HPLC Determination of Enantiomeric 2-Azabicyclo[2.2.1] hept-5-en-3-one on Chiral Stationary Phase

A simple high-performance liquid chromatography method has been developed and validated for determination of enantiomeric purity of 2-azabicyclo[2.2.1] hept-5-en-3-one with a run time of less than 10 min. Complete separation of enantiomers has been achieved on a Chiralcel OD-H analytical column (250 × 4.6 mm) using n-hexane-isopropanol (80:20, v/v) as the mobile phase at a flow rate of 1.0 mL/min under UV and optical rotation detection. The effect of the mobile phase and temperature on enantioselectivity for the enantiomers were further evaluated. The method was validated with respect to precision, accuracy, linearity, limit of detection (LOD), limit of quantification (LOQ), and robustness. The recoveries were between 99.1 % and 102.2%, with percentage relative standard deviation less than 1.14%. The LOD and LOQ for the (+)-2-azabicyclo[2.2.1]hept-5-en-3-one were 1.2 and 4.3 μg/mL and for the (-)-2-azabicyclo[2.2.1]hept-5-en-3-one were 1.3 and 4.4 μg/mL, respectively. This method is expected to be helpful for the determination of the enantiomeric purity of 2-azabicyclo[2.2.1]hept-5-en-3-one in bulk samples.