Amukkara choornam’ is a popular polyherbal Siddha formulation used for gastritis, spleen enlargement, leucorrhoea, hiccup, anemia, tuberculosis and kappa diseases. Eucalyptol in one of the prevalent phytoconstituent present in the ingredients of this formulation. A sensitive, selective and precise thin-layer chromatographic method has been developed and validated for the analysis of eucalyptol in Amukkara choornam. Separation and quantification was achieved by TLC using mobile phase of ethanol-ethyl acetatate (9:2, v/v), (RF 0.38) on precoated silica gel 60F254 aluminium plates and densitometric determination was carried out in reflection/absorption mode at 343 nm. The calibration curve was linear in the concentration range of 4.612ng-55.344ng spot -1. The method was validated for precision, repeatability and accuracy. The proposed method was found to be simple, precise, specific, sensitive and accurate for the quantification of eucalyptol.