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Low Endotoxin Recovery—Masking of Naturally Occurring Endotoxin

Endotoxins are cell wall components of Gram-negative bacteria. A release of endotoxins into the human blood stream results in an inflammation reaction that can lead to life-threatening conditions like sepsis. Therefore, control for endotoxin contamination of intravenously administered drugs is cruci...

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Autores principales: Reich, Johannes, Weyer, Felix Alexander, Tamura, Hiroshi, Nagaoka, Isao, Motschmann, Hubert
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6412962/
https://www.ncbi.nlm.nih.gov/pubmed/30781342
http://dx.doi.org/10.3390/ijms20040838
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author Reich, Johannes
Weyer, Felix Alexander
Tamura, Hiroshi
Nagaoka, Isao
Motschmann, Hubert
author_facet Reich, Johannes
Weyer, Felix Alexander
Tamura, Hiroshi
Nagaoka, Isao
Motschmann, Hubert
author_sort Reich, Johannes
collection PubMed
description Endotoxins are cell wall components of Gram-negative bacteria. A release of endotoxins into the human blood stream results in an inflammation reaction that can lead to life-threatening conditions like sepsis. Therefore, control for endotoxin contamination of intravenously administered drugs is crucial. Drugs are usually tested for putative endotoxin contamination with Limulus-based tests. However, validity of the compendial test procedures is questioned in the case of low endotoxin recovery (LER). To assure validity, regulatory authorities request hold-time studies of endotoxin in addition to pharmacopoeial requirements. Within these studies, endotoxin is added (spiked) to an undiluted product. The spiked product is held for a certain period of time and subsequently diluted for endotoxin determination. Due to the known heterogeneity of endotoxin the question has been raised as to which source represents the most adequate endotoxin spike. In the present study, endotoxin hold-time studies were analyzed by using different sources of endotoxin. Highly purified endotoxin, crude endotoxin extracts (Naturally Occurring Endotoxin) from different bacterial species and varied growth conditions as well as endogenous endotoxin contaminations were investigated. The results clearly demonstrate that endotoxin masking—an effect of LER—is dependent on the endotoxin source used. Various parameters such as bacterial strain and growth conditions lead to different masking susceptibilities. Due to these effects it is impossible to predict the susceptibility of bacterial endotoxin contamination to LER. In order to determine whether a sample is prone to LER, an endotoxin spike that is susceptible to LER is required.
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spelling pubmed-64129622019-03-29 Low Endotoxin Recovery—Masking of Naturally Occurring Endotoxin Reich, Johannes Weyer, Felix Alexander Tamura, Hiroshi Nagaoka, Isao Motschmann, Hubert Int J Mol Sci Article Endotoxins are cell wall components of Gram-negative bacteria. A release of endotoxins into the human blood stream results in an inflammation reaction that can lead to life-threatening conditions like sepsis. Therefore, control for endotoxin contamination of intravenously administered drugs is crucial. Drugs are usually tested for putative endotoxin contamination with Limulus-based tests. However, validity of the compendial test procedures is questioned in the case of low endotoxin recovery (LER). To assure validity, regulatory authorities request hold-time studies of endotoxin in addition to pharmacopoeial requirements. Within these studies, endotoxin is added (spiked) to an undiluted product. The spiked product is held for a certain period of time and subsequently diluted for endotoxin determination. Due to the known heterogeneity of endotoxin the question has been raised as to which source represents the most adequate endotoxin spike. In the present study, endotoxin hold-time studies were analyzed by using different sources of endotoxin. Highly purified endotoxin, crude endotoxin extracts (Naturally Occurring Endotoxin) from different bacterial species and varied growth conditions as well as endogenous endotoxin contaminations were investigated. The results clearly demonstrate that endotoxin masking—an effect of LER—is dependent on the endotoxin source used. Various parameters such as bacterial strain and growth conditions lead to different masking susceptibilities. Due to these effects it is impossible to predict the susceptibility of bacterial endotoxin contamination to LER. In order to determine whether a sample is prone to LER, an endotoxin spike that is susceptible to LER is required. MDPI 2019-02-15 /pmc/articles/PMC6412962/ /pubmed/30781342 http://dx.doi.org/10.3390/ijms20040838 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Reich, Johannes
Weyer, Felix Alexander
Tamura, Hiroshi
Nagaoka, Isao
Motschmann, Hubert
Low Endotoxin Recovery—Masking of Naturally Occurring Endotoxin
title Low Endotoxin Recovery—Masking of Naturally Occurring Endotoxin
title_full Low Endotoxin Recovery—Masking of Naturally Occurring Endotoxin
title_fullStr Low Endotoxin Recovery—Masking of Naturally Occurring Endotoxin
title_full_unstemmed Low Endotoxin Recovery—Masking of Naturally Occurring Endotoxin
title_short Low Endotoxin Recovery—Masking of Naturally Occurring Endotoxin
title_sort low endotoxin recovery—masking of naturally occurring endotoxin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6412962/
https://www.ncbi.nlm.nih.gov/pubmed/30781342
http://dx.doi.org/10.3390/ijms20040838
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