A comparative study of the sensitivity of the 3-induction and 9-induction Buehler test procedures for assessing skin sensitisation potential

Food Chem Toxicol. 2005 Jan;43(1):65-75. doi: 10.1016/j.fct.2004.08.013.


Assessment of skin sensitization potential is a mandatory requirement for the registration or notification of most types of chemicals and products. Until recently, two methods using the guinea pig as test model were the most widely accepted; the guinea pig maximisation test and the Buehler test. In the case of agrochemical formulations, which constitute the final end use product in contact with operators, industry and also some regulatory authorities consider the Buehler method more appropriate as the methodology is more relevant to likely exposure in the field. However, certain European regulatory authorities have become concerned about the sensitivity of the Buehler test for this purpose and have requested that a modified method is used in which additional applications of test materials are used during the induction phase of the protocol (a total of 9 rather than the normal 3). This study was designed to assess whether this modification was justified. Six reference substances (formaldehyde, alpha-hexylcinnamaldehyde, fragrance mix, thimerosal, mercaptobenzothiazole and phthalic anhydride); all mild to moderate skin sensitizing chemicals, were assessed in a study, which compared the use of 3 and 9 induction applications. The results of this study demonstrated that, although most of these sensitisers were detected by both protocols, the modified method (9 induction applications) was no more sensitive than the standard method (3 induction applications). As the modified protocol is also potentially more stressful to the animals, it is concluded that the use of additional induction applications in the Buehler test cannot be justified from either a scientific or an animal welfare perspective.

Publication types

  • Comparative Study

MeSH terms

  • Allergens / toxicity*
  • Animal Welfare
  • Animals
  • Dermatitis, Allergic Contact / diagnosis*
  • Disease Models, Animal
  • Female
  • Guinea Pigs
  • Humans
  • Local Lymph Node Assay
  • Male
  • Risk
  • Risk Assessment
  • Sensitivity and Specificity
  • Skin Tests / methods*
  • Time Factors
  • Toxicology / methods*


  • Allergens