In view of the lack of specific operation methods of single particle effect simulation in large capacity NOR Flash memory, this paper proposes three methods of single particle effect software fault injection in NOR Flash memory: single particle flip, single particle function interrupt and single particle latching. The board level test system is designed for large capacity devices and its function is verified. The single particle effect simulation experiment is carried out by fault injection method. The single particle effect test system of NOR Flash memory consists of FPGA control logic，Flash detection board and host computer software. Results indicate that single particle flip, single particle latching and single particle function interruption software fault injection methods are verified by single particle effect test system of NOR Flash memory. This paper can provide a reference for related single particle simulation and the electrical system reliability analysis for single particle effect.