## 3 Materials and Methods
### Physical Characterisation  
3.10.1. Scanning Electron Microscopy  
Polymer morphology was examined using a Phillips XL30 scanning electron microscope. The sample was deposited on a sticky carbon tab and coated with gold, using a SPI gold spotter coating unit. SE micrographs of the polymers were obtained at 20,000\(\times\) magnification at 15.0 kV. 3.10.2. Swelling Measurements  
A total of 30 (3) mg of each polymer was packed into an NMR tube, and the height of the dry polymer measured. A solution of **1** (1.00 mL of 0.0800 mM) in acetonitrile or chloroform was added and allowed to soak for 24 h. Polymers were allowed to settle, and the bed height of the swollen polymers was measured. The swelling factor was calculated from the ratio of the bed height of the swollen polymer to the dry polymer. 4. Conclusions  
Using self-assembly (non-covalent) and semi-covalent methods, we designed **1**-specific MIPs. In the case of the self-assembly MIPs, a range of potential functional monomers (FM) were screened using a combination of pre-synthetic interaction studies (by molecular modelling and NMR analysis) and binding assays. The best performing self-assembly **1**-MIPs gave IFs of 3 to 7 and were formulated from FM **7** with ethylene glycoldimethoxy-late (EGDMA) and trimethylolpropane trimethacrylate(TRIM) crosslinkers, using chloroform as porogen and rebinding solvent at T:FM ratios of 1:1 (E7\({}_{\text{1-MIPCHCI3}}\) and T7\({}_{\text{1-MIPCHCI3}}\)) and 1:2 (E7\({}_{\text{2-MIPCHCI3}}\) and T7\({}_{\text{2-MIPCHCI3}}\)). The binding parameters K\({}_{\text{d}}\) and B\({}_{\text{max}}\) were consistent with the MIPs exhibiting a stronger affinity towards **1** (lower K\({}_{\text{d}}\)), resulting in greater number of binding sites (higher B\({}_{\text{max}}\)) than their corresponding NIPs and observed to be higher for the 1:2 than the 1:1 T:FM formulations. The imprinting effect, as per IFs and NIP/MIP K\({}_{\text{d}}\) ratios, was observed to be higher for the EGDMA-based MIPs than for the TRIM-based MIPs. The semi-covalent **1**-MIPs were designed using O-4-vinyl 4-benzylpiperazine-1-carbothioate or benzylpiperazine (4-vinylphenyl) carbamate (**16**) as the template-monomer (TM) adduct copolymerized with either EDGMA (E16\({}_{\text{MIPCHCI3}}\)) or TRIM (T16\({}_{\text{MIPCHCI3}}\)). We found that **1** could be cleaved from **16** post-polymerisation, leaving a phenol moiety within the imprinted sites capable of hydrogen bonding with **1** upon re-exposure. T16\({}_{\text{MIPCHCI3}}\) exhibited a greater affinity for **1**--higher IFs, higher binding capacities and lower K\({}_{\text{d}}\) than E16\({}_{\text{MIPCHCI3}}\). The K\({}_{\text{d}}\) values for semi-covalent MIPs are significantly lower than their self-assembly equivalents, while their B\({}_{\text{max}}\) are, at the most, comparable. The self-assembly MIPs displayed high levels of cross-reactivity with **19** and **20**, low to moderate with **18** and marginal with **17**. The analytes **17** and **18** are both large molecules that differ in shape, potential and available functional groups to **1** and are therefore a poor fit for the imprinted cavities. In contrast, **19** and **20** are similar in size and chemical character to **1**, consistent with the observed high cross-reactivity. The semi-covalent MIPs showed similar cross-reactivity trend to the self-assembly MIPs, except for analyte **17**, which recorded high levels of cross-reactivity. The unexpected high cross-reactivity with **17** could possibly be due to its benzoyl group, which is similar in structure and size to the benzyl group of **1**, and could easily fit the imprinted cavities and interact with the phenol moiety. Consistent with the results of the non-competitive cross-reactivity assays, the results of the competitive (binary mixtures) binding studies showed both self-assembly and semi-covalent MIPs to be highly selective towards **1** in the presence of **17**, moderately selective in the presence of **18** but non-selective in the presence of **19**.