Immunoaffinity column purification-liquid chromatogram-tandem mass spectrometry domoic acid toxin measuring method

A liquid chromatography and tandem mass spectrometry technology, applied in the field of domoic acid toxoid detection, can solve the problems of unsatisfactory sample purification and shortening, and achieve the effects of reducing sample matrix interference, high sensitivity and shortening analysis time.

Active Publication Date: 2019-02-22
MARINE FISHERIES RES INST OF ZHEJIANG
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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for the determination of domoic acid toxoids by immunoaffinity column purification-liquid chromatography-tandem mass spectrometry with high sensitivity, precision and recovery that can meet the requirements of detection and analysis. The use of high-performance, strong recognition of DA toxin immunoaffinity column, combined with high sensitivity and accuracy of liquid chromatography-tandem mass spectrometry to detect the analysis and determination of DA toxins in different marine organisms, can effectively solve the problem of sample purification in pretreatment. problems, and reduce sample matrix interference, shorten analysis time, improve method recovery, and fill the gap in the field of toxin detection by immunoaffinity technology

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  • Immunoaffinity column purification-liquid chromatogram-tandem mass spectrometry domoic acid toxin measuring method
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  • Immunoaffinity column purification-liquid chromatogram-tandem mass spectrometry domoic acid toxin measuring method

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[0059] (1) Sample pretreatment: Accurately weigh 2.00 g of the fully homogenized sample into a 50 mL stoppered centrifuge tube, add 8 mL of 75% methanol aqueous solution, vortex for 2 min, ultrasonically extract for 10 min, and centrifuge at 7000 r / min for 5 min , move the supernatant to another 50mL centrifuge tube to obtain a primary extract; then add 8mL of methanol aqueous solution with a volume concentration of 75% to the remaining residue, and repeat the extraction once to obtain a secondary extract; combine the primary extract and After the second extraction, dilute to 20mL to obtain the extract; pipette 1mL of the extract into another 50mL centrifuge tube, add 5mL of PBS buffer for dilution, and obtain the sample solution to be purified;

[0060] (2) Immunoaffinity column purification: take a DA monoclonal antibody immunoaffinity column, remove the plug of the affinity column after it returns to room temperature, release the storage solution in the column, and load the ...

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Abstract

The invention relates to domoic acid toxin detecting methods, particularly to an immunoaffinity column purification-liquid chromatogram-tandem mass spectrometry domoic acid toxin measuring method. Theimmunoaffinity column purification-liquid chromatogram-tandem mass spectrometry domoic acid toxin measuring method helps analyze and measure DA (domoic acid) toxin of different marine organisms through high-performance and high-identifiability DA toxic immunoaffinity columns and by combining high-sensitivity and high-accuracy liquid chromatogram-tandem mass spectrometry to effectively solve the problem of unsatisfactory sample purification during pretreatment, to reduce interference of sample matrixes, to shorten the analyzing time and to improve the recovery rate, thereby filing the gap of lack of immunoaffinity technology in the field of domoic acid toxin detection.

Description

technical field [0001] The invention relates to a method for detecting domoic acid toxoids, in particular to a method for determining domoic acid toxoids by immunoaffinity column purification-liquid chromatography-tandem mass spectrometry. Background technique [0002] Amnestic shellfish poisoning (ASP) is a toxin secreted by red tide diatoms, and its main active ingredient, domoic acid (DA), is a neurogenic amino acid toxoid. The toxin was first identified in 1987 in poisoning deaths from blue mussels on the east coast of Prince Edward Island, Canada. Poisoning mainly manifests as abdominal pain, diarrhea, and vomiting, accompanied by severe headache, memory loss, confusion, coma and other symptoms. The study found that DA was mainly produced by Pseudonitzia multicolumns. When the content of domoic acid in shellfish tissue reaches 40mg / kg, it can cause poisoning to the eater, and when it is 150mg / kg, there is a risk of death. The maximum limit that humans can tolerate thr...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/06G01N30/88G01N2030/047
Inventor 陈思张小军严忠雍
Owner MARINE FISHERIES RES INST OF ZHEJIANG
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