By Laure Robert, Merck KGaA, Darmstadt Germany, Life Science
Product manager for Pyrogen Testing solutions
A pyrogen is a substance that produces a rise in temperature in a human or animal. Pyrogens constitute a heterogeneous group of contaminants, the most widely known pyrogen being endotoxin (LPS = Lipo-Polysaccharide) from Gram-negative bacteria. Other microbial substances include those derived from Gram-positive bacteria like Lipoteichoic Acid (LTA), particles from viruses and pyrogens originating from yeasts and fungi. Nonmicrobial pyrogenic substances include rubber particles, microscopic plastic particles or metal compounds in elastomers.
In pharmaceutical parenteral products, these pyrogenic substances can induce life-threatening fever reactions after injection into the human body. It is therefore a regulatory requirement to test such products for pyrogens to ensure product quality. The pyrogen test intends to check that the quantity of pyrogens contained in the product does not exceed a certain threshold, known as the contaminant limit concentration (CLC), that will guarantee patient safety.
The publication of "The Principles of Humane Experimental Technique" by W.M.S. Russel and R.L. Burch in 1959 marked the birth of the principle of the "Three Rs" which provides a framework for more humane animal research by Replacement, Reduction and Refinement of the use of animals in the lab.
The Monocyte Activation Test (MAT) is the human in vitro alternative to the rabbit pyrogen test and allows the detection of the full range of pyrogens, including endotoxins and non-endotoxin pyrogens (NEPs).
The use of MAT instead of the rabbit pyrogen test is therefore an interesting alternative to limit the use of animal testing from an ethical and regulatory perspective. Moreover, the MAT is more sensitive and robust than the rabbit pyrogen test, providing reproducible results for pyrogen testing.
The MAT has been qualified and validated for the detection of pyrogens by the European Center for the Validation of Alternative Methods (ECVAM) in 2005 leading to its implementation in the European Pharmacopeia as a compendial method for Pyrogen Testing (Chapter 2.6.30).
Even though the MAT has not yet been implemented in the USP, it is mentioned in the new version of USP <151> (Pyrogen test) as an alternative to in vivo Pyrogen test.
If you want to make the move and stop animal-based test for Pyrogen Detection, you can learn more about MAT by reading the Monocyte Activation Test white paper. Visit sigmaaldrich.com/pyromat to find out about available MAT solutions.