Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay

Barros, Jean‐Paul Pais de; Gautier, Thomas; Sali, Wahib; Adrie, Christophe; Choubley, Hélène; Charron, Émilie; Lalande, Caroline; Guern, Naïg Le; Deckert, Valérie; Monchi, Mehran; Quenot, Jean‐Pierre; Lagrost, Laurent

doi:10.1194/jlr.d059725

Cited by 101 publications

(107 citation statements)

References 43 publications

Supporting

Mentioning

103

Contrasting

Unclassified

Order By: Relevance

“…The FDA guidelines have received some criticism due to the fact, for example, that it is not easy to elute LPS from the test item. One study showed that only 40–70% of inoculated LPS was recovered from devices following a very stringent extraction protocol in ethanol …”

Section: Discussionmentioning

confidence: 99%

“…However, in this study, the authors used only small implants, due to the extremely costly LAL assay reagents. Lipscomb et al tested 260 surgical instruments, from nine primary care units, and used 4,6‐diamidino‐2‐phenylindole dihydrochloride to detect LPS and proteinaceous contaminations. Their results were negative for LPS, but 60% showed a high degree of protein soiling (0.4–4.2 μg protein/mm 2 ).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Characterization of endotoxins on orthopaedic fixation screws, using physicochemical surface analyses

França

Alfa

Olson

et al. 2016

Journal Orthopaedic Research

View full text Add to dashboard Cite

The objective of this study was to determine if surface analysis techniques could be used to detect endotoxin on stainless steel malleolus screws. New malleolus screws were compared to ones that had been coated in purified lipopolysaccharide (LPS) or Artificial Test Soil (ATS) containing lipopolysaccharide. X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), and time-of flight secondary ion mass spectrometry (TOF-SIMS) were used to assess the fixation screws surface. Organic material was visualized on the LPS and ATS-LPS inoculated screws but not on the new unsoiled screws. This was further supported by the peaks observed at masses between 40 and 100 D in TOF-SIMS spectra of the LPS and ATS-LPS inoculated screws. After deconvolution of N1s high resolution XPS spectra, the LPS inoculated screws showed amide groups whereas the ATS-LPS inoculated screws showed predominantly nitroso groups (C-NO). Our data demonstrate that surface analysis can be used to detect organic residuals present on fixation screws. The XPS data confirmed that LPS reacted predominantly with positively charged surface metallic ions (Fe and Cr), whereas proteins reacted with the surface oxide layer of fixation screws, forming C-NO groups. The application of these surface analysis techniques will be helpful in determining if the reprocessing of such items results in an accumulation of organic material that might lead to aseptic loosening, when implanted. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:240-247, 2017.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Characterization of endotoxins on orthopaedic fixation screws, using physicochemical surface analyses

França

Alfa

Olson

et al. 2016

Journal Orthopaedic Research

View full text Add to dashboard Cite

show abstract

“…Alternatively, a wide variety of mass spectrometry (MS)-based techniques has been applied for the detection of complex LPS or lipid A [27–30]. For example, Caroff et al reported direct micro-extraction and analysis of intact LPS by combined thin-layer chromatography and matrix-assisted laser desorption ionization (MALDI) MS [27].…”

Section: Introductionmentioning

confidence: 99%

“…Instead, other approaches for the measurement of LPS using MS are to fractionate intact LPS, extract specific fatty acids and derivatize them prior to analysis. Gas chromatography (GC)- or liquid chromatography (LC) coupled to MS is then applied for the separation, detection and quantitation of targeted fatty acid [30–32]. These methods can also be applied to carbohydrate-containing samples, which normally interfere with endotoxin biological assays but still suffer from multiple sample pre-treatment steps and tedious, time-consuming method development.…”

Section: Introductionmentioning

confidence: 99%

Rapid detection of bacterial endotoxins in ophthalmic viscosurgical device materials by direct analysis in real time mass spectrometry

Hitchins

Wickramasekara

2016

Analytica Chimica Acta

View full text Add to dashboard Cite

Bacterial endotoxins are lipopolysaccharides bound to the bacterial cell wall and released when bacteria rupture or disintegrate. Possible contamination of endotoxin in ophthalmic devices can cause a painful eye inflammation or result in toxic anterior segment syndrome after cataract surgery. Measurement of bacterial endotoxin in medical device materials is difficult since endotoxin binds with polymer matrix and some of the materials are very viscous and non-water soluble, where traditional enzyme-based Limulus amebocyte lysate (LAL) assay cannot be applied. Here we propose a rapid and high throughput ambient ionization mass spectrometric (MS) method using direct analysis in real time (DART) for the evaluation of endotoxin contamination in medical device materials. Large and structurally complex endotoxin instantaneously breaks down into low-mass characteristic fragment ions using DART and is detected by MS in both positive and negative ion modes. This method enables the identification and separation of endotoxin from medical materials with a detection limit of 0.03 ng mL−1 endotoxins in aqueous solution. Ophthalmic viscosurgical device materials including sodium hyaluronate (NaHA), non-water soluble perfluoro-n-octane (PFO) and silicone oil (SO) were spiked with different known concentrations of endotoxin and analyzed by DART MS, where the presence of endotoxin was successfully detected and featured small mass fragment ions were generated for NaHA, PFO and SO as well. Current findings showed the feasibility of measuring endotoxin contamination in medical device materials using DART-MS, which can lead to a one-step analysis of endotoxins in different matrices, avoiding any potential contamination during sample pre-treatment steps.

show abstract

“…It is increasingly used in metabolic studies and therefore was adapted to the plasma of different species to this aim (Laugerette et al, 2015). Of note, emerging methods aim at quantifying LPS molecules via the analysis of 3-hydroxymuristic acid from their lipid A in plasma (Pais de Barros et al, 2015).…”

Section: Endotoxins Are Proinflammatory Moleculesmentioning

confidence: 99%

Dietary lipid emulsions and endotoxemia

Michalski¹,

Vors²,

Lecomte³

et al. 2016

OCL

View full text Add to dashboard Cite

-The low-grade inflammation observed in obesity is a risk factor for cardiovascular diseases and insulin resistance. Among factors triggering such inflammation, recent works revealed the role of bacterial lipopolysaccharides (LPS), so-called endotoxins. LPS are naturally present in the gut via the intestinal microbiota. Recent studies show that they can induce in plasma a metabolic endotoxemia after the consumption of unbalanced hyperlipidic meals. This article reviews recent knowledge gained on the role of intestinal lipid absorption and the composition of dietary lipids on: (i) the induction of metabolic endotoxemia, (ii) the types of plasma transporters of LPS and (iii) associated low-grade inflammation. Notably, lipids are present in foods under various physicochemical structures and notably in emulsified form. Our recent works reveal that such structure and the type of emulsifier can modulate postprandial lipemia; recent results on the possible consequences on metabolic endotoxemia will be discussed.Keywords: Nutrition / fat / oil / emulsifier / inflammation Résumé -Émulsions lipidiques alimentaires et endotoxémie. L'inflammation à bas bruit observée en situation d'obésité représente un facteur de risque cardiovasculaire et d'insulinorésistance. Parmi les facteurs responsables de cette inflammation, des travaux récents ont révélé l'implication des lipopolysaccharides bactériens, aussi appelés endotoxines. Ces endotoxines sont naturellement présentes dans le tractus digestif via le microbiote intestinal. Des études récentes montrent qu'elles peuvent générer une endotoxémie métabolique dans le plasma suite à la consommation de repas déséquilibrés hyperlipidiques. L'objectif de cette revue est de faire le point sur les connaissances récentes concernant le rôle de l'absorption intestinale des lipides et de la composition de ces derniers sur : (i) l'établissement d'une endotoxémie métabolique, (ii) la qualité des transporteurs plasmatiques des LPS et (iii) l'inflammation à bas bruit associée. En particulier, les lipides sont présents dans les aliments sous différentes structures physico-chimiques et notamment sous forme émulsionnée. Nos travaux récents montrent que cette structure est à même de moduler la lipémie postprandiale. Des résultats récents sur les possibles conséquences sur l'endotoxémie métabolique sont discutés.

show abstract

Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay

Cited by 101 publications

References 43 publications

Characterization of endotoxins on orthopaedic fixation screws, using physicochemical surface analyses

Characterization of endotoxins on orthopaedic fixation screws, using physicochemical surface analyses

Rapid detection of bacterial endotoxins in ophthalmic viscosurgical device materials by direct analysis in real time mass spectrometry

Dietary lipid emulsions and endotoxemia

Contact Info

Product

Resources

About