玉米中伏马菌素测定免疫亲和柱的重复利用。

B Fazekas, A Koncz-Tar, E Tóth-Hajdu, M Zomborszky-Kovács
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引用次数: 7

摘要

采用免疫亲和柱-高效液相色谱法测定了18份玉米样品中伏马菌素B1 (FB1)和B2的含量。使用一次FumoniTest柱分离伏马菌素(一次性柱法)。在实验的第二部分,柱再生。用甲醇洗脱后,PBS溶液在室温下停留1天,使柱再生(再生柱法)。通过测定色谱柱的回收率、回收率和测定结果的重现性,考察了两次再生柱的效率。单柱法测定FB1的回收率为82% (RSD: 5.7%),再生柱法测定FB1的回收率为82.6% (RSD: 5.6%);在柱上加载500 ~ 8000 ng FB1对柱性能没有影响。两种方法测定含伏马菌素的玉米样品中FB1含量的结果几乎相同。结果表明,该方法可以在不降低柱性能的情况下再生两次以上的FumoniTest柱。再生柱的伏马菌素亲和力、容量和特异性没有变化。因此,用这种方法再生的色谱柱可用于测定玉米样品中伏马菌素的含量至少三次。
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Reusability of immunoaffinity columns for determination of fumonisins in maize.

Eighteen maize samples were assayed for fumonisin B1 (FB1) and B2 content by immunoaffinity column coupled with high performance liquid chromatography (HPLC). The FumoniTest columns were used once for the isolation of fumonisins (single-use column method). In the second part of the assay the columns were regenerated. After elution with methanol, PBS solution was left on the column for one day at room temperature to regenerate the columns (regenerated column method). The efficiency of columns regenerated twice was tested by determining FB, recovery and the reproducibility of the determinations. The recovery rate of FB1 proved to be 82% by the single-use column method (RSD: 5.7%) and 82.6% (RSD: 5.6 %) by the regenerated column method; 500-8,000 ng FB1 loaded onto the columns did not affect column performances. Nearly identical values were obtained when the FB1 content of fumonisin-containing maize samples was determined by both methods. The results indicate that the FumoniTest columns can be regenerated by the method applied at least twice without decrease in column performance. The fumonisin affinity, capacity and specificity of the regenerated columns were not changed. Thus, columns regenerated in this way can be used for determining the fumonisin content of maize samples at least three times.

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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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