A rapid and sensitive procedure for the quantitative detection of AL-toxin by fluorescence derivatization and separation by high performance liquid chromatography
1995
Kodama, M. (Tottori Univ. (Japan). Faculty of Agriculture) | Otani, H. | Kohmoto, K.
An improved high performance liquid chromatographic (HPLC) method was developed for the detection and quantitation of host-specific Al-toxin produced by Alternaria alternata tomato pathotype. Precolumn derivatization of the toxin with o-phthalaldehyde (OPA) plus mercaptoethanol or 4-fluoro-7-nitrobenzofurazan (NBD-F) yielded highly fluorescent products showing well-resolved peaks on reversed phase HPLC. The minimum amounts of AL-toxins detectable by this method were approximately 1 ng. This method seems to offer advantages over conventional techniques, because prechromatographic derivatization of the toxin with OPA or NBD-F is rapid and easy, and fluorescent derivatives permit excellent detection sensitivity. Toxin production in culture filtrates and spore-germinated fluids of the pathogen was quantitatively analyzed using this procedure
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