Large-scale production of selected type A trichothecenes: the use of HT-2 toxin and T-2 triol as precursors for the synthesis of d 3-T-2 and d 3-HT-2 toxin

Abstract

The type A trichothecenes T-2 and HT-2 toxins are toxic secondary metabolites produced by fungi of the Fusarium genus. Their occurrence in cereals, especially in oats, implies health risks for the consumer. Therefore, it is an important task to develop selective and sensitive methods for the analysis of T-2 and HT-2 toxins, and to undertake further studies on their stability and toxicity. Although most toxins are commercially available, their high prices are the limiting factor on the realization of these experiments. Thus, we developed a method for large-scale production of T-2 and HT-2 toxin as well as T-2 triol and T-2 tetraol. T-2 toxin was obtained in gram quantities by biosynthetic production with cultures of F. sporotrichioides. As HT-2 toxin was only formed as a by-product, and T-2 triol and T-2 tetraol were not generated, these compounds were produced by alkaline hydrolysis of T-2 toxin. Separation and isolation of crude toxins was achieved by fast centrifugal partition chromatography (FCPC), which is an efficient tool for the large-scale purification of natural products. Using this fast and yield effective technique, several hundred milligrams of HT-2 toxin, T-2 triol, and T-2 tetraol were obtained. Subsequent, HT-2 toxin and T-2 triol were used for the large-scale synthesis of isotope-labeled T-2 and HT-2 toxin, respectively. Using these standards, an isotope dilution-(ID)-HPLC-MS/MS method for the quantification of T-2 and HT-2 toxin in different matrices was developed.

Influence of T-2 and HT-2 Toxin on the Blood-Brain Barrier In Vitro: New Experimental Hints for Neurotoxic Effects

“…Both trichothecene mycotoxins, T-2 and HT-2 toxin, were biosynthetically prepared and isolated in our laboratory [30]. All chemicals including cell culture medium and supplements were purchased from Merck (Darmstadt, Germany), Sigma-Aldrich (Steinheim, Germany) or Biochrom AG (Berlin, Germany).…”

A comparative study of the human urinary mycotoxin excretion patterns in Bangladesh, Germany, and Haiti using a rapid and sensitive LC-MS/MS approach

“…EnB was from Enzo Life Sciences (Lörrach, Germany). ZEA, DON, FB 1 , OTA, OTα, T-2, and HT-2 were isolated and purified from fungal cultures (Beyer et al 2009;Cramer et al 2007Cramer et al , 2010Bittner et al 2013;Bretz et al 2006;Hübner et al 2012). Glucuronic acid conjugates (DON-3-GlcA, ZEA-14-GlcA, ZAN-14-GlcA, α-ZEL-14-GlcA, β-ZEL-14-GlcA, and HT-2-4-GlcA) were obtained from enzymatic synthesis using rat and pig liver microsomes (Welsch and Humpf 2012).…”

Determination of mycotoxin exposure in Germany using an LC‐MS/MS multibiomarker approach

“…EnB was from Enzo Life Sciences (Lörrach, Germany). ZEN, DON, FB 1, FB 2 , OTA, OT␣, T-2, HT-2 were isolated and purified from fungal cultures [14][15][16][17][18][19][20]. Glucuronic acid adducts (DON-3-GlcA, ZEN-14-GlcA, zearalanone-14-O-glucuronide, ␣-zearalenol-14-Oglucuronide, ␤-zearalenol-14-O-glucuronide, and HT-2-GlcA were synthesized enzymatically using rat and pig liver microsomes according to a recently published procedure [13].…”