The bacterial endotoxin test (BET) is a crucial assay used to detect the presence of endotoxins, which are toxic components found in the outer membrane of Gram-negative bacteria.
Endotoxins can trigger severe immune responses in humans and animals, making it essential to monitor their levels in pharmaceuticals, medical devices, and other products that come into contact with the human body.
๐๐ซ๐ข๐ง๐๐ข๐ฉ๐ฅ๐
Bacterial endotoxins are components of the outer membrane of Gram-negative bacteria. When endotoxins come in contact with Limulus Amebocyte Lysate (LAL) reagent, they activate a clotting reaction. The amount of clot formation or color/turbidity change is proportional to the amount of endotoxin present in the sample.
๐๐๐ญ๐ก๐จ๐๐ฌ ๐จ๐ ๐๐๐:
- ๐๐๐ฅ-๐๐ฅ๐จ๐ญ ๐ฆ๐๐ญ๐ก๐จ๐:
*A qualitative test based on the formation of a gel clot.
*If endotoxins are present above the specified limit, a firm gel clot is formed. - ๐๐ฎ๐ซ๐๐ข๐๐ข๐ฆ๐๐ญ๐ซ๐ข๐ ๐ฆ๐๐ญ๐ก๐จ๐:
*Measures the increase in turbidity caused by the endotoxin-LAL reaction.
*The endotoxin level is calculated based on the change in turbidity. - ๐๐ก๐ซ๐จ๐ฆ๐จ๐ ๐๐ง๐ข๐ ๐ฆ๐๐ญ๐ก๐จ๐:
*Measures the yellow color produced during the reaction.
*The intensity of color is proportional to the endotoxin concentration.
๐๐ซ๐จ๐๐๐๐ฎ๐ซ๐ (๐ ๐๐ง๐๐ซ๐๐ฅ):
*Prepare the sample and required dilutions.
*Add LAL reagent to the sample.
*Incubate at the specified temperature (commonly 37 ยฑ 1ยฐC).
*Observe the reaction and calculate endotoxin levels.
*Compare results with the Endotoxin Limit (EL).
The endotoxin level must be within the limit specified by pharmacopeias such as USP, EP, or IP.
