Effect of Some Parameters on Enantiomer Separation of Ephedrine, Methamphetamine and Selegiline Using HPLC with β-Cyclodextrin Stationary Phase

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Abstract
The influence of different parameters (mobile phase composition - pH, organic solvent, salt nature and concentration; flow rate, injected amount and temperature) on enantiomeric separation of three pharmacologically important compounds (ephedrine, methamphetamine and selegiline) was studied using β-cyclodextrin stationary phase. The evaluation of effect of these parameters allowed to optimize condition for optical purity determination. The following experimental conditions were chosen for separation of racemic mixture: stationary phase - ChiraDex, 5 μm; column - LiChroCart 250 × 4 mm I.D.; mobile phase - 500 mmol triethylamine/1 with H2SO4 in water, pH = 3.5, flow-rate - 0.8 mL/min; detection - UV absorption at 206 nm; temperature -ambient. The separation of minor (1%) enantiomer in excess of major one can be improved using flow-rate 0.2 mL/min and thermostated column (20°C for methamphetamine and selegiline, 5°C for ephedrine).
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