Abstract
Gas chromatography-mass spectrometry (GC-MS) can be applied to detect and characterize microorganisms in clinical and environmental samples, and microbial contaminants in biotechnological production cultures. With this approach, unique microbial monomeric compounds, known as chemical markers, are used as analytes. In the present article, two GC-MS-based techniques, viz. GC-ion trap tandem MS (GC-MS-MS) and conventional quadrupole GC-MS used in the selected ion monitoring mode, were compared regarding their ability to detect 3-hydroxy fatty acids, muramic acid, and ergosterol (markers for endotoxin, peptidoglycan, and fungal biomass, respectively) in complex matrices. When using GC-MS-MS, daughter ion spectra were obtained for all markers present in amounts close to the detection limit of the GC-MS. Ion-trap GC-MS-MS shows great promise as a chemical marker analysis technique for application in clinical diagnosis, occupational and public health care, and biotechnology.
Similar content being viewed by others
References
Alugupalli, S., Portaels, F., and Larsson, L. (1994) Systematic study of the 3-hydroxy fatty acid composition of mycobacteria.J. Bacteriol. 176, 2962–2969.
Sonesson, A., Jantzen, E., Bryn, K., Larsson, L., and Eng, J. (1989) Chemical composition of a lipopolysaccharide fromLegionella pneumophila.Arch. Microbiol. 153, 72–78.
Mayberry, W. R. and Lane, J. R. (1993) Sequential alkaline saponification/acid hydrolysis/esterification: a one-tube method with enhanced recovery of both cyclopropane and hydroxylated fatty acids.J. Microbiol. Methods 18, 21–32.
Alugupalli, S., Larsson, L., Slosarek, M., and Jaresova, M. (1992) Application of gas chromatography-mass spectrometry for rapid detection ofMycobacterium xenopi in drinking water.Appl. Environ. Microbiol. 58, 3538–3541.
Slosárek, M., Alugupalli, S., Kaustová, J., and Larsson L. (1996) Rapid detection ofMycobacterium kansasii in water by gas chromatography-mass spectrometry.J. Microbiol. Methods 27, 229–232.
Gilbart, J. (1984) Detection ofLegionella spp. in water samples by electron-capture gas chromatography, inProc. 2nd Int. Symp on Legionella, ASM, pp 296–298.
Donham, K., Haglind, P., Peterson, Y., Rylander, R., and Belin, L. (1989) Environmental and health studies of farm workers in Swedish swine confinement buildings.Br. J. Ind. Med. 46, 31–37.
Heederik, D., Brouwer, R., Bierstecker, K., and Boleij, J. (1991) Relationship of airborne endotoxin and bacteria levels in pig farms with the lung function and respiratory symptoms of farmers.Int. Arch. Environ. Health 62, 595–601.
Palchak, R. B., Cohen, R., Ainslie, M., and Hoerner, C. L. (1988) Airborne endotoxin associated with industrial-scale production of protein products in gramnegative bacteria.Am. Ind. Hyg. Assoc. J. 49, 420,421.
Rylander, R., Soerensen, S., Goto, H., Yuasa, K., and Tanaka, S. (1989) The importance of endotoxin and glucan for symptoms in sick buildings.Present and Future of Indoor Air Quality 219–226.
Sonesson, A., Larsson, L., Fox, A., Westerdahl, G., and Odham, G. (1988) 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. Biomed. Appl. 431, 1–15.
Mielniczuk, Z., Mielniczuk, E., and Larsson, L. (1993) Gas chromatography-mass spectrometry methods for analysis of 2- and 3-hydroxylated fatty acids: application for endotoxin measurement.J. Microbiol. Methods 17, 91–102.
Wilkinson, S. G. (1988) Gram-negative bacteria, inMicrobial Lipids, vol. 1 (Ratledge, C. and Wilkinson, S. G., eds.), Academic, London, pp. 299–488.
Mielniczuk, Z., Mielniczuk, E., and Larsson, L. (1995) Determination of muramic acid in organic dust by gas chromatography-mass spectrometry.J. Chromatogr. B 670, 167–172.
Fox, A., Rosario, R., and Larsson, L. (1993) Monitoring of bacterial sugars and hydroxy fatty acids in dust from air conditioners by gas chromatography-mass spectrometry.Appl. Environ. Microbiol. 59, 4354–4360.
Axelsson, B., Saraf, A., and Larsson, L. (1995) Determination of ergosterol in organic dust by gas chromatography-mass spectrometry.J. Chromatogr. B 666, 77–84.
Miller, J. D., Laflamme, A. M., Sobol, Y., Lafontaine, P., and Greenhalgh, R. (1988) Fungi and fungal products in some Canadian houses.Int. Biodeterioration 24, 103–120.
Zhiping, W., Malmberg, P., Larsson, B.-M., Larsson, K., Larsson, L., and Saraf, A. (1996) Exposure to bacteria in swine-house dust and acute inflammatory reactions in humans.Am. J. Respir. Crit. Care Med. 154, 1261–1266.
Walters, M., Milton, D., Larsson, L., and Ford, T. (1994) Airborne environmental endotoxin: a cross-validation of sampling and analysis techniques.Appl. Environ. Microbiol. 60, 996–1005.
Maitra, S., Nachum, R., and Pearson, F. (1986) Establishment of beta-hydroxy fatty acids as chemical marker molecules for bacterial endotoxin by gas chromatography-mass spectrometry.Appl. Environ. Microbiol. 52, 510–514.
Christensson, B., Gilbart, J., Fox, A., and Morgan, S. (1989) Mass spectrometric quantitation of muramic acid, a bacterial cell wall component, in septic synovial fluids.Arthritis Rheum. 32, 1268–1272.
Odham, G., Larsson, L., and Mårdh, P.-A. (1979) Demonstration of tuberculostearic acid in sputum from patients with pulmonary tuberculosis by selected ion monitoring.J. Clin. Invest. 63, 813–819.
Mårdh, P.-A., Larsson, L., Hoiby, N., Engbaek, H. C., and Odham, G. (1983) Tuberculostearic acid as a diagnostic marker in tuberculous meningitis.The Lancet 8320, 367.
Alugupalli, S., Olsson, B., and Larsson, L. (1993) Detection of 2-eicosanol by gas chromatographymass spectrometry in sputa from patients with pulmonary mycobacterial infections.J. Clin. Microbiol. 31, 1575–1578.
Bosquée, L., Böttger, E., de Beenhouwer, H., Fonteyne, P., Hirschel, B., Larsson, L., Meyers, W., Palomino, J., Realini, L., Rigouts, L., Silva, M., Teske, A., van der Auwera, P., and Portaels, F. (1995) Cervical lymphadenitis caused by a fastidious mycobacterium closely related toMycobacterium genavense in an apparently immunocompetent woman: diagnosis by culture-free microbiological methods.J. Clin. Microbiol. 33, 2670–2674.
Larsson, L., Persson, C., Wiebe, T., and Christensson, B. (1994) Gas chromatographic determination ofd-arabinitol/l-arabinitol ratios in urine: a potential method for diagnosis of disseminated candidiasis.J. Clin. Microbiol. 32, 1855–1859.
Christensson, B., Wiebe, T., Pehrson, C., Larsson, L. (1997) Diagnosis of invasive candidiasis in neutropenic children with cancer by determination ofd-arabinitol/l-arabinitol in urine.J. Clin. Microb. 35, 636–640.
Lehtonen, L., Anttila, V.-J., Ruutu, T., Salonen, J., Nikoskelainen, J., Eerola, E., and Ruutu, P. (1996) Diagnosis of disseminated candidiasis by measurement of urined-arabinitol/l-arabinitol ratio.J. Clin. Microbiol. 34, 2175–2179.
Elmroth, I., Valeur, A., Odham, G., and Larsson, L. (1990) Detection of microbial contamination in fermentation processes: mass spectrometric determination of gram-negative bacteria inLeuconostoc mesenteroides cultures.Biotechnol. Bioeng. 35, 787–792.
Elmroth, I., Fox, A., Holst, O., and Larsson, L. (1993) Detection of bacterial contamination in cultures of eucaryotic cells by gas chromatography-mass spectrometry.Biotechnol. Bioeng. 42, 421–429.
Fox, A., Krahmer, M., and Harrison, D. (1996) Monitoring muramic acid in air (after alditol acetate derivatization) using a gas chromatograph-ion trap tandem mass spectrometer.J. Microbiol. Methods 27, 129–138.
Saraf, A. and Larsson, L. (1996) Use of gas chromatography-ion trap tandem mass spectrometry for the determination of chemical markers of microorganisms in organic dust.J. Mass Spectrom. 31, 389–396.
Fox, A., Fox, K., Christensson, B., Krahmer, M., and Harrelson D. (1996) Absolute identification of muramic acid at trace levels in human septic fluids in vivo and absence in aseptic fluids.Infect. Immun. 64, 3911–3955.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Larsson, L., Saraf, A. Use of gas chromatography-ion trap tandem mass spectrometry for the detection and characterization of microorganisms in complex samples. Mol Biotechnol 7, 279–287 (1997). https://doi.org/10.1007/BF02740818
Issue Date:
DOI: https://doi.org/10.1007/BF02740818