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Open AccessArticle

Influence of Drying Method on NMR-Based Metabolic Profiling of Human Cell Lines

Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, USA
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Author to whom correspondence should be addressed.
Metabolites 2019, 9(11), 256; https://doi.org/10.3390/metabo9110256
Received: 30 August 2019 / Revised: 24 October 2019 / Accepted: 28 October 2019 / Published: 31 October 2019
(This article belongs to the Special Issue Sample Preparation in Metabolomics)
Metabolic profiling of cell line and tissue extracts involves sample processing that includes a drying step prior to re-dissolving the cell or tissue extracts in a buffer for analysis by GC/LC-MS or NMR. Two of the most commonly used drying techniques are centrifugal evaporation under vacuum (SpeedVac) and lyophilization. Here, NMR spectroscopy was used to determine how the metabolic profiles of hydrophilic extracts of three human pancreatic cancer cell lines, MiaPaCa-2, Panc-1 and AsPC-1, were influenced by the choice of drying technique. In each of the three cell lines, 40–50 metabolites were identified as having statistically significant differences in abundance in redissolved extract samples depending on the drying technique used during sample preparation. In addition to these differences, some metabolites were only present in the lyophilized samples, for example, n-methyl-α-aminoisobutyric acid, n-methylnicotimamide, sarcosine and 3-hydroxyisovaleric acid, whereas some metabolites were only present in SpeedVac dried samples, for example, trimethylamine. This research demonstrates that the choice of drying technique used during the preparation of samples of human cell lines or tissue extracts can significantly influence the observed metabolome, making it important to carefully consider the selection of a drying method prior to preparation of such samples for metabolic profiling. View Full-Text
Keywords: metabonomics; metabolomics; metabolic profiling; NMR; nuclear magnetic resonance spectroscopy; cell line; human cell line; MiaPaCa-2; Panc-1; AsPC-1 metabonomics; metabolomics; metabolic profiling; NMR; nuclear magnetic resonance spectroscopy; cell line; human cell line; MiaPaCa-2; Panc-1; AsPC-1
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Petrova, I.; Xu, S.; Joesten, W.C.; Ni, S.; Kennedy, M.A. Influence of Drying Method on NMR-Based Metabolic Profiling of Human Cell Lines. Metabolites 2019, 9, 256.

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