How do you detect pyrogen in the laboratory? Currently, the only available replacement for the RPT and for the assessment of non-endotoxin pyrogens is the Monocyte Activation Test (MAT). This in vitro test method can detect all pyrogens that activate the Toll-Like Receptor (TLRs) pathway, including endotoxins and non-endotoxin pyrogens (NEPs).
Why pyrogen test is performed?
Pyrogen test is performed to check the presence or absence of pyrogens in all aqueous parenterals. Rabbits are used to perform the test because their body temperature increases when pyrogen is introduced by the parenteral route. For this test, three healthy rabbits are selected each weighing at least 1.5 kg.
How do you test for pyrogens?
Two widely used pyrogen test methods are the rabbit pyrogen test (RPT) and the bacterial endotoxin test (BET) based on Limulus Amebocyte Lysate (LAL). Both tests have their limitations. The RPT can miss human-specific pyrogens, does not allow for quantification and requires the use of animals.
What is the principle of pyrogen test?
The pyrogen test on rabbits is based on the measurement of the increase in the rabbit's temperature upon being injected with a product that could contain a contaminant of the pyrogen type. The pyrogens, as their name suggests, refer to all the substances that cause an increase in fever, also known as pyrexia.
Why is Lal important?
The LAL (limulus amebocyte lysate) testing, also known as bacterial endotoxin testing, is an in vitro assay used to detect the presence and concentration of bacterial endotoxins in drugs and biological products, and is an important part of pharmaceutical microbiology.
Related guide for How Do You Detect Pyrogen In The Laboratory?
What is detected in pyrogen testing of pharma product?
Learn how to determine the Pyrogen in injection samples and water for injection using animals. The test involves measurement of the rise in body temperature of rabbits following the intravenous injection of a sterile solution of the substance under examination.
What is the effect of pyrogens on the body?
When bacterial pyrogens are injected in sufficient amounts, perhaps in microgram quantities, the fever produced is accompanied by chills, body aches, a rise in blood pressure, and possibly a state of shock and death.
How pyrogen contributes to hyperthermia in the human body?
Pyrogenic hyperthermia is a result of the hypothalamus being reset to a higher point than normal. Nonpyrogenic hyperthermia is caused by external or environmental factors, and the hypothalamus “set point” remains normal but the ability to regulate body temperature is lost.
What are some examples of pyrogens?
Good examples of pyrogen-producing gram-negative bacteria are Escherichia coli (E. Coli), Proteus, Pseudomonas, Enterobacter, and Klebsiella (Dixon, 2001).
Why are rabbits selected for pyrogen test?
If a pyrogen is present in the test substance, it will cause an elevation of the animal's body temperature from normal. Rabbits have similar temperature response to endotoxins as humans. For this reason, they are ideal for use in pyrogen detection.
Are pyrogens cytokines?
Pyrogenic cytokines review
The discovery that several proinflammatory cytokines act as endogenous pyrogens and that other cytokines can act as antipyretic agents provided a link between the immune and the central nervous systems and stimulated the study of the central actions of cytokines.
Why do we test for bacterial endotoxin?
All injectable pharmaceutical products and implantable medical devices need to be tested to ensure there is no presence of endotoxin, which can lead to a pyrogenic response (fever) and symptoms of septic shock. Endotoxins can be detected in these products and devices through bacterial endotoxin testing (BET).
How is LAL test performed?
Test procedure: A BET involves analyzing the liquid sample or sample extract using Limulus Amebocyte Lysate (LAL). LAL is a reagent made from the blood of the horseshoe crab. In the presence of bacterial endotoxins, the lysate reacts to form a clot or cause a color change depending on the technique.
How is pyrogen removed from water?
Ultrafiltration (UF) is an excellent way of removing pyrogen contamination from water. Ultrafilters (positively charged nylon 66 membranes) are recommended for the final “polishing” of water already treated by deionization (DI) or reverse osmosis RO.
Why LAL test is preferred?
LAL test is recommended in all international pharmacopeias as the method for finding bacterial endotoxins. Gram-negative bacteria produce endotoxins (pyrogen). Exceptionally Bacillus thuringiensis , a Gram-positive bacteria produce delta toxin as endotoxins  (Figures 1 and 2).