Research On The Preparation And Application Of Clenbuterol Artificial Antibody

Artificial antibody, i.e. molecularly imprinted polymers, is a kind ofmolecular recognition material with special recognition capacity fortarget molecular, prepared by molecular imprinting technology.Comparing with antibody, not only artificial antibody could be simplyprepared, but also it can keep high activity in bad circumstance and hasstable performance and cost less. Moreover, as a molecular recognitionelement in rapid determination methods, it will have a bright future in thefield of rapid determination of drug and drug residues. This thesisprepares the Clenbuterol artificial antibodies by bulk polymerization andprecipitation polymerization through the molecular imprinting technologyseperately, and researches theirs characters such as adsorption isotherm,static adsorption and selectivity performance and its primary application.The experimental results indicated that in the bulk polymerization, theartificial antibody played top in the binding and selective function performance when MAA was functional molecular and the molar rate offunctional molecular to template molecular reached 4:1. The amount ofbound (Q) of 1mmol/L Clenbuterol was 46.1μmol/g. In the precipitationpolymerization, the microsphere artificial antibody, using acetonitrile asporogen, played top performance in the binding and selectivityperformance when MAA was functional molecular and the molar rate offunctional molecular to template molecular reached 4:1. In this situation,the amount of bound (Q) of 1mmol/L Clenbuterol was 50.6μmol/g. Theartifical antibody prepared by bulk polymerization's adsorption balancetime for binding Clenbuterol (0.1/1mmol/L) was 6h/8h, however themicrosphere artificial antibody prepared by precipitation polymerization'sadsorption balance time was only 45min/1h. So the characters ofmicrosphere artificial antibody prepared by precipitation polymerizationwere much better, especially its absorption balance time for the substancemolecule.In the experiment through making use of 2% agarose sol-gel fixing theClenbuterol artificial antibody prepared by precipitation polymerizationon the surface of pt electrode, it would form a sensitive membrane of theClenbuterol determination electrode. Then the electrode's relatedperformance was studied by DPV (Clenbuterol has electrochemical redoxin the artificial antibody modified pt electrode). The experiment fordetermination electrode related performance could be divided into 2 steps: first, the recognition element (Clenbuterol artificial antibody) ofdetermination electrode adsorbed the Clenbuterol in solution. Then, whenputting the determination electrode with Clenbuterol to the backgroundsolution, the chemical bond between Clenbuterol to artificial antibodywas destroyed because of the polarity circumstance in acid liquid,whereafter, the Clenbuterol bound in first step was released from theartificial antibody. The results showed that when the concentration ofClenbuterol, ranging from 1μmol/L-10μmol/L and 20μmol/L-80μmol/L,it displayed good linear relation. The linear equations are separatelyy=1.35+71.3C (r=0.99901) and y=6.30+9.47E-9C (r=0.988). Theresearch, about the reproducibility of electrode, showed that RSD is1.38%. In the selectivity experiment, when Clenbuterol wascompetitively bound by using SM₂ and DMZ as fake molecules, the valueof peak current has few changes.