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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Project:

Project ID:	PR001238
Project DOI:	doi: 10.21228/M8PH6J
Project Title:	Analysis of ω-3 fatty acids and their derivatives in lungs from hypoxia-induced pulmonary hypertension (PH) mice.
Project Summary:	Pulmonary arterial hypertension (PAH) is a rare, fatal disease that causes idiopathic pulmonary artery stenosis, which leads to increased pulmonary artery pressure and this chronic pressure-overload eventually results in right heart failure and death. Although the available therapies for PH have notably improved the survival of patients with PAH, there is still a significant portion of patients who do not achieve the expected efficacy. Here, in order to identify bioactive lipids which are protective against pulmonary hypertension (PH), we performed comprehensive lipidomic analysis of lung samples from hypoxia-induced PH mice.
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

Subject:

Subject ID:	SU002029
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Gender:	Male

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id	local_sample_id	Time after hypoxia (days)
SA184038	Day0-1	-
SA184039	Day0-3	-
SA184040	Day0-2	-
SA184044	Day14-3	14
SA184045	Day14-1	14
SA184046	Day14-2	14
SA184047	Day28-3	28
SA184048	Day28-2	28
SA184049	Day28-1	28
SA184041	Day4-3	4
SA184042	Day4-1	4
SA184043	Day4-2	4

Showing results 1 to 12 of 12

Showing results 1 to 12 of 12

Collection:

Collection ID:	CO002022
Collection Summary:	Wild-type male mice (C57BL/6J) were anaesthetized and tissues were harvested. Isolated mouse tissues were rinsed with cold PBS and immediately frozen in liquid nitrogen.
Sample Type:	Lung

Treatment:

Treatment ID:	TR002041
Treatment Summary:	In chronic hypoxia mice model, the male mice were housed in a hypoxic chamber (10 % O2) maintained using a hypoxic air generator (TEIJIN) and monitored with an O2 analyzer (JIKO-255), for 4, 14 or 28 day. Animals were fed a standard diet.

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).

Combined analysis:

Analysis ID	AN003176
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Shimadzu Nexera UC/s system (Shimadzu)
Column	Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um)
MS Type	ESI
MS instrument type	Triple quadrupole
MS instrument name	ABI Sciex 4500 Qtrap
Ion Mode	NEGATIVE
Units	ng/mg tissue

Chromatography:

Chromatography ID:	CH002348
Instrument Name:	Shimadzu Nexera UC/s system (Shimadzu)
Column Name:	Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um)
Column Temperature:	40
Flow Gradient:	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).
Flow Rate:	(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)
Solvent A:	100% water; 0.1% acetic acid; 10 mM ammonium acetate
Solvent B:	80% acetonitrile/20% methanol; 10 mM ammonium acetate
Chromatography Type:	Reversed phase

MS:

MS ID:	MS002954
Analysis ID:	AN003176
Instrument Name:	ABI Sciex 4500 Qtrap
Instrument Type:	Triple quadrupole
MS Type:	ESI
MS Comments:	The instrument parameters of QTRAP4500 for negative ion mode were as follows: curtain gas, 10 psi; ionspray voltage, −4500 V; temperature, 600°C; ion source gas 1, 70 psi; ion source gas 2, 80 psi. and quantification was performed using MultiQuant™ software (SCIEX).
Ion Mode:	NEGATIVE