Moringa oleifera Lam. is mostly known as the „potent medicinal flora“ in terms of enormous therapeutic efficacy due to the availability of certain biologically active molecules. This has dual characteristics: it can be used as food and an active ingredient in many pharmaceutical formulations. Flavonoids, phenols, and alkaloids are some of Moringa‘s most abundant bioactive compounds, and they are responsible for its significant therapeutic properties. The current study is based on extracting and identifying major bioactive molecules from M. oleifera leaf and seed parts using a high-performance liquid chromatography technique to formalize its therapeutic potency of bioactive compounds. M. oleifera leaves and seed extracts in methanol were successfully analyzed in a single run using the HPLC method designed for this purpose. A Sunfire C18 column measuring 4.6 x 250 mm was used for the 220 nm HPLC analysis, and the solvent system consisted of 0.1% acetate buffered by acetonitrile (65:35 v/v) at a flow rate of 1.2 mL/min. The peak observed at 3.269 minutes in leaf extract and 3.289 minutes in seed extract shows the presence of niaziridin. The recovery values of niaziridin in M. oleifera leaves and seed extracts were 45.90 and 66.85%, respectively. Chromatograms show that seed extract has a larger amount of nitrile glycoside than leaf extract. The present investigation used an HPLC process to investigate the characteristics of niaziridin. This research is useful for standardizing M. oleifera leaf and seed extracts and producing pharmaceutical drugs from M. oleifera. © 2023, Dr. Yashwant Research Labs Pvt. Ltd.. All rights reserved.