Skin Sensitization Assay of Topical Occluded Heat Patches by Modified Buehler Method Using Rabbits

  • Dr. Muhi. N. Salmam
Keywords: Dose response, Guinea pig, OECD guideline, Allergic contact dermatitis, Contact Rabbits abdominal wall


Background:  Many heat patches are known, based on a reaction system comprising iron, carbon, vermiculite, potassium chloride, and water. The heat release from the inorganic oxidation of iron is initiated by exposure to oxygen, such as when the patch is removed from the plastic pouch. Severe burns have occurred when these products are used in conjunction with a heat source such as an analgesic heating patches.

Objectives: Concepts in Cutaneous toxicity is to determine the potential of heat patches to promote skin sensitization reactions after repeated applications using the although modified Buehler test protocols were used.  Buehler tests were conducted according to OECD guideline #406 using a multiple-dose design and test results were analyzed using a standard logistic dose-response model. Contact dermal sensitization is an immunological process where the host animal, through repeated skin exposure, acquires a specific allergic sensitivity to heat patches. In the modified Buehler model,   using rabbit contact dermal sensitivity is manifested as increased erythema and /or oedema.

 Materials and Methods: In the modified using rabbits Buehler protocol test, the induction group at 24 hours after challenge patch removal and again at 48 hours after patch removal with induction patches (1 patch per week). Two weeks after induction the animals are challenged by closed-patch tests to the rabbit's abdominal wall skin for 6h. All tests in the Buehler assay are performed on restrained animal we investigated the feasibility of using heat patches topically applied to abdominal wall skin (rabbits) clipped free of hair by experimental induction and challenge phases as an acceptable method to assess the potential of a occluded exposure to analgesic heat patches to promote skin sensitization reaction.

Ressuts: Depending on collected tables data of challenge  versus  rechallenge weeks (1,2,7) senisation reaction results ,shown  that  the potential cutaneous toxicity of the test  heat patches was non –irritating   to week sensitizer category using rabbit's abdominal wall skin-topical exposure in the test animals by the modified  Buehler test.

Conclusion: The safety use of topical occluded heat patches as pain analgesic if the end-user follows the instruction of application properly.


Download data is not yet available.


Abel, E.A and Wood, G.S (1986). Mechanism of contact dermatitis. Clin Exp Allergy, 4, 339.

Buehler, E. V. (1965). Delayed Contact Hypersensitivity In the Guinea Pig. Archives of Dermatology, 91(2), 171-177.

Organisation for Economic Cooperation Development (OECD). (1992). OECD Guideline for Testing of Chemicals. In Skin Sensitisation. Available at:

Thestrup-Pedersen, K., Larsen, C. G., & Rønnevig, J. (1989). The immunology of Contact dermatitis: A review with special reference to the pathophysiology of eczema. Contact Dermatitis, 20(2), 81–92.

Magnusson, B. (1980). Identification of contact sensitizers by animal assay. Contact Dermatitis, 6(1), 46–50.

K E Andersen, A Volund, & S Frankild. (1995). The guinea pig maximization test--with a multiple dose design. Acta Derm Venereol, 75(6), 463-469.

National Research Council, Committee for the Revision of NAS Publication 1138. (1977). Principles and Procedures for Evaluating the Toxicity of Household Substances. Washington, D.C.: National Academy of Sciences. pp. 23-59.

Griffith. J.F and Buchler, E.V. (1977). Prediction of Skin Irritancy and Sensitizing Potential by Testing Animals and Man, In: Cutananeous Toxicity (V.A. Drill and P. Lazar). Academic Press, New York, pp 155-173.

Finkelstein, P. Laden. K & Miechowski, W. (1965). Laboratory methods for evaluating skin irritating, toxical. Appl. Pharmacol, 7, 74-78.

Justice JD, Travers JJ, Vinson LJ (1961). The correlation between animal tests and human tests in assessing product mildness. Proc Sci Sect Toilet Goods Assoc, 35, 12–17

Bartek, M. J., Labudde, J. A., & Maibach, H. I. (1972). Skin Permeability In Vivo: Comparison in Rat, Rabbit, Pig and Man. Journal of Investigative Dermatology, 58(3), 114–123.

Ritz HL, Buehler EV. (1980). Planning, conduct and interpretation of guinea pig sensitization patch tests. In: Drill VA, Lazar P, eds. Current concepts in cutaneous toxicity. New York: Academic Press, pp. 25- 40.

Buehler EV. (1964). A new method for detecting potential sensitizers using the guinea pig. Toxicol Appl Pharmacol, 6, 341.

Buehler EV. Protocol. (1994) Delayed contact hypersensitivity study in guinea pigs (Buehler technique). Cincinnati, OH: Hill Top Biolabs.

Vùlund A. (1993). Statistical appendix. In: Andersen KE. Potency evaluation of contact allergens. Nordiske Seminar of Arbejdsrapporter. Copenhagen. Nordic Council of Ministers, Tema Nord, 570, 63-107.

Addison, J. (1995). Vermiculite: A Review of the Mineralogy and Health Effects of Vermiculite Exploitation. Regulatory Toxicology and Pharmacology, 21(3), 397–405.

Amandus, H. E., Wheeler, R., Jankovic, J., & Tucker, J. (1987). The morbidity and mortality of vermiculite miners and millers exposed to tremolite-actinolite: Part I. Exposure estimates. American Journal of Industrial Medicine, 11(1), 1–14.

How to Cite
Dr. Muhi. N. Salmam. (2020). Skin Sensitization Assay of Topical Occluded Heat Patches by Modified Buehler Method Using Rabbits. International Journal for Research in Applied Sciences and Biotechnology, 7(5), 1-6.