Development Of Dummy Template Molecularly Imprinted Polymer Targeted At Clenbuterol And Its Metabolites

Clenbuterol is the main constituent of lean meat powder,which are illegally used for animals to obtain more muscle tissue and to promote animal growth.Conventional detection methods are costly and time-consuming.So it is hotspot of research to establish a quick and accurate method to determine CLB residues in swine urine.Clenbuterol and its metabolites are determined according the reported paper,and 4-amino-3,5-dichloromandelic acid clenbuterol,and4-amino-3,5-dichlorohippuricacidaretheresearchtargets.4-hydroxymandelic acid(HMA),phenylephrine(PE)and 4-Aminohippuric acid are used as dummy template of 4-amino-3,5-dichloromandelic acid clenbuterol,and 4-amino-3,5-dichlorohippuric acid respectively.In the first part of present study,CLB,HMA and AHA were separated by high performance liquid chromatography with Inertsil ODS-3C18 column(4.6 mm ID×250mm,5)and the mobile phase consisting of0.05%trifluoroacetic acid in water and 0.05%trifluoroacetic acid in acetonitrile(73:17,v/v).At the same time,the standard curves of CLB,HMA and AHA were built in this condition,and the standard curve of CLB is y=17315.6734 x+112602.8427 and the regression coefficient(R~2)is0.9995,MHA is y=6386.5714 x-1762.0928 and the regression coefficient(R~2)is 0.9992,and AHA is y=13744.7429 x-89583.6575 and the regression coefficient(R~2)is 0.9998,all of the standard curve have good linear.In the second part of present study,we study the adsorption performance of dummy template molecularly imprinted polymer which were grafted on silica bead.The silica were activated with acid solution,then APTS-silica and ACPA-silica synthesized,the acid-activated silica,APTS-silica and ACPA-silica were characterized using FT-IR spectroscopy elemental analysis,the results show successfully grafting.The optimum reaction conditions of PE-MIPs,AHA-MIPs and HMA-MIPs molecularly imprinted membranes on silica gel were investigated and obtained(65?,12h).The adsorption rate of PE-MIPs to CLB,AHA-MIPs to AHA and HMA-MIPs to HMA was 92.34%,88.68%and 91.84%,respectively.Meanwhile,the adsorption rate of silica gel to CLB,AHA,HMA was18.36%,33.65%and 22.41%,respectively.And the adsorption rate of was NIP-MIPs was 56.99%?51.19%and 51.21%,respectively.In the last part of present study,we study the characterization of dummy template molecularly imprinted polymer grafting on quartz crystal microbalance(QCM).ABAH-QCM was prepared by grafting on the QCM.The results of microscopic infrared characterization of ABAH-QCM showed that the grafting was successful.And the MIPs-QCM was successful synthesized and was analyzed by circular dichroism showing MIPs having a similar structure to the protein secondary structure,the characterization of MIPs Bio-FastScan Scanning Probe Microscope show MIPs with three-dimensional network structure morphology.This study can provide experimental reference and theoretical support for QCM sensor array rapid detection technology and food safety electronic tongue development.At the same time,due to the general applicability of molecular imprinting technology,it also provides reference for food safety field,such as enrichment and detection of poison.