A new method to extract complex refractive indices of films with biaxial symmetry from polarized transmission and reflection spectra is described. Theoretical analysis demonstrates that the reflectance/transmittance ratio (R/T ratio) of two films of different thicknesses but with the same optical anisotropy is a simple function of the refractive index (n) and extinction coefficient (k). For films with biaxial symmetry, components of n and k on symmetric axes can be extracted from either s- or p-polarized R/T ratios if the film thickness values are known. The R/T ratio method can generate n and k spectra within a particular wavelength range without assuming a specific relationship among n, k, and wavelength, which is an advantage over many currently available techniques. The R/T ratio method is used to extract the anisotropic complex refractive indices of a polyimide film with known uniaxial symmetry. The resultant n and k spectra compare well with simulations based on known n and k values. The accuracy of n and k spectra is affected mostly by data error in reflection and transmission spectra collection, thickness variation across sample films, and error in sample alignment.