Capillary electrophoresis electrochemical enzyme linked immunoassay for detecting amnesic shellfish toxicity

A technology of capillary electrophoresis and shellfish toxins, which is applied in the direction of material analysis, material analysis, and measuring devices through electromagnetic means, can solve problems that have not been reported, and achieve the effects of less reagent consumption, high separation efficiency, and high sensitivity

Inactive Publication Date: 2011-01-19
QINGDAO UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, the detection method of amnesiac shellfish toxin based on capillary

Method used

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  • Capillary electrophoresis electrochemical enzyme linked immunoassay for detecting amnesic shellfish toxicity
  • Capillary electrophoresis electrochemical enzyme linked immunoassay for detecting amnesic shellfish toxicity
  • Capillary electrophoresis electrochemical enzyme linked immunoassay for detecting amnesic shellfish toxicity

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Example 1 Detection of Amnestic Shellfish Toxin in Standard Samples by Capillary Electrophoresis Electrochemical Enzyme-Linked Immunoassay

[0032] The instruments and reagents used are as described above.

[0033] Using non-competitive mode, add a series of amnestic shellfish toxin standards and HRP-labeled amnestic shellfish toxin antibody with different concentrations into the micro-enrichment tube, incubate in a water bath at 37°C for 30min, take it out and dilute to 200μL with buffer solution , pressure injection (9cm, 20s), run electrophoresis and record. In non-competitive mode, HRP-labeled amnestic shellfish toxin antibody (Ab*) was in excess, and the sample solution after incubation contained both HRP-labeled amnestic shellfish toxin antibody-antigen conjugate (Ag-Ab*) and untreated The reacted amnestic shellfish toxin antibody (Ab*), the amnestic shellfish toxin-enzyme-labeled antibody complex in the mixture and the remaining enzyme-labeled amnestic shellfish...

Embodiment 2

[0036] Example 2 Detection of Amnestic Shellfish Toxin in Shellfish Samples Using Capillary Electrophoresis Electrochemical Enzyme-Linked Immunoassay

[0037] Remove the shells from shellfish (scallops or oysters), soak the shellfish with distilled water for 3 times, homogenize for 10 minutes, sonicate in an ice-water bath for 10 minutes, adjust the pH to 3.4-4.0 with 6mol / L hydrochloric acid, centrifuge for 5 minutes, and pour off the supernatant. Add methanol and stir evenly, ultrasonically extract for 5 minutes, centrifuge for 5 minutes, collect the upper layer extract, evaporate the extract under reduced pressure in a water bath at 35-50°C until the methanol is volatilized, and dissolve the eluate with sodium lauryl sulfate micellar solution;

[0038] Add excess enzyme-labeled amnesic shellfish toxin antibody to the above-mentioned treated shellfish sample solution, incubate and react at 37°C for 30 minutes, amnestic shellfish toxin in the shellfish sample reacts with the e...

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Abstract

The invention discloses capillary electrophoresis electrochemical enzyme linked immunoassay for detecting amnesic shellfish toxicity, which belongs to the technical field of assay and detection. Solution of standard substances and shellfish samples with the amnesic shellfish toxicity are assayed by combining the capillary electrophoresis technology and electrochemical enzyme linked immunoassay technology. The capillary electrophoresis electrochemical enzyme linked immunoassay comprises the following steps of: reacting sample solution and an amnesic shellfish toxicity antibody marked by a horse radish peroxidase (HRP) in a noncompetitive mode; separating an amnesic shellfish toxicity-enzyme labeled antibody compound and the rest amnesic shellfish toxicity antibody marked by the HRP in the mixed solution by capillary electrophoresis; generating a 3-aminophenazine with electrochemical activities by using aminophenol which is the oxidogenic substrate of peroxide in the presence of a catalyst; and performing electrochemical detection. The method simplifies a sample treatment process and has high selectivity and high accuracy. The linear detection range of the solution of amnesic shellfish toxicity standard substances of the method is 0.5 to 50ng/mL and a detection limit of the method is 0.2ng/mL. The method is an ideal method for detecting the amnesic shellfish toxicity of shellfish samples.

Description

technical field [0001] The invention relates to capillary electrophoresis electrochemical enzyme-linked immunoassay technology, in particular to capillary electrophoresis electrochemical enzyme-linked immunoassay method for detecting amnestic shellfish toxin. Background technique [0002] The ever-increasing environmental pollution has led to an increase in the types and quantities of red tide toxins in the ocean. These red tide toxins are transmitted through the food chain and threaten human health. The inspection of red tide toxins in imported aquatic products by countries around the world is becoming more and more stringent, and it is urgent to develop a fast, sensitive and reliable method for detecting red tide toxins in the ocean. [0003] Red tide toxin is also known as "shellfish toxin". According to the poisoning symptoms of the human body, it can be further divided into paralytic shellfish poisoning (PSP), diarrhea shellfish poisoning (DSP), amnestic shellfish pois...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N27/447
Inventor 李雪梅葛安青李晓琳张召香梅振华张书圣
Owner QINGDAO UNIV OF SCI & TECH
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