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A Lipid Extraction and Analysis Method for Characterizing Soil Microbes in Experiments with Many Samples
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Modified Lipid Extraction Methods for Deep Subsurface Shale.

Rawlings N Akondi1, Ryan V Trexler2, Susan M Pfiffner3

  • 1Department of Geology and Geography, West Virginia UniversityMorgantown, WV, United States.

Frontiers in Microbiology
|August 10, 2017
PubMed
Summary
This summary is machine-generated.

Investigating microbial lipid extraction from deep shale, this study found that modified Bligh and Dyer (mBD) methods, particularly with intact phospholipids, offered higher yields and better reproducibility for phospholipid fatty acids (PLFAs) and diglyceride fatty acids (DGFAs).

Keywords:
DGFAPLFAdeep subsurfacemicrobial biomassshale ecosystem

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Area of Science:

  • Geomicrobiology
  • Organic Geochemistry
  • Analytical Chemistry

Background:

  • Deep subsurface shale ecosystems are gaining interest for hydrocarbon and environmental applications.
  • Microbial functional diversity in these environments is crucial but challenging to study.
  • Lipid biomarkers like phospholipid fatty acids (PLFAs) and diglyceride fatty acids (DGFAs) are key indicators of microbial biomass and community structure.

Purpose of the Study:

  • To evaluate and compare the efficiency and reproducibility of three lipid extraction methods (modified Bligh and Dyer, Folch, and microwave-assisted extraction) for analyzing microbial lipid biomarkers in deeply buried shales.
  • To identify the most suitable method for accurate lipid biomarker recovery from complex shale matrices.

Main Methods:

  • Three lipid extraction methods were tested: modified Bligh and Dyer (mBD), Folch (FOL), and microwave-assisted extraction (MAE).
  • Lipid biomarkers were analyzed as fatty acid methyl esters (FAMEs) using gas chromatography-mass spectrometry (GC-MS).
  • Yields and reproducibility of phospholipid fatty acids (PLFAs) and diglyceride fatty acids (DGFAs) were quantified and compared across methods.

Main Results:

  • Lipid biomarker yields (PLFAs: 67-400 pmol/g; DGFAs: 600-3,000 pmol/g) varied significantly across methods.
  • The intact phospholipid Bligh and Dyer (mBD + Phos + POPC), Folch, mBD-Citrate, and MAE treatments showed relatively higher and statistically similar yields for both PLFAs and DGFAs.
  • The mBD + Phos + POPC method demonstrated better reproducibility in fatty acid profiles compared to other treatments, despite overall variability.

Conclusions:

  • The complex nature of deeply buried shale matrices presents significant challenges for lipid biomarker extraction.
  • The modified Bligh and Dyer method, particularly with intact phospholipids, appears most promising for improving lipid biomarker recovery and reproducibility in shale ecosystems.
  • Further methodological development is necessary to optimize lipid biomarker analysis in challenging deep subsurface shale environments.