The quantity of fossil fuel is declining year by year, and the pollutant also released to the environment which insist on the exploration of new alternative fuel. Biodiesel from non-edible seeds is of primary concern for increasing prices of crude oil and thereof preventing greenhouse gas emissions. In the present study, the production of biodiesel by In-situ acid catalyzed transesterification from Sterculia foetida oil and seeds were reported. GC-FID characterization of oil sample revealed the presence of long-chain fatty acids with a higher proportion of sterculic acid (32%). The operating parameters in acid transesterification process viz., the weight of the oil and biomass, solvent volume, acid volume, temperature and time were optimized using Placket-Burman design. The maximum biodiesel yield obtained for sterculia oil and seeds were 91.58% and 95.91% exhibiting volume of oil being the critical factor based on Plackett-Burman design. One-way ANOVA analysis showed that chosen parameters were statistically significant (P < .05) except for temperature. FT-IR analysis confirms the presence of biodiesel (1741 cm–1) obtained from the transesterification process. © 2019, © 2019 Taylor & Francis Group, LLC.