Study On Rapid Screening Methods For Illegally Added Salbutamol In Chinese Patent Drugs For Relieving Cough And Asthma Based On Aptamer

ObjectiveThe aptamer is a short-chain nucleic acid(DNA or RNA)with a length of 20-100 bases,which has high specificity and affinity for targets.Chinese patent drugs for cough and asthma have a large share in the Chinese market.Some illegal businesses have illegally added a small amount of salbutamol(SAL)in order to gain benefits,which seriously harmed the interests of consumers.Rapid detection plays a very important role in the production and sales process.To fill the gaps in the rapid detection method for SAL in Chinese patent medicines,two novel aptasensors based on the competition effects were constructed in this study.Methods1.Firstly,we designed a rapid visual screening method for the illegally added SAL based on aptamer technologies.The aptamer against SAL was used as the detection element,and the horseradish peroxidase(HRP)and 3,3’,5,5’-tetramethylbenzidine(TMB)which could set off a color reaction were used as the output signal.Conveniently,the qualitative test results were visible to the naked eye and specific quantitative conclusions were achieved by ultraviolet-visible spectrophotometer detection.2.Based on the first sensor developed,a new fluorescent aptasensor for rapid detection of illegally added SAL in Chinese patent drugs was established.The recognition element was still the SAL aptamer,in contrast to the first sensor,the signal output component was quantumdots,and the magnetic microspheres were used for separation.Then,the results of test were determined by measuring the fluorescence values with a fluorescence spectrophotometer.Results1.Under the optimized experimental conditions,the color reaction catalyzed by HRP was quantitatively characterized by an ultraviolet-visible spectrophotometer,the absorbance was linear to the concentrations of SAL in the range from 0.1 to 10 μmol/L(r2=0.9959),with a detection limit of 0.1 μmol/L.The mean recoveries at the three spiked levels were all in the range of 86.0%to 106.0%,and the relative standard deviation ranged from 3.9%to 11.6%.2.Under the optimized experimental conditions of the sensor,the fluorescence value gradually increased with the increase of the concentration of SAL,and the detection limit was 0.1 μmol/L.The SAL spiked tests were performed on the cough and asthma Chinese patent medicine samples,and final results showed that the fluorescence intensity increased with the increase of SAL concentration.ConclusionThe visual aptasensor established is easy to operate and has strong anti-interference ability and high stability.The qualitative detection results can be obtained by observing the color depth of the test phenomenon with naked eyes.This sensor has a good application prospect in the field of rapid detection of actual Chinese patent medicine samples illegally added SAL.The fluorescent aptasensor is on the basis of the previous visible aptasensor simple in preparation,low in cost,and good in specificity.In addition,it can achieve qualitative detection,and can be used for actual detection of illegally added SAL.To sum up,this paper has established two methods for rapid detection of illegally added SAL in Chinese patent medicines.We open a new field for the rapid detection of salbutamol in Chinese patent medicines and enrich the application of aptamers in the field of rapid detection.