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Clenbuterol aptamer and electrochemical biosensor of aptamer for detecting clenbuterol

A biosensor and aptamer technology, applied in the detection field, can solve the problems of long-term pretreatment, expensive and complicated test procedures, and achieve the effects of strong affinity, low detection cost and high detection sensitivity

Active Publication Date: 2014-10-22
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods have high selectivity and sensitivity, they require expensive instruments or long sample pretreatment and complex testing procedures

Method used

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  • Clenbuterol aptamer and electrochemical biosensor of aptamer for detecting clenbuterol
  • Clenbuterol aptamer and electrochemical biosensor of aptamer for detecting clenbuterol
  • Clenbuterol aptamer and electrochemical biosensor of aptamer for detecting clenbuterol

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 clenbuterol aptamer solution with a concentration of 100 μmol / L and add it ...

Embodiment 2

[0043] Example 2: Electrochemical response of clenbuterol aptamer electrochemical biosensor to clenbuterol 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 ] / K 4 [Fe(CN) 6 ](1:1) In the electrolyte, the electrolyte contains KCl with a concentration of 0.1mol / L, AC impedance analysis, initial potential 0.22V, frequency: 0.1-1.0×10 5 Hz, the AC impedance spectrum and its impedance value Ret0 of the aptamer electrochemical biosensor were obtained.

[0045] (2) Rinse the aptamer electrochemical biosensor described in step (1) with ultrap...

Embodiment 3

[0049] Embodiment three: Determination and recovery rate experiment are carried out to muscle:

[0050] Treatment of muscle samples: take 2.00 g of muscle samples, add different concentrations of clenbuterol, and extract and purify clenbuterol in the samples according to the method described in GB / T 5009.192-2003. The obtained clenbuterol dry powder was dissolved in ultrapure water, passed through a 0.45 μM filter membrane, and then electrochemically detected.

[0051] 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 ] / K 4 [Fe(CN) 6 ](1:1) In the electrolyte, the electrolyte contains KCl with a concentration of 0.1mol / L, AC impedance analysis, initial potential 0.22V, frequency: 0.1-1.0×10 5 Hz, the AC impedance spectrum and its impedance value Ret0 of the aptamer electrochemical biosensor were obtained.

[0052] Rins...

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Abstract

The invention relates to a clenbuterol aptamer and an electrochemical biosensor of the aptamer for detecting clenbuterol, and a preparation method and a detection method of the sensor. The clenbuterol aptamer disclosed by the invention is a single-stranded deoxyribonucleic acid (DNA) probe, which contains a sequence GGATATTGG, and the length is 13-18 basic groups; and 3'-NH2-(CH2)7- is modified at the 3'terminal. By adopting the sensor, the sensitivity for detecting the clenbuterol is improved, 1.0pg / mL of clenbuterol can be detected to the lowest extent, the detection cost of the clenbuterol is greatly lowered, the operation is simple and convenient, and quick detection of the clenbuterol can be achieved.

Description

technical field [0001] The invention belongs to the technical field of detection, and relates to a clenbuterol aptamer, an aptamer electrochemical biosensor for detecting clenbuterol, and a preparation method and a detection method of the sensor. Background technique [0002] Clenbuterol (Clenbuterol, CLB), chemically named α-[(tert-butylamino)methyl]-4-amino-3,5-dichlorobenzyl alcohol hydrochloride, molecular weight 277.19, molecular formula C 12 h 18 Cl 2 N2O is a synthetic chemical substance with the following chemical structure: [0003] [0004] Clenbuterol has stable physical and chemical properties and is not easy to decompose at higher temperatures. After animal ingestion, it mainly remains in the muscle tissue and internal organs in the form of the original drug. Clenbuterol residues in livestock and poultry products are enriched in the human body through the food chain. Studies have shown that after long-term consumption of food with clenbuterol residues, ad...

Claims

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Application Information

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IPC IPC(8): C12N15/115G01N27/26G01N27/327
Inventor 刘大岭陈丹姚冬生谢春芳
Owner JINAN UNIVERSITY
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