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

Published

December 11, 2018

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

Pub Type

Journals

Download Paper

https://doi.org/10.1016/j.aca.2018.08.004

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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 December 5, 2024)