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Endotoxins and their Relevance in R&D ApplicationsLPS Detection & Removal Methods

has been also given admittance by the FDA for the identification of patients with high sepsis risk.

Both, the FDA & US Pharmacopoeia regulate that medical equipment or drugs, which enter the circulating blood or the central nervous system, may not cause any pyrogenic reactions. Therefore everything declared as "pyrogenic free" has to be checked.

The advantage of the rabbit test (Ph. Eur. 2002, 2.6.8) is that all known and unknown pyrogenics can be detected. Therefore it is indispensable for the drug study till now. But it represents a timeconsuming animal experiment and not all substances that seem pyrogenic do cause fever in rabbits. In the LAL test (Ph. Eur. 2002, 2.6.14) a special property of the haemolymph of Limulus polyphernus is used: in the presence of LPS it starts to coagulate.

The following LAL test methods were developed meanwhile:
Gel Clot
Chromogenic End-Point Test
Kinetic Turbidimetric Test
Kinetic Chromogenic Test


The LAL test has the advantage that it is sensitive and can be better standardized than the rabbit test. Moreover, it allows a quantitative test evaluation. However it is very susceptible and it is also an animal experiment thats why it can be transferred only with restriction to the human system. A further disadvantage is that only unbound LPS molecules can be detected. Therefore it can be possible that up to 30-50 lower values10 are measured with the LAL test than with detection systems which consider only the chemical connection and not the molecule activity. Thus there are different new approaches for the LPS detection. In the indirect method (e.g. "monocyte activation cytokine assay") the
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