GLP Research Unit – Pyrogenic – Rabbit pyrogen test according to European Pharmacopoeia

The pyrogen test is an in vivo test using New Zealand albino rabbits. The test methodology presented in the European Pharmacopoeia, 6th edition, chapter 2.6.8, specifies the method of conducting the pyrogen test of test materials, including those meeting the definition of medical devices.

Pyrogenicity is the ability of a chemical or other substance to cause a febrile reaction. The pyrogenic response can be caused by endotoxins from Gram-negative bacteria (endotoxin-mediated pyrogenicity) and by factors unrelated to endotoxins (material-mediated pyrogenicity).

The in vivo pyrogen test involves analyzing the change in rabbit body temperature caused by intravenous injection of a solution of the tested substance (picture 1). The temperature is measured using an electronic thermometer with a resolution of 0.1°C.

The procedure for testing the pyrogenic effect according to the European Pharmacopoeia is carried out in an animal house for small animals. All animals submitted for testing are quarantined and acclimatized in accordance with applicable legal regulations and internal procedures.

The in vitro pyrogen test is used for the biological assessment of medical devices using albino New Zealand rabbits (Oryctolagus cuniculus) in a small animal facility. The test is a two-stage one, consisting of a preliminary test during which a substance that does not cause a pyrogenic effect (raising body temperature) is administered to rabbits. The preliminary test is intended to eliminate individuals giving false positive results in the test.

The pyrogenic effect of the tested material on animals is determined on the basis of the difference in temperatures measured before and after administration of the tested material to the animal during the main test. Each rabbit’s response is the difference between the maximum temperature obtained from measurements carried out for 3 hours after administration of the tested material every 30 minutes, and the initial temperature. When this difference is negative, the result is counted as a zero response.

The biological assessment of a medical device in terms of in vivo pyrogenicity is based on temperature values depending on the number of rabbits used in the test. The European Pharmacopoeia provides permissible values for product acceptance and values, exceeding which result in non-approval of the tested product.

Please contact us directly to select the appropriate test method for your medical device.

Our laboratory has a Good Laboratory Practice certificate in the biocompatibility section (scope 9) for the biological assessment of medical devices for in vitro and in vivo tests.