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Method for determining tetrodotoxin in marine organisms by utilizing immunoaffinity column purification-liquid phase chromatography-tandem mass spectrometry

A liquid chromatography, tetrodotoxin technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve the effect of high-efficiency determination

Active Publication Date: 2015-03-04
MARINE FISHERIES RES INST OF ZHEJIANG +1
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

At present, there is no relevant report on the detection of tetrodotoxin by using immunoaffinity column as a pretreatment and purification method

Method used

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  • Method for determining tetrodotoxin in marine organisms by utilizing immunoaffinity column purification-liquid phase chromatography-tandem mass spectrometry
  • Method for determining tetrodotoxin in marine organisms by utilizing immunoaffinity column purification-liquid phase chromatography-tandem mass spectrometry
  • Method for determining tetrodotoxin in marine organisms by utilizing immunoaffinity column purification-liquid phase chromatography-tandem mass spectrometry

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Embodiment

[0045] A method for the determination of tetrodotoxin in marine organisms by immunoaffinity column purification-liquid chromatography-tandem mass spectrometry, comprising the following steps:

[0046] (1) Sample treatment: Accurately weigh 2.00 g of the fully homogenized sample into a 50 mL stoppered centrifuge tube, add 10 mL of methanol solution containing 1% acetic acid, vortex for 2 min, ultrasonically extract in a 60°C water bath for 15 min, and cool to room temperature , centrifuge at 6000r / min for 5min, transfer 5mL supernatant to another 50mL centrifuge tube, add 20ml PBS solution for dilution, adjust pH to 7-8 with 1mol / L sodium hydroxide solution, and obtain sample solution.

[0047] (2) Immunoaffinity column purification: the sample solution on the immunoaffinity column, after loading the sample, rinse the affinity column with water, squeeze out the residual liquid in the column, and discard all the above effluents, and finally wash the column with 1% acetic acid Th...

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Abstract

The invention discloses a method for determining tetrodotoxin in marine organisms by utilizing immunoaffinity column purification-liquid phase chromatography-tandem mass spectrometry. The method is characterized in that the tetrodotoxin in the marine organisms can be rapidly, simply and high efficiently determined by utilizing the unique selection identifiability of the immunoaffinity column on the basis of the high sensitivity and accuracy of the liquid phase chromatograph tandem mass spectrometry.

Description

technical field [0001] The invention relates to a detection method of tetrodotoxin, in particular to an immunoaffinity column purification-liquid chromatography-tandem mass spectrometry method for determining tetrodotoxin in marine organisms. Background technique [0002] Tetrodotoxin (TTX) is a natural marine biological neurotoxin that is widely distributed and mainly exists in puffer fish, but also in other animals such as horseshoe crabs, blue-ringed octopuses, and snails. Tetrodotoxin is extremely toxic, and eating 0.5mg can cause death. Its positively charged guanidinium group can extend into the ion-selective filter of the sodium channel, and combine with the free carboxyl group on the inner wall of the channel to prevent the entry of sodium ions, thereby inhibiting nerves. Impulse conduction causes sensory nerve paralysis, quadriplegia, dyspnea, and finally death due to respiratory depression. Every year in our country, there are poisoning incidents caused by eating ...

Claims

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Application Information

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IPC IPC(8): G01N30/06
Inventor 张小军王莹伦丽丽
Owner MARINE FISHERIES RES INST OF ZHEJIANG
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