Veolia Water Technologies & Solutions

Bacterial Endotoxins Testing (BET)

We know you want simpler, faster endotoxin testing so you can release products sooner while still ensuring safety and compliance. In order to do that, you need technology that makes BET assay setup easy, automated, and less prone to errors – and the technology has to be easy to validate and maintain. You shouldn’t have to dedicate so much analyst time to manual pipetting, training, and re-tests, when microfluidic automation can do the work for you – eliminating hundreds of pipetting steps and hours of unnecessary time, while reducing opportunities for error along the way.  

Bacterial Endotoxins Testing (BET)

We know you want simpler, faster endotoxin testing so you can release products sooner while still ensuring safety and compliance. In order to do that, you need technology that makes BET assay setup easy, automated, and less prone to errors – and the technology has to be easy to validate and maintain. You shouldn’t have to dedicate so much analyst time to manual pipetting, training, and re-tests, when microfluidic automation can do the work for you – eliminating hundreds of pipetting steps and hours of unnecessary time, while reducing opportunities for error along the way.  

Make endotoxin testing easier, compliant, and more sustainable

We believe in making complex measurements simple.  

For years, endotoxin testing has required well-trained technicians to carefully prepare samples and standards for gel clot and 96-well plate-based assays. These analytical approaches are slow to prepare, prone to error and retests, and don’t meet the latest data integrity guidelines. And while these methods benefit from the sensitivity and specificity of Limulus amebocyte lysate (LAL) for the detection of endotoxins, there is a desire to decrease the use of LAL reagent. This is because LAL is created from the blood of horseshoe crabs, and we all want to conserve and optimize the use of this resource as much as possible.  

Now, there’s an easier way to achieve compliant endotoxin testing and improve sustainability with a new compendial endotoxin assay using microfluidics. It simplifies test setup, decreases retest rates, fully complies with the latest data integrity guidelines, and reduces LAL usage by up to 90%.  

Portfolio

Despite the need for easier, automated, and more sustainable solutions for endotoxin testing, many platforms fall short of meeting key requirements or streamlining installation and validation for easy adoption. With the Sievers Eclipse, you get the benefits of significantly easier BET assay setup and full compliance, without the complexity of robotics or difficult implementation. 

The Sievers Eclipse BET Platform and enterprise software provide: 

  • 9-minute setup for 21-sample assay (time-savings of up to 85%)  
  • Reduction in LAL reagent by up to 90% 
  • Compliance with all requirements of the harmonized pharmacopeia: USP <85>, EP 2.6.14, and JP 4.01, as well as ChP 1143
  • Highly customizable software with full compliance with 21 CFR Part 11 and ALCOA+ 

Looking for support along the way? Here's how you'll be supported throughout implementation: 

  • Get validated fast - IQ/OQ/PQ Validation Support Package gets your system validated and installed in days
  • Receive comprehensive documentation - Full platform validation report delivered to your team
  • Streamline product validation - Specific software protocol simplifies your validation process
  • Switch methods seamlessly - Easily transition from Kinetic Turbidimetric → Kinetic Chromogenic
  • Bridge platforms smoothly - Structured bridge study protocol helps you move from your current kinetic chromogenic platform to Eclipse
  • Compare with confidence - Side-by-side comparison studies between Eclipse and the 96-well plate platform

Products

Endotoxin Testing Products and Services

Testing Requirements Sievers Eclipse BET Platform 
Requirements of USP <85>, EP 2.6.14, JP 4.01, and ChP 1143  ✔️
Samples and PPCs in at least duplicate  ✔️
Minimum 3-point standard curve in at least duplicate using standardized endotoxin (RSE/CSE)  ✔️
Negative controls in at least duplicate  ✔️
Analyst and lysate lot qualification in at least triplicate  ✔️
Use of FDA licensed LAL  ✔️
Compliance with 21 CFR Part 11 and data integrity guidelines  ✔️

Resources

Frequently Asked Questions for Bacterial Endotoxins Testing

Why is endotoxin testing required for pharmaceuticals?

Endotoxin testing ensures product safety by detecting harmful bacterial components that can cause severe immune reactions. This quality control procedure is mandatory across pharmaceutical, medical device, and biotech industries for products contacting human blood or body fluids - including injectable drugs (parenteral drugs) like vaccines and insulin, as well as medical devices such as catheters, implants, and dialysis equipment. Water for injection (WFI) and other pharmaceutical water systems also require regular endotoxin monitoring as they're critical raw materials in drug manufacturing. Even trace amounts of endotoxins can be dangerous, making testing essential for patient safety and regulatory compliance.

What is the LAL bacterial endotoxin test procedure?

The LAL (Limulus amoebocyte lysate) test has been FDA-approved for 40 years as an in vitro alternative to rabbit pyrogen testing. Using horseshoe crab blood lysate, it detects bacterial endotoxins through three methods: gel-clot (qualitative), turbidimetric, and chromogenic (both quantitative). Results are measured via gel formation, turbidity, or color change against standard curves. Testing requirements are outlined in USP <85>, EP 2.6.14, JP 4.01, and ChP 1143, with new recombinant methods (using rFC or rCR) now available.

What is the US Pharmacopoeia guidance on recombinant BET methods?

USP Chapter <86> “Bacterial Endotoxins Test Using Recombinant Reagents”  became official in May 2025. USP <86> provides additional techniques for endotoxin testing using non-animal derived reagents, specifically the recombinant Factor C (rFC) protein or a recombinant cascade reagent (rCR). These methods offer additional techniques to the current bacterial endotoxins tests described in USP <85>. 

USP Chapter <86> describes how users can use traditional LAL or recombinant reagents for endotoxin testing by leveraging the recombinant reagent’s Primary Validation Package to demonstrate the method is suitable for its intended use. With the publication of this chapter and the use of the Sievers Eclipse, users can achieve results in significantly less time than with traditional methods and with a greater emphasis on sustainability. The plate set up takes approximately 5 minutes using the Sievers Eclipse, including the standard curve, and uses up to 90% less reagent.

What are barriers to the use of recombinant bacterial endotoxins test methods, and how can any hurdles be streamlined for labs?

Prior to adopting recombinant reagents such as rFC or rCR, the user should first evaluate the reagent manufacturer’s Primary Validation Package with their Quality team to determine whether there are any gaps in testing that need to be remedied prior to adoption. Similar to adopting any new method, it is a normal requirement to demonstrate that the method is fit for use. The use of recombinant reagents is still considered to be an alternative method, so users must prove that the reagent is suitable for its intended purpose under the conditions of use for the material, drug substance, and/or drug product. If there are gaps with the Primary Validation testing of the reagent, further testing may be required. Additional testing can be done with autochthonous organisms that are commonly present in the manufacturer’s facility, as well as purified endotoxin standards. 

A side-by-side protocol or a comparability study comparing current methods to rCR reagents will provide the necessary data to demonstrate comparability. While transitioning existing products to rCR may seem challenging, analysts only need to complete alternative method testing followed by a single-lot verification to confirm the rCR reagent performs similarly to existing methods like chromogenic assays.

A microfluidic platform like the Eclipse streamlines method validation through rapid assay setup. With integrated liquid handling plus embedded endotoxin standards and PPCs, assays can be configured in minutes using LAL or rCR.

What are endotoxins and why are they dangerous?
Endotoxins are lipopolysaccharide components from gram-negative bacterial cell walls, specifically the Lipid A portion that triggers fever and immune responses. These potent toxins become life-threatening when present in bloodstream-contacting products above safe concentrations. They can contaminate pharmaceutical water systems, raw materials, and finished products throughout the manufacturing process. Water systems are particularly susceptible to endotoxin contamination as bacteria can form biofilms in pipes and storage tanks. Global regulators including the FDA mandate endotoxin testing for all life sciences products due to their severe impact on patient safety. Compendial test methods for pharmaceutical waters include requirements for bacterial endotoxin limits in waters for parenteral (injectable) use, such as Water for Injection and Sterile Water for Injection.