In this study of the orofacial formalin test in rats, the effects of different formalin concentrations (0.2%, 0.5%, 1.5%, 2.5%, 5% and 10%) on the behavioural nociceptive response (face rubbing) was investigated. The histological responses of the skin were also evaluated. Increasing the concentration of formalin caused a parallel aggravation of histological signs of tissue inflammation and injury. All concentrations provoked an early phase of nociceptive response, but its intensity was not concentration-dependent. The 2nd phase of response to formalin only occurred for concentrations of 1.5% and higher. A positive relationship between the formalin concentration and the amplitude of the rubbing activity measured between 12 and 45 min after injection could be observed until 2.5% but with the highest concentrations (5 and 10%), the amplitude of the response decreased. Our findings indicate that the orofacial formalin test should be carried out using concentration between 0.5 and 2.5%. This is essential to assess increase as well as decrease in pain intensity. Moreover, this will have the effect of minimizing the suffering of the experimental animal.