Separation of d4T-P-N-PheOMe Diastereoisomers by Supercritical Fluid Chromatography

The separation of d4T-P-N-PheOMe diastereoisomers has been carried out by supercritical fluid chromatography on Hypersil ODS2 column (250 mm × 4.6 mm, 5 μm). Supercritical CO2 modified with methanol, ethanol and 2-propanol were used as mobile phase. The influence of type and concentration of alcohol modifier, back pressure and column temperature on retention factor, diastereoselectivity and resolution were comprehensively studied. Among methanol, ethanol and 2-propanol, the methanol was proved to be the most favorable modifier, and 7% (V/V) of methanol was the preferred concentration at which d4T-P-N-PheOMe diastereoisomers could be separated with a resolution of 3.35. The optimal separation conditions were 7% (V/V) methanol as modifier, 2.0 mL min−1 of flow rate for mobile phase, 308.15 K of column temperature, 15 MPa of back pressure. The d4T-P-N-PheOMe was separated baselinely in the running time of 15 min under these optimal conditions.