Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1997 Jul;63(7):2554–2559. doi: 10.1128/aem.63.7.2554-2559.1997

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.

A Saraf 1, L Larsson 1, H Burge 1, D Milton 1
PMCID: PMC168553  PMID: 9212406

Abstract

Ergosterol and 3-hydroxy fatty acids, chemical markers for fungal biomass and the endotoxin of gram-negative bacteria, respectively, may be useful in studies of health effects of organic dusts, including domestic house dust. This paper reports a method for the combined determination of ergosterol and 3-hydroxy fatty acids in a single dust sample and a comparison of these chemical biomarkers determined by gas chromatography-mass spectrometry with results from fungal culture and Limulus assay. Analyses of replicate house dust samples resulted in correlations of 0.91 (ergosterol in six replicates; P < 0.01) and 0.94 (3-hydroxy fatty acids in nine replicates; P < 0.001). The amounts of ergosterol (range, 2 to 16.5 ng/mg of dust) correlated with those of total culturable fungi (range, 6 to 1,400 CFU/mg of dust) in 17 samples, (r = 0.65; P < 0.005). The amounts of endotoxin (range, 11 to 243 endotoxin units/mg of dust) measured with a modified chromogenic Limulus assay correlated with those of lipopolysaccharide (LPS) determined from 3-hydroxy fatty acid analysis of 15 samples. The correlation coefficient depended on the chain lengths of 3-hydroxy acids used to compute the LPS content. The correlation was high (r = 0.88 +/- 0.01; P < 0.001) when fatty acid chains of 10 to 14 carbon atoms were included; the correlation was much lower when hydroxy acids of 16- or 18-carbon chains were included. In conclusion, the results of the described extraction and analysis procedure for ergosterol and 3-hydroxy fatty acids are reproducible, and the results can be correlated with fungal culture and endotoxin activity of organic dust samples.

Full Text

The Full Text of this article is available as a PDF (137.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Axelsson B. O., Saraf A., Larsson L. Determination of ergosterol in organic dust by gas chromatography-mass spectrometry. J Chromatogr B Biomed Appl. 1995 Apr 7;666(1):77–84. doi: 10.1016/0378-4347(94)00553-h. [DOI] [PubMed] [Google Scholar]
  2. Börjesson T., Stöllman U., Schnürer J. Volatile metabolites produced by six fungal species compared with other indicators of fungal growth on cereal grains. Appl Environ Microbiol. 1992 Aug;58(8):2599–2605. doi: 10.1128/aem.58.8.2599-2605.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Flaherty D. K., Deck F. H., Cooper J., Bishop K., Winzenburger P. A., Smith L. R., Bynum L., Witmer W. B. Bacterial endotoxin isolated from a water spray air humidification system as a putative agent of occupation-related lung disease. Infect Immun. 1984 Jan;43(1):206–212. doi: 10.1128/iai.43.1.206-212.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fox A., Rosario R. M., Larsson L. Monitoring of bacterial sugars and hydroxy Fatty acids in dust from air conditioners by gas chromatography-mass spectrometry. Appl Environ Microbiol. 1993 Dec;59(12):4354–4360. doi: 10.1128/aem.59.12.4354-4360.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Heederik D., Brouwer R., Biersteker K., Boleij J. S. Relationship of airborne endotoxin and bacteria levels in pig farms with the lung function and respiratory symptoms of farmers. Int Arch Occup Environ Health. 1991;62(8):595–601. doi: 10.1007/BF00381114. [DOI] [PubMed] [Google Scholar]
  6. Helander I., Saxén H., Salkinoja-Salonen M., Rylander R. Pulmonary toxicity of endotoxins: comparison of lipopolysaccharides from various bacterial species. Infect Immun. 1982 Feb;35(2):528–532. doi: 10.1128/iai.35.2.528-532.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Horner W. E., Helbling A., Salvaggio J. E., Lehrer S. B. Fungal allergens. Clin Microbiol Rev. 1995 Apr;8(2):161–179. doi: 10.1128/cmr.8.2.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Maitra S. K., Nachum R., Pearson F. C. Establishment of beta-hydroxy fatty acids as chemical marker molecules for bacterial endotoxin by gas chromatography-mass spectrometry. Appl Environ Microbiol. 1986 Sep;52(3):510–514. doi: 10.1128/aem.52.3.510-514.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Michel O., Ginanni R., Duchateau J., Vertongen F., Le Bon B., Sergysels R. Domestic endotoxin exposure and clinical severity of asthma. Clin Exp Allergy. 1991 Jul;21(4):441–448. doi: 10.1111/j.1365-2222.1991.tb01684.x. [DOI] [PubMed] [Google Scholar]
  10. Mielniczuk Z., Mielniczuk E., Larsson L. Determination of muramic acid in organic dust by gas chromatography-mass spectrometry. J Chromatogr B Biomed Appl. 1995 Aug 4;670(1):167–172. doi: 10.1016/0378-4347(95)00152-9. [DOI] [PubMed] [Google Scholar]
  11. Milton D. K., Amsel J., Reed C. E., Enright P. L., Brown L. R., Aughenbaugh G. L., Morey P. R. Cross-sectional follow-up of a flu-like respiratory illness among fiberglass manufacturing employees: endotoxin exposure associated with two distinct sequelae. Am J Ind Med. 1995 Oct;28(4):469–488. doi: 10.1002/ajim.4700280404. [DOI] [PubMed] [Google Scholar]
  12. Milton D. K., Gere R. J., Feldman H. A., Greaves I. A. Endotoxin measurement: aerosol sampling and application of a new Limulus method. Am Ind Hyg Assoc J. 1990 Jun;51(6):331–337. doi: 10.1080/15298669091369754. [DOI] [PubMed] [Google Scholar]
  13. Milton D. K., Wypij D., Kriebel D., Walters M. D., Hammond S. K., Evans J. S. Endotoxin exposure-response in a fiberglass manufacturing facility. Am J Ind Med. 1996 Jan;29(1):3–13. doi: 10.1002/(SICI)1097-0274(199601)29:1<3::AID-AJIM2>3.0.CO;2-V. [DOI] [PubMed] [Google Scholar]
  14. Olsen J. H., Dragsted L., Autrup H. Cancer risk and occupational exposure to aflatoxins in Denmark. Br J Cancer. 1988 Sep;58(3):392–396. doi: 10.1038/bjc.1988.226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rylander R., Bake B., Fischer J. J., Helander I. M. Pulmonary function and symptoms after inhalation of endotoxin. Am Rev Respir Dis. 1989 Oct;140(4):981–986. doi: 10.1164/ajrccm/140.4.981. [DOI] [PubMed] [Google Scholar]
  16. Schnürer J. Comparison of methods for estimating the biomass of three food-borne fungi with different growth patterns. Appl Environ Microbiol. 1993 Feb;59(2):552–555. doi: 10.1128/aem.59.2.552-555.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schwartz D. A., Thorne P. S., Yagla S. J., Burmeister L. F., Olenchock S. A., Watt J. L., Quinn T. J. The role of endotoxin in grain dust-induced lung disease. Am J Respir Crit Care Med. 1995 Aug;152(2):603–608. doi: 10.1164/ajrccm.152.2.7633714. [DOI] [PubMed] [Google Scholar]
  18. Sonesson A., Larsson L., Fox A., Westerdahl G., Odham G. Determination of environmental levels of peptidoglycan and lipopolysaccharide using gas chromatography with negative-ion chemical-ionization mass spectrometry utilizing bacterial amino acids and hydroxy fatty acids as biomarkers. J Chromatogr. 1988 Sep 23;431(1):1–15. doi: 10.1016/s0378-4347(00)83064-9. [DOI] [PubMed] [Google Scholar]
  19. Sonesson A., Larsson L., Schütz A., Hagmar L., Hallberg T. Comparison of the limulus amebocyte lysate test and gas chromatography-mass spectrometry for measuring lipopolysaccharides (endotoxins) in airborne dust from poultry-processing industries. Appl Environ Microbiol. 1990 May;56(5):1271–1278. doi: 10.1128/aem.56.5.1271-1278.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Takada H., Kotani S. Structural requirements of lipid A for endotoxicity and other biological activities. Crit Rev Microbiol. 1989;16(6):477–523. doi: 10.3109/10408418909104475. [DOI] [PubMed] [Google Scholar]
  21. Teeuw K. B., Vandenbroucke-Grauls C. M., Verhoef J. Airborne gram-negative bacteria and endotoxin in sick building syndrome. A study in Dutch governmental office buildings. Arch Intern Med. 1994 Oct 24;154(20):2339–2345. [PubMed] [Google Scholar]
  22. Walters M., Milton D., Larsson L., Ford T. Airborne environmental endotoxin: a cross-validation of sampling and analysis techniques. Appl Environ Microbiol. 1994 Mar;60(3):996–1005. doi: 10.1128/aem.60.3.996-1005.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Zhiping W., Malmberg P., Larsson B. M., Larsson K., Larsson L., Saraf A. Exposure to bacteria in swine-house dust and acute inflammatory reactions in humans. Am J Respir Crit Care Med. 1996 Nov;154(5):1261–1266. doi: 10.1164/ajrccm.154.5.8912733. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES