Enantioselective method development and validation of proline by using high performance liquid chromatography

Chirality is a major concern in the modern pharmaceutical industry. The separation of chiral compounds has been of great interest because the majority of bioorganic and synthetic molecules are chiral. Aim of the present investigation was to develop a stereo specific, simple and precise normal phase high performance liquid chromatography (NP - HPLC) method for the separation and enantiopurity of Dextro (D) and Levo (L) enantiomers of proline (PRO) by using Lux 5m Amylose 1 LC column (250.6mm) by using n- Hexane: Iso propyl alcohol (IPA)as mobile phase in the ratios of 90:10 v/v at flow rate of 1.2 ml/ min. D and L forms of PRO was detected at 210nm with retention time of 8.1min and 9 min respectively with correlation coefficient (R 2 ) of 0.999.The method was validated with reference to International conference of harmonization (ICH) in terms of linearity, accuracy, precision (Inter - day and intra - day precision), limit of detection (LOD), limit of quantification (LOQ), stability of test solutions, specificity, system suitability, robustness and ruggedness