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Lipidomics and Transcriptomics in Neurological Diseases
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Evaluation of tissue-specific extraction protocols for comprehensive lipid profiling.

Dana Hicks1, Monique J Ryan1, Amira Allahham2

  • 1Australian National Phenome Centre, Health Futures Institute, Harry Perkins Institute, Murdoch University, Perth, WA, Australia; Centre for Computational and Systems Medicine, Health Futures Institute, Harry Perkins Institute, Murdoch University, Perth, WA, Australia.

Analytica Chimica Acta
|March 2, 2025
PubMed
Summary

Optimizing lipid extraction for metabolic phenotyping requires tissue-specific protocols. This study identifies the best lipid extraction methods for adipose, liver, and heart tissues, enhancing lipidomic analysis and understanding of metabolic changes.

Keywords:
Extraction solventsLipidomics∗Lipids∗Mass spectrometryMurine tissueSample preparation

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Area of Science:

  • Lipidomics
  • Metabolic Phenotyping
  • Biochemistry

Background:

  • Robust tissue pre-treatment and lipid extraction are vital for accurate metabolic phenotyping and lipid profile interpretation.
  • Numerous lipid extraction methods exist, with technique choice significantly impacting analysis outcomes.
  • This study comprehensively evaluates six liquid-liquid extraction methods for lipidomic tissue analysis using liquid chromatography-mass spectrometry.

Purpose of the Study:

  • To evaluate six liquid-liquid extraction methods for comprehensive lipid profiling across diverse tissue types (adipose, liver, heart).
  • To determine tissue-specific optimal extraction protocols for enhanced lipidomic analysis.
  • To validate optimized methods in a diet-induced metabolic change study in mice.

Main Methods:

  • Six liquid-liquid extraction methods (three biphasic, three monophasic) were tested on lyophilized and fresh frozen adipose, liver, and heart tissues.
  • Extraction efficiency, lipid coverage, yield, and reproducibility were assessed.
  • Optimized methods were validated in C57BL/6 mice subjected to high-fat diet (HFD) and normal diet (ND) interventions.

Main Results:

  • Optimal lipid extraction methods were tissue-specific: butanol:methanol (BUME) (3:1) for adipose, methyl tert-butyl ether (MTBE) with ammonium acetate for liver, and BUME (1:1) for heart.
  • Significant differences in lipid species coverage and reliability were observed based on tissue type.
  • Validated methods revealed distinct, tissue-specific lipid profiles differentiating HFD and ND groups in adipose and liver tissues.

Conclusions:

  • Tailored, tissue-specific lipid extraction protocols significantly improve lipid species detection sensitivity and reliability in comprehensive lipidomics.
  • These optimized protocols provide robust tools for identifying lipid changes and understanding tissue-specific metabolic processes.
  • The findings support the development of bespoke protocols for diverse research and clinical applications in metabolic studies.