{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003142","ANALYSIS_ID":"AN005155","VERSION":"1","CREATED_ON":"March 21, 2024, 5:54 pm"},

"PROJECT":{"PROJECT_TITLE":"Global Lipidomics of Serum Samples from control and ACOX1-LKO Mouse","PROJECT_SUMMARY":"In this study, we use a Acox1 liver specific knock mouse to explore the role of liver peroxisomal fatty acids beta-oxidation in whole body metabolic homeostasis. As the key enzyme of peroxisomal fatty acid beta-oxidation, knock out Acox1 in liver will affect the very long chain fatty acid accoumaltion in liver and their secretion into circulating.","INSTITUTE":"Washington University School of Medicine","LAST_NAME":"Lu","FIRST_NAME":"Dongliang","ADDRESS":"660 S. Euclid Ave., St. Louis, Missouri, 63110, USA","EMAIL":"ludong-liang@wustl.edu","PHONE":"3147476766"},

"STUDY":{"STUDY_TITLE":"Effect of liver Acox1 knockout on serum lipidome in mice","STUDY_SUMMARY":"The liver gene expression of the peroxisomal β-oxidation enzyme acyl-coenzyme A oxidase 1 (ACOX1), which catabolizes very long chain fatty acids (VLCFA), increases in the context of obesity. To check if liver peroxisomal fatty acids beta-oxidation deficiency will affect whole body metabolic homeostasis through circulating lipids. We analyzed serum samples from 5 WT and 5 Acox1-LKO mice.","INSTITUTE":"Washington University in St. Louis","LAST_NAME":"Lu","FIRST_NAME":"Dongliang","ADDRESS":"660 S. Euclid Ave.","EMAIL":"ludong-liang@wustl.edu","PHONE":"3147476766"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"Sample1",
"Factors":{"Sample_ID":"Wild-type","Sample source":"mouse serum","Sample source":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sample2",
"Factors":{"Sample_ID":"Wild-type","Sample source":"mouse serum","Sample source":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sample3",
"Factors":{"Sample_ID":"Wild-type","Sample source":"mouse serum","Sample source":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sample4",
"Factors":{"Sample_ID":"Wild-type","Sample source":"mouse serum","Sample source":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sample5",
"Factors":{"Sample_ID":"Wild-type","Sample source":"mouse serum","Sample source":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sample6",
"Factors":{"Sample_ID":"Acox1-LKO","Sample source":"mouse serum","Sample source":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sample7",
"Factors":{"Sample_ID":"Acox1-LKO","Sample source":"mouse serum","Sample source":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sample8",
"Factors":{"Sample_ID":"Acox1-LKO","Sample source":"mouse serum","Sample source":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sample9",
"Factors":{"Sample_ID":"Acox1-LKO","Sample source":"mouse serum","Sample source":"-"}
},
{
"Subject ID":"-",
"Sample ID":"Sample10",
"Factors":{"Sample_ID":"Acox1-LKO","Sample source":"mouse serum","Sample source":"-"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"After collecting the whole blood of mice, allow the blood to clot by leaving at room temperature for 30 minutes. Then serum were collected by centrifugation at 2000 g for 10 minutes, and save at -80C","SAMPLE_TYPE":"Blood (serum)"},

"TREATMENT":{"TREATMENT_SUMMARY":"WT and Acox1-LKO mice were maintained under constant temperature (24°C), circulating air and humidity (45-65%) with 12h:12h light/dark cycle. All mice had free access to chew diet and water. 10 weeks age mice were used for analysis."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"The lipids extraction was performed strictly following the SOP established based on a modified Folch liquid-liquid extraction protocol. Briefly, an aliquot of 4.5 μL of each sample was vortexed with 1.5 μL of internal standard solution and methanol, followed by adding dichloromethane and vortexing for another 20 s. A clean-up step was performed with water and 10 seconds of vortex. Samples were allowed to equilibrate at room temperature for 10 min and centrifuged at 16,000 g for 10 min at 4°C. An aliquot of the organic layer was evaporated to dryness with a nitrogen blowdown evaporator. The residue was re-suspended in 4.5 μL of NovaMT MixB, vortexed for 1 min, and diluted with 40.5 μL of NovaMT MixA"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)","SOLVENT_A":"10 mM NH4COOH in 50:40:10 MeOH/ACN/Water","SOLVENT_B":"10 mM NH4COOH in 95:5 IPA/Water","FLOW_GRADIENT":"t = 0 min, 5% B; t = 10 min, 40% B; t = 18.8 min, 98% B; t = 20.5 min, 98% B.","FLOW_RATE":"250 μL/min.","COLUMN_TEMPERATURE":"42 °C"},

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

"MS":{"INSTRUMENT_NAME":"Bruker Impact HD","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"The acquisition rate was 1.44 Hz for MS acquisition and 4 – 10 Hz for MS/MS spectra acquisition, with an m/z range from 150 to 1500. For both positive and negative ionization. Intensity threshold:2500 cts for positive ionization. Lipid features were extracted and aligned using software LipidScreener 1.1.0","CAPILLARY_VOLTAGE":"4200 V","COLLISION_ENERGY":"10-70 eV","DRY_GAS_FLOW":"5.0 L/min","DRY_GAS_TEMP":"240°C","NEBULIZER":"1.2 Bar","MS_RESULTS_FILE":"ST003142_AN005155_Results.txt UNITS:peak area Has m/z:Yes Has RT:Yes RT units:Minutes"}

}