Facile detection of anatoxin-a in algal material by thin-layer chromatography with Fast Black K salt

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Abstract
A method for facile high-capacity screening of algal samples for anatoxin-a (ANTX-a), a potent neurotoxin of Anabaena flos-aquae, is presented. The method is based on in situ colour reaction of algal extracts containing ANTX-a on a thin-layer chromatographic plate with the diazonium reagent Fast Black K salt, and subsequent separation of the orange–red product. The product, shown to be a stable 3,3-dialkyltriazene, is derived from a reaction involving the aliphatic secondary amino group of ANTX-a. The detection limit for ANTX-a is 10 µg g–1 of lyophilized algal material, which is comparable to earlier methods using more complex instrumentation.