## 2 Synthesis and Characterization of MIP
### Polymerization Parameter in MIP Synthesis
#### 2.1.2 Template Molecule  
The selection of template molecules is based on several variables such as its stability, cost, solubility, toxicity, and functional groups that can interact with monomers [28]. MIP selectivity is greatly influenced by functional groupings, whereby its molecular recognition is based on the pre-organization and shape selectivity of the functional group. Previous studies have recommended the generation of binding sites that possess better specificity and affinity with numerous functional group interactions [29]. [PERSON] et al. [30] demonstrated the remarkable selectivity of templates with two or fewer functional groups, indicating that shape selectivity is the most important mechanism for molecular recognition. This might be due to a conflict between shape selectivity and functional group pre-organization during imprinting, or a rebinding behavior that did not appear to be executed. The imprinting performance of the templates with three or more functional groups significantly improves with distance between the functional groups. [PERSON] et al. [31] discovered two notable observations towards MIP selectivity that were influenced by cavity shape. To begin with, steric limitations occur when a molecular structure that is too big cannot fit into an imprinted site that was produced using a minimal template molecule. In addition, to maximize Van der Waals interactions inside the MIP binding sites, the selected molecular structures must be equivalent to or smaller than the template molecule. Furthermore, diverse branching topologies outperform straight-chain hydrocarbons in terms of selectivity, with straight-chain groups possessing eight or more carbons completely losing recognition. Researchers have explored the potential of increasing the template concentration while maintaining the monomer concentration, however, this approach does not increase MIP performance [32]. One of the reported adverse effects of introducing too many templates is the loss of site credibility [33]. When the template/monomer ratio improves, high-selective two-point binding sites should be replaced by a point binding site. Similar findings were subsequently achieved for templates with a single functional group that were anticipated to interact with the functional monomer once. The finding shows that the pre-polymer complex solution phase does not significantly influence the number of functional groups at the binding site of the polymer. Multi-functional binding sites result in the assembly of binding sites of polar functional groups, which occur during polymerization in the phase separation processes. Thus, the high-affinity binding sites of the imprinted polymer which is non-covalent are independent towards the pre-polymer stoichiometry due to the interactions between several monomers. In most applications, a starting working concentration of about 5% of the total monomer is utilized, then changed according to the experimental parameters [34]. To enhance recognition, the imprinting molecule should be structurally similar to the analyte [35]. While the target analytes appear to be a good fit for the template, numerous investigations have demonstrated that completely removing the template is challenging [36]. As a result, there are fewer accessible binding sites, but the remaining template is also released slowly, especially during trace analysis, ultimately influencing the final findings [37]. The usage of a dummy template (a template having a similar structure to the target analyte) potentially prevents problems caused by template leakage. Aside from that, a preliminary amount of the template (typically 1 mmol) must be present for MISPE preparation in a normal scale polymerization [28]. [PERSON] et al. [38] and [PERSON] et al. [39] reported that 1 mmol of template was used during preparation of MIPs is sufficient to ensure the formation of the defined sites with polymers. However, the selection on the concentration of the template usually depending on the overall ratio and scale of MIP production.