Introduction: Rajakoshataki (Luffa acutangula(L.) Roxb. is a succulent trailing climber that belongs to cucurbitaceae family. The present study attempts to evaluate macroscopy, microscopy, physio-chemical, phytochemical and HPTLC studies of different extracts of fruit of Luffa acutangular (L.)Roxb. Materials and Methods: Fresh Rajakoshataki fruit was collected from the local vegetable market, Udupi. Sample was preserved in fixative solution. The fixative used was FAA (Formalin-5ml + Acetic acid-5ml + 70% Ethyl alcohol-90ml) for pharmacognostical study. Left sample were shade dried for preliminary phytochemical test, physiochemical and HPTLC. Results and discussion: Microscopic image showed the presence of normal fruit structure like Epicarp, Mesocarp and Endocarp. The study proved the presence of Alkaloids, Tannins, Saponins and carboxylic acid and HPTLC densitometric graph showed the peaks. Conclusion: These parameters help in the identification and standardization of fruit of Rajakoshataki.