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Comparing the extraction performance in mouse plasma of different biphasic methods for polar and nonpolar compounds

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Item Type:Article
Title:Comparing the extraction performance in mouse plasma of different biphasic methods for polar and nonpolar compounds
Creators Name:Gutmann, F. and Fritsche-Guenther, R. and Dias, D.B. and Kirwan, J.A.
Abstract:Many metabolomic studies are interested in both polar and nonpolar analyses. However, the available sample volume often precludes multiple separate extractions. Therefore, there are major advantages in performing a biphasic extraction and retaining both phases for subsequent separate analyses. To be successful, such approaches require the method to be robust and repeatable for both phases. Hence, we determined the performance of three extraction protocols, plus two variant versions, using 25 μL of commercially available mouse plasma. The preferred option for nonpolar lipids was a modified diluted version of a method employing methyl tert-butyl ether (MTBE) suggested by Matyash and colleagues due to its high repeatability for nonpolar compounds. For polar compounds, the Bligh-Dyer method performs best for sensitivity but with consequentially poorer lipid performance. Overall, the scaled-down version of the MTBE method gave the best overall performance, with high sensitivity for both polar and nonpolar compounds and good repeatability for polar compounds in particular.
Keywords:Lipidomics, Central Carbon Metabolism, Targeted Methods, Biphasic Extraction, Validation, Animals, Mice
Source:Journal of Proteome Research
ISSN:1535-3893
Publisher:American Chemical Society
Date:6 February 2024
Official Publication:https://doi.org/10.1021/acs.jproteome.3c00596
PubMed:View item in PubMed

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