Enantiomeric Separation of Tenofovir on an Achiral C18 Column by HPLC Using L‐Phenylalanine as a Chiral Mobile Phase Additive

Abstract

A high‐performance liquid chromatographic method for chiral separation of tenofovir enantiomers was developed. The (R) and (S) isomers were separated on intertsil ODS 3V column (150 mm×4.6 mm i.d., 5 µm) at 25°C. The mobile phase contained the complex of Cu(II) with the optical selector L‐phenylalanine (L‐PheA). Satisfactory results were achieved with the mobile phase consisting of buffer (3 mM of copper sulfate, 1 mM of L‐PheA, and 20 mM of ammonium dihydrogen phosphate, pH adjusted to 4.0) and acetonitrile in the ratio of 95∶5. The method was validated for linearity, repeatability, LOD, LOQ, and robustness. The solution stability was studied and found to be stable for the period of 7 days.