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目的建立雪卡毒素的细胞毒性试验检测和小鼠生物法检测方法,对比评价两种方法的优缺点。方法利用不同浓度的雪卡毒素标准品(P-CTX-1)结合乌苯苷和黎芦定处理小鼠神经母细胞瘤细胞(N-2a),研究和建立毒素剂量与细胞活性的毒性曲线关系式。同时,通过测定昆明系小白鼠的雪卡毒素半数致死剂量LD50和毒素剂量与小鼠死亡时间的关系方程式,建立小鼠生物法检测标准,再分别以两种方法对32份不同月份采集的鱼样品的毒素提取物进行检测,对两种方法检测结果做分析比较。结果细胞毒性试验法检测结果与小鼠生物法检测结果具有一定的相关性,但细胞毒性试验法检测雪卡毒素的检测灵敏度远高于小鼠生物法。结论细胞毒性试验检测法适用于大量样品的初步筛选方法,而小鼠生物法可作为雪卡毒素检测的重要参考方法。
OBJECTIVE To establish a method for the detection of ciguatoxin cytotoxicity and mouse bioassay, and to compare the advantages and disadvantages of the two methods. Methods The neurotoxicity of toxin neurotrypsin (N-2a) was treated with different concentrations of Ciguatoxin standard (P-CTX-1) and viprofloxacin and rutin. The toxicity curves of toxin dosage and cell viability Relationship. At the same time, the bioassay standard of mice was established by measuring the LD50 and the dose of LD50 of dexamethasone in Kunming mice and the death time of mice. The toxin extract of the sample was tested, and the results of the two methods were analyzed and compared. Results The results of cytotoxicity assay showed a certain correlation with the bioassay results of mice. However, the cytotoxicity assay was more sensitive than that of mouse bioassay. Conclusion The cytotoxicity test method is suitable for the preliminary screening of a large number of samples, while the mouse biological method can be used as an important reference method for the determination of ciguatoxin.