Development of the sensor for the determination of Na channel blockers and its application to the determination of fish and shellfish toxins and the survey of the physiological active products

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The sensor for the determination of NaィイD1+ィエD1 channel blockers consisted of frogs bladder, NaィイD1+ィエD1 electrode, flow cell, peristaltic pump and recorder. The basic conditions for the determination of the Na channel blockers were established as follows.Preparation of Na channel electrode : The tip of NaィイD1+ィエD1 electrode was covered by the frogs bladder membrane sandwiched between 2 sheets of cellulose acetate membrane. This electrode was integrated within a flow cell connected to a peristaltic pump and injection port.Some conditions for the determination of a channel blockers : The initial carrier solution contained 8% NaCl and adjusted to pH 4.8 was transferred into the flow cell with 0.8 ml/min at 30℃. Once the background signal had stabilized, 50 ul of TTX sample solution was injected into the sensor system and the response was measured.Preparation of sample : Toxins of swellfish and shellfish was extracted from each 1 g of them with 0.1% acetic acid and hydrochloric acid at 100℃ for 10 min, respectively and planktons toxin was also extracted from the filter paper which the plankton was corrected on with 0.01 M acetic acid at 100 ℃ for 20 min. NaィイD1+ィエD1 channel blockers contained into seaweed and herb of Chinese medicine was extracted from 1 g of dried seaweed with 0.1 N hydrochrolic acid at 100℃ for 10 min and 20 to 30 g of the dried herb after refluxing with hot water at 90℃ for 1 hr, respectively.Toxins of swellfish and plankton were determined under the conditions mentioned above. NaィイD1+ィエD1 channel blockers contained in seaweed and herb of chinese medicine were also searched.From these results, It had become feasible to determine NaィイD1+ィエD1 channel blockers rapidly, simply, high sensitively and continuously.