To solve the inefficiency and complete autonomy issues for Deflection of the Vertical (DOV) determination in geomatics, navigation, and military weapon launching fields, a calculation method based on path compensation has been proposed. After conducting the discretization of the Vening-Meinesz formula in the spherical surface, a novel path gravity anomaly compensation based on the “remove-restore” algorithm was formulated to satisfy the requirement of regional high-precision gravity anomaly data. Meanwhile, utilizing the gravity field spherical harmonic model EIGEN-6c4, the deflection of the vertical was calculated using the Vening-Meinesz formula based on the “remove-restore” method, unlike the previous regional direct integral calculations. Finally, to verify the reliability and feasibility of the algorithm, the gravity measurement system, consisting of the high-precision gravimeter and Strapdown Inertial Navigation System, was constructed to conduct the simulation and shipborne experiments. The simulation and experiment results of the proposed method show that the root mean squares, relative to the real values, of DOV calculation components, are 0.89" and 1.67", while standard deviation values of the DOV components are the same within 2" in experiments, which satisfies the precision requirement of the DOV.