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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 detect residual toxins in puffed detoxified castor meal. Methods The castor meal was crushed, homogenized and centrifuged to obtain the homogenate supernatant. The enzyme-linked immunosorbent assay (ELISA), Western blotting and gold standard immunochromatographic test strip were used to test the effect of castor meal on the surface of PBST homogenate Of ricin. Cell cytotoxicity of castor meal in supernatant of saline solution was measured by cytostatic assay. Results Double-sandwich ELISA showed that ricin-loaded raw meal homogenate contained a high concentration of ricin, which reached 0.25 mg / g, and had a significant inhibitory effect on cell growth. The ricin test strip showed strong positive bands. The application of test strips to heat puffed samples showed only weak bands. The results of ELISA showed that the toxin content in castor meal decreased obviously after heating and puffing, and the detoxification rate reached more than 99%. The homogenate supernatant of the sample after heating and puffing treatment still inhibited the cell growth at high concentration, but the cytotoxicity was obviously decreased compared with the original meal. The result of the immunoblot assay of the castor meal was consistent with ELISA and cell experiment. Conclusion In this study, ELISA, Western blotting and immunochromatographic test strips were combined to establish an effective method for the determination of residual ricin in castor meal, which provided an experiment for the quality control of castor meal detoxification samples and their application in accordance with.