Sarasvata ghrita (SG) is an ayurvedic polyherbal formulation prescribed for improvement of intelligence and memory, treatment of speech delay, speaking difficulties and better digestion in children. Literature survey revealed that there is lack of standardisation aspects of SG. Standardisation of the SG was done by newly developed high performance thin layer chromatography (HPTLC) method for determination of Bebeerine (BEB), Piperine (PIP), 6- Shogaol (SHO) and β- Asarone (ASA) in commercially available marketed and in-house prepared formulations of SG. The simultaneous estimation of these markers was carried out on silica gel precoated thin layer chromatography plates with 60F254 as the stationary phase and eluted using Toluene: Methanol: Triethylamine (9.2:0.5:0.3) as mobile phase. Camag TLC scanner III was used for densitometric scanning. Optimum wavelength 282 nm was selected for detection and quantification. The method was validated as per ICH guidelines. The retention factor for BEB, PIP, 6-SHO and ASA were found to be 0.11, 0.39, 0.51 and 0.81 respectively. The developed HPTLC method was found to be linear with correlation coefficient 0.999 for BEB, PIP and SHO; and 0.998 for ASA. The limits of detection and limit of quantification were determined to be 10.89 and 33.04 ng/spot, 8.12 and 24.62 ng/spot, 27.38 and82.98 ng/spot, 18.13 and 54.95 ng/spot for BEB, PIP, SHO and ASA respectively. The developed HPTLC method was found to be simple, specific, accurate, precise and robust, thus can be used for routine analysis of SG for standardization with respect to selected active markers.