Andrographis paniculata [(Burm. f.) Wall. ex Nees, family Acanthaceae], commonly known as kalmegh is used both in Ayurvedic and Unani system of medicines for a number of ailments related to digestion, hepatoprotection, hypoglycaemic and as anti-bacterial, analgesic, anti-inflammatory, vermicidal and antipyretic. Although kalmegh is extensively used in Indian Systems of Medicine, but it is not yet cultivated on a large-scale. The supply of herb is met from the wild growing populations which is now under constrain due to over-collection. Considering its value as a drug raw material, genetic variability of A. paniculata was augmented from different geographical regions of India by explorations and was conserved at the National Gene Bank, NBPGR, New Delhi. The therapeutic activity of the herb has been attributed to andrographolide, a bicyclic diterpenoid lactone present in the whole herb. Thirty germplasm collections of A. paniculata were evaluated for bioactive compound andrographolide to find out superior genotype for high drug yield. The andrographolide content in the germplasm was quantitatively analyzed by high performance thin-layer chromatography (HPTLC) at 232 nm. Results indicated wide variability among thirty accessions of A. paniculata for andrographolide content which ranged from 0.72 to 2.99 % on dry weight basis. DIVA-GIS analysis of A. paniculata germplasm for andrographolide content showed that diverse accessions of A. paniculata exist in three Indian states of Orissa, Tamil Nadu and Kerala. Promising accessions having high andrographolide content were identified; IC520361 (2.99 %), IC520395 (2.86 %), IC399125 (2.81 %), IC369404 (2.77 %) and IC520394 (2.61 %). Due to the economic importance of the drug, these promising collections can be exploited for large-scale cultivation for their pharmacologically important constituents mainly andrographolide content and also can serve as potential source for breeding and improvement of cultivars for obtaining higher drug yield.