Development and Validation of HPTLC Method for Estimation of Acenocoumerol in Bulk Drug and Dosage form

A simple, accurate and precise high performance thin layer chromatographic (HPTLC) method has been developed for the estimation of Acenocoumerol in bulk drug and dosage form. The method employed silica gel 60 F254 precoated plates as stationary phase and mixture of toluene: isopropyl alcohol:formic acid (7.5:2:0.5) as mobile phase. Densitometric scanning was performed at 290 nm using Camag TLC scanner 3 with WINCAT software of version 1.4.3 Camag. Beer’s law was obeyed in the concentration range of 30ng/spot-90ng/spot. The Retention factor for Acenocoumerol was found to be 0.68. The limit of detection and limit of quantitation were found to be 5 ng/spot & 15 ng/spot respectively. The % RSD of intra-day variation and inter day variation were found to be 1.10 and 1.12 respectively. As per ICH guidelines the results of the analysis were validated in terms of linearity, precision, accuracy, limit of detection and limit of quantification, and were found to be satisfactory. The proposed method can also be used for routine quality control to accurately determine Acenocoumerol in bulk and dosage form.