Clenbuterol aptamer and an aptamer electrochemical biosensor for detecting clenbuterol
A biosensor and aptamer technology, applied in the detection field, can solve the problems of expensive, complicated test process, long-term pretreatment, etc., and achieve the effect of low detection cost, high detection sensitivity and strong affinity
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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 / T5009.192-2003. 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] Rinse...
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