A Novel Reverse Phase High Performance Liquid Chromatography Method For Enantiomeric Separation of Pregabalin With Immobilized Zwitterionic Stationary Phase

Background: The objective of present investigation was to develop simple, sensitive, specific, precise and accurate reversed phase high performance liquid chromatographic (RP-HPLC) method and subsequent validation of the method for the determination of R-isomer in Pregabalin. Method: The chromatographic method was carried out using Chiral pak ZWIX (+) column (250mm x 4.0mm i.d., 3μm particle size) and mobile phase consisting of methanol and ammonium dihydrogen orthophosphate buffer in the ratio of 10:90 v/v and 0.2% trifluoro acetic acid. Mobile phase was delivered at a flow rate of 0.5ml/min. Ultra violet detection was carried out at 210nm. The elution technique was based on isocratic mode and the sample volume 25 μL was used. The method was validated in terms of linearity, range, specificity, accuracy, precision, limit of detection (LOD) and limit of quantitation (LOQ). Results: The retention time of R-isomer and Pregabalin was found to be 7.2 and 10.7 min respectively. The developed method illustrated excellent linearity (r2 > 0.99) in the concentration range of 1.4 μg/ml – 11.25 μg/ml for Pregabalin. No chromatographic interference from the tablets excipients was found. The mean recoveries were found in the range of 99.2-100.8% for Pregabalin which shows accuracy of the method. Conclusion: The developed method was found to be accurate, precise, reproducible and specific and can be successfully applied for the determination of R-isomer in Pregabalin in pharmaceutical formulation and routine analysis in quality control laboratories.