Supercritical fluid extraction (SFE) was used in the analysis of bacterial respiratory quinone (RQ), bacterial phospholipid fatty acid (PLFA), and archaeal phospholipid ether lipid (PLEL) from anaerobically digested sludge. Bacterial RQ were determined using ultra performance liquid chromatography (UPLC). Determination of bacterial PLFA and archaeal PLEL was simultaneously performed using gas chromatography-mass spectrometry (GC-MS). The effects of pressure, temperature, and modifier concentration on the total amounts of RQ, PLFA, and PLEL were investigated by 23 experiments with five settings chosen for each variable. The optimal extraction conditions that were obtained through a multiple-response optimization included a pressure of 23.6 MPa, temperature of 77.6 °C, and 10.6% (v/v) of methanol as the modifier. Thirty nine components of microbial lipid biomarkers were identified in the anaerobically digested sludge. Overall, the SFE method proved to be more effective, rapid, and quantitative for simultaneously extracting bacterial and archaeal lipid biomarkers, compared to conventional organic solvent extraction. This work shows the potential application of SFE as a routine method for the comprehensive analysis of microbial community structures in environmental assessments using the lipid biomarkers profile.
References
[1]
Sekiguchi, Y.; Kamagata, Y.; Harada, H. Recent advances in methane fermentation technology. Curr. Opin. Biotechnol 2001, 12, 277–282.
[2]
Rittmann, B.E. Microbial ecology to manage processes in environmental biotechnology. Trends Biotechnol 2006, 24, 261–266.
[3]
Narihiro, T.; Sekiguchi, Y. Microbial communities in anaerobic digestion processes for waste and wastewater treatment: A microbiological update. Curr. Opin. Biotechnol 2007, 18, 273–278.
[4]
White, D.C.; Davis, W.M.; Nickels, J.S.; King, J.D.; Bobbie, R.J. Determination of the sedimentary microbial biomass by extractable lipid phosphate. Oecologia 1979, 40, 51–62.
[5]
Zelles, L. Fatty acid patterns of phospholipids and lipopolysaccharides in the characterisation of microbial communities in soil: A review. Biol. Fertil. Soils 1999, 29, 111–129.
[6]
White, D.C.; Ringelberg, D.B.; Hedrick, D.B.; Nivens, D.E. Rapid Identification of Microbes from Clinical and Environmental Matrices by Mass Spectrometry. In Mass Spectrometry for the Characterization of Microorganisms; Fenselau, C., Ed.; ACS symposium series 541; American Chemical Society: Washington, DC, USA, 1993; pp. 8–17.
[7]
Green, C.T.; Scow, K.M. Analysis of phospholipid fatty acids (PLFA) to characterize microbial communities in aquifers. Hydrogeol. J 2000, 8, 126–141.
[8]
Hedrick, D.B.; White, D.C. Microbial respiratory quinones in the environment: A sensitive liquid chromatographic method. J. Microbiol. Methods 1986, 5, 243–254.
[9]
Hiraishi, A. Isoprenoid quinones as biomarkers of microbial populations in the environment. J. Biosci. Bioeng 1999, 88, 449–460.
[10]
Collins, M.D.; Jones, D. Distribution of isoprenoid quinone structural types in bacteria and their taxonomic implications. Microbiol. Rev 1981, 45, 316–354.
[11]
Joergensen, R.G.; Wichern, F. Quantitative assessment of the fungal contribution to microbial tissue in soil. Soil Biol. Biochem 2008, 40, 2977–2991.
[12]
Smith, C.A.; Phiefer, C.B.; Macnaughton, S.J.; Peacock, A.; Burkhalter, R.S.; Kirkegaard, R.; White, D.C. Quantitative lipid biomarker detection of unculturable microbes and chlorine exposure in water distribution system biofilms. Water Res 2000, 34, 2683–2688.
[13]
Gattinger, A. Phospholipid etherlipid and phospholipid fatty acid fingerprints in selected euryarchaeotal monocultures for taxonomic profiling. FEMS Microbiol. Lett 2002, 213, 133–139.
[14]
Bligh, E.G.; Dyer, W.J. A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol 1959, 37, 911–917.
[15]
Geyer, R.; Peacock, A.D.; White, D.C.; Lytle, C.; Van Berkel, G.J. Atmospheric pressure chemical ionization and atmospheric pressure photoionization for simultaneous mass spectrometric analysis of microbial respiratory ubiquinones and menaquinones. J. Mass Spectrom 2004, 39, 922–929.
[16]
Nishihara, M.; Koga, Y. Extraction and composition of polar lipids from the archaebacterium, Methanobacterium thermoautotrophicum: Effective extraction of tetraether lipids by an acidified solvent. J. Biochem 1987, 101, 997–1005.
[17]
White, P.M. Pressurized liquid extraction of soil microbial phospholipid and neutral lipid fatty acids. J. Agric. Food Chem 2009, 57, 7171–7177.
[18]
Pinkart, H.C.; Devereux, R.; Chapman, P.J. Rapid separation of microbial lipids using solid phase extraction columns. J. Microbiol. Methods 1998, 34, 9–15.
[19]
Cescut, J.; Severac, E.; Molina-Jouve, C.; Uribelarrea, J.L. Optimizing pressurized liquid extraction of microbial lipids using the response surface method. J. Chromatogr. A 2011, 1218, 373–379.
[20]
Pourmortazavi, S.M.; Hajimirsadeghi, S.S. Supercritical fluid extraction in plant essential and volatile oil analysis. J. Chromatogr. A 2007, 1163, 2–24.
[21]
Irvan; Atsuta, Y.; Saeki, T.; Daimon, H.; Fujie, K. Supercritical carbon dioxide extraction of ubiquinones and menaquinones from activated sludge. J. Chromatogr. A 2006, 1113, 14–19.
[22]
Hanif, M.; Atsuta, Y.; Fujie, K.; Daimon, H. Supercritical fluid extraction of microbial phospholipid fatty acids from activated sludge. J. Chromatogr. A 2010, 1217, 6704–6708.
[23]
White, D.C.; Lytle, C.A.; Gan, Y.M.; Piceno, Y.M.; Wimpee, M.H.; Peacock, A.D.; Smith, C.A. Flash detection/identification of pathogens, bacterial spores and bioterrorism agent biomarkers from clinical and environmental matrices. J. Microbiol. Methods 2002, 48, 139–147.
[24]
Hawthorne, S.B.; Miller, D.J.; Nivens, D.E.; White, D.C. Supercritical fluid extraction of polar analytes using in situ chemical derivatization. Anal. Chem 1992, 64, 405–412.
[25]
Gharaibeh, A.A.; Voorhees, K.J. Characterization of lipid fatty acids in whole-cell microorganisms using in situ supercritical fluid derivatization/extraction and GC/MS. Anal. Chem 1996, 68, 2805–2810.
[26]
Hedrick, D.B.; Guckert, J.B.; White, D.C. Archaebacterial ether lipid diversity analyzed by supercritical fluid chromatography: Integration with a bacterial lipid protocol. J. Lipid Res 1991, 32, 659–666.
[27]
Hoogerbrugge, R.; Stolker, A.A.M.; Barendregt, A.; Hogendoorn, E.A. A systematic approach for optimisation of supercritical-fluid extraction of polycyclic aromatic hydrocarbons from earthworms. Anal. Bioanal. Chem 2003, 377, 715–722.
[28]
Marsili, R.; Callahan, D. Comparison of a liquid solvent extraction technique and supercritical fluid extraction for the determination of α- and β-carotene in vegetable. J. Chromatogr. Sci 1993, 31, 422–428.
[29]
Roop, R.K.; Akgerman, A.; Dexter, B.J.; Irvin, T.R. Extraction of phenol from water with supercritical carbon dioxide. J. Supercrit. Fluids 1989, 2, 51–56.
IUPAC-IUB Commission on Biochemical Nomenclature. The nomenclature of lipids. Biochem. J. 1978, 171, 21–35.
[32]
IUPAC-IUB Commission on Biochemical Nomenclature. The nomenclature of quinones with isoprenoid side-chains. Pure Appl. Chem. 1974, 38, 439–447.
[33]
Virtue, P.; Nichols, P.D.; Boon, P.I. Simultaneous estimation of microbial phospholipid fatty acids and diether lipids by capillary gas chromatography. J. Microbiol. Methods 1996, 25, 177–185.
[34]
McKinley, V.L.; Peacock, A.D.; White, D.C. Microbial community PLFA and PHB responses to ecosystem restoration in tallgrass prairie soils. Soil Biol. Biochem 2005, 37, 1946–1958.
[35]
Katayama, A.; Funasaka, K.; Fujie, K. Changes in the respiratory quinone profile of a soil treated with pesticides. Biol. Fertil. Soils 2001, 33, 454–459.
[36]
Koga, Y.; Nishihara, M.; Morii, H.; Matsushita, M.A. Ether polar lipids of methanogenic bacteria: Structures, comparative aspects, and biosynthesis. Microbiol. Rev 1993, 57, 164–182.
[37]
MacNicol, R.D.; Beckett, P.H.T. The distribution of heavy metals between the principal components of digested sewage sludge. Water Res 1989, 23, 199–206.
[38]
Okabe, A.; Toyota, K.; Kimura, M. Seasonal variations of phospholipid fatty acid composition in the floodwater of a Japanese paddy field under a long-term fertilizer trial. Soil Sci. Plant Nutr 2000, 46, 177–188.
[39]
Montgomery, D.C. Design and Analysis of Experiment, 6th ed. ed.; John Wiley & Sons: New York, NY, USA, 2005; pp. 427–439.
