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A study of ten toxins associated with paralytic shellfish poison using prechromatographic oxidation and liquid chromatography with fluorescence detection.

Ten paralytic shellfish toxins [saxitoxin, neosaxitoxin, B-1, B-2, gonyautoxin 1, 2, and 3 (i.e., GTX-1, GTX-2, and GTX-3), C-1, C-2, and C-3] were oxidized at room temperature under mildly basic conditions with hydrogen peroxide or periodic acid. The products were then analyzed by liquid chromatography (LC). The N-1-hydroxylated toxins (neosaxitoxin, B-2, GTX-1, and C-3) formed fluorescent products after periodate oxidation at ca pH 8.7, but did not form fluorescent derivatives with peroxide oxidation. The non-N-1-hydroxylated toxins (saxitoxin, B-1, GTX-2, GTX-3, C-1, and C-2) formed highly fluorescent derivatives with both peroxide and periodate oxidations. Individual toxins produced mainly single fluorescent peaks by reverse-phase LC. However, all GTX toxins eluted with the same retention time. Also, C-1 and C-2 eluted together, as did neosaxitoxin and B-2. The non-N-1-hydroxylated toxins could be detected in quantities as low as 20-50 pg/injection, while the N-1-hydroxy analogues could be detected at levels as low as 100-500 pg/injection. UV absorption and fluorescence emission spectra were similar for the oxidation products of all toxins examined (max. 333 +/- 2 nm absorption, 389 +/- 4 nm fluorescence emission).

Animals↗
PubMed1983

Release of tetrodotoxin and paralytic shellfish poison from puffer liver by RNase.

A nontoxic high-molecular fraction was separated from the extracts of toxic liver of a puffer, Takifugu poecilonotus, by Sephadex G-50 gel filtration. This fraction became toxic when digested with RNase T2. The toxins were partially purified by activated charcoal treatment, followed by chromatography on Bio-Gel P-2 and Bio-Rex 70, and were analyzed by TLC and electrophoresis. The results showed that most of the toxicity is accounted for by tetrodotoxin, and the remainder by saxitoxin and other unidentified toxins. The corresponding high-molecular fraction separated from nontoxic liver of another puffer, T. rubripes, did not release any toxin on RNase digestion.

Animals↗