Summary of Study ST001951

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 PR001238. The data can be accessed directly via it's Project DOI: 10.21228/M8PH6J This work is supported by NIH grant, U2C- DK119886.

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

Study ID	ST001951
Study Title	Quantification of ω-3 fatty acids and their derivatives in lungs from hypoxia-induced pulmonary hypertension (PH) mice.
Study Summary	Using a liquid chromatography tandem mass spectrometry (LC-MS/MS) system, we quantified the ω-3 fatty acid (EPA and DHA) metabolites in the lung tissues of mice with PH induced by chronic hypoxia (10% oxygen concentration).
Institute	University of Tokyo
Last Name	Kono
First Name	Nozomu
Address	7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Email	nozomu@mol.f.u-tokyo.ac.jp
Phone	+81-3-5841-4723
Submit Date	2021-10-17
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	LC-MS
Release Date	2022-03-31
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP002035
Sampleprep Summary:	Lipids of tissues were extracted by the method of Bligh and Dyer. The extracted solutions were dried up with centrifugal evaporator, dissolved in methanol : isopropanol=1:1, and stored at −20°C. Fatty acid metabolites were further purified from tissues by solid-phase extraction using InertSep NH2 columns (GL Science) with deuterium-labelled internal standard (11(12)-EET-d11). Briefly, InertSep NH2 columns were precon- ditioned with 6mL of hexane and lipids extracted from tissues by the method of Bligh and Dyer were applied with 500μL of chloroform. Columns were then washed with 6mL of chloro- form/isopropanol (2/1, v/v), followed by the elution with diethyl ether/acetic acid (98/2, v/v).The extracted solutions were dried up with centrifugal evaporator, dissolved in methanol:isopro- panol=1:1, and stored at −20°C. For the detection of fatty acid metabolites, chromatographic separation was performed on a ACQUITY UPLC HSS T3 column (2.1×100 mm, 1.8 µm; Waters) maintained at 40°C using mobile phase A (water/acetic acid (100/0.1, v/v) containing 10 mM ammonium acetate) and mobile phase B (acetonitrile/methanol (4/1, v/v) containing 10 mM ammonium acetate) in a gradient programme (0–2 min: 90% A; 2–10 min: 90% A →30% A; 10–24 min: 30% A →27% A; 24–27 min: 1% A; 27–32 min: 90% A) with a flow rate of 0.2 mL/min(0–10 min), 0.1 mL/min(10–15 min),0.2 mL/min(15–24 min) and 0.5 mL/min(24–32 min).

Sampleprep ID:

SP002035

Sampleprep Summary:

Lipids of tissues were extracted by the method of Bligh and Dyer. The extracted solutions were dried up with centrifugal evaporator, dissolved in methanol : isopropanol=1:1, and stored at −20°C. Fatty acid metabolites were further purified from tissues by solid-phase extraction using InertSep NH2 columns (GL Science) with deuterium-labelled internal standard (11(12)-EET-d11). Briefly, InertSep NH2 columns were precon- ditioned with 6mL of hexane and lipids extracted from tissues by the method of Bligh and Dyer were applied with 500μL of chloroform. Columns were then washed with 6mL of chloro- form/isopropanol (2/1, v/v), followed by the elution with diethyl ether/acetic acid (98/2, v/v).The extracted solutions were dried up with centrifugal evaporator, dissolved in methanol:isopro- panol=1:1, and stored at −20°C. For the detection of fatty acid metabolites, chromatographic separation was performed on a ACQUITY UPLC HSS T3 column (2.1×100 mm, 1.8 µm; Waters) maintained at 40°C using mobile phase A (water/acetic acid (100/0.1, v/v) containing 10 mM ammonium acetate) and mobile phase B (acetonitrile/methanol (4/1, v/v) containing 10 mM ammonium acetate) in a gradient programme (0–2 min: 90% A; 2–10 min: 90% A →30% A; 10–24 min: 30% A →27% A; 24–27 min: 1% A; 27–32 min: 90% A) with a flow rate of 0.2 mL/min(0–10 min), 0.1 mL/min(10–15 min),0.2 mL/min(15–24 min) and 0.5 mL/min(24–32 min).