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Interlaboratory evaluation of endotoxin analyses in agricultural dusts - comparison of LAL assay and mass spectrometry

Författare

  • SJ Reynolds
  • DK Milton
  • D Heederik
  • PS Thorne
  • KJ Donham
  • EA Croteau
  • KM Kelly
  • J Douwes
  • D Lewis
  • M Whitmer
  • I Connaughton
  • S Koch
  • P Malmberg
  • BM Larsson
  • J Deddens
  • A Saraf
  • Lennart Larsson

Summary, in English

Endotoxin exposure is associated with wheeze and asthma morbidity, while early life exposure may reduce risk of allergy and asthma. Unfortunately, it is difficult to compare endotoxin results from different laboratories and environments. We undertook this study to determine if lipopolysaccharide ( LPS) extraction efficiency could account for differences among laboratories. We generated and collected aerosols from chicken and swine barns, and corn processing. We randomly allocated side-by-side filter samples to five laboratories for Limulus assay of endotoxin. Lyophilized aliquots of filter extracts were analyzed for 3-hydroxy fatty acids ( 3-OHFAs) as a marker of LPS using gas chromatography-mass spectrometry. There were significant differences in endotoxin assay and GC-MS ( LPS) results between laboratories for all dust types ( p < 0.01). Patterns of differences between labs varied by dust type. Relationships between assay and GC/MS results also depended on dust type. The percentages of individual 3-OHFA chain lengths varied across labs ( p < 0.0001) suggesting that each lab recovered a different fraction of the LPS available. The presence of large amounts of particle associated LPS and absence of a freezing thawing cycle were associated with lower correlations between LPS and bioactivity, consistent with an absence of Limulus response to cell-bound endotoxin. These data suggest that extraction methods affect endotoxin measurements. The LAL methods may be most suitable when comparing exposures within similar environments; GC-MS offers additional information helpful in optimizing sample treatment and extraction. GC-MS may be of use when comparing across heterogeneous environments and should be considered for inclusion in future studies of human health outcomes.

Publiceringsår

2005

Språk

Engelska

Sidor

1371-1377

Publikation/Tidskrift/Serie

Journal of Environmental Monitoring

Volym

7

Issue

12

Dokumenttyp

Artikel i tidskrift

Förlag

Royal Society of Chemistry

Ämne

  • Microbiology in the medical area

Status

Published

ISBN/ISSN/Övrigt

  • ISSN: 1464-0325