How do you test for bacterial endotoxin?

How do you test for bacterial endotoxin?

Test procedure: A BET involves analyzing the liquid sample or sample extract using Limulus Amebocyte Lysate (LAL). LAL is a reagent made from the blood of the horseshoe crab. In the presence of bacterial endotoxins, the lysate reacts to form a clot or cause a color change depending on the technique.

What is the permissible limit of bacterial endotoxin?

Water for Injection, Sterile Water for Injection and Sterile Water for Irrigation have an allowable endotoxin limit of 0.25 Endotoxin Units (EU)/ml. (EU=Unit of measurement for endotoxin activity).

How do you calculate bacterial endotoxin limits?

If the dose is 1 mg/kg/hr, the endotoxin limit is (5 EU/kg/hr) ÷ (1 mg/kg/hr) = 5 EU/mg. If the dose is 10 mg/kg/hr, the endotoxin limit is (5 EU/kg/hr) ÷ (10 mg/kg/hr) = 0.5 EU/mg. If the dose is 100mg/kg/hr, the endotoxin limit is (5 EU/kg/hr) ÷ (100 mg/kg/hr) = 0.05 EU/mg.

Why is bacterial endotoxin test important?

Bacterial endotoxin testing (BET) is a fundamental safety requirement in the pharmaceutical and biomedical industries. If bacterial endotoxins enter patients’ bloodstream in sufficient concentrations, this can cause harmful symptoms such as fever and septic shock and can be fatal in the most severe cases.

Can endotoxin samples be frozen?

Endotoxin levels are relatively stable when samples are stored frozen with test variations between 1 and 38% among different aliquots. However, about 25% of the samples showed significant differences between testing modalities and/or reagents.

How are endotoxins measured?

Endotoxin is measured in endotoxin units per milliliter (EU/mL). One EU equals approximately 0.1 to 0.2 ng endotoxin/mL of solution.

What is bacterial endotoxin?

An endotoxin is a lipopolysaccharide (LPS) found in the cell wall of gram-negative bacteria. It exists in the environment (e.g. water, air) inhabited by gram-negative bacteria, and bacterial endotoxins (LPS) remain even after the bacteria die.

What is bacterial endotoxin test?

The bacterial endotoxins test (BET) is a test to detect or quantify endotoxins from Gram- negative bacteria using amoebocyte lysate from the horseshoe crab (Limulus polyphemus or. Tachypleus tridentatus).

Is there a test to detect bacterial endotoxins?

This chapter provides a test to detect or quantify bacterial endotoxins that may be present in or on the sample of the article (s) to which the test is applied.

What’s the minimum temperature to test for endotoxins?

A commonly used minimum time and temperature is 30 minutes at 250°C. If employing plastic apparatus such as microplates and pipet tips for automatic pipetters, use apparatus shown to be free of detectable endotoxin and which does not interfere in the test.

When was the last endotoxin testing guideline published?

These tests were described in a series of draft and final guidance documents. The last guidance document, Guideline on Validation of the Limulus Amebocyte Lysate Test as an End-Product Endotoxin Test for Human and Animal Parenteral Drugs, Biological Products, and Medical Devices, was published in 1987 (the 1987 Guidance).

When did the USP change the bacterial endotoxin test?

The USP’s subcommittee responsible for revisions of compendial test methods and/or product monographs has, in the past few years, made some significant changes in the bacterial endotoxin test and product monograph requirements. In 1984, five USP water products were given specific bacterial endotoxin limits.