不同配比微生物毒性传感器对河豚毒素毒性的测定及与HPLC法比较.docVIP

不同配比微生物毒性传感器对河豚毒素毒性的测定及与HPLC法比较 　　摘要：河豚毒素传感器响应性能与固定化菌体的量和辅料的量都有一定关系。设计微生物量为10 μL，不加辅料及加入1%、2%辅料3种微生物膜，以夹层法固定微生物膜，以氧电极为换能器，分别组装微生物传感器，通过测定传感器的线性范围、线性相关系数以及加标回收率等获知固定化膜的通透性的好坏和响应信号的大小，最后确定3种组合中较优膜为10 μL菌悬液加入1%的辅料。以最优膜组装毒性传感器与高效液相色谱方法的对照发现两者的差异不显著，表明试验建立的毒性微生物传感器方法可以很好地测定河豚毒素的毒性。 　　关键词：河豚毒素;毒性传感器;HPLC法;微生物膜 　　中图分类号：S852.4+4 文献标识码：A 文章编号：0439-8114（2014）20-4952-03 　　DOI：10.14088/j.cnki.issn0439-8114.2014.20.048 　　Biosensor Monitoring Tetrodotoxin with Toxicity Sensor of Different Ratio Microbes 　　and Comparison with HPLC Methods 　　CHEN Jing， YANG Zhuo 　　（Environmental Management College of China， Qinhuangdao 066004， Hebei，China） 　　Abstract： Tetrodotoxin sensor response performance is closely related with the amount of immobilized bacteria and accessories. Three kinds of microbial membrane were designed. The first one was 10 μL microbial without no accessory material. The second one was 10 μL microbial with 1% accessory material. The third one was 10 μL microbial with 2% accessory material immobilized by sandwich immobilization. A biosensor was fabricated based on oxygen electrode. By measuring the linear range， the linear correlation and the reclaim rate of the two sensor， the property of membrane and value of the response signal was found out. Results showed that 10 microbial with 1% auxiliary materials was the optimal combination of the two membrane. The toxicity sensor was assembled with the optimal membrane and compared with HPLC method. The results showed that the difference between the two methods was not significant. The toxicity microbial sensor method could accurately measure TTX. 　　Key words： tetrodotoxin; toxicity sensor; HPLC; biofilm 　　河豚毒素（Tetrodotoxin，简称TTX）是一种强神经毒素，目前河豚毒素检测的标准方法是小鼠生物测定法[1]，其他的检测方法如高效液相色谱法[2]、免疫学方法[3]也日渐成熟。微生物传感器方法[4]由于其方法快捷迅速、价格低廉，越来越成为河豚毒素检测方法研究的重点。有文献报道[5-8]，以地衣芽孢杆菌作为响应菌株，采用夹层法制备微生物传感器测试其对河豚毒素的响应情况，在浓度10～100 μg/L范围内线性较好。 　　传感器响应性能与固定化菌体的量和辅料的量都有一定关系。在提高微生物膜的重现性及测量范围的可行性问题上设计微生物量为10 μL和辅料量分别占湿菌体0%、1%、2% 3种组合的微生物膜的性能测试。通