Vasicine is one of the important alkaloid obtained from the plant Adhatoda vasica. In the present study, the drug was collected from five accessions namely Bangalore, Shimoga, Trichy, Kolhapur and Palakkad (Kerala) between August and November. The Methanolic extracts were used for HPLC analysis. The results of the HPLC analysis showed variations in all the accessions. The extract from Bangalore accession showed significantly higher amount of Vasicine (0.93%) and that of Palakkad (Kerala) showed least amount (0.59%) of Vasicine.