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Clonpamine aptamer and an aptamer electrochemical biosensor for detecting clonbamine

A biosensor, the technology of clumbamine, applied in the field of detection, can solve the problems of low penetration rate and high price of liquid chromatography-tandem mass spectrometer, etc., and achieve the effect of wide target molecule range, good tissue permeability and small molecular weight

Active Publication Date: 2016-07-06
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high price of liquid chromatography-tandem mass spectrometry, the penetration rate in general testing institutions is not high

Method used

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  • Clonpamine aptamer and an aptamer electrochemical biosensor for detecting clonbamine
  • Clonpamine aptamer and an aptamer electrochemical biosensor for detecting clonbamine
  • Clonpamine aptamer and an aptamer electrochemical biosensor for detecting clonbamine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Construction of an aptamer electrochemical biosensor

[0036] (1) Polishing and activation treatment of bare gold electrode surface

[0037]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.

[0038] (2) Modification of aptamer electrochemical sensors

[0039] Take 2 μL of clonbamine aptamer solution with a concentration of 100 μmol / L and add it t...

Embodiment 2

[0042] Example 2: Electrochemical response of clombamine aptamer electrochemical biosensor to clombamine standard solution

[0043] (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.

[0044] (2) Rinse the aptamer electrochemical biosensor described in step (1) with ultrapur...

Embodiment 3

[0048] Embodiment three: carry out assay and recovery rate experiment to feed:

[0049] Feed sample processing: take 2.00 g feed sample, add different concentrations of clonbamine, and extract and purify the clonbamine in the sample according to the method described in Announcement No. 1486-1-2010 of the Ministry of Agriculture. The obtained dry clonbamine powder was dissolved in ultrapure water, passed through a 0.45 μM filter membrane, and then electrochemically detected.

[0050] 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. ...

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Abstract

The invention relates to a clumpamine aptamer and an aptamer electrochemical biosensor for detecting clonbamine, as well as a preparation method and a detection method of the sensor. The clonbamine aptamer of the present invention is a single-stranded DNA probe containing GCGTAGCTACTCATAT sequence and a length of 20-25 bases, and the 3' end is modified with 3'-NH 2 -(CH 2 ) 7 -. The sensor improves the sensitivity of detecting clombamine, and can detect a minimum of 1.0 pg / mL of clombamine; greatly reduces the cost of detecting clombamine, is simple and convenient to operate, and can realize rapid detection of clombamine.

Description

technical field [0001] The invention belongs to the technical field of detection, and relates to a clonbamine aptamer and an aptamer electrochemical biosensor for detecting clonbamine, as well as a preparation method and a detection method of the sensor. Background technique [0002] Phenylethanolamine (PHL), chemically named 2-[4-(4-nitrophenyl)butyl-2-amino]-1-(4-methoxyphenyl)ethanol, also known as phenylethanolamine A, molecular weight 344.17, molecular formula C 19 h 24 N 2 o 4 , is a synthetic chemical substance, the chemical structure is as follows: [0003] [0004] Clonbamine is an isomer of formoterol, has the same function and effect as clenbuterol and ractopamine, belongs to a kind of β-agonist, and has the effect of nutrient redistribution. In December 2010, the Ministry of Agriculture of my country included clonbamine in the list of substances prohibited from being used in feed and animal drinking water. Therefore, it is very important to establish a ra...

Claims

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Application Information

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