A kind of preparation method of tetrodotoxin standard biological sample
A technology of tetrodotoxin and biological samples, which is applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problems of insufficient research on artificially synthesized TTX technology, difficulty in large-scale application, difficulty in artificial synthesis, etc.
Active Publication Date: 2021-07-06
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology
This method can avoid the problem of wild puffer fish being hunted and killed in large quantities due to the acquisition of raw materials, and at the same time avoids the destruction of puffer fish resources and ecological balance, and is not limited by seasons and regions. However, the content of TTX produced by bacteria is low, which is currently practical. Difficult to apply on a large scale in production
[0004] The chemical structure of TTX is relatively complex (see attached figure 1 ), artificial synthesis is relatively difficult, and there are not many studies on artificial synthesis of TTX technology
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preparation example Construction
[0032] A method for preparing a standard biological sample of tetrodotoxin, which is to extract the snail sample with acetic acid methanol solution, purify the crude extract with a carbon nanotube solid-phase extraction column, purify the purified venom by high-performance liquid chromatography, and then freeze-dry Concentrate and crystallize to obtain a standard biological sample of tetrodotoxin.
[0033] The preparation method of this tetrodotoxin standard biological sample comprises the following steps:
[0034] 1) Extraction of snail samples: Take the fully homogenized snail samples and mix them with acetic acid-methanol solution, extract in an ultrasonic water bath, then pipette the supernatant, add acetic acid-methanol solution to the residue, repeat the extraction once, and combine the two samples. The second crude extract, for subsequent use;
[0035] 2) Carbon nanotube solid phase extraction column purification: pipette the crude extract of step 1), adjust the pH wit...
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The invention discloses a method for preparing a tetrodotoxin standard biological sample. This preparation method is to extract the snail sample with acetic acid methanol solution, purify the crude extract with a carbon nanotube solid-phase extraction column, purify the purified venom by high-performance liquid chromatography, freeze-dry and concentrate, and precipitate crystals to obtain puffer fish Toxin standard biological samples. The present invention cuts in from the aspects of TTX extraction, purification and preparation of pure products, uses snails as raw materials, and purifies the crude extract through acetic acid methanol extraction and large-volume carbon nanotube solid-phase extraction, then concentrates the venom, and prepares high-efficiency liquid phase Sample injection for purification preparation. The sample preparation method and process of the invention are simple and efficient to operate, and are suitable for batch production and technical preparation of tetrodotoxin standard biological samples.
Description
technical field [0001] The invention relates to a method for preparing a tetrodotoxin standard biological sample. Background technique [0002] Tetrodotoxin (TTX) is a highly toxic non-protein neurotoxin that exists in pufferfish (Tetraodontidae) and other marine organisms. As an analgesic and anesthetic, it is widely used in medicine, and TTX standards are also required in the field of aquatic animal food testing. Due to the difficulties in purification and preparation technology and process, high-purity TTX standard products are in short supply. The preparation of TTX mainly relies on the method of extraction from biological materials, and undergoes multi-step purification and concentration steps to obtain high-purity products. [0003] The traditional TTX standard sample preparation process mostly uses the viscera of puffer fish as the extraction raw material, extracts it with an acidic solvent, removes impurities through various chromatography methods, and combines mul...
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IPC IPC(8): G01N30/14
CPCG01N30/14
Inventor 赵东豪王旭峰王强孙博伦黎智广
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI