/g digestive gland, and the highest total toxicity per specimen was 37, 000MU.Attempts were made to isolate the toxin from the toxic digestive glands by a TTX isolation method, which consisted of activated charcoal treatment and chromatography on CM-Sephadex C-25 and Bio-Rex 70. The toxin thus isolated possessed a specific activity of 3, 700MU/mg. It showed thin-layer chromatographic and electrophoretic properties indistinguishable from those of authentic TTX. Both toxins exhibited an identical 1H-NMR spectrum.It was concluded from these results that the toxin responsible for this incident of paralytic poisoning was TTX.

Occurrence of tetrodotoxin as the major toxin in a xanthid crab Atergatis floridus.

The toxins of a xanthid crab Atergatis floridus from Miura Peninsula were purified by a method which consisted of activated charcoal treatment, and chromatography on Amberlite IRC-50, Bio-Gel P-2 and Bio-Rex 70 columns. In the Bio-Rex 70 column chromatography, the major toxin equivalent to about 90% of the total toxicity was eluted with 0-0.03 N acetic acid, whereas the minor toxin, with 0.03-1 N acetic acid. The major toxin showed a single spot in TLC and electrophoreis, whose Rf and Rm values coincided well with those of tetrodotoxin run for reference. Furthermore, the major toxin agreed well with tetrodotoxin in NMR spectrum. From these results, the major toxin was identified as tetrodotoxin. On the other hand, the minor toxin was demonstrated to consist of saxitoxin and some unidentified toxic components.

Occurrence of tetrodotoxin in the Japanese ivory shell Babylonia japonica.

Specimens of the Japanese ivory shell Babylonia japonica from Sakajiri Bay, Fukui Prefecture, were all found to be toxic throughout the survey period, Aug. to Nov. 1980; the maximum toxicity (as tetrodotoxin) was 53 MU/g digestive gland. The responsible toxin was isolated from the digestive gland by a method which essentially consisted of the adsorption onto activated charcoal, chromatography on Amberlite IRC-50 and CM-Sephadex C-25, and finally crystallization from an acidified methanolic solution. The toxin and its derivatives were indistinguishable from those of tetrodotoxin (TTX) in thin-layer chromatography using 3 solvent systems. The toxin showed the same mobility as that of TTX in cellulose acetate membrane electrophoresis. The C9-base-trimethylsilyl derivative from the toxin gave the same mass spectrum as that of the corresponding TTX derivative. The dose-death time curves of this toxin and TTX coincided well with each other. It was concluded from those results that the Japanese ivory shell toxin isolated was indeed TTX.

Occurrence of tetrodotoxin in a trumpet shell, "boshubora" Charonia sauliae.

A paralytic poisoning occurred in Deccember, 1979, due to ingestion of a trumpet shell, “boshubora” Charonia sauliae which was caught off Miho, Shizuoka Prefecture. The toxicity study showed that most of the boshubora specimens collected around there were toxic at the digestive gland. The boshubora toxin (abbreviated as BST below) was purified from the digestive gland by a method which consisted of the treatment with activated charcoal, chromatography on Amberlite IRC-50 and CM-Sephadex C-25, and finally crystallixation. The recrystallixed toxin shwed a toxicity of 3, 700 MU/mg. Thin-layer chromatographic and electrophoretic results for BST were the same as those for tetrodotoxin (TTX) analyzed for reference. Furthermoree. Furthermore, both toxins coincided well in their IR and NM Rspectra, the mass spectra of the C9-base-trimethylsilyl derivative, and the dose-death time curves. Based on these results, BST was identified as TTX.

Occurrence of tetrodotoxin in the ivory shell Babylonia japonica from wakasa bay.

During the surveillance on paralytic shellfish toxins in Fukui Prefecture, the Japanese ivory shell Babylonia japonica collected at Sakajiri of Wakasa Bay was found to contain toxin (s) of strong paralytic action. The symptoms in mice were distinctly different from those reported for surugatoxin, which had been isolated from the ivory shells of Suruga Bay as the toxic principle responsible for the food poisonings which were prevalent in the 1960s in the region. The newly found toxin (s) was extracted from the viscera (162g) with 75% ethanol acidified with acetic acid; the extracts were purified successively on columns of activated charcoal. Bio-Gel P-2, and Bio-Rex70. Two toxins (BJT-1 and BJT-2) having specific activities of 3300 MU/mg and 590 MU/mg, respectively, were obtained. The yields were 0.7mg for BJT-1 and 1.0mg for BJT-2. Comparison of BJT-1 with tetrodotoxin and saxitoxin by tlc, color reactions, dose-survival time curve, and 1HNMR spectroscopy led to the conclusion that BJT-1 is tetrodotoxin. The reaction in mice and to color reagents suggested that BJT-2 also is a metabolite of tetrodotoxin but it differed from tetrodotoxin in its tlc properties and in its positive reaction to Sakaguchi and ninhydrin reagents.

The occurrence of tetrodotoxin (tarichatoxin) in amphibia and the distribution of the toxin in the organs of newts (Taricha)

Abstract Among amphibia tetrodotoxin was found by bioassay only in the family Salamandridae. It was present in substantial concentration (mean value of about 25 μg per g) only in the genus Taricha: T. torosa, T. rivularis, T. granulosa; smaller amounts (1–4 μg per g) were found in Notophthalmus viridescens, Cynops pyrrhogaster, C. ensicauda and trace amounts (less than 0·1 μg per g) in Triturus vulgaris, T. cristatus, T. alpestris, T. marmoratus. This toxin was not detected in fourteen species of the orders Caudata and Salientia. In adult Taricha torosa, tetrodotoxin is concentrated in the skin, ovaries and ova, muscle and blood and is present in trace amounts in the liver, viscera and testes