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Classification of bacteria by simultaneous methylation–solid phase microextraction and gas chromatography/mass spectrometry analysis of fatty acid methyl esters

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Abstract

Direct methylation and solid-phase microextraction (SPME) were used as a sample preparation technique for classification of bacteria based on fatty acid methyl ester (FAME) profiles. Methanolic tetramethylammonium hydroxide was applied as a dual-function reagent to saponify and derivatize whole-cell bacterial fatty acids into FAMEs in one step, and SPME was used to extract the bacterial FAMEs from the headspace. Compared with traditional alkaline saponification and sample preparation using liquid–liquid extraction, the method presented in this work avoids using comparatively large amounts of inorganic and organic solvents and greatly decreases the sample preparation time as well. Characteristic gas chromatography/mass spectrometry (GC/MS) of FAME profiles was achieved for six bacterial species. The difference between Gram-positive and Gram-negative bacteria was clearly visualized with the application of principal component analysis of the GC/MS data of bacterial FAMEs. A cross-validation study using ten bootstrap Latin partitions and the fuzzy rule building expert system demonstrated 87 ± 3% correct classification efficiency.

 

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Acknowledgements

The Center for Chemical Instrumentation and the Department of Chemistry and Biochemistry at Ohio University are acknowledged for their financial support. Xiaobo Sun, Zhanfeng Xu, and Weiying Lu are thanked for their helpful comments and suggestions.

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Authors and Affiliations

  1. Clippinger Laboratories, Center for Intelligent Chemical Instrumentation, Department of Chemistry and Biochemistry, Ohio University, Athens, OH, 45701-2979, USA

    Yao Lu & Peter B. Harrington

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  1. Yao Lu
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  2. Peter B. Harrington
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Correspondence to Peter B. Harrington.

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Lu, Y., Harrington, P.B. Classification of bacteria by simultaneous methylation–solid phase microextraction and gas chromatography/mass spectrometry analysis of fatty acid methyl esters. Anal Bioanal Chem 397, 2959–2966 (2010). https://doi.org/10.1007/s00216-010-3840-3

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  • DOI: https://doi.org/10.1007/s00216-010-3840-3

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