Development and Validation of Newer High Performance Thin Layer Chromatographic Method for Quantification of Eflornithine Hydrochloride in Pharmaceutical Formulations

A new, simple, sensitive, precise and robust high performance thin layer chromatography (HPTLC) method was developed for the estimation of eflornithine hydrochloride (DFMO) in pharmaceutical dosage forms. Estimation was performed on TLC aluminum plates precoated with silica gel 60F₂₅₄ as stationary phase. Linear ascending development was carried out in twin trough glass chamber saturated with mobile phase consisting of methanol: chloroform: acetic acid: 1% triethylamine (4:6:0.1:0.5 v/v/v) at room temperature (25±2°C). Analysis of the plate in absorbance mode at 220nm was carried out. The system was found to give compact spots for DFMO with R_f (Retardation factor) value of 0.55 respectively. The data for calibration plots showed good linear relationship with correlation coefficient of 0.999 in the concentration range of 300–800ng mL^-1 for DFMO respectively. The values of limit of detection (LOD) were 0.6238ng mL^-1 and limit of quantification (LOQ) were 1.8903ng mL^-1 for DFMO respectively. The accuracy of the method was 100.44%. The precision demonstrated a relative standard deviation of less than 2.5%. The results were satisfactory when compared with the literature. This new method was validated according to the International Conference on Harmonization (ICH) guidelines which include linearity, precision, accuracy, specificity, robustness, detection and quantitation limits. The developed methods found to be sufficiently precise and reproducible for established conditions and after validation may be used for routine analysis of eflornithine hydrochloride in pharmaceutical formulations.