The invention discloses a separation estimation method of multiple motion parameters in an X-ray angiographic image. Firstly a cardiac motion signal cycle and a sequence of a translational motion change frame are determined according to an angiographic image sequence, and then the motion sequence of a point is tracked through the vascular structure feature point in the angiographic image sequence. Secondly, the motion sequence is processed through multi-variable parameter optimization and Fourier frequency domain filtering, and a best translational motion curve, a best cardiac motion curve, a best respiratory motion curve and a best high frequency motion curve can be separated according to the sequence of the translational motion change frame, the cardiac motion signal cycle, the range of a respiratory motion signal cycle and the range of a high frequency motion signal cycle. According to the method, since the fact that the integration of multiple motion signals is reflected by the X-ray angiographic image is considered, the separation estimation of the multiple motion parameters is provided, each separated motion can be more precise, and accurate and effective help is provided for the diagnosis of a doctor.