{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001951","ANALYSIS_ID":"AN003176","VERSION":"1","CREATED_ON":"October 23, 2021, 3:27 pm"},

"PROJECT":{"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"},

"STUDY":{"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"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENDER":"Male"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"Day0-1",
"Factors":{"Time after hypoxia (days)":"-"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-0d1"}
},
{
"Subject ID":"-",
"Sample ID":"Day0-2",
"Factors":{"Time after hypoxia (days)":"-"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-0d2"}
},
{
"Subject ID":"-",
"Sample ID":"Day0-3",
"Factors":{"Time after hypoxia (days)":"-"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-0d3"}
},
{
"Subject ID":"-",
"Sample ID":"Day4-1",
"Factors":{"Time after hypoxia (days)":"4"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-4d1"}
},
{
"Subject ID":"-",
"Sample ID":"Day4-2",
"Factors":{"Time after hypoxia (days)":"4"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-4d2"}
},
{
"Subject ID":"-",
"Sample ID":"Day4-3",
"Factors":{"Time after hypoxia (days)":"4"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-4d3"}
},
{
"Subject ID":"-",
"Sample ID":"Day14-1",
"Factors":{"Time after hypoxia (days)":"14"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-14d1"}
},
{
"Subject ID":"-",
"Sample ID":"Day14-2",
"Factors":{"Time after hypoxia (days)":"14"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-14d2"}
},
{
"Subject ID":"-",
"Sample ID":"Day14-3",
"Factors":{"Time after hypoxia (days)":"14"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-14d3"}
},
{
"Subject ID":"-",
"Sample ID":"Day28-1",
"Factors":{"Time after hypoxia (days)":"28"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-28d1"}
},
{
"Subject ID":"-",
"Sample ID":"Day28-2",
"Factors":{"Time after hypoxia (days)":"28"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-28d2"}
},
{
"Subject ID":"-",
"Sample ID":"Day28-3",
"Factors":{"Time after hypoxia (days)":"28"},
"Additional sample data":{"RAW_FILE_NAME":"Data180501 lung oxFFA-28d3"}
}
],
"COLLECTION":{"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_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."},

"SAMPLEPREP":{"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)."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Shimadzu Nexera UC/s system (Shimadzu)","COLUMN_NAME":"ACQUITY UPLC HSS T3 column (2.1×100 mm, 1.8 µm; Waters)","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)","COLUMN_TEMPERATURE":"40","SOLVENT_A":"water/acetic acid (100/0.1, v/v) containing 10 mM ammonium acetate","SOLVENT_B":"acetonitrile/methanol (4/1, v/v) containing 10 mM ammonium acetate"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"ABI Sciex 4500 Qtrap","INSTRUMENT_TYPE":"Ion trap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","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)."},

"MS_METABOLITE_DATA":{
"Units":"ng/mg tissue",

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}

}