| Intoxication by paralytic shellfish poisoning (PSP) toxins in seafood was one of potential food safety threaten, relates directly to human health, so it has the important meaning to study the PSP tolerance mechanism. A post-column derivatization high performance liquid chromatography method was developed to detect PSP toxins with fluorescence detector(FLD) in this paper. Taking crabs inhabited near Zhanjiang leizhou peninsula as the research objects, analyze the PSP toxicity and resistance to PSP toxins, and extract and purify the PSP-binding protein in the body fluid of crab T. crenata, further expound its physical and chemical properties, and establish the dynamic model of combination with PSP. The conclusion will provide scientific theoretical basis for the PSP resistance medicine and mechanism.The experimental results showed that, crabs T. crenata, L. exaratus, M. latifrons, lived in Zhanjiang leizhou peninsula contained little PSP toxins and high tolerance to PSP. Preparated crabs’body fluid and mixed with PSP, after intraperitoneal injection of mice found that the group contained body fluid compared with controls, the mouse was significantly longer survival. In contrast to that of the low molecular fraction (<10KDa), the death time of mouse for high molecular fraction (>10KDa) prolonged2times, implying there was substance with toxicity neutralization effect in the high molecular fraction. Protein and sugar analysis showed that the protein content was72.06%, and99.63%of the protein mainly focus in>10KDa. In the addition, after treated by protease, the PSP toxicity neutralization effect of the body fluid disappeared, implying the functional composition with PSP toxicity neutralization effect was protein. These results indicate there is PSP-binding protein in the body fluid, and result a PSP toxicity neutralization effect.Using the affinity resin AF-Epoxy-650M to purify PSP-binding protein, collected the eluent and concentration, the PSP-binding protein yield is1.32%finally. Through the electrophoresis analysis indicated the PSP-binding protein a single channel with a high purity, should be a protein with high molecular weight more than200KDa.Based on the establishment platform of HPLC, detected the combine rates of PSP-binding protein to PSP toxins, the results showed different combine rate with GTXs〠STXs, CTXs. Among them STXs showed a high rate, the rate of neoSTX, dcSTX, STX are72.27%,69.20%and56.92%, respectively; followed by GTX4,1,2,3, combined with the rate in61.45％,60.56％,33.56％,29.78％；And for CTX1and CTX2combination of low rate,to14.22％and15.04％,respectively. |
