Cortisol is an important steroid hormone in human physiology. Variations or abnormalities in the physiological cortisol levels control acute and chronic stress response, as well as contribute to diseases and syndromes including Addison's disease and Cushing syndrome. The ability to monitor cortisol levels in the physiological range is key in diagnosis and monitoring of these conditions, where current methodology for determination of cortisol levels relies on instrumentation that requires extensive sample preparation, long run times, and is destructive to the sample. Raman spectroscopy provides rapid sample analysis with relatively simple instrumentation; however, Raman spectroscopy is an inherently weak technique. To provide an enhanced Raman signal, we use surface enhanced Raman spectroscopy (SERS) which utilizes oscillating electric fields of metal nanoparticles, enhancing the overall electric field and therefore resulting in an enhanced signal. We demonstrate SERS-based detection of cortisol in the physiologically relevant range using colloidal silver nanoparticles in ethanolic solutions and bovine serum albumin. The SERS spectra obtained in an ethanol matrix demonstrate a sigmoidal concentration response over the physiologically relevant concentration range, with a limit of detection established at 177 nM. Analysis of cortisol solutions in a complex matrix (bovine serum albumin in phosphate buffered saline) is also demonstrated through the use of principal components analysis, a multivariate technique, which shows the separation of cortisol in a linear fashion with respect to cortisol concentration.