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
Email	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.

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.