Precolumn derivatization and high-efficiency liquid-phase fluorescence detection method for tetrodotoxin and kit based on the method

Abstract

The invention discloses a precolumn derivatization and high-efficiency liquid-phase fluorescence detection method for tetrodotoxin. The method includes following steps: (1) preparing a sample solution of a sample to be measured and a standard substance solution of tetrodotoaxin; (2) adding 0.2 ml of a pH maintenance solution to 0.4 ml of the sample solution for controlling pH of the sample solution at a range between 11.6 and 11.9, then adding 0.1 ml of a derivatization agent 1 and 0.1 ml of a derivatization agent 2, mixing uniformly, carrying out a water-bath heating process for 20 min and then carrying out a cooling process to room temperature, adding 0.1 ml of a stopping solution for enabling the pH of the sample solution to be neutral, and then carrying out a then diluting to volume with ultrapure water, adding the standard substance solution of the tetrodotoxin to the sample solution, mixing uniformly, and then carrying out a fluorescence derivatization process to the sample solution; and (3) with a liquid-phase chromatograph instrument equipped with a fluorescence detector, analyzing the sample solution which is subjected to the fluorescence derivatization process in the step (2) and the standard substance solution of the tetrodotoxin to detect the tetrodotoxin qualitatively and quantitatively in the sample solution. The invention also discloses a kit which is suitable for the method. The method is used for detecting the tetrodotoxin in aquatic products qualitatively and quantitatively and is reliable in result and low in detection limit.

Description

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AI Technical Summary

Benefits of technology

This patented technology allows us to detect certain chemical compounds called tetraodontoxynosum through their ability to change color when exposed to light or another stimulus. By doing this we have developed an assay system where there was only one type of molecular species involved but multiple types were possible due to its unique properties. These techniques could make our analysis faster and more accurate compared to previous methods like enzyme catalysis.

Problems solved by technology

The technical problem addressed in this patented text relating to the use of drugs called tetraodontotrichloroethane or tricincholinum ethylene dicholate (TETOCHLOR®)) during treatments on seafoods caused by harmful chemical substances like triphenol A (TrPA). This can lead to serious health issues if taken carelessly without being able to identify any specific contaminants that may have been present beforehand. Current testing techniques involve extracting samples containing these materials through various means followed by analysis based upon their properties. However, there exist concerns over potential safety hazards associated with animal sampling procedures. Additionally, current tests require complicated equipment and complex analytic processes which make it challenges to develop effective fluorochemistry assays suitable for identifying small amounts of drug residue left behind after treatment.

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