{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000393","ANALYSIS_ID":"AN000629","VERSION":"1","CREATED_ON":"May 10, 2016, 12:16 pm"},

"PROJECT":{"PROJECT_TITLE":"Long-Chain Fatty Acid Combustion Rate Is Associated with Unique Metabolite Profiles in Skeletal Muscle Mitochondria","PROJECT_SUMMARY":"Incomplete or limited long-chain fatty acid (LCFA) combustion in skeletal muscle has been associated with insulin resistance. Signals that are responsive to shifts in LCFA β-oxidation rate or degree of intramitochondrial catabolism are hypothesized to regulate second messenger systems downstream of the insulin receptor. Recent evidence supports a causal link between mitochondrial LCFA combustion in skeletal muscle and insulin resistance. We have used unbiased metabolite profiling of mouse muscle mitochondria with the aim of identifying candidate metabolites within or effluxed from mitochondria and that are shifted with LCFA combustion rate. This proof-of-principle study establishes that large-scale metabolomics methods can be applied to organelle-level models to discover metabolite patterns reflective of LCFA combustion, which may lead to identification of molecules linking muscle fat metabolism and insulin signaling. Our results suggest that future studies should focus on the fate of effluxed TCA cycle intermediates and on mechanisms ensuring their replenishment during LCFA metabolism in skeletal muscle.","INSTITUTE":"University of California, Davis","DEPARTMENT":"Genome and Biomedical Sciences Facility","LABORATORY":"WCMC Metabolomics Core","LAST_NAME":"Fiehn","FIRST_NAME":"Oliver","ADDRESS":"1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616","EMAIL":"ofiehn@ucdavis.edu","PHONE":"(530) 754-8258","FUNDING_SOURCE":"NIH U24DK097154","PUBLICATIONS":"http://dx.doi.org/10.1371/journal.pone.0009834"},

"STUDY":{"STUDY_TITLE":"Long-Chain Fatty Acid Combustion Rate Is Associated with Unique Metabolite Profiles in Skeletal Muscle Mitochondria","STUDY_SUMMARY":"Incomplete or limited long-chain fatty acid (LCFA) combustion in skeletal muscle has been associated with insulin resistance. Signals that are responsive to shifts in LCFA β-oxidation rate or degree of intramitochondrial catabolism are hypothesized to regulate second messenger systems downstream of the insulin receptor. Recent evidence supports a causal link between mitochondrial LCFA combustion in skeletal muscle and insulin resistance. We have used unbiased metabolite profiling of mouse muscle mitochondria with the aim of identifying candidate metabolites within or effluxed from mitochondria and that are shifted with LCFA combustion rate. This proof-of-principle study establishes that large-scale metabolomics methods can be applied to organelle-level models to discover metabolite patterns reflective of LCFA combustion, which may lead to identification of molecules linking muscle fat metabolism and insulin signaling. Our results suggest that future studies should focus on the fate of effluxed TCA cycle intermediates and on mechanisms ensuring their replenishment during LCFA metabolism in skeletal muscle.","INSTITUTE":"University of California, Davis","DEPARTMENT":"Genome and Biomedical Sciences Facility","LABORATORY":"WCMC Metabolomics Core","LAST_NAME":"Fiehn","FIRST_NAME":"Oliver","ADDRESS":"1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, CA 95616","EMAIL":"ofiehn@ucdavis.edu","PHONE":"(530) 754-8258","PUBLICATIONS":"http://dx.doi.org/10.1371/journal.pone.0009834"},

"SUBJECT":{"SUBJECT_TYPE":"Animal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"C57Bl/6J","AGE_OR_AGE_RANGE":"4 months","GENDER":"Female","HUMAN_SMOKING_STATUS":"Former/Current","ANIMAL_ANIMAL_SUPPLIER":"Jackson Laboratories","ANIMAL_HOUSING":"housed in a facility at 23°C","ANIMAL_LIGHT_CYCLE":"12∶12 hr light cycle, lights on 0700","ANIMAL_FEED":"with free access to chow (4.5% fat/weight), for at least 1 week before being studied"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"103879",
"Sample ID":"061215byusa1119_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"103884",
"Sample ID":"061215byusa1127_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"103889",
"Sample ID":"061215byusa1088_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 3"}
},
{
"Subject ID":"103894",
"Sample ID":"061215byusa1096_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 4"}
},
{
"Subject ID":"103899",
"Sample ID":"061215byusa1092_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 5"}
},
{
"Subject ID":"103904",
"Sample ID":"061215byusa1091_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 6"}
},
{
"Subject ID":"103910",
"Sample ID":"061215byusa1066_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"103915",
"Sample ID":"061215byusa1083_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"103920",
"Sample ID":"061215byusa1081_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 3"}
},
{
"Subject ID":"103925",
"Sample ID":"061215byusa1072_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 4"}
},
{
"Subject ID":"103930",
"Sample ID":"061215byusa1107_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 5"}
},
{
"Subject ID":"103935",
"Sample ID":"061215byusa1089_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 6"}
},
{
"Subject ID":"103941",
"Sample ID":"061215byusa1129_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"103946",
"Sample ID":"061215byusa1105_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"103951",
"Sample ID":"061215byusa1128_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 3"}
},
{
"Subject ID":"103956",
"Sample ID":"061215byusa1074_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 4"}
},
{
"Subject ID":"103961",
"Sample ID":"061215byusa1116_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 5"}
},
{
"Subject ID":"103966",
"Sample ID":"061215byusa1123_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 6"}
},
{
"Subject ID":"103972",
"Sample ID":"061215byusa1130_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"103977",
"Sample ID":"061215byusa1120_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"103982",
"Sample ID":"061215byusa1067_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 3"}
},
{
"Subject ID":"103987",
"Sample ID":"061215byusa1097_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 4"}
},
{
"Subject ID":"103997",
"Sample ID":"061215byusa1104_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitic acid 2.4 uM"},
"Additional sample data":{"Mouse #":"# 6"}
},
{
"Subject ID":"104003",
"Sample ID":"061215byusa1065_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"104008",
"Sample ID":"061215byusa1103_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"104013",
"Sample ID":"061215byusa1086_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 3"}
},
{
"Subject ID":"104018",
"Sample ID":"061215byusa1134_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 4"}
},
{
"Subject ID":"104023",
"Sample ID":"061215byusa1079_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 5"}
},
{
"Subject ID":"104028",
"Sample ID":"061215byusa1090_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 6"}
},
{
"Subject ID":"104034",
"Sample ID":"061215byusa1115_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"104039",
"Sample ID":"061215byusa1095_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"104044",
"Sample ID":"061215byusa1113_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 3"}
},
{
"Subject ID":"104054",
"Sample ID":"061215byusa1099_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 5"}
},
{
"Subject ID":"104059",
"Sample ID":"061215byusa1094_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 6"}
},
{
"Subject ID":"104065",
"Sample ID":"061215byusa1093_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"104070",
"Sample ID":"061215byusa1100_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"104075",
"Sample ID":"061215byusa1085_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 3"}
},
{
"Subject ID":"104080",
"Sample ID":"061215byusa1080_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 4"}
},
{
"Subject ID":"104085",
"Sample ID":"061215byusa1078_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 5"}
},
{
"Subject ID":"104090",
"Sample ID":"061215byusa1076_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 6"}
},
{
"Subject ID":"104096",
"Sample ID":"061215byusa1102_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"104101",
"Sample ID":"061215byusa1082_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"104111",
"Sample ID":"061215byusa1071_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 4"}
},
{
"Subject ID":"104116",
"Sample ID":"061215byusa1135_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 5"}
},
{
"Subject ID":"104121",
"Sample ID":"061215byusa1131_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitic acid 9.6 uM"},
"Additional sample data":{"Mouse #":"# 6"}
},
{
"Subject ID":"104127",
"Sample ID":"061215byusa1112_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"104132",
"Sample ID":"061215byusa1114_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"104137",
"Sample ID":"061215byusa1077_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 3"}
},
{
"Subject ID":"104142",
"Sample ID":"061215byusa1075_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 4"}
},
{
"Subject ID":"104147",
"Sample ID":"061215byusa1087_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 5"}
},
{
"Subject ID":"104152",
"Sample ID":"061215byusa1124_1",
"Factors":{"Organ":"cytosol","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 6"}
},
{
"Subject ID":"104158",
"Sample ID":"061215byusa1101_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"104163",
"Sample ID":"061215byusa1111_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"104168",
"Sample ID":"061215byusa1122_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 3"}
},
{
"Subject ID":"104173",
"Sample ID":"061215byusa1136_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 3"}
},
{
"Subject ID":"104178",
"Sample ID":"061215byusa1110_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 5"}
},
{
"Subject ID":"104183",
"Sample ID":"061215byusa1098_1",
"Factors":{"Organ":"mitochondria","Time":"0 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 6"}
},
{
"Subject ID":"104189",
"Sample ID":"061215byusa1109_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"104194",
"Sample ID":"061215byusa1133_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"104199",
"Sample ID":"061215byusa1068_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 3"}
},
{
"Subject ID":"104204",
"Sample ID":"061215byusa1084_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 4"}
},
{
"Subject ID":"104209",
"Sample ID":"061215byusa1106_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 5"}
},
{
"Subject ID":"104214",
"Sample ID":"061215byusa1125_1",
"Factors":{"Organ":"cytosol","Time":"20 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 6"}
},
{
"Subject ID":"104220",
"Sample ID":"061215byusa1070_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 1"}
},
{
"Subject ID":"104225",
"Sample ID":"061215byusa1121_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 2"}
},
{
"Subject ID":"104235",
"Sample ID":"061215byusa1132_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 4"}
},
{
"Subject ID":"104240",
"Sample ID":"061215byusa1118_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 5"}
},
{
"Subject ID":"104245",
"Sample ID":"061215byusa1069_1",
"Factors":{"Organ":"mitochondria","Time":"20 min","Dose":"palmitoyl carnitine 9.6 uM"},
"Additional sample data":{"Mouse #":"# 6"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Skeletal muscle mitochondria were isolated essentially according to Chappell and Perry [Chappell JB, Perry SV (1954) Biochemical and osmotic properties of skeletal muscle mitochondria. Nature 173: 1094–1095.]. All media were ice-cold, and procedures done on ice or at 4°C. Briefly, pectoral, forelimb and hindlimb muscles were rapidly dissected and placed in basic medium [BM (mM): KCl (140), HEPES (20), MgCl2 (5), EGTA (2); pH 7.0]. Together, these muscle groups comprise a mixed population of mainly type II oxidative and glycolytic fibers. Muscle was cleaned of connective tissue and fat, minced and placed in 15 vol of homogenizing medium (HM: BM with 1 mM ATP and 1% BSA (w/v)) containing one unit of protease (Subtilisin A) per g muscle wet weight.","COLLECTION_PROTOCOL_FILENAME":"Long-Chain_Fatty_Acid_Metabolite_Profiles_in_Skeletal_Muscle_Mitochondria.PDF"},

"TREATMENT":{"TREATMENT_SUMMARY":"Mitochondria (0.6 mg/ml) were supplied with three concentrations of palmitate corresponding to rates of β-oxidation: 1. low (2 µM) 2. medium (9 µM) 3. high (19 µM) Three ml aliquots of incubation medium [IM, (mM): KCl (120), HEPES (5), KH2PO4 (5), MgCl2 (5) and EGTA (1); pH 7.4] were supplemented with (mM) ATP (1), malate (0.05), coenzyme A (0.025), and carnitine (0.5) and added to 20-ml glass reaction vials. Solutions of low, medium and high palmitate concentrations were added to vials in a 6∶1 FA:BSA complex. Two additional incubations were performed as controls: 1. 0 µM palmitate 2. 9 µM palmitate + inhibitors","TREATMENT_PROTOCOL_FILENAME":"Long-Chain_Fatty_Acid_Metabolite_Profiles_in_Skeletal_Muscle_Mitochondria.PDF","TREATMENT_PROTOCOL_COMMENTS":"The first control condition evaluated the metabolic profile of mitochondria oxidizing only malate, and included ATP, carnitine and CoA and ethanol (0.5%). The second control condition assessed effects of FA in the absence of complete oxidative catabolism, and consisted of malate, 9 µM palmitate, ATP, carnitine and CoA, and supplemented with the TCA cycle inhibitor malonate (10 mM) and the electron transport chain complex I inhibitor rotenone (5 µM)"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Tissue was homogenized using a glass/Teflon Potter-Elvehjem tissue grinder (240 rpm) and fractionated by centrifugation at 800 g (10 min), and the supernatant collected and spun at 12000 g (9 min). The pellet was resuspended in 20 ml BM and incubated on ice for 5 min (myofibrillar repolymerization). Samples were spun at 800 g (8 min) to pellet actin-myosin polymers. The supernatant was then spun at 12000 g (9 min). The final pellet was resuspended in 220 µl of BM.","SAMPLEPREP_PROTOCOL_FILENAME":"Long-Chain_Fatty_Acid_Metabolite_Profiles_in_Skeletal_Muscle_Mitochondria.PDF","SAMPLEPREP_PROTOCOL_COMMENTS":"This isolation procedure yields mitochondria with high respiratory control ratios (state 3/state 4; ∼8–10 when supplied with 10 mM pyruvate/5 mM malate), and which are capable of activating palmitate [29], a process dependent on the integrity of enzymes on the mitochondrial outer membrane. Protein concentration was determined by a modified Lowry method with BSA as standard.","PROCESSING_METHOD":"Homogenized"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Leco Pegasus III GC","COLUMN_NAME":"Restek Corporation Rtx-5Sil MS","FLOW_RATE":"1 ml/min","COLUMN_TEMPERATURE":"50-330C","COLUMN_PRESSURE":"7.7 PSI","INJECTION_TEMPERATURE":"50 C ramped to 250 C by 12 C/s","SAMPLE_INJECTION":"0.5 uL","TRANSFERLINE_TEMPERATURE":"230C","WASHING_BUFFER":"Ethyl Acetate","SAMPLE_LOOP_SIZE":"30 m length x 0.25 mm internal diameter","RANDOMIZATION_ORDER":"Excel generated"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"WCMC Metabolomics Core","DETECTOR_TYPE":"TOF","SOFTWARE_VERSION":"ChromaTOF","DATA_FORMAT":".peg, .txt, .cdf"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Leco Pegasus III GC TOF","INSTRUMENT_TYPE":"GC-TOF","MS_TYPE":"EI","ION_MODE":"POSITIVE","ION_SOURCE_TEMPERATURE":"250 C","IONIZATION":"Pos","IONIZATION_ENERGY":"70 eV","MASS_ACCURACY":"Nominal","SOURCE_TEMPERATURE":"250 C","SCAN_RANGE_MOVERZ":"85-500 Da","SCANNING_CYCLE":"17 Hz","SCANNING_RANGE":"85-500 Da","SKIMMER_VOLTAGE":"1850 V"},

"MS_METABOLITE_DATA":{
"Units":"counts",

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}

}