The chromatographic performances of four coated and immobilized amylose phenylcarbamate-based chiral columns were evaluated and compared under normal phase (NP) elution conditions by using chiral 4,4'-bipyridine derivatives as analytes . n-Hexane/2-propanol 90:10 and n-hexane/2-propanol/methanol 90:5:5 mixtures were employed as mobile phases (MPs), and the effect of adding methanol in the MP on retention and selectivity was considered . The effect of temperature on retention and selectivity was also evaluated, and overall thermodynamic parameters associated with the analyte adsorption onto the CSP surface were derived from van't Hoff plots . Interesting cases of enantiomer elution order (EEO) reversal, which are dependent on the nature of polar modifier, analyte structure, column-type, and temperature, were observed . The impact of substitution pattern and electronic properties of analytes and selectors on the separation behaviour was investigated by correlating chromatographic parameters and molecular properties determined by using density functional theory (DFT) calculations . Both coated and immobilized amylose tris (3,5-dimethylphenylcarbamate) columns allowed for the baseline enantioseparation (2.0 ≤ RS ≤ 4.9) of all 4,4'-bipyridines considered in this study . These results appear particularly useful because both enantiomers of these 4,4'-bipyridine derivatives are currently under investigation as new inhibitors of transthyretin fibrillogenesis, a biochemical phenomenon which is implicated to cause amyloid diseases.