In spectroscopic optical coherence tomography, it is important and useful to separately estimate the absorption and the scattering properties of tissue. In this paper, we propose a least-squares fitting algorithm to separate absorption and scattering profiles when near-infrared absorbing dyes are used. The algorithm utilizes the broadband Ti:sapphire laser spectrum together with joint time-frequency analysis. Noise contribution to the final estimation was analyzed using simulation. The validity of our algorithm was demonstrated using both single-layer and multi-layer tissue phantoms.