Investigations into the use of gold nanorods (Au-NRs) for biological applications are growing exponentially due to their distinctive physicochemical properties, which make them advantageous over other nanomaterials. Au-NRs are particularly renowned for their plasmonic characteristics, which generate a robust photothermal response when stimulated with light at a wavelength matching their surface plasmon resonance. Numerous reports have explored this nanophotonic phenomenon for temperature driven therapies; however, to date there is a significant knowledge gap pertaining to the kinetic heating profile of Au-NRs within a controlled physiological setting. In the present study, the impact of environmental composition on Au-NR behavior and degree of laser actuated thermal production was assessed. Through acellular evaluation, we identified a loss of photothermal efficiency in biologically relevant fluids and linked this response to excessive particle aggregation and an altered Au-NR spectral profile. Furthermore, to evaluate the potential impact of solution composition on the efficacy of nano-based biological applications, the degree of targeted cellular destruction was ascertained in vitro and was found to be susceptible to fluid-dependent modifications. In summary, this study identified a diminution of Au-NR nanophotonic response in artificial physiological fluids that translated to a loss of application efficiency, pinpointing a critical concern that must be considered to advance in vivo, nano-based bio-applications.