LAL Validation assay
LAL Validation Assay
Non-Pyrogenic Validation is a requirement of the FDA in order to advertise a product as being Pyrogen Free or Non-Pyrogenic. This test consists of testing in quadruplicate three separate lots of the same product. Product extracts are made using the three separate lots. The product extracts are used to prepare three sets of standard series of the Control Standard Endotoxin (CSE). Each dilution of the standard series is run in quadruplicate. The product extract series are incubated along with a standard series of the CSE. After the incubation period, the tubes containing the controls and the extract are observed for the presence of the gel clot.
If the unspiked product samples are all negative and the product series Endotoxin concentration is between 0.03 Eu/ml and 0.12 Eu/ml for all three lots then the product is not introducing any inhibitory or enhancing effects. The LAL validation is a one time procedure and does not need to be repeated unless there are changes in manufacturing procedures, raw materials or product formulations.
Once it is determined that the product does not have any inhibitory or enhancing effects the product is validated. After product validation, each lot that will be advertised as Non-Pyrogenic must be routinely tested in accordance with FDA guidelines.