Cymbopogon citratus (DC) Stapf belongs to the genus Cymbopogon (Poaceae) and are known to possess essential oil rich in Citral content. During the present investigation different accessions of C. citratus were collected from various locations of South India, assigned with different accession numbers and maintained at Departmental garden, Bangalore University, Bangalore for further studies. The essential oils were extracted by steam distillation and the essential oils were subjected to GC-MS analysis. A total of nine accessions (Cc1-Cc9) were selected for study and the total essential oil and Citral content were assessed. The Citral content was found to be highest in Cc5 accession. The study showed that there is variation existing with total percentage and composition of the essential oil among the accessions of C. citratus collected from various ecological locations.