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Sample Set Information

Title Grobal triacylglycerol analysis in mouse liver and white adipose tissue (WAT) by high resolution LC/ESI-QTOF MS/MS
Description An effective method was established for global analysis of triacylglycerol (TAG) molecular species from complex lipid mixtures of mouse liver and white adipose tissue (WAT) using reverse-phase high resolution liquid chromatography (LC) coupled with electrospray ionization (ESI)-quadrapole/time of flight hybrid mass spectrometer (QTOF-MS).
Authors Ikeda K, Oike Y, Shimizu T, Taguchi R
Reference Ikeda K, Oike Y, Shimizu T, Taguchi R (2009) Journal of Chromatography B 877: 2639-2647
Comment The metadata is prepared by the Metabolonote administrator (Sakurai N).

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Sample Information

Title Mouse white adipose tissue (WAT)
Organism - Scientific Name Mus musculus
Organism - ID NCBI taxonomy:10090
Compound - ID
Compound - Source
Preparation Mice (C57BL/6) were purchased from SLC, Shizuoka, Japan. White adipose tissue was extracted and used.
Sample Preparation Details ID

Analytical Method Information

Title UPLC-Q-TOF-MS, ESI Positive analysis
Method Details ID MS1
Sample Amount 20 ug
Comment The amount of white apidose tissue contained in the 2 ul solution typically injected to the RPLC.

Analytical Method Details Information

Title ESI positive LC-MS analysis of triacylglycerols (TAG)
Instrument ACQUITY UPLC system (Waters Corporation), QTOF micro (Micromass)
Instrument Type UPLC-QTOF-MS
Ionization ESI
Ion Mode Positive
Description Sample Preparation

Tryacylglycerols (TAGs) in C57BL/6 mouse (purchased from SLC, Shizuoka, Japan) liver (100 mg) and white adipose tissue (WAT) (100 mg) were extracted using Bligh and Dyer's method in the following procedure: the liver and WAT were homogenized with 6 mL chloroform/methanol (1:2) for 10 strokes and left for 1 h at room temperature. Each phase separation was achieved by adding 2 mL chloroform and 2 mL water. After vortexing, the mixture was centrifuged at 3000 rpm for 10 min. The bottom organic layer containing the total lipid extract was collected and dried under a gentle stream of nitrogen, and then dissolved in chloroform / methanol (2:1) at a concentration of 10 ug (tissue weight) /uL.

LC conditions

Reverse-phased LC separation was achieved using ACQUITY UPLC BEH column (1.0 mm×150 mm i.d., particle size 1.7 um, Waters Corporation, Milford, MA, USA) at 45C. The mobile phase was acetonitrile / methanol / water : 19 / 19 / 2 (0.1% acetic acid + 0.028% ammonia) (A), and isopropanol (0.1% acetic acid + 0.028% ammonia) (B), and the composition was produced by mixing these solvents. The gradient consisted of holding solvent (A/B:90/10) for 5 min, then linearly converting to solvent (A/B:60/40) for 35 min and then linearly converting solvent (A/B:45/55) for 50 min. The mobile phase was pumped at a flow rate of 50 uL/min and the column pressure was adjusted lower than 5000 psi. Typically, 2 uL of sample solution was injected.

ESI-MS conditions

The ESI-MS analysis was performed using a quadrapole/time of flight hybrid mass spectrometer (QTOF microTM, Micromass, Manchester, UK) with an ACQUITY UPLC system (Waters Corporation). The mass range of the instrument was set at m/z 200–1100 and scan duration of MS and MS/MS at 0.5 s in positive ion mode. The capillary voltage was set at 3.5 kV, cone voltage at 30V, and source block temperature at 100C. The collision gas used for MS/MS experiments was argon (7.5×10E5 mbar), and the collision energy was set at 30 V.


Data Analysis Information

Title Identification of triacylglycerols with 2D maps
Data Analysis Details ID DS1
Recommended decimal places of m/z default

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The mass spectrum data are available at Bio-MassBank.

Data Analysis Details Information

Title Identification of triacylglycerols with 2D maps
Description Two-dimensional (2D) maps were constructed for effective profiling of triacylglycerols (TAGs), and individual structures of TAGs were correctly identified by the elution profile and MS/MS.
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