Summary of study ST001691

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001087. The data can be accessed directly via it's Project DOI: 10.21228/M8698W This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST001691
Study TitleLC-MS untargeted lipidomics of primary human fibroblasts
Study TypeLC-MS untargeted lipidomics of fibroblasts
Study SummaryWe did LC-MS untargeted lipidomics of primary human fibroblasts to have a comprehensive overview of their lipidome in positive ion mode
Institute
École polytechnique fédérale de Lausanne (EPFL)
Last NameCapolupo
First NameLaura
AddressRoute Cantonale
Emaillaura.capolupo@epfl.ch
Phone+41 21 693 42 79
Submit Date2021-02-10
Raw Data AvailableYes
Raw Data File Type(s).raw
Analysis Type DetailLC-MS
Release Date2021-05-01
Release Version1
Laura Capolupo Laura Capolupo
https://dx.doi.org/10.21228/M8698W
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP001774
Sampleprep Summary:Total lipid extracts were prepared using a standard MTBE protocol followed by a methylamine treatment for total lipid analysis by mass spectrometry 7. Briefly, cell pellet was resuspended in 100 μL H2O. 360 μL methanol and 1.2 mL of MTBE were added and samples were placed for 10 min on a vortex at 4 °C followed by incubation for 1 h at room temperature on a shaker. Phase separation was induced by addition of 200 μL of H2O. After 10 min at room temperature, samples were centrifuged at 1000 g for 10 min. The upper (organic) phase was transferred into a glass tube and the lower phase was re-extracted with 400 μL artificial upper phase [MTBE/methanol/H2O (10:3:1.5, v/v/v)]. The combined organic phases were dried in a vacuum concentrator. Lipids where then resuspended in 500 μL of CHCl3 and divided in two aliquots for a further methylamine treatment for sphingo- and glycosphingolipids analysis. In details, 500 μL of freshly prepared monomethylamine reagent [methylamine/H2O/nbutanol/ methanol (5:3:1:4, (v/v/v/v)] was added to the dried lipid extract and then incubated at 53 °C for 1 h in a water bath. Lipids were cooled to room temperature and then dried. The dried lipid extract was then extracted by n-butanol extraction using 300 μL water-saturated nbutanol and 150 μL H2O. The organic phase was collected, and the aqueous phase was reextracted twice with 300 μL water-saturated n-butanol. The organic phases were pooled and dried in a vacuum concentrator.
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