Agilent Technologies has produced an application note demonstrating the use of the Agilent ChemStation Method Scouting Wizard for the development of a chiral separation method on the Agilent 1260 Infinity II Analytical SFC system. The SFC system was equipped with a four-column selection valve and four different chiral columns for scouting runs against different isocratic separation conditions.
The separation of enantiomers is one of the main application areas for modern SFC instruments. On a SFC instrument, chiral separations are typically 10 to 20 times faster than their classical separation on normal phase HPLC. In addition, the solvents used for the SFC separation are less harmful, and waste disposal is less expensive than for normal phase solvents.
Results and Discussion
In the described chiral screening campaign, four different chiral columns were used with the strong eluting solvent, methanol, as the modifier. The goal of the campaign was the identification of a fast
method with a good baseline separation of the propranolol enantiomers within a maximum 6 minutes run time. Therefore, four isocratic compositions containing 15, 20, 25, and 30 % methanol were used.
After a quick optimization of the identified conditions, a separation of two enantiomers in under 4 minutes could be achieved. This is typically a factor of 10-times faster than the classical separation of enantiomers by normal phase chromatography. Finally, it is shown that the identified separation method could also be used for the analytical preparative separation of enantiomers.