Talanta 2008-02-15

Improvement of detection sensitivity of T-2 and HT-2 toxins using different fluorescent labeling reagents by high-performance liquid chromatography.

Vincenzo Lippolis, Michelangelo Pascale, Chris M Maragos, Angelo Visconti

Index: Talanta 74(5) , 1476-83, (2008)

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Abstract

T-2 and HT-2 toxins are Fusarium mycotoxins that can occur in cereals and cereal-based products. Three fluorescent labeling reagents, i.e. 1-naphthoyl chloride (1-NC), 2-naphthoyl chloride (2-NC) and pyrene-1-carbonyl cyanide (PCC), were used for the determination of T-2 and HT-2 toxins by high-performance liquid chromatography (HPLC) with fluorescence detection (FD). Pre-column derivatization of T-2 and HT-2 toxins was carried out under mild conditions (50 degrees C, 10 min) in toluene with 4-dimethylaminopyridine (DMAP) as catalyst. All fluorescent derivatives were identified and characterized by HPLC-tandem mass spectrometry (HPLC-MS/MS). Optimal stoichiometric ratios (toxin:derivatizing reagent:catalyst), linear range and repeatability of the reaction, stability and sensitivity of the derivatives were determined. A wide linear range (10-1000 ng of either derivatized T-2 or HT-2 toxin), good stability (up to 2 weeks at -20 degrees C or 5 days at room temperature) of the fluorescent derivatives and good repeatability of the reaction (RSD