Wedelolactone, a characterizing compound present in the plant Eclipta alba is used in viral hepatitis, liver diseases, skin and hair care. Wedelolactone was extracted from the plant using Soxhlet and supercritical fluid extraction technique and the compound was isolated using column chromatography. The compound was then characterized by spectroscopic techniques. The method employed HPTLC aluminum plates precoated with silica gel 60 F254 as the stationary phase. The solvent system consisted of toluene: ethyl acetate: formic acid (5:5:0.1v/v). Densitometric analysis of wedelolactone was carried out in the absorption mode at 351 nm. Quantitative evaluation was achieved by measurement of peak intensity. The method was validated for precision, accuracy, robustness and recovery. The HPTLC method can be applied for identification and quantitation of wedelolactone in herbal extracts of Eclipta alba.