Abstract: The long lived radioisotope of ⁵⁹Ni has wide application in many research fields, such as nuclear physics and astrophysics, the management of radioactive waste, biomedical tracer and so on. In order to carry out the applications based on ⁵⁹Ni, the method of the accelerator mass spectrometry (AMS) was developed into the best way to measure ⁵⁹Ni with high sensitivity. Interfering isobars are the main problem for ⁵⁹Ni measurement with AMS. Based on the HI-13 tandem accelerator at the China Institute of Atomic Energy, high precision Q3D detection system and chemical separation technology, the interfering isobar of ⁵⁹Co is greatly eliminated. ⁵⁹Ni-AMS highly sensitive measurement method is established with NiCl₂ as the target substance. A series of laboratory standard samples and blank sample were measured to check the performance of ⁵⁹Ni measurement. The results show that the ⁵⁹Co can be excluded clearly, and the measurement sensitivity of about 5×10^-13 for ⁵⁹Ni/Ni is obtained based on the measurement of the blank sample, which can be used in the measurement of actual samples.