反相高效液相色谱法(HPLC)测定河豚毒素的方法研究

Study on the Method of Detecting Tetrodotoxin by Reverse phase HPLC

本文介绍反相HPLC法测定河豚毒素(TTX),用美国waters公司产HPLC仪,配带M486紫外检测器,检测波长210nm,C_(16)径向加压柱(8×100mm,10μm),流动相选用pH6.8磷酸盐缓冲溶液,流速0.4ml/min,U_6K手动进样阀,2个510型高压泵,M810微机数据处理。该法快速,仅用12min4种异构体便可分离定量。精密度高,标准差为0.025～0.223,相对标准偏差为0.92％～4.36％。检出限为10ng。标准曲线在5～20ng范围内线性关系良好,回归方提Y=0.0001+0.28x,回归系数0.9999。

The authors reported a reverse phase HPLC method to detect tetrodotoxin using Waters' HPLC instrument with M486 UV detector at 210nm, Cis radial pressured column (8?100mm, 10μm).U5K manual injection vatlve, two-type 510 re- ciprocation pumps and type 810 Computer. Buffer solution at pH6. 8 was used as mobile phase at the rate of 0. 4ml/min. The method is quite quick. The 4 derivatives of tetrodotoxin can be isolated and detected within 12 minutes. The precision is high with standard deviation between 0. 025 - 0. 223 and relative standard deviation between 0. 92%~4. 36%. Detection limit is l0ng . A good linearity was shown while the amount of tetrodotoxin was ranging from 0. 5 to 4200μg. Regression e-quation is given as Y=0. 0001 + 0. 28X and regression coefficient is 0. 9999