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Deoxytetrodotoxin extraction method

A technology of tetrodotoxin and extraction method, which is applied in the field of marine biology, can solve the problems of unreported and difficult extraction of deoxyTTX, and achieve the effects of easy implementation, low production cost, and improved purity

Inactive Publication Date: 2014-05-07
河北省海洋与水产科学研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the molecular weight, molecular structure, and physical and chemical properties of deoxyTTX are very close to those of various derivatives contained in TTX, so it is very difficult to single-extract deoxyTTX
At present, there is no report on the preparation method of deoxyTTX

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] (1), preparation of tetrodotoxin mother liquor:

[0037] Put 1g of crude tetrodotoxin in a glass beaker, add 20mL of 12% acetic acid to dissolve to obtain "A" solution, add 70mL of absolute ethanol to the "A" solution, stir evenly, and obtain "A" after dissolving "B" solution, separate the "B" solution through a centrifuge, remove the insoluble matter therein, and obtain the tetrodotoxin supernatant; pour the tetrodotoxin supernatant into a beaker, move it to an airtight container, and then place it in the airtight container Place a cup of ether solution inside, and control the temperature in the airtight container at 20°C. Due to the strong volatility of the ether solution, the ether solution in the bottle will naturally volatilize and diffuse. When the ether volatiles are compatible with the tetrodotoxin supernatant After 50 hours of reaction, tetrodotoxin crystals can be precipitated to obtain mother liquor "C";

[0038] (two), high performance liquid chromatography...

Embodiment 2

[0043] (1), preparation of tetrodotoxin mother liquor

[0044] Put 2g of crude tetrodotoxin in a glass beaker, add 30mL of 15% acetic acid for dissolution to obtain "A" solution, add 120mL of absolute ethanol to the "A" solution, stir evenly, and obtain "A" after dissolving "B" solution, the "B" solution is separated and processed by a centrifuge to remove the insoluble matter therein to obtain the tetrodotoxin supernatant; after pouring the tetrodotoxin supernatant into a beaker, move it to an airtight container, and then Put a glass of ether solution in the container, and control the temperature in the airtight container at 15°C. Due to the strong volatility of the ether solution, the ether solution in the bottle will naturally volatilize and diffuse. When the ether volatiles and the tetrodotoxin supernatant phase After 60 hours of reaction, tetrodotoxin crystals can be precipitated to obtain the mother liquor "C";

[0045] (two), high performance liquid chromatography sepa...

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Abstract

The invention discloses a deoxytetrodotoxin extraction method. The deoxytetrodotoxin extraction method comprises the following steps of dissolving a tetrodotoxin crude product in acetic acid, adding anhydrous ethanol into the solution of the tetrodotoxin crude product, stirring uniformly, carrying out centrifugation to remove insoluble substances and to obtain a tetrodotoxin supernatant, heating the tetrodotoxin supernatant in a closed container containing ether so that tetrodotoxin crystals are precipitated and a mother liquor is obtained, concentrating the mother liquor, adding ammonia water into the concentrated mother liquor to adjust a pH value so that precipitates are separated out, treating the precipitates to obtain deoxytetrodotoxin-containing precipitates, dissolving the deoxytetrodotoxin-containing precipitates in acetic acid or phosphoric acid, carrying out liquid chromatographic separation by a preparative high performance liquid chromatograph, carrying out detection by a differential refraction detector or an ultraviolet detector, collecting a deoxytetrodotoxin fraction according to a peak map, adding the deoxytetrodotoxin fraction into a chromatographic column, carrying out elution by an acetum, collecting a fraction, adjusting a pH value of the fraction by ammonia water so that white solids are precipitated, and treating the white solids to obtain deoxytetrodotoxin crystals.

Description

technical field [0001] The invention relates to a method for extracting deoxytetrodotoxin, belonging to the technical field of marine organisms. Background technique [0002] Tetrodotoxin (TTX) is one of the highly toxic non-protein substances extracted from the viscera of puffer fish. It has a unique chemical structure and pharmacological properties and is a well-known neurotoxin; its toxicity 1250 times higher than sodium cyanide. Modern medical research shows that tetrodotoxin is also widely present in marine organisms such as gobies, salamanders, octopuses, shellfish, snails, starfish, and crabs. The crystal of tetrodotoxin is an aminohydroquinazoline type compound with the molecular formula: C 11 h 17 N 3 o 8 , Molecular weight: 319.27. Only 0.3mg-0.5mg of this dreadful toxin can cause death, because it can specifically block the nerve cell membrane Na + Therefore, tetrodotoxin has the effects of analgesia, detoxification, antispasmodic, antipruritic, and local a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D491/22
Inventor 黄海枫赵振良黄致强吴新民赵海涛冯子龙郝志毅
Owner 河北省海洋与水产科学研究院
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