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Optimization of Folch, Bligh-Dyer, and Matyash Sample-to-Extraction Solvent Ratios for Mammalian Cell- and Plasma-Based Lipidomics Studies

Published

Author(s)

John Bowden, Candice Z. Ulmer, Christina Jones, Richard A. Yost, Timothy J. Garrett

Abstract

In order to investigate changes in the lipidome, analysis by ultra-high performance liquid chromatography - high resolution mass spectrometry (UHPLC-HRMS) typically requires the extraction of lipid content from sample matrices using optimized, matrix-specific conditions. The Folch, Bligh-Dyer, and Matyash methods, while promising approaches, were originally tailored to specific matrices (brain tissue, fish muscle, and E. coli, respectively). Thus, the sample-to-solvent ratios for these methods should be vetted for the sample matrix of interest prior to employment. This study evaluated the appropriate sample-to-extraction solvent ratios for mammalian cellular and plasma-based lipidomics studies. This work also evaluated the multi- omic capability of each biphasic lipid extraction method in an effort to provide a workflow capable of increasing analyte coverage in a single analysis, thus providing a more complete understanding of complex biological systems. Results showed that the Folch method was better suited for the extraction of polar and non-polar lipids from mammalian cells, more specifically, Jurkat T-lymphocyte cells. An increase in the plasma sample-to-solvent ratio from 1:4, 1:10, 1:20, to 1:100 (v/v) resulted in a gradual increase in the peak area of metabolite (aqueous layer) and lipid (organic layer) species for each extraction method interrogated in this study. The Bligh-Dyer and Folch methods yielded comparable results for the 1:20 and 1:100 (v/v) plasma sample-to-solvent ratios for both, metabolite and lipid species.
Citation
Analytica Chimica ACTA

Keywords

lipidomics, metabolomics, multi-omics, liquid-liquid extraction, sample preparation, LC-MS, mammalian cells, plasma, SRM 1950

Citation

Bowden, J. , Ulmer, C. , Jones, C. , Yost, R. and Garrett, T. (2018), Optimization of Folch, Bligh-Dyer, and Matyash Sample-to-Extraction Solvent Ratios for Mammalian Cell- and Plasma-Based Lipidomics Studies, Analytica Chimica ACTA, [online], https://doi.org/10.1016/j.aca.2018.08.004, https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=924616 (Accessed March 4, 2024)
Created December 10, 2018, Updated October 12, 2021