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Journal article
Evaluation of Tissue-Specific Extraction Protocols for Comprehensive Lipid Profiling
Analytica chimica acta, Vol.1347, 343791
2025
Abstract
Background
Robust tissue pre-treatment and lipid extraction workflows are crucial to metabolic phenotyping studies and accurate interpretation of lipid profiles. Numerous methods for lipid extraction from tissues have been developed, and the choice of technique influences analysis. This study provides a comprehensive evaluation of six liquid-liquid extraction methods (three biphasic and three monophasic) used for lipidomic tissue analysis by liquid chromatography-mass spectrometry. Extraction methods were assessed for their suitability for comprehensive lipid profiling across diverse tissue types: adipose, liver, and heart. These techniques were compared using lyophilised and fresh frozen samples.
Results
The study revealed significant differences in the coverage and reliability of lipid species extracted using each technique, dependent on the tissue type. The optimal extraction method for adipose tissue was butanol:methanol (BUME) (3:1) which achieved the highest lipid coverage, yield and reproducibility (886 lipids with a coefficient of variation (CV) < 30 %); methyl tert-butyl ether (MTBE) with ammonium acetate was most effective for liver tissue (707 lipids CV < 30 %) and BUME (1:1) for heart tissue (311 lipids CV < 30 %). These findings showed that the most effective lipid extraction methods are highly tissue-specific, underscoring the critical need for bespoke protocols tailored to each tissue type. The optimised tissue-specific methods were validated using an intervention study in C57BL/6 mice to investigate diet-induced metabolic changes. The results demonstrated distinct discriminating lipid profiles unique to each tissue type, with 374 lipid species from 13 subclasses significantly different between high-fat diet (HFD) and normal diet (ND) in adipose tissue, while 485 lipid species from 17 subclasses were significantly different between HFD and ND in liver tissue.
Significance and novelty
This study presents a new approach to studying lipid profiles derived from diverse tissues that substantially improve comprehensive lipid species’ detection sensitivity and reliability. Our systematic evaluation provides evidence that tailored tissue-specific extraction protocols are highly valuable in comprehensive lipidomics studies, offering robust tools for reliably identifying lipid changes and facilitates a deeper understanding of tissue-specific metabolic processes in diverse research and clinical applications.
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Details
- Title
- Evaluation of Tissue-Specific Extraction Protocols for Comprehensive Lipid Profiling
- Authors/Creators
- Dana Hicks - Australian National Phenome Centre, Health Futures Institute, Harry Perkins Institute, Murdoch University, Perth, WA, AustraliaMonique J. Ryan - Murdoch University, Australian National Phenome CentreAmira Allahham - Burn Injury Research Unit, School of Biomedical Sciences, University of Western Australia, Crawley, WA, AustraliaLucy W. Barrett - Burn Injury Research Unit, School of Biomedical Sciences, University of Western Australia, Crawley, WA, AustraliaSilvia Lee - Department of Advanced Clinical and Translational Cardiovascular Imaging, Harry Perkins Institute of Medical Research, University of Western Australia, WA, AustraliaBenjamin Bartlett - Department of Advanced Clinical and Translational Cardiovascular Imaging, Harry Perkins Institute of Medical Research, University of Western Australia, WA, AustraliaHerbert Ludewick - Heart and Lung Research Institute, Perth, WA, AustraliaNatalie C. Ward - The University of Western AustraliaRuey Leng Loo - Murdoch University, Centre for Computational and Systems MedicineGirish Dwivedi - School of Medicine, University of Western Australia, Perth, WA, AustraliaJeremy K. Nicholson - University of LondonJulien Wist - Murdoch University, Centre for Computational and Systems MedicineElaine Holmes - Murdoch University, Centre for Computational and Systems MedicineNicola Gray - Murdoch University, Centre for Computational and Systems Medicine
- Publication Details
- Analytica chimica acta, Vol.1347, 343791
- Publisher
- Elsevier B.V.
- Number of pages
- 13
- Grant note
- Australian Government Research Training Program Scholarship - Commonwealth of AustraliaWestern Australian State GovernmentDepartment of Jobs, Tourism, Science and Innovation (WA) , Government of Western Australian Premier's FellowshipAustralian National Phenome Centre (ANPC)ARC Laureate Fellowship
This research is supported by an Australian Government Research Training Program Scholarship awarded by the Commonwealth of Australia to Higher Degree Research students for DH. We thank the Western Australian State Government, and the MRFF for funding the Australian National Phenome Centre (ANPC) ; Department of Jobs, Tourism, Science and Innovation (WA) , Government of Western Australian Premier's Fellowship for funding RLL and EH; ARC Laureate Fellowship for funding EH. We acknowledge the ANPC technical and academic team for their support, with particular thanks to Assoc Prof Berin Boughton.
- Identifiers
- 991005743939907891
- Copyright
- © 2025 The Authors.
- Murdoch Affiliation
- Australian National Phenome Centre; Centre for Computational and Systems Medicine
- Language
- English
- Resource Type
- Journal article
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- 2.211 Mass Spectrometry
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