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Ractopamine aptamers and aptamer electrochemical biosensors for the detection of ractopamine

A ractopamine and biosensor technology, applied in the detection field, can solve the problems of high ractopamine detection cost, complex detection scheme, long detection time, etc., so as to reduce the detection cost and the complexity of the detection scheme, the modification is convenient and fast, and the detection cost is reduced. low effect

Active Publication Date: 2015-12-02
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the shortcomings of existing ractopamine detection costs, complex detection schemes, and long detection time, the first purpose of the present invention is to provide a ractopamine aptamer

Method used

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  • Ractopamine aptamers and aptamer electrochemical biosensors for the detection of ractopamine
  • Ractopamine aptamers and aptamer electrochemical biosensors for the detection of ractopamine
  • Ractopamine aptamers and aptamer electrochemical biosensors for the detection of ractopamine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036]Example 1: Construction of an aptamer electrochemical biosensor

[0037] (1) Polishing and activation treatment of bare gold electrode surface

[0038] Grind the bare gold electrode with 0.3 μm alumina powder for 10 minutes, then polish it with 0.05 μm alumina powder for 20 minutes, and then ultrasonically clean it twice with ultrapure water for 5 minutes each time to completely remove the non-specific adsorption on the bare gold electrode. Aluminum oxide powder on the electrode surface. Immerse the cleaned bare gold electrode in freshly prepared hot Piranha solution (concentrated sulfuric acid: 30% H 2 o 2 =7:3, V:V) activated at room temperature for 10 minutes to obtain activated bare gold electrodes. Then ultrasonic cleaning was performed twice with ultrapure water and absolute ethanol, 5 minutes each time.

[0039] (2) Modification of aptamer electrochemical sensors

[0040] Take 2 μL of ractopamine aptamer solution with a concentration of 100 μmol / L and add it ...

Embodiment 2

[0043] Example 2: Electrochemical response of ractopamine aptamer electrochemical biosensor to ractopamine standard solution

[0044] (1) The electrochemical nucleic acid aptamer electrochemical biosensor produced in Example 1 was used as a working electrode, the Ag / AgCl (saturated KCl) electrode was used as a reference electrode, and the platinum wire electrode was used as a counter electrode. The electrochemical nucleic acid aptamer electrochemical biosensor made in Example 1 was rinsed with ultrapure water to remove unbound aptamers, and then placed in a concentration of 5 mmol / L K 3 [Fe(CN) 6 ] In the electrolyte, the electrolyte contains KCl with a concentration of 0.1mol / L. Differential pulse voltammetry analysis shows that the high potential of scanning is 0.7V, the low potential is 0.3V, the scanning speed is 0.02V / s, and the amplitude is 0.05V. DPV spectrum of ligand sensor and its peak current value Ip 0 .

[0045] (2) Rinse the aptamer sensor described in step (1...

Embodiment 3

[0048] Example 3: Quantitative detection of compound feed containing ractopamine

[0049] Treatment of compound feed samples: Take 5.00 g of compound feed samples, add different concentrations of ractopamine, and extract and purify the ractopamine in the compound feed according to the method described in GB / T22147-2008. The obtained ractopamine dry powder was dissolved in ultrapure water, passed through a 0.45 μM filter membrane, and then electrochemically detected.

[0050] The electrochemical nucleic acid aptamer sensor made in Example 1 was rinsed with ultrapure water to remove unbound aptamers, and then placed in K with a concentration of 5 mmol / L. 3 [Fe(CN) 6 ] In the electrolyte, the electrolyte contains KCl with a concentration of 0.1mol / L. Differential pulse voltammetry analysis shows that the high potential of scanning is 0.7V, the low potential is 0.3V, the scanning speed is 0.02V / s, and the amplitude is 0.05V. DPV spectrum of ligand sensor and its peak current val...

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Abstract

The invention relates to a ractopamine aptamer, an electrochemical biosensor for detecting the ractopamine aptamer same, a method for manufacturing the electrochemical biosensor and a method for detecting the ractopamine aptamer by using the electrochemical biosensor. The ractopamine aptamer is a single-chain DNA (deoxyribonucleic acid) probe which contains an AGTGCGGGC sequence and has a length of 13-20bp, and the amino group of 5'-NH2-(CH2)6- is modified on the 5' terminal. The electrochemical biosensor for detecting the ractopamine aptamer comprises the ractopamine aptamer, has higher ractopamine detection sensitivity, can be used for detecting 0.1ng / mL of ractopamine at least, ensures that the ractopamine detection cost is lowered greatly, is easy and convenient to operate and can realize the rapid detection of the ractopamine.

Description

technical field [0001] The invention belongs to the technical field of detection, and relates to a ractopamine aptamer, an aptamer electrochemical biosensor for detecting ractopamine, and a preparation method and a detection method of the sensor. Background technique [0002] Ractopamine (RAC) is a phenylethanolamine beta 2 - Adrenaline stimulants can significantly increase the lean meat rate of animal carcasses and reduce the deposition rate of body fat, so they are widely used illegally in animal husbandry. Eating animal viscera with RAC residues often causes acute or chronic food poisoning in humans. In addition, RAC is also a food-derived stimulant, which is an important item for the safety monitoring of animal-derived food in large-scale sports events. The World Health Organization Joint Committee on Food Additives (JECFA) stipulates that the maximum residue limits (maximalresiduelimits, MRLs) of RAC in liver, kidney, and muscle are 40.0, 90.0, and 10.0ng / mL, respecti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/115C12Q1/68G01N27/30G01N27/26
Inventor 刘大岭陈丹周妮谢春芳姚冬生
Owner JINAN UNIVERSITY
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