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Enzyme-linked immunosorbent method for determining malachite green oxalate in aquatic product

A technology of malachite green and aquatic products, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of complex sample pretreatment, high detection cost, expensive instruments, etc., and achieve low test cost, high sensitivity and strong specificity Effect

Inactive Publication Date: 2016-01-27
JIANGSU WISE SCI & TECH DEV
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  • Description
  • Claims
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Problems solved by technology

Although the accuracy of these methods is high, the instruments are expensive, the sample pretreatment is complicated, time-consuming, and the detection cost is high.

Method used

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  • Enzyme-linked immunosorbent method for determining malachite green oxalate in aquatic product

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specific Embodiment

[0046] 1. Preparation of coating solution: 1.465g sodium bicarbonate and 0.795g sodium carbonate were mixed with 500mL deionized water to make a solution, and stored at 4°C;

[0047] Preparation of washing solution: use 8g sodium chloride, 2.9g disodium hydrogen phosphate dodecahydrate, 0.2g potassium dihydrogen phosphate, 0.2g potassium chloride, 0.5mL of 0.05% Tween 20, and 1000mL of high-pressure distilled water to prepare a solution , pH7.4, stored at 4°C;

[0048] Preparation of stop solution: prepare a solution with 21.7mL concentrated sulfuric acid and 178.3mL high-pressure distilled water;

[0049] Preparation of blocking solution: 8g sodium chloride, 2.9g disodium hydrogen phosphate dodecahydrate, 0.2g potassium dihydrogen phosphate, 0.2g potassium chloride, 0.5mL 0.05% Tween 20, high pressure distilled water 1000mL, 10g bovine serum white Protein, prepared as a solution, pH7.4;

[0050] Preparation of substrate solution: use 25.7mL, 0.2moL / L disodium hydrogen phosp...

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Abstract

The present invention relates to an indirect competitive enzyme-linked immunosorbent method for determining malachite green (MG) in an aquatic product or water sample. According to the method, malachite green molecules are modified, the modified malachite green molecules and a carrier protein are subjected to cross-linking to obtain immunogen and coating antigen, and multiple animal immunizations are performed to prepare the anti-malachite green polyclonal antibody, wherein the IC50 value (the corresponding determined substance concentration when the absorbance in the standard curve is inhibited to achieve 50% of the maximum absorbance value) is 0.9-2.6 [mu]g / L and the detection limit is 0.02-0.10 [mu]g / L under the optimized experiment conditions, the recovery rate of the malachite green in the water sample and the aquatic product is 76.2-95.0%, and the cross-reaction rate of the malachite green and the leucomalachite green is 95.25%. According to the present invention, the method comprises: coating liquid preparing, washing liquid preparing, termination liquid preparing, blocking liquid preparing, substrate solution preparing, antibody immobilization, immune reaction, detection signal obtaining, and other step; and with the application of the characteristics of the competitive enzyme-linked immunosorbent method to detect the malachite green content in the aquatic product, the advantages of high sensitivity, low interference, convenience, rapidness, safe operation, strong specificity, and low detection cost are provided.

Description

technical field [0001] Malachite Green (MG) is a triphenylmethane industrial dye, commonly used in ceramics, textiles, leather, food colorants and cytochemical stains, etc.? It can also be used as an insect repellant, insecticide, and preservative. It is a class with strong antibacterial effect among medicinal dyes, and is used to prevent and treat saprolegniasis, gill mold and melon worm disease of various aquatic animals. Wait. In recent years, it has been found that malachite green (MG), especially its metabolite colorless malachite green (LMG), has obvious accumulation residues in aquatic animals, and its chemical functional group, triphenylmethane, is a kind of carcinogen. The immune system and reproductive system are greatly affected after the aquatic products are collected. Specifically, it is a detection method of malachite green content in aquatic products. Background technique [0002] At present, there are more than 450 kinds of compounds that can cause cancer. ...

Claims

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Application Information

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IPC IPC(8): G01N33/543
Inventor 洪霞邓榕榕
Owner JIANGSU WISE SCI & TECH DEV
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