The covalent self-assembly process was essentially solvent-free and with a very low process mass intensity (PMI, the mass of all used reagents per formed product ). The most significant chemical and technical factors affecting macrocyclization were identified with response surface methodology (RSM), and thorough analysis of intermediates by HPLC-MS provided mechanistic insight to this complex process. The affinity of mixHC [8] for selected anions and the subtle differences in the binding properties of the two new chimeric diastereomers were determined by isothermal titration calorimetry (ITC) and their structures characterised by single-crystal X-ray crystallography (SC-XRD) and modelling studies. Furthermore, a practical example of the selective anion capture by silicaimmobilized mixHC [8] was demonstrated in the efficient removal of perchlorate from a spiked mineral matrix.