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Sievers Eclipse Bacterial Endotoxins Testing (BET) Platform

Breakthrough BET technology that delivers automation and compliance

The innovative Sievers Eclipse automated endotoxin detection 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 pharmacopeia: USP <85>, EP 2.6.14 and JP 4.01. Through groundbreaking?technology, the Eclipse endotoxin testing solution significantly decreases pipetting steps, reduces operator-to-operator variability, and simplifies BET test 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 LAL test.

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Compliant, consistent, and conscious bacterial endotoxin testing equipment and services

At Veolia, 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. 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 reagents. 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%.

The Sievers Eclipse Endotoxin Testing System

Features & Benefits

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
  • Highly customizable software with full compliance with 21 CFR Part 11 and ALCOA+

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Testing Requirements Sievers Eclipse BET Platform?
Requirements of USP <85>, EP 2.6.14, and JP 4.01? ??
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? ??

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Looking for support along the way? Here is how the Sievers Eclipse team supports the implementation process:

  • IQ/OQ/PQ Validation Support Package (VSP) to comprehensively validate the system, with Installation and validation completed in days
  • Our team performs a full platform validation report
  • Our Specific Software Protocol assists in Product Validation
  • We help you switch methods (ex: Kinetic Turbidimetric → Kinetic Chromogenic)
  • We offer a bridge study protocol to assist in bridging from your current kinetic chromogenic platform to the Eclipse platform
  • Our comparability protocol executes a comparison study between the Sievers automated platform and the 96-well plate platform

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System Specifications

System Specifications


BET Method

Kinetic chromogenic

Detection Mode

Absorbance

Range

0.005–50 EU/mL

Precision

≤ 15% CV onset time

Accuracy

50–200% of actual

Sample Type

Aqueous, injected by pipette

Calibration

Up to 12 months

Analysis Time

Up to 2 hours

Sample Temperature

37 ± 1 °C

Ambient Temperature

17–30 °C

Capacity

Up to 21 samples in duplicate with positive product controls

Temperature Control

37 ± 0.5 °C

Light Source

LED emitter

Fluidic Failure Detection

1450 nm emitter

Optical Accuracy

≤ 5% deviation from expected value

Optical Linearity

R-Value ≥ 0.980

Optical Wavelength Filter

405 nm

Read Interval

5 Seconds

Analyzer Specifications


Outputs

Digital via USB

Display

OLED

Power Requirement

100–240 Volts AC @50/60 Hz

Fuses

T 8 A 250 VAC Fuse, size 5 x 20 mm. ONLY Littelfuse 218008 or Cooper Bussmann S506-8-R.

Dimensions

H: 17.5 cm (6.9 in); W: 35.1 cm (13.8 in); D: 50.3 cm (19.8 in)

Weight

10 kg (22.1 lbs)

Safety Certifications

UL 61010-1:2012Ed.3+R:20Apr2016 ? ? ? ? ? ? CSA C22.2#61010-2-020

CSA C22.2#61010-1-12:2012Ed.3+U1;U2? ? IEC 61010-1:2010 Ed.3+C1;C2

UL 61010-2-010:2015Ed.3 ? ? ? ? ? ? ? ? ? ? ? ? ? ? IEC 61010-2-010:2003Ed.2

UL 61010-2-020:2016Ed.3 ? ? ? ? ? ? ? ? ? ? ? ? ? ? IEC 61010-2-020:2016Ed.3

CSA C22.2#61010-2-010:2015Ed.3


Maximum Relative Humidity

85% non-condensing

Maximum Altitude

3,000 m (9,800 ft)

Pollution Degree
2

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Options & Accessories

  • Analyzer: Absorbance analyzer with consistent incubation control at 37 °C, centrifugal technology, and secure data transmission.

  • Microplate: Precise liquid handling device that achieves automation through innovative microfluidic technology with embedded endotoxin standards and PPCs.

  • Software: Highly customizable enterprise solution with 21 CFR Part 11 and ALCOA+ Data Integrity compliance features.

The Eclipse microplate, in conjunction with the Eclipse analyzer and software, uses centrifugal force and pneumatic chambers to measure and evenly distribute precise amounts of LAL reagent water, samples, and LAL across 104 optical wells. The optical wells comprise segments for standards curves, samples, and positive product controls (PPCs). The reaction is controlled at 37 +/-1 C, and a high definition view of each well is obtained through a reading at 405 nm every five seconds.

Resources

Frequently Asked Questions (FAQs) for Bacterial Endotoxins Testing?

What is bacterial endotoxin testing?

Bacterial endotoxin testing is a critical quality control procedure used in the pharmaceutical, medical device, and biotechnology industries to detect and quantify the presence of endotoxins in various products. Endotoxins are components of the outer cell membrane of gram-negative bacteria and can cause severe immune responses in humans and animals, even in small amounts.

What are bacterial endotoxins?

Bacterial endotoxins are found in the outer membrane of the cell wall of Gram-negative bacteria. The endotoxin 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 endotoxins in water or final products?

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 does the bacterial endotoxin test method work?

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 bacterial endotoxin test principle 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 pharmacopeia, 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.

How much LAL reagent is used with the Sievers Eclipse BET Platform?

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 of this valuable natural?resource and delivers a fully compliant BET assay that the global HSC population can sustain.

Is the Sievers Eclipse BET Platform compliant?

Yes. The Eclipse platform uses commercially 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.

Are standard curves automated with the Sievers Eclipse?

Yes.?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.??

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