酵母中毛霉烯的生物测定。

J Binder
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引用次数: 9

摘要

与所有真核生物细胞一样,酵母对毛霉烯,特别是T-2毒素和疣胞素a敏感,基于这种敏感性,开发了酵母生物测定法来评估玉米样品的毒性。采用加标玉米提取物对生物测定法进行优化。样品的毒性定义为毒性相当于一定浓度的T-2毒素标准。该方法可用于粗提物,但净化柱后的结果更为精确。一般来说,该试验也可用于筛选天花粉的毒性。在85%生长抑制(EC85)下,T-2毒素标准(n = 8)的相对标准偏差(RSD)为4.5%。这对应于玉米样品中T-2毒素的初始浓度约为58 ppb。含有188和113 ppb T-2毒素的样品对生长的抑制作用大于85%,而毒素浓度为56和19 ppb的样品对生长的抑制作用小于85%。因此,该测试可用于定性评价玉米样品高达58 ppb +/- 2.8 ppb的水平。该生物测定法易于操作,对设备的要求最低。可在24小时内获得结果,每天可分析大量样品。成本低,所得结果可重复。经过一些修改,本试验可用于对毛霉烯代谢物的毒性研究,以及具有与毛霉烯性质相似的未知化合物的提取物。
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A yeast bioassay for trichothecenes.

Like all eucaryotic cells, yeasts are sensitive to trichothecenes, especially T-2 toxin and verrucarin A. Based on this sensitivity, a yeast bioassay was developed to evaluate the toxicity of corn samples. The bioassay was optimized using spiked maize extracts. The toxicity of samples was defined as toxicity equivalent to a certain concentration of T-2 toxin standards. The assay can be performed on crude extracts, but the results are more precise after column clean-up. The test can also be used for the screening of trichothecene toxicity in general. The relative standard deviation (RSD) at 85 % growth inhibition (EC85) was 4.5% for the T-2 toxin standards (n = 8). This corresponds to an initial T-2 toxin concentration of approximately 58 ppb in the corn sample. Samples containing 188 and 113 ppb T-2 toxin caused a growth inhibition higher than 85%, whereas samples with toxin concentrations of 56 and 19 ppb had a growth inhibition less than 85%. Therefore the test can be used for the qualitative evaluation of corn samples up to a level of 58 ppb +/- 2.8 ppb. The bioassay is easy to perform with minimum requirements for equipment. Results can be obtained within 24 h and a large number of samples can be analysed daily. The costs are low and the results obtained are repeatable. With some modifications this test can be used for toxicity studies on trichothecene metabolites as well as for extracts with unknown compounds with properties similar to trichothecenes.

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Erratum: Alfonso D, Johnson HA, Colman-Saizarbitoria T, Presley CP, McCabe GP, McLaughlin JL (1996): SARs of annonaceous acetogenins in rat liver mitochondria. Nat Toxins 4:181-188. Advances in detection methods for fungal and algal toxins. HPLC/MS analysis of fusarium mycotoxins, fumonisins and deoxynivalenol. Neuronal binding of tetanus toxin compared to its ganglioside binding fragment (H(c)). A new type sandwich immunoassay for microcystin: production of monoclonal antibodies specific to the immune complex formed by microcystin and an anti-microcystin monoclonal antibody.
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