Validated High Performance Thin Layer Chromatography Method for the Simultaneous Determination of Amitriptyline, Atenolol and Propranolol

A new simple, precise, rapid and selective High-Performance Thin-Layer Chromatographic (HPTLC) method has been developed for the simultaneous determination of amitriptyline (AMTRL), atenolol (ATL) and propranolol (PRPL) using benzene: methanol: acetone: ammonia solution (25% NH3) (5:3:2:0.2; v:v) as the mobile phase and Merck HPTLC plates (0.2 mm thickness) pre coated with 60F254 silica gel on aluminium sheet as the stationary phase. Detection was carried out densitometrically using a UV detector at 254 nm. The retention factors (Rf value) of AMTRL, ATL and PRPL were 0.80, 0.37 and 0.69 respectively. The developed method was fully validated as per the guidelines given by the International conference on Harmonisation (ICH) for precision, recovery, accuracy, robustness, ruggedness and specificity and determination of the limits of detection and quantification. The amount of AMTR, PRPL and ATL present in marketed tablets was quantified and estimated using the developed method which was found to be in close agreement with the labelled content. The linear regression analysis for calibration plots showed good linear relationship in the concentration range of 0.1ug/spot to 10ug/spot with respect to peak area.