A sensitive, selective, precise and stability indicating high-performance thin layer chromatographic method (HPTLC) was developed and validated for analysis of eugenol from methanolic extract of Myristica fragrans seed. The chromatography study was performed on aluminium foil-backed silica gel 60 F-254 HPTLC plates using toluene-ethyl acetate-formic acid (2:5:0.2, v/v/v) as the mobile phase. The densitometric determination was carried out by TLC scanner (CAMAG) at 560 nm in reflectance/absorbance mode. The developed method was validated for different parameters like linearity, precision, recovery, robustness, detection limits, quantification limits and stressed stability study in accordance with ICH guidelines. The developed analytical method were found to be linear in the concentration range of 100-1000 ng band -1 with regression value closer to unity (r2=0.998). The developed system was found to give compact spots for eugenol (Rf 0.55) with the limit of detection and limit of quantification (21 and 63 ng band -1 ) respectively. Further the study showed recovery of eugenol in the range of 99.3-99.8% with the precision value ≤ 1.85% and ≤ 1.71%. The all validation parameters result showed satisfactory result in comparison to pure eugenol. The stress degradation study of eugenol showed well separated degraded peak from the pure eugenol. This developed stability indicating HPTLC method was found to ideal method to separate the eugenol from its degradation products. Further this method will be successfully used in the analysis and routine quality control of eugenol loaded formulations.