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Zukunft, S. ; Prehn, C. ; Röhring, C.* ; Möller, G. ; Hrabě de Angelis, M. ; Adamski, J. ; Tokarz, J.

High-throughput extraction and quantification method for targeted metabolomics in murine tissues.

Metabolomics 14:18 (2018)

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	Open Access Gold (Paid Option)

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Introduction Global metabolomics analyses using body fluids provide valuable results for the understanding and prediction of diseases. However, the mechanism of a disease is often tissue-based and it is advantageous to analyze metabolomic changes directly in the tissue. Metabolomics from tissue samples faces many challenges like tissue collection, homogenization, and metabolite extraction. Objectives We aimed to establish a metabolite extraction protocol optimized for tissue metabolite quantification by the targeted metabolomics AbsoluteIDQ (TM) p180 Kit (Biocrates). The extraction method should be non-selective, applicable to different kinds and amounts of tissues, monophasic, reproducible, and amenable to high throughput. Methods We quantified metabolites in samples of eleven murine tissues after extraction with three solvents (methanol, phosphate buffer, ethanol/phosphate buffer mixture) in two tissue to solvent ratios and analyzed the extraction yield, ionization efficiency, and reproducibility. Results We found methanol and ethanol/phosphate buffer to be superior to phosphate buffer in regard to extraction yield, reproducibility, and ionization efficiency for all metabolites measured. Phosphate buffer, however, outperformed both organic solvents for amino acids and biogenic amines but yielded unsatisfactory results for lipids. The observed matrix effects of tissue extracts were smaller or in a similar range compared to those of human plasma. Conclusion We provide for each murine tissue type an optimized high-throughput metabolite extraction protocol, which yields the best results for extraction, reproducibility, and quantification of metabolites in the p180 kit. Although the performance of the extraction protocol was monitored by the p180 kit, the protocol can be applicable to other targeted metabolomics assays.

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Publication type Article: Journal article

Document type Scientific Article

Corresponding Author

Keywords Metabolomics ; Tissue Extraction ; Lipids ; Amino Acids ; Biogenic Amines ; Acylcarnitines; Chromatography/tandem Mass-spectrometry; Electrospray-ionization; Insulin-resistance; Breast-cancer; Liquid; Suppression; Standards; Taurine; Sample; Mice

ISSN (print) / ISBN 1573-3882

e-ISSN 1573-3890

Journal Metabolomics

Quellenangaben Volume: 14, Issue: 1, Article Number: 18

Publisher Springer

Publishing Place New York, NY

Non-patent literature Publications

Reviewing status Peer reviewed

Institute(s) Molekulare Endokrinologie und Metabolismus (MEM)
Institute of Experimental Genetics (IEG)