Methods Of Extraction, Isolation And Detection Of Paralytic Shellfish Poisoning Toxins In Bivalve

PSP ( Paralytic shellfish poisoning) toxins, distributing widely in marine environment, are potent neurotoxins which can block voltage-gated sodium channels on excitable cells and finally result in signs of paralysis. As one of the most toxic natural toxins, PSP has gained more and more attention. In this thesis, the extraction, isolation and detection of PSP have been studied.The optimal condition was established for extraction of PSP from shellfish using boiling extraction with HAC and HCl in the concentration range 0.04-1.Omol/L. Hydrochloric acid in the concentration range 0.25-1.0mol/L caused the amount of C1/2 and B1 toxins to decreased sharply and the concomitant increase of GTX2/3. However the amount of the three unstable toxins did not show any change using the extraction with HAC. Ultrasonic extraction of PSP were also studied. The results indicated that there was not any difference in the extraction with 0.2mol/L HC1 and 0.3mol/L HAC.PSP toxins were purified from the extraction of shellfish by BIO-GEL P2 column. Fluorescence detector and HPLC were used in the whole process to monitor the toxins. The optimal derivatization condition for fluorescence detection of PSP toxins was investigated according to a L₉(3⁴) orthogonal test design that was also used to optimize the post-column oxidant condition in HPLC method.A rapid and accurate high-performance liquid chromatography (HPLC) system was set up to simultaneously analyse GTX1, GTX4, GTX2, GTX3, dcGTX2, dcGTX3, neoSTX, STX, dcSTX, B1(GTX5). Three-step elution are performed on a 25-cm RP-C8 chromatographic column with two eluents containing phosphate buffer and sodium heptanesulfonate as ion-pair former and the latter also containing acetonitrile. This method gave a satisfied resolution and sensitivity; all of the 10 PSP toxins had high linear correlation with the coefficient >0.99A simple and efficient method for determination of paralytic shellfish poisoning (PSP) was developed. Only one buffer is used and the gradients with the buffer and pure acetonitrile are performed to determine all GTXs and NEO/dcSTX/STX. The...