Diisononyl phthalate (DINP) is a widely used industrial plasticizer. People come into contact with this chemical by using plastic products made with it. Human health can be adversely affected by long-term DINP exposure. However, because the body rapidly excretes DINP metabolites, the use of single-point urine analysis to assess long-term exposure may produce inconsistent results in epidemiologic studies. Hair analysis has a useful place in biomonitoring, particularly in estimating long-term or historical exposure for some chemicals. Several studies have reported using hair analysis to assess the concentrations of heavy metals, drugs and organic pollutants in humans. As a biomarker, DINP metabolites were measured in rat hair in animal experiments to evaluated long-term exposure to DINP. In addition, we evaluated the correlation between the levels of DINP metabolites in hair and in urine. The levels of DINP metabolites in rat hair were significantly higher in the exposure group, relative to the control group (p<0.05). DINP metabolites had a positive correlation with increasing administered dose. Significant positive correlations for MINP, MOINP and MHINP were found between hair and urine (r=0.86, r=0.79 and r=0.74, respectively, p<0.05). Several metabolites in urine showed earlier saturation than in hair. In this report, we detected eight metabolites in hair and demonstrate that hair analysis has potential applications in the assessment of long-term exposure to DINP.