Indolinospirobenzoxazine was synthesized by direct condensation of 4-amino-2-nitrosophenol with 1,3,3-trimethyl-2-methylene indoline. Its biological effects on bacteria species, Salmonella typhi, Escherichia coli, Staphylococcus aureus and fungi species, Candida albican have been determined. Indolinospirobenzoxazine was found to be highly active on all the bacteria species tested, with inhibition zone >12 mm. Moderate activity was observed against Candida albican (inhibition zone 11 mm) which represented the lowest sensitivity among the microorganisms tested.