{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001117","ANALYSIS_ID":"AN001814","VERSION":"1","CREATED_ON":"December 26, 2018, 12:51 pm"},

"PROJECT":{"PROJECT_TITLE":"A Metabolomic study of hibernating Syrian hamster brain: in search of neuroprotective agents","PROJECT_SUMMARY":"hamster brain samples, divided in 3 groups: torpor, arousal, control group were compared via metabolomics analysis","INSTITUTE":"Universidad CEU San Pablo","LAST_NAME":"Gonzalez-Riano","FIRST_NAME":"Carolina","ADDRESS":"Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte, Boadilla del Monte, Madrid, 28668, Spain","EMAIL":"car.gonzalez@ceindo.ceu.es","PHONE":"00 34 91 3724753"},

"STUDY":{"STUDY_TITLE":"A Metabolomic study of hibernating Syrian hamster brain: in search of neuroprotective agents","STUDY_TYPE":"Multiplatform non-targeted metabolomics","STUDY_SUMMARY":"hamster brain samples, divided in 3 groups: torpor, arousal, control group were compared via metabolomics analysis","INSTITUTE":"Universidad CEU San Pablo","LABORATORY":"CEMBIO (Centre for Metabolomics and Bioanalysis)","LAST_NAME":"Gonzalez-Riano","FIRST_NAME":"Carolina","ADDRESS":"Facultad de Farmacia, Universidad CEU San Pablo, Campus Monteprincipe, Boadilla del Monte, Boadilla del Monte, Madrid, 28668, Spain","EMAIL":"car.gonzalez@ceindo.ceu.es","PHONE":"00 34 91 3724753"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mesocricetus Auratus","TAXONOMY_ID":"10036"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"A1",
"Factors":{"hibernation":"arousal"}
},
{
"Subject ID":"-",
"Sample ID":"A3",
"Factors":{"hibernation":"arousal"}
},
{
"Subject ID":"-",
"Sample ID":"A6",
"Factors":{"hibernation":"arousal"}
},
{
"Subject ID":"-",
"Sample ID":"A10",
"Factors":{"hibernation":"arousal"}
},
{
"Subject ID":"-",
"Sample ID":"A11",
"Factors":{"hibernation":"arousal"}
},
{
"Subject ID":"-",
"Sample ID":"C1",
"Factors":{"hibernation":"control"}
},
{
"Subject ID":"-",
"Sample ID":"C2",
"Factors":{"hibernation":"control"}
},
{
"Subject ID":"-",
"Sample ID":"C3",
"Factors":{"hibernation":"control"}
},
{
"Subject ID":"-",
"Sample ID":"C4",
"Factors":{"hibernation":"control"}
},
{
"Subject ID":"-",
"Sample ID":"C5",
"Factors":{"hibernation":"control"}
},
{
"Subject ID":"-",
"Sample ID":"T2",
"Factors":{"hibernation":"torpor"}
},
{
"Subject ID":"-",
"Sample ID":"T4",
"Factors":{"hibernation":"torpor"}
},
{
"Subject ID":"-",
"Sample ID":"T5",
"Factors":{"hibernation":"torpor"}
},
{
"Subject ID":"-",
"Sample ID":"T10",
"Factors":{"hibernation":"torpor"}
},
{
"Subject ID":"-",
"Sample ID":"T11",
"Factors":{"hibernation":"torpor"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"A total of 15 male 3-month-old Syrian hamsters had free access to food and water and were kept at 23C with an 8:16-h light/dark cycle for a four to six-week acclimatization period in our animal facility. All animals were euthanized by decapitation. Brains were then removed and immediately transferred to a N2(l)-containing recipient to freeze the tissues.","SAMPLE_TYPE":"Brain"},

"TREATMENT":{"TREATMENT_SUMMARY":"the whole right hemisphere (300 mg approx.) was analyzed to decrease possible biological variability due to the brain region employed. Brain homogenate was prepared by adding cold (-20°C) methanol:water (1:1, v/v), (1:10 tissue:solvent). Tissue disruption was achieved with TissueLyser LT homogenizer (Qiagen, Germany) for metabolite extraction. Subsequently, 100 μL of brain tissue homogenate was vortex-mixed with 320 μL of methanol for 2 min, followed by the addition of 80 μL of MTBE for the extraction of non-polar compounds. Then, vials were rapidly capped and placed on a shaker for 1 h at room temperature. The extracted samples were centrifuged at 4000 g for 20 min at 20°C. For GC-MS analysis, 300 μL of supernatant was evaporated to dryness (SpeedVac Concentrator System, Thermo Fisher Scientific, Waltham, MA, USA). Methoxymation was then performed with 20 µL O-methoxyamine hydrochloride (15 mg/mL in pyridine) and vigorously vortex-mixed for 5 min. Vials were then incubated in darkness at room temperature for 16 hours. For silylation, 20 μL of BSTFA:TMCS (99:1) was added, vortex-mixed for 5 min, and capped vials were placed in the oven at 70°C for 1 h. Finally, 100 μL of heptane containing C18:0 methyl ester (10 ppm) as Internal Standard (IS) was added to each vial prior to injection. For LC-MS analysis, 90 μL of supernatant was transferred to an Ultra-High Performance Liquid Chromatography-Mass (UHPLC-MS) chromatography vial with insert and was directly injected into the system. For CE-MS analysis, 200 μL of initial brain homogenate was centrifuged separately at 16000 g for 30 min at 15°C. 150 μL of supernatant was evaporated to dryness using the SpeedVac, and re-suspended in 150 μL of 0.2 mM Methionine Sulfone (IS) in 0.1 M formic acid. Samples were vortex-mixed, sonicated, and then centrifuged at 16000 g for 20 min at 4°C. Finally, 100 μL of supernatant was transferred to a CE-MS vial for the analysis."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"the whole right hemisphere (300 mg approx.) was analyzed to decrease possible biological variability due to the brain region employed. Brain homogenate was prepared by adding cold (-20°C) methanol:water (1:1, v/v), (1:10 tissue:solvent). Tissue disruption was achieved with TissueLyser LT homogenizer (Qiagen, Germany) for metabolite extraction. Subsequently, 100 μL of brain tissue homogenate was vortex-mixed with 320 μL of methanol for 2 min, followed by the addition of 80 μL of MTBE for the extraction of non-polar compounds. Then, vials were rapidly capped and placed on a shaker for 1 h at room temperature. The extracted samples were centrifuged at 4000 g for 20 min at 20°C. For GC-MS analysis, 300 μL of supernatant was evaporated to dryness (SpeedVac Concentrator System, Thermo Fisher Scientific, Waltham, MA, USA). Methoxymation was then performed with 20 µL O-methoxyamine hydrochloride (15 mg/mL in pyridine) and vigorously vortex-mixed for 5 min. Vials were then incubated in darkness at room temperature for 16 hours. For silylation, 20 μL of BSTFA:TMCS (99:1) was added, vortex-mixed for 5 min, and capped vials were placed in the oven at 70°C for 1 h. Finally, 100 μL of heptane containing C18:0 methyl ester (10 ppm) as Internal Standard (IS) was added to each vial prior to injection. For LC-MS analysis, 90 μL of supernatant was transferred to an Ultra-High Performance Liquid Chromatography-Mass (UHPLC-MS) chromatography vial with insert and was directly injected into the system. For CE-MS analysis, 200 μL of initial brain homogenate was centrifuged separately at 16000 g for 30 min at 15°C. 150 μL of supernatant was evaporated to dryness using the SpeedVac, and re-suspended in 150 μL of 0.2 mM Methionine Sulfone (IS) in 0.1 M formic acid. Samples were vortex-mixed, sonicated, and then centrifuged at 16000 g for 20 min at 4°C. Finally, 100 μL of supernatant was transferred to a CE-MS vial for the analysis"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent 1290 Infinity","COLUMN_NAME":"Agilent Zorbax Eclipse Plus C8 (150 x 2.1mm, 1.8 um)"},

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

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Agilent 6550 QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_RESULTS_FILE":"ST001117_AN001814_Results.txt UNITS:peak area Has m/z:Yes Has RT:Yes RT units:Minutes"}

}