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Determination of metolcarb residues by a biotin-streptavidin-amplified enzyme-linked immunosorbent assay in vegetables and edible fungus

机译:通过生物素-链霉亲和素扩增的酶联免疫吸附法测定蔬菜和食用菌中的灭多威残留量

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摘要

An effective biotin-streptavidin-amplified enzyme-linked immunosorbent assay (BA-ELISA) to rapidly detect metolcarb (MTMC) residues is reported. Nonspecific adsorption was minimized by using 0.5% skimmed milk powder as a blocking buffer and 0.5% bovine serum albumin/phosphate-buffered saline (PBS) as a buffer for streptavidin-horse radish peroxidase conjugates. The established method is four times sensitive than direct competitive enzyme-linked immunosorbent assay, with the IC50 of 10.0 ng mL(-1). The samples were prepared from mustard, cucumber and mushroom with simple extraction and dilution methods, including use of PBS without concentration or cleanup steps. The samples prepared from spinach and shiitake were quantified after 2-fold methanol extraction and 20-fold dilution with 2.0% fish glutin/PBS. Good accuracy and precision were obtained with mean recoveries between 80.0% and 95.6%, and mean coefficients of variation below 12.1%. A good correlation (R-2=0.9931) between the BA-ELISA and high-performance liquid chromatography was observed for MTMC analysis in different samples. This method could be potentially useful for high-throughput food inspection.
机译:据报道,一种有效的生物素-链霉亲和素扩增的酶联免疫吸附测定(BA-ELISA)可快速检测灭多威(MTMC)残留。通过使用0.5%脱脂奶粉作为封闭缓冲液和0.5%牛血清白蛋白/磷酸盐缓冲盐水(PBS)作为链霉亲和素-马萝卜过氧化物酶结合物的缓冲液,可以最大程度地减少非特异性吸附。建立的方法的灵敏度是直接竞争酶联免疫吸附法的四倍,IC50为10.0 ng mL(-1)。样品是从芥菜,黄瓜和蘑菇中提取的,采用简单的提取和稀释方法,包括使用没有浓缩或纯化步骤的PBS。在用2%甲醇提取和20%用2.0%鱼胶/ PBS稀释后,对从菠菜和香菇制备的样品进行定量。获得了良好的准确度和精密度,平均回收率在80.0%至95.6%之间,平均变异系数低于12.1%。在不同样品的MTMC分析中,BA-ELISA与高效液相色谱法之间观察到良好的相关性(R-2 = 0.9931)。此方法可能对高通量食品检查有用。

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