Summary of Study ST001332
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 PR000904. The data can be accessed directly via it's Project DOI: 10.21228/M8TX11 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.
Study ID | ST001332 |
Study Title | LCMS lipid and acyl-carnitine analysis |
Study Summary | LCMS Lipidomics and acyl-carnitine analysis. |
Institute | University of Cambridge |
Laboratory | CMaLL |
Last Name | Jenkins |
First Name | Benjamin |
Address | Department of Biochemistry, University of Cambridge, c/o Level 4, Pathology Building |
bjj25@medschl.cam.ac.uk | |
Phone | 07731103718 |
Submit Date | 2020-01-09 |
Raw Data Available | Yes |
Raw Data File Type(s) | raw(Thermo) |
Analysis Type Detail | LC-MS |
Release Date | 2020-03-30 |
Release Version | 1 |
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Sample Preparation:
Sampleprep ID: | SP001414 |
Sampleprep Summary: | C. elegans was prepared for LC-MS lipidomics and acyl-carnitine analysis as previously described (3) with minor modifications. Briefly, ~40 µL of concentrated embryos were re-suspended in 100 µL of water, then 0.4 mL of chloroform was added to each sample followed by 0.2 mL of methanol containing the stable isotope labelled acyl-carnitine internal standards (Butyryl-L-carnitine-d7 at 5 µM and Hexadecanoyl-L-carnitine-d3 at 5 µM). The samples were then homogenised by vortexing then transferred into a 2 mL Eppendorf screw-cap tube. The original container was washed out with 0.5 mL of chloroform: methanol (2: 1, respectively) and added to the appropriate 2 mL Eppendorf screw-cap tube. This was followed by the addition of 150 µL of the following stable isotope labelled internal standards (approximately 10 to 50 µM in methanol): Ceramide_C16d31, LPC_(C14:0d42), PC_(C16:0d31 / C18:1), PE_(C16:0d31 / C18:1), PG_(C16:0d31 / C18:1), PI_(C16:0d31 / C18:1), PS_(C16:0d62), SM_(C16:0d31), TG_(45:0d29) and TG_(48:0d31). Then, 400 µL of sterile water was added. The samples were vortexed for 1 min, and then centrifuged at ~20,000 rpm for 5 minutes. For the intact lipid sample preparation, 0.3 mL of the organic layer (the lower chloroform layer) was collected into a 2 mL amber glass vial (Agilent Technologies, Santa Clara California, USA) and dried down to dryness in an Eppendorf Concentrator Plus system (Eppendorf, Stevenage, UK) run for 60 minutes at 45 °C. The dried lipid samples were then reconstituted with 100 µL of 2:1:1 solution of propan-2-ol, acetonitrile and water, respectively, and then vortexed thoroughly. The lipid samples were then transferred into a 300 μL low-volume vial insert inside a 2 mL amber glass auto-sample vial ready for liquid chromatography separation with mass spectrometry detection (LC-MS) of intact lipid species. For the acyl-carnitine sample preparation, 0.2 mL of the organic layer (the lower chloroform layer) and 0.2 mL of the aqueous layer (the top water layer) were mixed into a 2 mL amber glass vial and dried down to dryness. The dried acyl-carnitine samples were then reconstituted with 100 µL of water and acetonitrile (4: 1, respectively) and thoroughly vortexed. The acyl-carnitine samples were then transferred into a 300 μL low-volume vial insert inside a 2 mL amber glass auto-sample vial ready for liquid chromatography separation with mass spectrometry detection (LC-MS) of the acyl-carnitine species. |