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Method for preparing immuno biosensor for measuring ractopamine (RAC)

A technology of ractopamine and biosensors, which is applied in the field of devices in the field of chemical detection technology, can solve the problems of inability to realize on-site detection and cumbersome operation process, and achieve the effect of promoting electron transfer, high activity, and easy fixation

Inactive Publication Date: 2011-02-23
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these analytical methods require large-scale analytical instruments, the operation process is cumbersome, and on-site detection cannot be realized.

Method used

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  • Method for preparing immuno biosensor for measuring ractopamine (RAC)
  • Method for preparing immuno biosensor for measuring ractopamine (RAC)
  • Method for preparing immuno biosensor for measuring ractopamine (RAC)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Step 1, the construction of the immunosensor: first carboxylate the multi-walled carbon nanotubes, and then immobilize the BSA-coupled ractopamine antigen on the functionalized carbon nanotubes in the form of conjugate bonding to prepare carbon nanotubes - Ractopamine antigen-BSA conjugate. After the glassy carbon electrode was polished and cleaned, gold nanoparticles, carbon nanotube-ractopamine antigen-BSA conjugate, and Prussian blue were deposited sequentially, and the modified electrode was soaked in 5% BSA solution for 30 min at 37°C.

[0026] Step 2, detection of ractopamine by immunosensor: immerse the modified electrode in a total volume of 50 μL of phosphate buffer solution containing different concentrations of free ractopamine and 8 μg / mL of ractopamine polyclonal antibody, at 37°C Incubate for 40 min, rinse with phosphate buffer solution and in 2 mM K 3 [Fe(CN) 6 ] solution for differential pulse voltammetry (DPV) scanning. The experimental results showe...

Embodiment 2

[0035] Determination of spiked ractopamine in real samples of animal feed

[0036] Step 1, processing of animal feed samples: take 1g of pig feed samples and grind them into fine pieces, accurately weigh them into 10mL sample tubes, prepare a blank sample not containing ractopamine and 3 blank samples containing different ractopamine concentrations by standard addition method. For the spiked sample, add 8 mL of fresh phosphoric acid-methanol extract (0.2M), ultrasonically shake the mixture for 30 min, centrifuge at 3000 r / m for 10 min, transfer the supernatant to a 25 mL volumetric flask, and use 8 mL, 5 mL Repeat the extraction 3 times with 4mL of the same extract, and combine the supernatant in a volumetric flask. Transfer 1 mL of the supernatant to a 5 mL sample tube and concentrate and evaporate at 55°C under nitrogen blowing. The concentrate is dissolved in 1 mL of phosphate buffer solution with a pH of 7.4 and then used for electrochemical analysis.

[0037] Step 2, the...

Embodiment 3

[0042] Determination of spiked ractopamine in actual pork samples

[0043] Step 1, the processing of actual pork samples: Take 2g pork samples and crush them, accurately weigh them into 10mL sample tubes, prepare blank samples without ractopamine by standard addition method, and prepare 3 spiked samples containing different concentrations of ractopamine. Standard sample, add 7mL ethyl acetate, the mixture is ultrasonically oscillated for 30min, centrifuged at 4000r / m for 15min, the supernatant is transferred to a volumetric flask with a volume of 25mL, and the residue is extracted three times with 6mL ethyl acetate, and the The supernatant was combined in a volumetric flask. Transfer 1 mL of the supernatant to a 5 mL sample tube and concentrate and evaporate at 55°C under nitrogen blowing. The concentrate is dissolved in 1 mL of phosphate buffer solution with a pH of 7.4 and then used for electrochemical analysis.

[0044] Step 2, the construction of the immunosensor: firstly...

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Abstract

The invention discloses a method for preparing an immuno biosensor for measuring ractopamine (RAC) in the technical field of chemical detection. Nanogold, a carbon nano tube having conjugated bonding coupled RAC-bovine serum albumin, and prussian blue are modified on a glassy carbon electrode sequentially to obtain the electrochemical immuno biosensor. By the method, the electrochemical immuno biosensor which has high sensitivity, high stability and can quickly test the ractopamine can be prepared.

Description

technical field [0001] The present invention relates to a device and method in the technical field of chemical detection, in particular to an immunobiosensor using ternary composite nanomaterials for measuring β-agonist hormone ractopamine (RAC) in solution and a preparation method thereof . Background technique [0002] Ractopamine (RAC), which belongs to β-stimulant, can make the nutrients in the animal body transfer from fat to muscle, showing a nutrient redistribution effect, and then regulate the material metabolism of the animal body, enhance fat decomposition, and promote protein synthesis. Significantly improve carcass lean meat rate and feed remuneration. However, after humans eat animal foods with ractopamine residues, they will be poisoned to varying degrees. Their abuse and residues in animal foods seriously endanger people's health and life safety, and seriously affect my country's livestock and poultry. Product export trade. Therefore, my country, the Europea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N27/26B82B1/00
Inventor 王正武赵波陈昌云颜妍米芹
Owner SHANGHAI JIAO TONG UNIV
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