A three-dimensional (3D) printing apparatus, a printing calibration board and a three dimensional printing calibration method thereof are provided. The three-dimensional printing apparatus is adapted to spray the printing material. The three-dimensional printing apparatus includes a nozzle module, a printing platform and a control unit. The printing platform has a carrying surface, where the calibration pattern is disposed on the carrying surface. The calibration pattern at least includes a datum path and a first auxiliary path. The control unit controls the nozzle module coupled to the control unit to spray the printing material along the datum path, and the printing speed of the nozzle module is adjusted in response to the width of coverage of the printing material on the calibration pattern.