eng Oriental Scientific Publishing Company Biosciences Biotechnology Research Asia 0973-1245 2026-10-08 23 3 59878 article Samta Amratbhai Mahyavanshi 1 Kalpesh Babubhai Ishnava 2 Hiren Babubhai Soni 1 Department of Environmental Science and Technology (EST), Institute of Science and Technology for Advanced Studies and Research (ISTAR), The Charutar Vidya Mandal (CVM) University, Vallabh Vidyanagar, Gujarat, India P G Department of Biosciences, Sardar Patel University, Vallabh Vidyanagar, Gujarat, India The present study aimed to evaluate the in vitro anti-denaturation activity of crude leaf extracts of Nyctanthes arbor-tristis L. using bovine serum albumin (BSA) as a protein denaturation model. Dried leaves were extracted by the cold extraction method using four solvents, namely water, methanol, ethyl acetate, and chloroform. The highest extractive yield was obtained with the methanolic extract (1.34%), followed by the water (1.25%), ethyl acetate (0.61%), and chloroform (0.41%) extracts. Qualitative phytochemical screening revealed the presence of various secondary metabolites, with the methanolic extract showing alkaloids, phenols, saponins, and terpenoids. Quantitative analysis of the methanolic extract revealed a total alkaloid content of 1.70% (17.0 mg/g extract) and a total phenolic content of 11.20 mg GAE/g extract. In the BSA protein denaturation assay, the water extract exhibited the highest anti-denaturation activity at the highest tested concentration of 50 mg/mL (82.02%), followed by the methanolic (75.00%), ethyl acetate (74.43%), and chloroform (64.66%) extracts. HPTLC analysis revealed five distinct bands, while GC-MS analysis tentatively identified several fatty acid constituents, including undecanoic acid, tridecanoic acid, pentadecanoic acid, hexadecenoic acid, heptadecanoic acid, octadecanoic acid, and nonadecanoic acid. Overall, the findings provide preliminary evidence of inhibition of heat-induced BSA protein denaturation by N. arbor-tristis leaf extracts under the tested conditions, with the water extract showing the highest observed inhibition at 50 mg/mL. These preliminary findings support further investigation of N. arbor-tristis leaves to elucidate their potential properties and the phytoconstituents potentially associated with the observed activity. https://www.biotech-asia.org/vol23no3/in-vitro-anti-denaturation-activity-and-phytochemical-analysis-of-nyctanthes-arbor-tristis-l-leaf-extracts-using-a-bsa-protein-denaturation-assay/ Anti-denaturation; GC-MS; HPTLC; In vitro; Leaves; Nyctanthes arbor-tristis; Phytochemicals