Related Experiment Video
Updated: Jul 6, 2026

Detection of Toxin Translocation into the Host Cytosol by Surface Plasmon Resonance
Published on: January 3, 2012
Improvement of detection sensitivity of T-2 and HT-2 toxins using different fluorescent labeling reagents by
Vincenzo Lippolis1, Michelangelo Pascale, Chris M Maragos
1Institute of Sciences of Food Production (ISPA), National Research Council (CNR), Via G. Amendola 122/O, 70126 Bari, Italy.
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=8%) were observed. Detection limits (based on a signal-to-noise ratio of 3:1) were 10.0, 6.3 and 2.0 ng for derivatized T-2 toxin and 6.3, 2.3 and 2.8 ng for derivatized HT-2 toxin with 1-NC, 2-NC and PCC, respectively. In terms of sensitivity and repeatability, PCC and 2-NC reagents showed better performance than 1-anthroylnitrile (1-AN), a previously reported labeling reagent for T-2- and HT-2 toxins. Preliminary studies also showed the applicability of PCC and 2-NC as fluorescent labeling reagents for the simultaneous determination of T-2 and HT-2 toxins in cereal grains by HPLC/FD following immunoaffinity column clean-up.
Related Concept Videos
Labeling DNA Probes
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
High-Performance Liquid Chromatography: Types of Detectors
