Abstract: Standardization of herbal drugs is the need of the hour as the use and practice of traditional herbal drugs has increased tremendously. The main objective of the present study is to standardize the root tubers of Aconitum heterophyllum as per pharmacopoeial testing protocol which include powder microscopy, physico-chemical screening, HPTLC fingerprinting and GC-MS analysis. Preliminary phytochemical tests indicate the presence of alkaloids, sugars, flavonoids, steroids, quinones and tannins. HPTLC profiling of the ethanol extract using Toluene/ Ethyl acetate (8: 1) as mobile phase revealed the presence of phytochemicals with different Rf values. The GC-MS analysis of the diethyl ether fraction showed the presence of 39 compounds of which 21 were identified.