[A new test for the detection of pyrogens in pharmaceutical products. Examinations for the validation of the human whole blood assay]
The human whole blood assay utilises the natural fever response to detect pyrogens by determination of the release of IL-1beta. In order to replace the official method, the rabbit pyrogen test, a validation of the whole blood assay is necessary. A comparison of the results obtained from many blood samples has revealed the following: 1) Blood not stimulated by LPS does not produce IL-1beta. 2) Stimulation by LPS induces a concentration-dependent release of IL-1( beginning at a concentration of between 2-5 pg/mL LPS. 3) The amount of IL-1beta released varies greatly between samples obtained from different individuals. 4) Storing blood samples results in a right shifted LPS/IL-1beta curve with a steeper gradient and higher maximum value of IL-1beta. In this paper we suggest an experimental method for the determination of pyrogens based on the established semi-quantitative LAL gelation method as detailed in the European Pharmacopeia. Using this methodology, we were able to show that the amount of endotoxin in a number of different infusion solutions was below the LAL-endotoxin limit concentration. LPS was quantitatively determined from spiked samples.
Weigandt
FIRST_NAME
F
Hygiene-Institut der Universität D-Heidelberg.
Lexa
Peter
P
Sonntag
Hans-Günther
HG