[A new test for the detection of pyrogens in pharmaceutical products] [Article in German]

Abstract

The human whole blood assay utilises the natural fever response to detect pyrogens by determination of the release of IL-1b.
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-1b.
2) Stimulation by LPS induces a concentration-dependent release of IL-1b beginning at a concentration of between 2-5 pg/mL LPS.
3) The amount of IL-1b released varies greatly between samples obtained from different individuals.
4) Storing blood samples results in a right shifted LPS/IL-1b curve with a steeper gradient and higher maximum value of IL-1b.
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.