ISOLATION, IDENTIFICATION, AND CHARACTERIZATION OF NATURAL CHEMICAL COMPOUNDS AND GC-MS ANALYSIS OF METHANOL EXTRACT OF STEM BARK OF MORINGA OLEIFERA WITH ANTI-HYPERGLYCEMIA AND ANTI-MICROBIAL ACTIVITIES

Objectives: The study aimed to isolate and identify novel and known phytochemical compounds from the methanol extract of Moringa oleifera stem bark, to carry out comprehensive phytochemical profiling using gas chromatography-mass spectrometry (GC-MS), and to evaluate the α-amylase and α-glucosidase inhibitory activities of the isolated compounds for potential anti-diabetic effects, along with assessing their antibacterial and antifungal activities. Methods: The methanol extract of M. oleifera stem bark was prepared and subjected to various chromatographic techniques for compound isolation. The structures of the isolated compounds were confirmed through spectral analysis, including proton nuclear magnetic resonance and carbon-13 nuclear magnetic resonance, infrared, and mass spectrometry spectroscopy. GC-MS analysis was conducted to identify the phytochemical constituents based on peak area, retention time, molecular weight, and molecular formula. The isolated compounds were tested for inhibitory effects on α-amylase and α-glucosidase enzymes, and their antibacterial and antifungal activities were evaluated using standard in vitro assays. Results: Two novel compounds, 4-hydroxy-7-methylmellein and 8-methylluteolin, along with four known compounds, benzyl laurate, 4-hydroxymellein, kaempferol, and quercetin, were isolated from the methanol extract. GC-MS analysis revealed the presence of eighty-five compounds in the extract in which 07 compounds identified as common laboratory contaminants or plasticizers, 70 compounds are identified as bioactive phytochemicals, and eight compounds are repeated at different R. Time. Total relative abundance by common laboratory contaminants or plasticizers is 22.61%. Alcohol, alkane, ester, fatty acid, phenolic, sterol, and miscellaneous bioactive phytochemicals are also identified with their total relative abundance, that is, 22.35%, 2.05%, 5.36%, 15.19%, 2.06%, 14.7%, and 15.7% respectively. Sterol, that is, dl-α-Tocopherol and γ-Sitosterol, and alcohols such as isocitronellol, cis-sinapyl alcohol, n-tetracosanol-1, and humulane-1,6-dien-3-ol are identified. The isolated six compounds exhibited inhibitory activity against α-amylase and α-glucosidase enzymes, indicating potential anti-hyperglycemia properties. In addition, antibacterial and antifungal assays demonstrated significant antimicrobial potential for certain isolated compounds. Conclusion: The methanol extract of M. oleifera stem bark contains diverse bioactive phytochemicals with notable pharmacological activities, including enzyme inhibition relevant to hyperglycemia management and antimicrobial properties. The discovery of two novel compounds expands the phytochemical knowledge of M. oleifera and supports its traditional medicinal applications.