Fluorescent detection of -estradiol

Polymers were formed by heating a solution of the template, MAA (functional monomenteraplate ratio of 8 1), EDM A and initiator in MeCN at 40°C for 16 h. Polymers were then ground, particles sized to 25-45 jum were washed with EtOH until the template could no longer be detected and then packed into 100 x 4.6 mm (i.d.) HPLC columns. 

Displacement technique [44] Initially, the polymer was equilibrated in MeCN and the template and a variety of compounds, including potential reporters (Fig. 20.17), were chromatographed. All of the reporter compounds examined displayed a low affinity for the MIP, giving k values of at most 1.0, while that of )8-estradiol was 3.5 (kimp / -estradiol dansylate = 0.73). 

The polymer was then equilibrated in MeCN containing )3-estradiol dansylate 

Further attempts to produce a peak upon injection of j8-estradiol solutions were made using various concentrations of j6-estradiol dansylate in the mobile phase, but these were also unsuccessful. Thus, it appears that our designed reporter was not appreciably retained by the polymer and thus could not be displaced. Subsequently, we examined a variety of other non-specific reporters, varying their concentrations in the mobile phase from their fluorescence detection limit to levels at which they could easily be detected using UV spectroscopy ca. 1-100 nM). Unfortunately, no peaks were observed. 

Nevertheless, it may be possible to construct a displacement type system for the detection of j8-estradiol by labelling the molecule at C-17 and using DEAEMA as the functional monomer. Alternatively, the reporter itself could be used as the template molecule. The smaller jS-estradiol molecule should enjoy easy access to the binding sites and be able to compete effectively with the reporter. Finally, this technique may find more success for the analysis of corticosteroids [45], which possess 

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