4/5 April 2023
In the near past, the topic of pyrogen testing has been covered in the newsletter several times. In 2020 we discussed which test to use and in 2021 we summarized the developments from the classical rabbit test to alternative methods in a review. In 2022 we informed about the revision of "PharmEuropa - Revision of Chapter 2.6.30, Monocyte Activation Test". Particularly through the initiatives to reduce or replace animal testing, the issue has been driven forward in Europe and worldwide.
But especially the activities related to the European Pharmacopoeia are progressing. For example, the EDQM has recently published Chapter 5.1.13 Pyrogenicity for comment. It can be found in Pharmeuropa 35.1 under the texts for public comment, with a deadline of 31 March 2023.
The new chapter is intended to provide information on the selection and performance of appropriate testing for pyrogenicity, e.g., the bacterial endotoxin test or the Monocyte Activation Test (MAT). This draws on the content of Chapter 5.1.10. "Guidance on the use of the text for bacterial endotoxins", and in particular on the considerations for risk assessment and method performance. The goal of the new chapter 5.1.13. is to bring together the information from several chapters to support the intent of the Ph.Eu. Commission to abolish the rabbit test. The EDQM points out at this point: "It should be noted that the intention will eventually lead to the abolition of the general chapter 2.6.8. Manufacturers still using the rabbit pyrogen test are urged to take the necessary steps to replace it with an appropriate in vitro alternative (e.g., the monocyte activation test)."
It is not a goal to question already existing pyrogen controls with the test for bacterial endotoxins or to change them for approved products. It is also not doubted that the widely used tests for bacterial endotoxins described in the general chapters 2.6.14 and 2.6.32 are suitable in principle. This only applies, though, if the presence of pyrogenic non-endotoxins can be excluded.
In cases where this cannot be assumed to be certain, to exclude the presence of non-endotoxin pyrogens in substances or products, it is recommended that the monocyte activation test (2.6.30) is performed during the development of the manufacturing process. As in many cases in the field of pharmaceutical quality assurance and quality control, risk assessment plays an important role in the selection of an appropriate procedure. The chapter says the following under the heading "Risk Assessment in Selecting the Test for Bacterial Endotoxins as the Sole Method for Assessing Pyrogenicity":
"The decision to use the test for bacterial endotoxins as the sole pyrogenicity test should be made after careful evaluation of the risk that the substance or product contains non-endotoxin pyrogens. The risk assessment should be repeated if important process parameters have changed, since possible contamination by non-endotoxin pyrogens cannot be excluded."
The risk assessment should consider all factors that could lead to the ingestion of pyrogens not detected by the bacterial endotoxin test. The following is a non-exhaustive list of factors to be considered in the risk assessment." More detailed information can be found on the Pharmeuropa website. Registration is required for access.
In this context, please note the Monocyte Activation Test Laboratory Course of the ECA Academy on 09/10 March in Bernried at Lake Starnberg in Germany.