Optimization and Evaluation of Column Efficiency in RP-HPLC Method for Pharmaceutical and Forensic Drug Analysis

Abstract

Reverse Phase High Performance Liquid Chromatography (RP-HPLC) is an efficient and reliable technique employed in pharmaceutical and forensic analysis. The assay determination in pharmaceutical and certain molecular detection is performed using this technique. Performance of chromatographic columns play a central role in development of more efficient separation methods. It is one of the main component of system suitability testing in a chromatographic system. The column efficiency can be assessed by using various traditional methods and reproducible results can be evaluated using liquid chromatography We studied the column efficiency using RP-HPLC technique for pharmaceutical drug product Aceclofenac in a validated method. Two mobile phases combinations Acetonitrile: Methanol (80:20 v/v) and Acetonitrile: Methanol: Ammonia (225:50:1 v/v) were compared on C-8 Lichrospher RP-HPLC column for Aceclofenac and three tablet samples (30 ppm). The height equivalent to theoretical plate (HETP) referred as direct measure of column efficiency produced efficient results (530-545 and 190-230) for respective mobile phases. The lower HEPT is an indication of efficient elution in second mobile phase. The change in polarity as per pKa of the analyte may enhance elution and resolution. helpful in method development for pharmaceutical and forensic drug analysis using liquid chromatography.