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The roots of Clerodendrum phlomidis L.f. is known to be used as Agnimantha in Ayurvedic system of medicine. Whereas literature emerged from various texts reports other species of Premna as Agnimantha that leads to controversies in usage of authentic drug source. Hence the study aims to develop pharmacognosy characteristics to identify the authentic and substitute sources of Agnimantha. The Ayurvedic verses prescribed for Agnimantha compliance with Clerodendrum phlomidis, Premna serratifolia and Premna mollissima. Whereas the vertically fissured stem found prominent in Clerodendrum phlomidis. Likewise cork region is composed of homogenous cells with grouped and scattered calcium oxalate crystals, without oils. Whereas heterogeneous cells with oils and scattered calcium oxalate crystals are found in Premna serratifolia and Premna mollissima. Similarly phytosterols found unique in Clerodendrum phlomidis roots. The developed HPTLC method serves as tool to identify and distinguish the authentic from substitute and adulterants. Enthrallingly quantitative HPLC analysis revealed the presence of verbascoside in all sources of Agnimantha and comparatively more in substitutes. Hence the study concludes that roots of Clerodendrum phlomidis is authentic source of Agnimantha, where Premna serratifolia and Premna mollissima forms the equivalent substitutes
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