超声波提取-固相萃取联用检测河鲀毒素的研究

Detection of tetrodotoxin in aquatic products using HPLC-UV

使用热处理、震摇、超声波等手段改善水产品中的河鲀毒素提取率,利用多种固相萃取柱纯化提取液,采用高效液相色谱-紫外检测法进行检测,重点对河鲀毒素的提取和固相萃取纯化进行了研究。试验发现,结合振荡和加热双重作用的索氏提取法和超声波加热辅助提取法具有较高的提取能力,其中超声波(35kHz,强度100%,60℃)提取10min即可达到最好效果,耗时短,效率更高;比较SCX、PRS、WCX、C18、SAX 5种固相萃取柱对河鲀毒素的吸附作用,SCX、PRS、WCX柱则能够将河鲀毒素完全保留在其上。样品纯化试验结果显示,C18-WCX小柱的组合纯化样品的效果最好,回收率在75%—82%之间,相对标准偏差小于15%。研究结果对河鲀毒素的提取、纯化和检测工作具有重要的参考意义。

Tetrodotoxin （TTX） accumulated in some aquatic products can be extremely dangerous to human beings. Detecting the content of TTX in aquatic products can help to avoid consuming the toxic products. Due to the complexity of sample composition, TTX detection method has not been developed. The new methods of extracting TTX in puffer fish were studied and presented here. A variety of solid phase extraction column for purification were compared, the high performance liquid chromatography （HPLC） coupled with a UV detector was adopted in the detection. The results showed that both the Soxhlet extraction method and ultrasound treatment method have higher extraction efficiency. Solid-phase extraction （SPE） cartridges were compared in the TTX adsorption experiments, which revealed that the strong cation exchanger （SCX）, the propanesulfonic acid SPE cartridge （PRS）, and the weak cation exchanger （WCX） had good capacity of retaining TTX. The combination of the octadecyl silane SPE cartridge （C18） and WCX cartridge is even better. The best result can be achieved by extracting 10 min at 35 kHz, 100% intensity, 60~C ultrasound treatments. The recovery rate is 75%--82%, with the relative standard deviation （RSD） of less than 15%. This newly developed method is efficient and ready to be used in the laboratory.