Nanoscale resonant devices based on optical tweezers are widely used in the field of precision sensing. In the process of driving the nanoresonator based on the Coulomb force, the real-time, precise regulation of the charge carried by the charged resonator is essential for continuous manipulation. However, the accuracy of the existing charge measurement methods for levitated particles is low, and these methods cannot meet the needs of precision sensing. In this study, a novel net charge measurement protocol for levitated particles based on spatial speed statistics is proposed. High-precision mass measurement based on Maxwell’s rate distribution law is the basis for improving the accuracy of charge measurement, and accurate measurement of net charge can be achieved by periodic electric field driving. The error of net charge measurement is less than 7.3% when the pressure is above 0.1 mbar, while it can be less than 0.76% at 10 mbar. This proposed method features real-time, high-precision, non-destructive, and in situ measurement of the net charge of particles in the medium vacuum, which provides new solutions for practical problems in the fields of high-precision sensing and nano-metrology based on levitated photodynamics.