Sievers Eclipse Bacterial Endotoxins Testing (BET) Platform
Breakthrough technology that delivers automation and compliance
The innovative Sievers Eclipse platform decreases assay setup time by up to 85% and reduces Limulus Amebocyte Lysate (LAL) reagent use by up to 90% while meeting all requirements of the harmonized pharmacopoeia: USP <85>, EP 2.6.14 and JP 4.01. Through groundbreaking technology, the Eclipse platform significantly decreases pipetting steps, reduces operator-to-operator variability, and simplifies assay setup. The Eclipse platform leverages precise microfluidic liquid handling and embedded endotoxin to automate kinetic chromogenic assays. Throughput of 21 samples per plate is maintained without the complexity of robotics or the time and technique demands of a traditional assay.
Compliant, consistent, conscious bacterial endotoxins testing
The Eclipse platform achieves automation and precision in BET assays without sacrificing compliance, reaction biochemistry, or footprint in the lab. Using embedded endotoxin standards and positive product controls (PPCs) with only <30 pipetting steps, the patented Sievers Eclipse microplate delivers automation without the complexity or expense of robotics.
What is bacterial endotoxin?
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.
Why would you test for bacterial endotxins?
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.
How is endotoxin testing performed?
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.