Cu(II)-azamacrocyclic complexes are of current interest due to their potential application as copper-64 radioisotope-based imaging and therapeutic agents. The development of separation conditions is important in evaluating their purity as well as assessment of their physical properties including conditional formation constants, acid dissociation constants and lipophilicity, which can have a major bearing on the in vivo behavior of these complexes. This study evaluates the chromatographic properties of a series of copper (II) complexes of chelating cross-bridged tetraamine ligands with methanephosphonate arms having different ring sizes (12- and 14-membered) and different pendant arm moetics. Cu-CB-DO2P^OEt and Cu-CB-TE2P^OEt are two complexes with different ring size but the same pendant arm. Cu-CB-TE2P^OEt and Cu-CB-TE2P are two complexes with different pendant arms but the same ring size. A model study proved the viability of chromatographic methods as a means of measuring the conditional formation constant of copper(II) complexes. The conditional formation constant for Cu-cyclen was evaluated through a competition reaction with cyclam. The estimation of the pK_as for a copper (II) complex with methanephosphonate pendant arms is reported. The role of the stationary phase in the determination of the pK_a values was investigated using different reversed phase stationary phases. In addition the lipophilicity parameter for a series of methanephosphonate copper (II) complexes (Cu-CB-TE2P, Cu-CB-TE1A1P, Cu-CB-DO2P^OEt, and Cu-CB-TE2P^OEt) was estimated. The chiral separation for polycyclic tetramines including dibenzocyclam bisaminal, CB-cyclam, and dimethyl dibenzo CB-cyclam was achieved on chiral polysaccharide stationary phases. Successful enantiomeric separation methodology allows for evaluation of the enantiopurity of the resolved ligands. In addition the evaluation of thermodynamic data such as the barrier of racemization for CB-cyclam was investigated.