Construction And Application Of Clenbuterol Aptasensor

Clenbuterol hydrochloride(CLB)is a type of clenbuterol,which can accelerate the growth of lean meat and reduce fat in livestock.Once CLB enters the body,it can promote the decomposition of fat,and a large number of fatty acids enter the blood vessels,which increases blood pressure and dilates blood vessels,compresses nerves and leads to muscle tremor,which is extremely harmful to the human body.Therefore,it is very important to strengthen the detection of CLB in meat products to protect human health.So far,chromatographic and immunoassay methods have been used to detect CLB.These methods have high cost,long detection time and complex operation.Aptamer sensors are more and more widely used in food detection due to their advantages of high sensitivity,low cost,wide detection range and strong specificity.In this thesis,three kinds of simple and fast aptamer sensors were constructed with CLB as the detection target and aptamer as the recognition element to achieve efficient and rapid detection of CLB.The main research contents are as follows:(1)Using SYBR Green I(SGI)as fluorescent probe and CLB-adapter as recognition element,a SYBR Green I based unlabeled aptamer fluorescence sensor was constructed for the detection of clenbuterol hydrochloride In order to improve the detection sensitivity,the concentration of solution p H reactant and reaction time were optimized.Under the optimal experimental conditions,the linear range of CLB detection was 50 n M ～ 450 n M,the detection limit was 18.8 n M,and the selectivity was good.The detection of CLB in pork was successfully achieved with the recovery of 90.37 % ～ 104.98 %(2)Based on nucleic acid adaptation body-a gold nanoparticles colorimetric method for rapid detection of clenbuterol hydrochloride Nucleic acid adaptation body as recognition elements Gold nanoparticles as a colorimetric probe Sodium chloride as inducers,established a rapid sensitive method of clenbuterol hydrochloride detection In order to improve the detection sensitivity,the reactant concentration,the optimization of experimental conditions such as reaction time Under the optimal experimental conditions,the linear range of CLB was 50 n M ～350 n M,the detection limit was 18.7 n M,and the selectivity was good.The detection of CLB in water samples was successfully achieved,and the recovery was 93.8 % ～ 108.3 %(3)Sensitive detection of clenbuterol hydrochloride based on go quenching rhodamine B.An unlabeled fluorescence sensor based on go quenching rhodamine B was constructed by using an aptamer that could specifically bind clenbuterol hydrochloride(CLB)as recognition unit,rhodamine B as fluorescence probe,and go as quenching agent,so as to achieve rapid and sensitive detection of CLB.In order to improve the detection sensitivity,the buffer solution,p H,reactant concentration,reaction time and other experimental conditions were optimized.Under the optimal experimental conditions,the linear range of CLB was 100 n M ～ 700 n M,the detection limit was 9.6 n M,and the selectivity was good.The detection of CLB in pork was successfully achieved,and the recovery was 96.75% ～ 104.91%.