Next Article in Journal
Urinary Elimination of Ecdysterone and Its Metabolites Following a Single-Dose Administration in Humans
Previous Article in Journal
Towards Standards for Human Fecal Sample Preparation in Targeted and Untargeted LC-HRMS Studies
Previous Article in Special Issue
Untargeted Lipidomic Profiling of Dry Blood Spots Using SFC-HRMS
Article

Accurate Lipid Quantification of Tissue Homogenates Requires Suitable Sample Concentration, Solvent Composition, and Homogenization Procedure—A Case Study in Murine Liver

Institute of Clinical Chemistry and Laboratory Medicine, University Hospital of Regensburg, Franz-Josef-Strauß-Allee 11, 93053 Regensburg, Germany
*
Author to whom correspondence should be addressed.
Academic Editors: Leonardo Tenori and Peter Meikle
Metabolites 2021, 11(6), 365; https://doi.org/10.3390/metabo11060365
Received: 12 May 2021 / Revised: 25 May 2021 / Accepted: 6 June 2021 / Published: 8 June 2021
(This article belongs to the Special Issue Mass Spectrometry-Based Lipidomics)
Lipidomics aim to quantify lipid species in all kinds of samples, including tissues. To subject a fixed amount of sample to various workflows, tissue homogenates were frequently prepared at defined concentrations in water or by addition of organic solvents. Here, we investigated this first step of tissue lipidomics by quantitative flow injection analysis coupled to Fourier-Transform mass spectrometry (FTMS). The influence of sample concentration, solvent composition, and homogenization procedure on the recovery of lipids was studied in murine liver. Liver homogenates were prepared either by grinding tissue in liquid nitrogen or by bead-based homogenization. Ground samples were dissolved at different concentrations in water, methanol, and water/methanol = 1/1 (v/v). Here, lipid recovery depends on solvent composition and sample concentration. The recovery of nonpolar lipid classes, including triglycerides and cholesteryl ester, was decreased in methanolic homogenates. In contrast, due to superior dispersion of precipitates, bead-based homogenization resulted in efficient lipid recovery independent of the solvent composition. However, lipid distribution within samples, i.e., lipid content of supernatant and pellet following centrifugation, was altered substantially by solvent composition. In conclusion, accurate lipid quantification of tissue homogenates requires evaluation of solvent composition, sample concentration, as well as the homogenization method to guarantee efficient lipid recovery. Due to a potential loss of lipids, removal of precipitates by centrifugation prior to lipid extraction should be avoided. View Full-Text
Keywords: lipidomics; lipids; extraction; recovery; solvent; quantification; preanalytics; tissue homogenization; mass spectrometry lipidomics; lipids; extraction; recovery; solvent; quantification; preanalytics; tissue homogenization; mass spectrometry
Show Figures

Graphical abstract

MDPI and ACS Style

Höring, M.; Krautbauer, S.; Hiltl, L.; Babl, V.; Sigruener, A.; Burkhardt, R.; Liebisch, G. Accurate Lipid Quantification of Tissue Homogenates Requires Suitable Sample Concentration, Solvent Composition, and Homogenization Procedure—A Case Study in Murine Liver. Metabolites 2021, 11, 365. https://doi.org/10.3390/metabo11060365

AMA Style

Höring M, Krautbauer S, Hiltl L, Babl V, Sigruener A, Burkhardt R, Liebisch G. Accurate Lipid Quantification of Tissue Homogenates Requires Suitable Sample Concentration, Solvent Composition, and Homogenization Procedure—A Case Study in Murine Liver. Metabolites. 2021; 11(6):365. https://doi.org/10.3390/metabo11060365

Chicago/Turabian Style

Höring, Marcus, Sabrina Krautbauer, Louisa Hiltl, Verena Babl, Alexander Sigruener, Ralph Burkhardt, and Gerhard Liebisch. 2021. "Accurate Lipid Quantification of Tissue Homogenates Requires Suitable Sample Concentration, Solvent Composition, and Homogenization Procedure—A Case Study in Murine Liver" Metabolites 11, no. 6: 365. https://doi.org/10.3390/metabo11060365

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop