Bio-switch is one of the new innovations in active packaging in order to extend shelf life of food products and preserve their best quality. In this research, the possibility of using curcumin, bixin and norbixin as a natural and safe preservatives in developing antimicrobial film for food packaging was investigated. First of all, curcumin and bixin were extracted from turmeric and annatto seeds respectively by maceration. To prepare the water-soluble norbixin, bixin was saponified. In order to study the antibacterial efficacy of curcumin, bixin and norbixin against S.aureus, E.coli and L. Innocoa, the extracts were identified and their inhibition zones, MIC and MBC were determined against selected bacteria. The probable synergistic effects were evaluated using fractional inhibitory concentration (FIC) index and isobologram curves. Bio switch technique was used to protect the preservative from oxidation, light and undesirable interaction with other food components. In order to formulate the composite, single component films from different modified starch, two components films from starch with Zein, PVA and Olibanum and finally, three components films of starch/zein/PVA were made. All physical, optical, mechanical and enzymatic sensitivity tests were performed and results compared to determine the optimum formulation of composite. Bixin was then incorporated to composite and antimicrobial efficacies of films against selected bacteria were evaluated. At the end, composite was mixed with industrial polymer of LDPE and final film packaging was produced. Supplementary tests including FTIR, TGA, XRD and microstructure studies using SEM were performed on optimum composite and final films. According to the results, bixin showed more significant inhibitory effects against selected bacterial strains than other natural pigments. Synergetic effects were observed by curcumin and bixin combination against S.aureus (FICI 0.5), but they acts as antagonist against E.coli and L. Innocoa. Simple corn starch films showed better performance due to higher stability in water and acidic and alkaline solutions and stronger mechanical properties among other kinds of starch films. Incorporating PVA into starch improved its mechanical properties, also zein at the starch film formulation can reduce the water solubility of mixed films. Optimum formulation of three components films was determined as 85% starch, 10% zein and 5% PVA, respectively with desirability of %98. Formulated bio-switch showed appropriate performance at realease of antimicrobial agents and films containing 200 ppm of bixin formed inhibition zones against all three microorganisms. Final films formulated with LDPE were effective against S.aureus and showed significant antibacterial activity.