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This paper studies the problem of separation of two harmonic-based source signals from a single-channel mixture signal based on employment of a sinusoidal model. The sinusoidal model represents the signal as a sum of sine-waves, whose parameters (i.e., frequencies, amplitudes, and phases) are estimated by a least-square method that minimizes the reconstruction error between the model and the mixture signal. A comprehensive evaluation of the performance of the sinusoidal model for separation of simulated harmonic signals with various fundamental frequencies is presented. Very good performance, in terms of signal-to-distortion ratio, is observed without any a-priori knowledge about F0s of individual signals. The studied model is then demonstrated for separation of a mixture of two speech signals.