A high-performance liquid chromatographic method for determining [3H]T-2 and its metabolites in biological fluids of the cynomolgus monkey
| Autor: | Syed M. Naseem, Judith G. Pace, Robert W. Wannemacher |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
Male
Chemical Health and Safety Chromatography Toxin Health Toxicology and Mutagenesis Metabolite Radioimmunoassay Urine Toxicology medicine.disease_cause High-performance liquid chromatography Thin-layer chromatography Analytical Chemistry chemistry.chemical_compound Macaca fascicularis T-2 Toxin chemistry medicine Environmental Chemistry Animals Mycotoxin Quantitative analysis (chemistry) Chromatography High Pressure Liquid |
| Zdroj: | Journal of analytical toxicology. 19(3) |
| ISSN: | 0146-4760 |
| Popis: | High-performance liquid chromatography (HP/C) was used to separate, identify, and quantitate the trichothecin mycotoxin, T-2 (4~,15-diacetoxy-30c-hydroxy-8oc [3-methyl-butyryloxy]-I 2,13epoxy Ag-trichothecin), and its metabolites in plasma and urine samples from cynomolgus monkeys treated with the toxin. A 15rain gradient elution system was developed to separate and measure radiolabeled T-2 mycotoxin and its metabolites. The HPLC technique for separating 1-2 and its metabolites was compared with thin-layer chromatography. Samples from the in vitro metabolism of T-2 by plasma and urine were included as controls and as a measure of the toxin's stability in biological samples. Within 5 min, 22% of the plasma radiolabeled 1"-2 toxin was detected as metabolites after an intravenous administration of [3H] T-2 toxin to cynomolgus monkeys. By 24 h post-exposure, there was no parent 1-2 toxin detected in plasma or urine. T-2 tetraol was the major metabolite detected in the plasma and urine of monkeys. Other metabolites observed in urine up to 5 days after exposure were 3'OH-T-2 and 3'OH-HT-2. We conclude that 1-2 toxin was rapidly metabolized to more polar metabolites, which were eliminated in urine. Several assay methods have been developed for detection and quantitation of T-2 toxin and other mycotoxins. Biological samples are usually analyzed by thin-layer chromatography (TLC) (8), immunological methods, radioimmunoassays (9), gas-liquid chromatography, and mass spectrometry (10-15). Biological methods can also be used, including mouse bioassay (16), skin test (17), and assays of inhibition of protein synthesis (18) and inhibition of cell growth in culture (19). There is also an enzyme method (20) (binding with reduced glutathione in the presence of glutathione-s-epoxide transferase) that is currently used. The disadvantages of these methods include low detection limits, la(:k of specificity and sensitivity, or they are excessively expensive for assaying T-2 and metabolites in biological samples. In this study, we compared high-performance liquid chromatography (HPLC) and thin-layer radio chromatography to isolate, separate, identify, and quantitate T-2 toxin and its metabolites in biological samples of cynomolgus monkeys. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|