Summary of Study ST001210

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 PR000813. The data can be accessed directly via it's Project DOI: 10.21228/M8KX27 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001210
Study TitleComprehensive UHPLC-MS/MS lipidomics profiling to study effects of betulin on keratinocytes
Study SummaryLipidomics analysis of betulin in human primary keratinocytes to monitor alterations in the lipid profiles induced by treatment with betulin
Institute
Eberhard Karls University of Tübingen
DepartmentInstitute of Pharmaceutical Sciences
LaboratoryPharmaceutical (Bio-)Analysis
Last NameCalderon
First NameCarlos
AddressAuf der Morgenstelle 8 (Haus B), D-72076, Tuebingen, baden württemberg, 72076, Germany
Emailcarlos.calderon@uni-tuebingen.de
Phone+49 (0)7071 29 74009
Submit Date2019-07-05
Num Groups2
Total Subjects20
Study CommentsEberhard-Karls-University Tuebingen
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2019-09-23
Release Version1
Carlos Calderon Carlos Calderon
https://dx.doi.org/10.21228/M8KX27
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP001285
Sampleprep Summary:Lipid extraction was performed with IPA:H2O (90:10 v/v). Briefly, a 5% (v/v) SPLASH Lipidomix solution was prepared in MeOH and then 50 µL of the diluted solution was added to the cell pellet. Next, 4.95 mL of IPA:H2O (90:10 v/v) were added. Samples were vortexed (30 s), sonicated (2 min) and vortexed again (30 s) to disrupt the pellet. Incubation on ice was continued on a shaker for 1h (500 rpm, 60 min). Samples were centrifuged (3500 rcf, 10 min) and the supernatant was transferred to 15 mL falcon tubes. Samples were dried in an evaporator for 10 hours under nitrogen protection. Dried extract was resuspended in 100 µL of methanol containing odd-chained lipid standards LPC 17:1 and PC 17:0-20:4 at 500 ng/mL and 125 ng/mL, respectively. Sonication (2 min) and vortexing (30 s) were applied to ensure that lipids were not stuck to the surface of the extraction container. Then, samples were centrifuged (3500 rcf, 10 min) and the supernatant was transferred to vials for LC-MS measurements. Quality control (QC) sample was prepared by pooling together 15 µL aliquot of each sample. Samples were measured randomly and QC samples were run at the beginning, at the end and every fifth sample during the sequence.
Sampleprep Protocol Filename:ccalcas_Extraction_and_treatment.pdf
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