Abstract We are developing a new MA/Ln separation process with TPEN ( N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine) and its derivatives for P&T technology of HLW from spent nuclear fuel reprocessing plants. TPEN is a hexadentate ligand that has six soft-donor sites as a kind of podand type molecule and can encapsulate a metal ion. TPEN has good selectivity of Am(III) from Ln(III) and has potential to establish partitioning of MA. However, there is a serious problem for the practical application. This is to the dissolution of a slight amount of TPEN (about 10 −4 mol/L) to water. High enrichment of Am(III) will be restricted by the dissolution of TPEN to water. In this study, the hydrophobicity of TPEN is improved by introducing alkyl groups and the effect of the introduction of alkyl groups on the separation of Am(III) and Eu(III) is examined. We tried to synthesize four hydrophobic derivatives of TPEN, and three derivatives were synthesized successfully. The derivatives were examined both the extractability and selectivity of Am(III) and Eu(III). One of them, tpdben, showed good selectivity and the maximum separation factor, SF Am/Eu, was 34 at pH 5.06. A hydrophobic derivative of TPEN that has potential of application to the MA/Ln separation process was synthesized successfully.