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.

Keywords:

• Column liquid chromatography
• Enantiomeric separation
• Chiral mobile phase
• Tenofovir
• L‐Phenylalanine
• Achiral column

Acknowledgment

The authors take this opportunity to thank the management of Matrix's group for support of this work. The author also wishes to acknowledge the colleagues in process research for their fruitful discussions and the quality laboratory and analytical development laboratory for their constant support extended during this work.