膨化脱毒处理后蓖麻籽粕中蓖麻毒素的检测

发布时间：2020-09-05 11:47:50 来源：文档文库

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膨化脱毒处理后蓖麻籽粕中蓖麻毒素的检测

武军华;李倩;董娜;王玉霞;贾培媛;梁晓;赵元忠

【期刊名称】《军事医学》

【年(卷),期】2013(037)001

【摘要】目的检测膨化脱毒蓖麻籽粕中的残余毒素.方法蓖麻籽粕经过粉碎、匀浆及离心后得到匀浆上清,应用酶联免疫吸附分析(ELISA)、免疫印迹、金标免疫层析试纸检测蓖麻籽粕在PBST匀浆上清中的蓖麻毒素;应用细胞生长抑制实验测定蓖麻籽粕在生理盐水匀浆上清的细胞毒性.结果经双夹心ELISA检测蓖麻原粕匀浆含有较高浓度的蓖麻毒素,达到0.25 mg/g;对细胞生长有明显的抑制作用;应用蓖麻毒素检测试纸显示强阳性条带.应用试纸条检测经加热膨化处理后的样品只显示微弱条带.ELISA检测结果表明加热膨化处理后蓖麻籽粕中的毒素含量明显下降,脱毒率达到99%以上.加热膨化处理后样品的匀浆上清在高浓度时对细胞生长仍有一定抑制作用,但与原粕比较细胞毒性明显下降;应用免疫印迹检测蓖麻籽粕结果与ELISA和细胞实验结果一致.结论本研究将ELISA、免疫印迹和免疫层析试纸相结合,建立了有效检测蓖麻籽粕中残余蓖麻毒素的方法,为蓖麻籽粕脱毒处理样品的质量检测及其应用提供了实验依据.%Objective To establish a method for residual ricin detection in the meal of castor beans. Methods Castor bean meal was shattered and then homogenized in PBST. The residual toxin in the supernatant of homogenate in PBST was analyzed by three immunoassay methods,!, e. ELISA, Western blotting and immunochromatographic colloid gold strip assay. The toxin in the supernatant of homogenate in saline was measured via cell growth inhibition assay. Results High-concentration ricin ( up to 0. 25 mg/g ) in the original meal was detected by sandwich ELISA. The supernatant showed a strong positive band on the colloid gold strip and displayed a significant inhibitory effect on cell growth. Castor bean meal could be detoxified through extrusion-expansion at a high temperature and high shearing force. The ricin in the supernatant from the detoxified meal was hardly detectable via the colloid gold strip. The detoxification efficiency exceeded 99% according to ELISA result, as with Western blotting. The homogenate supernatant of detoxified meal was still able to inhibit cell growth at a high concentration, but was much weaker than that from the untreated meal. Conclusion A combined immunodetection method, including ELISA, Western blotting and immunochromagraphic colloid gold strip assay, is estab lished for residual ricin detection in castor bean meal. It is a promising method for quality control of castor bean detoxification.