nsc-141537 and verrucarol

A reliable, sensitive and selective method was developed to determine different Fusarium mycotoxins (trichothecenes Type A and B, zearalenone) simultaneously in cereals and cereal-based samples using liquid chromatography with tandem mass spectrometry (LC-ESI-MS/MS). Sample preparation is based on a standard solvent extraction step followed by two different kinds of solid-phase clean-up procedures: using a multifunctional MycoSep material for trichothecenes and zearalenone. The average recoveries for trichothecenes ranged from 65% for nivalenol (NIV) up to 96% for deoxynivalenol (DON) and 89% for zearalenone (ZON). The limit of quantification varied between 0.02 and 10 ppb for each substance. In addition, a screening survey with 685 samples was carried out to compare contents of T-2 toxin and deoxynivalenol and to investigate potential coherence in contamination pattern.

Topics: Chromatography, Liquid; Edible Grain; Flour; Food, Fortified; Fusarium; Mass Spectrometry; T-2 Toxin; Trichothecenes; Triticum; Zearalenone

Mycotoxicoses have been recognized worldwide to cause problems in animal production. Trichothecene mycotoxins, for the most part produced by Fusarium spp., have obtained particular importance. Between 1982 and 1984, 295 samples of feedstuffs (cereals and mixed feeds) have been analyzed on natural occurrence of type-A trichothecenes. The skin toxicity test with guinea pigs was used as a screening method. Analysis of trichothecenes by capillary gaschromatography with flame ionization detection was complicated by interfering substances from the complex matrix of the sample-material. Definite results were only obtained by the use of gaschromatography-mass spectrometry. In comparison to the electron impact ionization the chemical ionization technique showed to be advantageous. Forty-two of the 295 samples analyzed were found to be positive in the biological assay. Nine of these positive samples contained trichothecenes as determined by mass spectrometry: T-2 toxin in mixed feed (65 micrograms/kg); oats (80 and 86 micrograms/kg) and wheat (100 micrograms/kg); diacetoxyscirpenol in mixed feed (125 micrograms/kg) and wheat (50 micrograms/kg); neosolaniol in oats (310 and 350 micrograms/kg); HT-2 toxin in oats (700 micrograms/kg). Oats proved to be contaminated more frequently as compared to the other cereals and the analyzed mixed feeds.

Topics: Animal Feed; Animals; Biological Assay; Chemical Phenomena; Chemistry; Flame Ionization; Gas Chromatography-Mass Spectrometry; Guinea Pigs; Sesquiterpenes; Skin; T-2 Toxin; Trichothecenes

Immunization of rabbits with deoxyverrucarol (DOVE) conjugated to bovine serum albumin resulted in antibodies bound with either tritiated DOVE or diacetoxyscirpenol (DAS), but not with T-2 toxin. The affinity of antibodies with DOVE was found to be much higher than with DAS. When [3H] DOVE was used as a marking ligand in the competitive radioimmunoassay, the concentrations causing 50% inhibition of binding radioactivities by unlabeled DOVE, verrucarol, verrucarin A, and 4-monoacetoxyscirpenol were found to be 0.32, 1,070, 9,500, and 10,000 ng per assay, respectively. T-2 toxin, 15-monoacetoxyscirpenol, and deoxynivalenol gave less than 20% inhibition at 10 micrograms per assay. However, when [3H] DAS was used as the marking ligand, the concentrations causing 50% inhibition by DOVE, DAS, and verrucarol were found to be in the 50 to 60 ng per assay range. The antibodies are thus highly specific to DOVE rather than a common trichothecene backbone. The possible use of this antiserum for assay of macrocyclic trichothecenes is discussed.

Topics: Animals; Antibodies; Antibody Specificity; Immunization; Mycotoxins; Rabbits; Serum Albumin, Bovine; Sesquiterpenes; Trichothecenes