Sievers Eclipse Bacterial Endotoxins Testing (BET) Platform

Bacterial endotoxins are found in the outer membrane of the cell wall of Gram-negative bacteria. The endotoxic effect from Gram-negative bacteria is primarily due to the Lipid A component of lipopolysaccharides (LPS), which can exert pyrogenic (fever-inducing) responses that can be dangerous and even fatal when found in the bloodstream and other body fluids above certain concentrations. Because certain pharmaceuticals and medical devices enter the bloodstream, endotoxin testing is required in those industries. Due to its potent toxicity and effect on patient safety, endotoxin is globally regulated by the US FDA and international pharmacopoeia for any life sciences product that will come in contact with human and/or animal blood.

Within the pharmaceutical, medical device, and other life science industries, testing for endotoxins is critical for patient safety and quality control. Endotoxin tests are performed on pharmaceutical grade water systems, including water for injection (WFI) that is used as ingredient water, and finished products such as drug, animal drug, biological, and medical device products. Pharmaceutical and medical products tested include parenteral drug products (administered through injection, such as intravenous, subcutaneous, intramuscular, and intradermal) and medical devices that have direct or indirect contact with the blood, cardiovascular system, lymphatic system, or cerebrospinal fluid. Global pharmacopoeia (USP <85>, EP 2.6.14 and JP 4.01) describe requirements for Bacterial Endotoxins Testing (BET) and endotoxin limits. FDA guidance provides recommendations for biological product, drug, and device manufacturers on current thinking concerning testing recommendations and acceptance criteria covered in compendial procedures.

For almost 40 years, the US FDA has accepted the Limulus amoebocyte lysate (LAL) test as a finished product test for endotoxins. This is in lieu of rabbit pyrogen testing and is an in vitro assay that can detect presence and concentration of bacterial endotoxins. In LAL tests, the lysate from the blood of horseshoe crabs (Limulus Polyphemus) reacts with bacterial endotoxins to indicate the presence of endotoxins in a sample. There are three principal methodologies for LAL testing: gel-clot, turbidimetric, and chromogenic. In the gel-clot method, LAL and sample are mixed, followed by the operator screening for gel formation in the reaction tube. The results are from a subjective interpretation of the clot formation, and thus the gel-clot method is a qualitative test. In turbidimetric and chromogenic methodologies, samples are mixed with LAL reagent, and the color change or turbidity are measured over time. Chromogenic and turbidimetric LAL assays are quantitative and demonstrate the amount of endotoxin present. Results are calculated from a standard curve. Endotoxin limits and requirements for conducting BET are described in global pharmacopoeia, USP <85>, EP 2.6.14 and JP 4.01. A new general chapter in the European Pharmacopoeia describes a method for BET using recombinant factor C (rFC) instead of the classic LAL-based method.

With just 1 mL LAL reagent, 21 samples can be run on the Eclipse platform. By decreasing horseshoe crab (HSC) lysate use by up to 90%, the Eclipse reduces the demand on this valuable natural resource and delivers a fully compliant BET assay that the global HSC population can sustain.

是的。Eclipse平台使用商业available, FDA licensed LAL and meets all requirements of the harmonized global pharmacopoeia, USP <85>, EP 2.6.14 and JP 4.01. Bacterial endotoxin testing with the Eclipse platform includes: minimum three-point standard curve in duplicate using standardized endotoxin; samples and PPCs in duplicate; negative controls in duplicate; analyst and lysate lot qualification in triplicate; use of FDA licensed LAL. It also complies with 21 CFR Part 11 and data integrity guidelines.

是的。Traditionally, for compliant endotoxin assays, the end user must construct at least a three-point standard curve in duplicate from a stock vial of standardized endotoxin; must have duplicate negative controls; and must run each sample in duplicate with a PPC, also in duplicate. With the Sievers Eclipse BET Platform, these steps are automated using preloaded endotoxin standards spanning up to a five-point standard curve and preloaded PPCs. Therefore, all the end user must do is load Water for BET and samples onto the plate with no additional prep work. The result is the ability to set up the assay in 9 minutes, compared to upwards of 60 minutes that other platforms require.