What is the difference between CSE and RSE?

CSE is used as an economic alternative to the RSE. CSEs are standardized against the RSE as indicated on the Certificate of Analysis, so that results can be reported in Endotoxin Units (EU) and International Units (IU). CSE can be used for all LAL testing.

What is endotoxin spiking?

Endotoxins are cell wall components of Gram-negative bacteria. A release of endotoxins into the human blood stream results in an inflammation reaction that can lead to life-threatening conditions like sepsis. Within these studies, endotoxin is added (spiked) to an undiluted product.

What is reference standard endotoxin?

USP Endotoxin RS (RSE) is the primary reference material used in the compendial BET tests <85>. The current RSE is lyophilized, and each vial has an assigned activity Page 4 value of 10,000 endotoxin units (EU).

What is low endotoxin?

Low endotoxin recovery (LER), also known as endotoxin masking, is a recently described but long-observed phenomenon. LER is the inability to recover known amounts of purified endotoxin from biological formulations.

What causes endotoxin?

The lipid A portion of LPS is the cause of the molecule’s endotoxin activity. While lipid A does not directly harm any tissue, the immune cells of humans and animals alike see it as an indicator for the presence of bacteria. Thus, these cells stimulate a response that is meant to fend off the unwelcome intruders.

What is the bacterial endotoxin test?

The bacterial endotoxins test (BET) is a test to detect or quantify endotoxins from Gram- negative bacteria using amoebocyte lysate from the horseshoe crab (Limulus polyphemus or. Tachypleus tridentatus).

What is endotoxin spike recovery?

Spike recovery: The spike is an aliquot of test sample together with a known amount of endotoxin. The spike serves as the check for interference (inhibition and enhancement). Inhibition and enhancement are conditions that adversely alter the recovery of endotoxin in a test sample.

How do I remove endotoxin?

There are different methods for endotoxin removal. These include depyrogenation,2 such as dry-heat processes applied to glassware, and rinsing,3 as might be applied to closures. These areas receive reasonable coverage within the pharmaceutical sector.

What is an example of endotoxin?

Endotoxin: Examples In bacteriology, this complex compound is also known as lipopolysaccharide and can be found on the outer membranes of bacteria like Escherichia coli, Salmonella shigella, Vibrio cholerae, and Haemophilus influenzae.

How is bacterial endotoxin calculated?

If the dose is 1 mg/kg/hr, the endotoxin limit is (5 EU/kg/hr) ÷ (1 mg/kg/hr) = 5 EU/mg. If the dose is 10 mg/kg/hr, the endotoxin limit is (5 EU/kg/hr) ÷ (10 mg/kg/hr) = 0.5 EU/mg. If the dose is 100mg/kg/hr, the endotoxin limit is (5 EU/kg/hr) ÷ (100 mg/kg/hr) = 0.05 EU/mg.

Can you remove endotoxin?

Ultrafiltration. Since endotoxin molecules tend to form micelles or vesicles in aqueous solution these can be removed from a solution by filtration.

How do I remove endotoxin from API?

The common methods for removal of endotoxin (LPS) from solutions are ultrafiltration, adsorptive matrices such as chromatographic resins, and ion-exchange resins (2–4).

Is there a naturally occurring endotoxin ( Noe )?

In response to stakeholder requests, USP (US Pharmacopeial Convention; www.usp.org) is proposing a new reference standard, Naturally Occurring Endotoxin (NOE), to be prepared from cell wall extracts of a well characterized Gram negative bacterium.

How does low endotoxin recovery ( ler ) work?

Low Endotoxin Recovery (LER) has been one of the most controversial recent topics in the BET for biopharmaceuticals. LER is a phenomenon of a loss of spiked endotoxin activity in a neat sample, and is observed with samples containing a chelating agent and a detergent [ 3 ].

Is the LPS preparation the real endotoxin?

While LPS preparations (Endotoxin RS) may be well suited for analytical calibration and in most cases as a spike in recovery studies, it is important to note that these LPS preparations bear minimal resemblance to the real world endotoxin contaminant that might be present during pharmaceutical manufacturing.

How is the expiration date of endotoxin determined?

If the endotoxin preparation is stored, storage parameters including the concentration of the preparation in Endotoxin Units (EU), the composition of the vessel, the temperature of storage, and the length of storage, should be defined. An expiration date should be assigned based on determined stability.