Development and Validation of a High-Performance Thin-Layer Chromatographic Method for the Quantitative Assessment of Topiramate from Human Serum: Application in Therapeutic Drug Monitoring

A simple and reliable high-performance thin-layer chromatographic (HPTLC) method has been developed and validated for the determination of topiramate in human serum which will help in therapeutic drug monitoring that aids in the clinical management of patient therapy. This method uses dipping method of derivatization to make the non-chromophore topiramate visible for detection. Pre-coated silica gel F-254 HPTLC plates were used to carry out chromatographic separation using a mixture of toluene-ethanol (9:1) as the mobile phase. Derivatization was carried out using dipping method. Betamethasone was used as internal standard. Densitometric detection was carried out at 417 nm. The method was validated for linearity, precision, selectivity, limit of detection and limit of quantification and accuracy. Linear calibration curves in the range of 0.25 to 40 mu g band(-1) gave a correlation coefficient of 0.9963. The intra-day (n = 6) and inter-day (n = 18) precision, expressed as the relative standard deviation, were in the range of 2.89% to 3.41% and from 2.97% to 4.20%. The limit of detection and the limit of quantification were found to be 0.36 mu g band(-1) and 1.10 mu g band(-1). The accuracy was calculated as percentage recovery and was found to be 97.41% and 110.7%. The method was specific and showed no matrix interference. Compared to other methods employed in the determination of topiramate from blood, this method is cost-effective, simple, and reliable to carry out therapeutic drug monitoring of topiramate.