Isotope fractionation of Zn was studied via liquid-liquid extraction with aqueous phase of ZnCl 2 /HCl and organic phase of crown ethers(CEs), including benzo15-crown-5(B15C5), benzo-18-crown-6(B18C6) and Dicyclohexano-24-crown-8(DC24C8)/ 1,2-dichloroethane. The isotope enrichment factor (ɛ 66), expressed in permil notation, of the isotope pair 66 ZnÀ 64 Zn was found to be between À 0.24 and À 0.52 % in the order of enrichment: B15C5 � B18C6 > DC24C8. The magnitude of ɛ 66 was well explained by theoretical isotope fractionation factors obtained by density functional theory method for zinc chloride and ZnCl 2 /CE model molecules, and a thermochemical description that correlates ɛ 66 with the differences in Gibbs free energy(ΔG) of the exchange reaction, here termed δΔG, based on the mass differences of Zn isotopes involved. Finally, results suggest that ɛ 66 may be used as a diagnostic and metric of Zn metabolic derangement, such as in porphyrias.