Objective: To perform phytochemical screening, estimate total saponins and to study the secondary metabolite profiling of Chlorophytum borivilianum by ESI-MS (Electron spray ionisation-mass spectroscopy) and GC-MS (Gas chromatography-mass spectroscopy) techniques.Methods: The powdered samples of leaves and roots (nature grown and freshly harvested and hardened in vitro regenerated) and callus of Chlorophytum borivilianum (2 g) were extracted exhaustively by soxhlet by refluxing with petroleum ether (60-80 °C) and then with ethanol (85%). Alternatively, the methanol extract was prepared for the samples by the cold maceration method and the filtered extract was used. This extract was further analyzed by gas chromatography-mass spectrometry to identify and characterize the chemical compounds present in the crude extracts. Vanillin sulphuric acid assay was done to estimate the total saponin content of extracts.Results: Phytochemical analysis of ethanolic extract showed the presence of major classes of phytochemicals. The spectrophotometric analysis as well as the GC-MS results revealed the remarkably high percentage of saponin content in the in vitro roots. Besides this, many important secondary metabolites viz. β-Sitosterol and Taraxerone were also revealed by GC-MS.Conclusion: Results of this study showed enhanced production of many important secondary metabolites, especially saponins in tubers of in vitro regenerated plantlets vis a vis that of the nature grown plants establishing the fact that micropropagation can pave new vistas for the research in phytopharmaceuticals and bioprospecting of rare and important medicinal plants.Â