background

Gram negative bacteria (GNB) have caused one of the most serious problems to global public health because they are highly resistant to antibiotics, and can cause pneumonia, blood infection, wound and surgical site infection, meningitis and other infectious diseases. 2. Bacterial endotoxins are a key component of GNB cell membrane, and when the concentration is high enough, they can trigger a thermogenic reaction (i.e. high fever). The US Food and Drug Administration (FDA) requires pharmaceutical companies to conduct Bacterial Endoxins Testing (BET), with a focus on measuring the degree of GNB contamination of all injectable drugs, including water for injection (WFI), pharmaceutical raw materials, finished drugs, medical devices, etc. There are three main methods to detect bacterial endotoxin: 1. gel Clot, 2. KTA, Kinetic Turbidimetric Assay, 3. KCA, Kinetic Chromogenic Assay.

Limulus Amebocyte Lysate (LAL) is extracted from the blood of horseshoe crabs and used for bacterial endotoxin detection. When endotoxins are detected, the horseshoe crab reagent cascade will undergo a series of enzymatic reactions, which will amplify the appearance of endotoxins. When endotoxins react with horseshoe crab reagent, endotoxins activate the first factor in the enzyme cascade - factor C, which then activates factor B. Factor B acts on the coagulation enzyme, making it a coagulation enzyme. The final detection steps are coagulation (using gel method), turbidity (using dynamic turbidity method) or color development (using dynamic color method).

Sievers Eclipse bacterial endotoxin detector

The Eclipse detector consists of three parts: instrument, microfluidic microplate, and software. The instrument is an absorbance analyzer with 37 ° C constant temperature cultivation control and centrifugation technology. The microplate is pre-set with Reference Standard Endoxin (RSE) and Positive Product Controls (PPC) for bacterial endotoxins, allowing users to run up to 5 standard curves (concentrations ranging from 50-0.005 EU/mL), as well as up to 21 samples and negative and positive controls during each assay. Eclipse software is easy to operate and complies with the provisions of 21 CFR Part 11.

Purpose and Scope

The horseshoe crab reagent (LAL) assay is susceptible to interference. The sample may contain substances that inhibit or enhance endotoxin detection, resulting in false results. It is necessary to conduct method suitability testing and inhibition/enhancement testing on the test sample to determine the dilution factor (or other effective method) required to overcome the aforementioned interference.

This application literature provides an overview of how to conduct inhibition/enhancement testing, how to run this test to demonstrate the applicability of the Sievers Eclipse bacterial endotoxin detector, and how to provide a method for running samples. The method information includes: the horseshoe crab reagent assay method used, assay sensitivity (λ), validated dilution factor, PPC (positive product control) spiked concentration, diluent,% CV (coefficient of variation) standard, and average PPC recovery rate.

The sample information includes: sample concentration (in mg/mL), endotoxin limit (EL, endotoxin limit; in EU/mL), maximum effective dilution (MVD, Maximum Valid Dilution), minimum effective concentration (MVC, Minimum Valid Concentration), average sample results (in EU/mL), and sample storage/processing conditions.

Sievers Eclipse bacterial endotoxin detector: an efficient and convenient solution

The Sievers Eclipse bacterial endotoxin detector can efficiently run inhibition/enhancement tests, as well as the validation procedures for the three batches of products mentioned above. This detector adopts advanced centrifugation technology and dynamic methods, simplifying the endotoxin detection process. The above advantages enable this detector to report detection results faster and more accurately, and provide accurate quantitative and interference analysis.

The advantages of Sievers Eclipse detector also include:

• Efficient: The automation function reduces the steps of laboratory settings and pipetting operations, improving throughput.

• Intuitive: The intuitive user interface simplifies the operation steps, reduces errors, and improves data quality.

Compliance: This detector meets strict regulatory requirements for pharmacopoeia testing and data reliability.

The Sievers Eclipse bacterial endotoxin detector is a significant technological advancement in the field of bacterial endotoxin testing, providing a comprehensive, efficient, and compliant solution for ongoing product validation needs, and improving product safety and quality.

More exciting content can be found in the attached materials!