Limulus amoebocyte lysate (LAL) test - an alternative method for detection of bacterial endotoxins

Abstract

The Limulus amebocyte lysate (LAL) test is an alternative method to the rabbit pyrogen test focussed on detection of pyrogenic substaces in sterile parenteral drugs. The aim of this work is the evaluation and introduction to common day use of LAL test gel-clot method for assay of bacterial endotoxins (the most common pyrogens) in examined product. A total number of 15 samples were tested for bacterial endotoxins to verify the method in our laboratory conditions. In 6 products, the presence of pyrogens was examined using simultaneously the LAL test and the rabbit pyrogen test. The replacement of the rabbit pyrogen test by the LAL test gel-clot method is possible when the endotoxin limit for the observed drug product is defined, the set maximal endotoxin concentration level in such material is acceptable and standardised test procedures and validation techniques are established. There are many advantages of LAL test over the rabbit pyrogen test, however, one of the most important aspects of LAL test is that LAL test is in accordance with the latest demand of the European Pharmacopoeia Commission for the replacement of the animal-based tests in favour of alternative methods where possible. The tests carried out have proved that the LAL test could replace the rabbit pyrogen test on condition that the validation parameters are fulfilled.