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Airborne environmental endotoxin: a cross-validation of sampling and analysis techniques

Walters, M., Milton, D., Larsson, L. and Ford, T.
1994
Applied and Environmental Microbiology, 60(3): 996-1005, 1994


Walters, M., Milton, D., Larsson, L. and Ford, T., (1994), "Airborne environmental endotoxin: a cross-validation of sampling and analysis techniques", Applied and Environmental Microbiology, 60(3): 996-1005, 1994.
Abstract:
A standard method for measurement of airborne environmental endotoxin was developed and field tested in a fiberglass insulation-manufacturing facility. This method involved sampling with a capillary-pore membrane filter, extraction in buffer using a sonication bath, and analysis by the kinetic-Limulus assay with resistant-parallel-line estimation (KLARE). Cross-validation of the extraction and assay method was performed by comparison with methanolysis of samples followed by 3- hydroxy fatty acid (3-OHFA) analysis by gas chromatography-mass spectrometry. Direct methanolysis of filter samples and methanolysis of buffer extracts of the filters yielded similar 3-OHFA content (P = 0.72); the average difference was 2.1%. Analysis of buffer extracts for endotoxin content by the KLARE method and by gas chromatography-mass spectrometry for 3-OHFA content produced similar results (P = 0.23); the average difference was 0.88%. The source of endotoxin was gram- negative bacteria growing in recycled washwater used to clean the insulation-manufacturing equipment. The endotoxin and bacteria become airborne during spray cleaning operations. The types of 3-OHFAs in bacteria cultured from the washwater, present in the washwater and in the air, were similar. Virtually all of the bacteria cultured from air and water were gram negative composed mostly of two species, Deleya aesta and Acinetobacter johnsonii. Airborne countable bacteria correlated well with endotoxin (r2 = 0.64). Replicate sampling showed that results with the standard sampling, extraction, and Limulus assay by the KLARE method were highly reproducible (95% confidence interval for endotoxin measurement +/- 0.28 log10). These results demonstrate the accuracy, precision, and sensitivity of the standard procedure proposed for airborne environmental endotoxin.

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Author Information and Other Publications Notes
Walters, M.
  1. Risk of sick leave associated with outdoor air supply rate, humidification, and occupant complaints  
Milton, D.
  1. Populations and determinants of airborne fungi in large office buildings
  2. Quantification of ergosterol and 3-hydroxy fatty acids in settled house dust by gas chromatography-mass spectrometry: Comparison with fungal culture and determination of endotoxin by a Limulus amebocyte lysate assay
  3. Risk of sick leave associated with outdoor air supply rate, humidification, and occupant complaints  
Larsson, L.
  1. Determination of ergosterol in organic dust by gas chromatography-mass spectrometry
  2. Determination of microbial chemical markers by gas chromatography-mass spectrometry - potential for diagnosis and studies on metabolism in situ
  3. Determination of microbial colonisation in water-damaged buildings using chemical marker analysis by gas chromatography-mass spectrometry
  4. Quantification of ergosterol and 3-hydroxy fatty acids in settled house dust by gas chromatography-mass spectrometry: Comparison with fungal culture and determination of endotoxin by a Limulus amebocyte lysate assay
  5. Use of gas chromatography-mass spectrometry/solid phase microextraction for the identification of MVOCs from moldy building materials  
Ford, T.
     



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