LAL Bacterial Endotoxin Test – USP <85>

Bacterial Endotoxins are pyrogens that are produced in bacteria which cause fever in humans and other animals. The most common pyrogen is Lipopolysaccharide (LPS) which is a component of the cell membrane of many gram negative organisms.

Limulus Amebocyte Lysate (LAL) is the most common reagent used to detect bacterial endotoxin contamination. The gold standard per USP for testing is the gel clot method.

The control and detection of endotoxins is a key component for quality release of parenteral drug formulations, medical devices, raw materials, excipients, water, water for injection, and active pharmaceutical ingredients (API). Endotoxin testing is required for many products, including medical devices with direct blood or cerebrospinal fluid (CSF) contact.

Testing should be performed on every production lot prior to release. Endotoxin Limits are typically established based on formulation, the dosage per hour, route of administration, and average patient weight, but may also incorporate trending data as well as monograph specifications.

Endotoxin method validation is performed on every product and/or formulation. Validation, or inhibition and enhancement (I/E) testing, ensures the sample does not interfere with the method, which could lead to false positive or negative data. Validation also ensures proper sensitivity, diluent selection, and dilution factor.

 

Guidance documents include:

“Guideline on the Validation of the Limulus Amebocyte Lysate Test for Human and Animal Parenteral Drugs, Biological Products and Medical Devices”, published December 1987 by FDA.

“Bacterial endotoxins – Test methodologies, routine monitoring, and alternatives to batch testing”, ST72 published 2002 by ANSI/AAMI.

USP <85> – Bacterial Endotoxins Test

USP <161> – Transfusion and Infusion assemblies and similar medical devices