SYNTHESIS AND BINDING-PROPERTIES OF A NONCOVALENT MOLECULARLY IMPRINTED TESTOSTERONE-SPECIFIC POLYMER

In this study, molecular imprinting was used to develop a method based on noncovalent interactions for synthesis of a testosterone-specific polymer. The effect of the different template-monomer ratios, the part icle sizes of polymers, and chromatographic mobile phases on steroid-p olymer interactions are discussed. The polymer obtained was found to i nteract specifically with testosterone, while other steroids under stu dy were eluted close to the void volume in the HPLC experiments. Batch rebinding studies in acetonitrile were undertaken to quantitatively e valuate the affinity of the polymer for testosterone. During this expe riment, the testosterone concentration was measured in two ways: spect rophotometrically and by HPLC on a column with testosterone-specific i mprinted polymer synthesized by us. Both methods resulted in similar v alues of association constants and the number of binding sites. Howeve r, the second method has obvious advantages when the analyzed solution contains a mixture of optically dense compounds. The results obtained focus on the two-point binding nature of the imprinted polymer-testos terone interaction and the significant role of hydrogen bonds between the OH group of testosterone and carboxy group of methacrylic acid res idues inside specific recognition sites of the imprinted polymer. (C) 1998 John Wiley & Sons, Inc.