Objective：This study is proposed to clarify the biological conversion process of p，p’-dichloro-diphenyl-trichloroethane（DDT）metabolized by cytochrome P450（CYP）2B6. The result will provide basis for the risk assessment and prevention of persistent organic pollutant. Methods：The detection method based on gas chromatography-mass spectrometry（GC-MS/MS）was established to detect DDT in biological samples. The metabolic model in vitro was established，using recombinant enzyme CYP2B6 and p，p’-DDT. The enzyme activity was calculated by Michaelis-Menten equation. The 8-MOP as enzyme inhibitor was used to verify the metabolical ability of CYP2B6 on p，p’-DDT. Meanwhile，the metabolic model in vivo was established in SD rat（rat CYP2B1 is homologous enzymes of human CYP2B6） by tail intravenous injection of p，p’-DDT and CYP2B6 specific chemical inhibitor KR-Ⅱ. The changes of p，p’-DDT prototypes and metabolites（p，p’-DDE，p，p’-DDD）and the changes of enzyme activity were assessed in the serum（0 min，5 min，10 min，20 min，30 min）and liver microsomes. Results：A detection method for DDT was successfully established and tested. Metabolic experiments in vitro confirmed that p，p’-DDT could metabolized by CYP2B6，and the main metabolite was p，p’-DDE. Kinetic parameter Km was 15.12 μmol/L，and the Vmax of per nanomolar P450 is 12.8 nmol/min. Inhibitor 8-MOP significantly inhibited the CYP2B6 activity and metabolites p，p’-DDE content significantly decreased. After CYP2B1 was inhibited，the metabolic enzyme activity was significantly decreased and the serum p，p’-DDE and p，p’-DDD product content was lowered. Conclusion：CYP2B6 can metabolize p，p’-DDT to produce metabolites dominated by p，p’-DDE，and it should be the dominant enzyme of DDT in the human body. Considering that there are differences in the expression and activity of CYP2B6. This study can provide a basis for screening of susceptible populations exposed to DDT and the optimization of risk assessment.