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ELISA adsorption analysis method for measuring gross amount of malachite green and colorless malachite green in water sample and aquatic products

A colorless malachite green and enzyme-linked immunosorbent adsorption technology, which is applied in measuring devices, analysis materials, material separation, etc., can solve the problems of expensive kits and achieve low test costs, high sensitivity, and simple sample processing.

Inactive Publication Date: 2009-07-15
SICHUAN UNIV
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  • Abstract
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Problems solved by technology

Recently, an ELISA kit for the determination of the total amount of malachite green or colorless malachite green has also been released (BioscientificCom), but this kit is expensive and requires an additional sample processing step, that is, the colorless malachite green must be removed before the measurement. Oxidation of stone green to malachite green

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  • ELISA adsorption analysis method for measuring gross amount of malachite green and colorless malachite green in water sample and aquatic products
  • ELISA adsorption analysis method for measuring gross amount of malachite green and colorless malachite green in water sample and aquatic products
  • ELISA adsorption analysis method for measuring gross amount of malachite green and colorless malachite green in water sample and aquatic products

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Embodiment

[0031] 1. Preparation of Colorless Malachite Green Derivatives

[0032] Take 5.5~7.5g p-nitrobenzaldehyde and dissolve it in 14~18g N,N-dimethylaniline, add 4.8~6.8g ZnCl 2 , Stir the reaction at a temperature of 80-100°C until the green solution forms a solid, cool to room temperature, the solid is dissolved in acetone, and at the same time filter to remove ZnCl 2Granules, the crude product is purified by column and dried to obtain nitro-colorless malachite green-yellow powder. Take 0.17-0.27g of nitro-colorless malachite green and dissolve it in 6-10mL of methanol, use 5% palladium carbon as a catalyst, and add it to the reaction kettle , feed hydrogen, react at a temperature of 100-120°C for 1-5 hours, filter the reactant, collect the filtrate, spin the solution with a rotary evaporator, and recrystallize the crude product with a methanol:benzene mixture with a volume ratio of 95:5 , to obtain amino colorless malachite green white powder;

[0033] 2. Preparation of Immuno...

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Abstract

The invention discloses an enzyme linked immunosorbent assay method (ELISA) for detecting the total amount of diamond green and blank diamond green in measurement water sample and aquatic products, characterized in that: the modified compound of amido blank diamond green is synthesized and linked with the protein to produce immunogen and envelope antigen, and the polyclonal antibody of rabbit-anti colorless diamond green is obtained by immuning animal and the concentration range of ELISA standard curve is 0.1-100ng / mL, IC50 is 0.9-2.6ng / mL and the recovery of standard addition is 76.2-95%, the correlation coefficient of ELISA and HPLC is 0.975, n=7; because the cross-reaction rates of the prepared antibody and diamond green are respectively 95.25%, the ELISA can be used for measuring the total amount of the diamond green and blank diamond green without any oxidation step.

Description

1. Technical field [0001] The invention relates to an enzyme-linked immunosorbent assay method (ELISA) for measuring the total amount of malachite green and colorless malachite green in water samples and aquatic products, belonging to the research field of food safety supervision or food analysis. 2. Background technology [0002] Malachite Green (Malachite Green, MG), also known as basic green, base block green, malachite green and Chinese green, is a green crystal with metallic luster, chemical name: tetramethyldiaminotriphenylmethane, molecular formula is C 23 h 25 N 2 Cl, with a molecular weight of 364.92, is easily soluble in water, methanol and ethanol, and its solution is blue-green. Malachite green is a kind of artificially synthesized triphenylmethane industrial dye, which is mainly used as a color agent in pottery, textile, leather and food industries, and as a dye in cell chemistry. Malachite green has antibacterial and insecticidal effects. It is a kind of me...

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Application Information

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IPC IPC(8): G01N33/53G01N30/74G01N33/531G01N33/543G01N33/535
Inventor 邓安平贺莉邢玮玮杨红王玉珍李大伟
Owner SICHUAN UNIV
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