{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000604","ANALYSIS_ID":"AN000925","VERSION":"1","CREATED_ON":"May 11, 2017, 10:44 am"},

"PROJECT":{"PROJECT_TITLE":"Impact Of High Sugar Diet On L-Arginine Metabolism In The Lung","PROJECT_SUMMARY":"Asthma is a progressive inflammatory airways disease that leads to structural airway changes and debilitating symptoms in many severely affected adults. We need novel therapeutic agents that are affordable, can decrease the reliance on steroids, and can improve quality of life. This clinical and mechanistic study has the potential to impact treatment of a subset of adult severe asthmatics and to further our understanding of the mechanisms of L-arginine metabolism and NO biology in the airways of asthmatics. We will pursue a clinical trial in subjects not well controlled on standard drug therapy; this strategy will address whether L-arginine is efficacious in patients receiving standard of care medications. In studies using animal models, we and others have shown that interventions that augment NO levels, through either supplementation of L-arginine or inhibition of arginase, decrease allergic airway inflammation and hyperresponsiveness-the two hallmarks of asthma. Overall, we hypothesize that a responder subset of adult severe asthma patients will derive clinical benefit from supplemental L-arginine therapy and that these patients will have a lower exhaled NO concentrations (<20 ppb) and a higher NOS2/Arg1 mRNA and protein ratio in their airway epithelial cells than non-responders. We aim to: 1) test the hypothesis that uncontrolled, adult severe asthma patients with exhaled breath NO concentrations <20 ppb will have fewer asthma exacerbations over 3 months when treated with L-arginine compared to patients with FeNO > 25, 2) determine the mechanisms by which L-arginine affects the regulation of NOS and arginase enzymes in primary airway epithelial cell cultures from severe asthmatic subjects, and 3) test the hypothesis that inhaled nanoparticle carrier formulations of L-arginine will decrease airway inflammation, airway hyperresponsiveness, and airway fibrosis at lower doses than systemically administered L-arginine. The major impact of our study will be to identify the adult severe asthma cohort that will benefit from supplemental L-arginine therapy. Our ultimate goal is to develop novel therapeutic agents to treat adult severe asthma patients better. PUBLIC HEALTH RELEVANCE: Asthma is a progressive inflammatory airways disease that leads to structural airway changes and debilitating symptoms in many severely affected adults. This clinical study has the potential to improve the care of adult severe asthmatics and to further our understanding of the mechanisms of L-arginine metabolism and nitric oxide biology in the lung. If we demonstrate that L-arginine supplementation can decrease asthma attacks in a subset of severe asthmatics, it will have great implications for future research as well as for the daily lives of patients with asthma.","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"},

"STUDY":{"STUDY_TITLE":"Impact Of High Sugar Diet On L-Arginine Metabolism In The Lung","STUDY_SUMMARY":"Asthma is a progressive inflammatory airways disease that leads to structural airway changes and debilitating symptoms in many severely affected adults. We need novel therapeutic agents that are affordable, can decrease the reliance on steroids, and can improve quality of life. This clinical and mechanistic study has the potential to impact treatment of a subset of adult severe asthmatics and to further our understanding of the mechanisms of L-arginine metabolism and NO biology in the airways of asthmatics. We will pursue a clinical trial in subjects not well controlled on standard drug therapy; this strategy will address whether L-arginine is efficacious in patients receiving standard of care medications. In studies using animal models, we and others have shown that interventions that augment NO levels, through either supplementation of L-arginine or inhibition of arginase, decrease allergic airway inflammation and hyperresponsiveness-the two hallmarks of asthma. Overall, we hypothesize that a responder subset of adult severe asthma patients will derive clinical benefit from supplemental L-arginine therapy and that these patients will have a lower exhaled NO concentrations (<20 ppb) and a higher NOS2/Arg1 mRNA and protein ratio in their airway epithelial cells than non-responders. We aim to: 1) test the hypothesis that uncontrolled, adult severe asthma patients with exhaled breath NO concentrations <20 ppb will have fewer asthma exacerbations over 3 months when treated with L-arginine compared to patients with FeNO > 25, 2) determine the mechanisms by which L-arginine affects the regulation of NOS and arginase enzymes in primary airway epithelial cell cultures from severe asthmatic subjects, and 3) test the hypothesis that inhaled nanoparticle carrier formulations of L-arginine will decrease airway inflammation, airway hyperresponsiveness, and airway fibrosis at lower doses than systemically administered L-arginine. The major impact of our study will be to identify the adult severe asthma cohort that will benefit from supplemental L-arginine therapy. Our ultimate goal is to develop novel therapeutic agents to treat adult severe asthma patients better. PUBLIC HEALTH RELEVANCE: Asthma is a progressive inflammatory airways disease that leads to structural airway changes and debilitating symptoms in many severely affected adults. This clinical study has the potential to improve the care of adult severe asthmatics and to further our understanding of the mechanisms of L-arginine metabolism and nitric oxide biology in the lung. If we demonstrate that L-arginine supplementation can decrease asthma attacks in a subset of severe asthmatics, it will have great implications for future research as well as for the daily lives of patients with asthma.","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"},

"SUBJECT":{"SUBJECT_TYPE":"Animal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"9606","GENDER":"Male"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"QC",
"Sample ID":"Inj011_CSH_QC_01.d",
"Factors":{"Source":"Plasma","Species":"Human","Treatment":"None"}
},
{
"Subject ID":"QC",
"Sample ID":"Inj022_CSH_QC_02.d",
"Factors":{"Source":"Plasma","Species":"Human","Treatment":"None"}
},
{
"Subject ID":"QC",
"Sample ID":"Inj033_CSH_QC_03.d",
"Factors":{"Source":"Plasma","Species":"Human","Treatment":"None"}
},
{
"Subject ID":"QC",
"Sample ID":"Inj039_CSH_QC_04.d",
"Factors":{"Source":"Plasma","Species":"Human","Treatment":"None"}
},
{
"Subject ID":"QC",
"Sample ID":"Inj050_CSH_QC_05.d",
"Factors":{"Source":"Plasma","Species":"Human","Treatment":"None"}
},
{
"Subject ID":"QC",
"Sample ID":"Inj061_CSH_QC_06.d",
"Factors":{"Source":"Plasma","Species":"Human","Treatment":"None"}
},
{
"Subject ID":"QC",
"Sample ID":"Inj067_CSH_QC_07.d",
"Factors":{"Source":"Plasma","Species":"Human","Treatment":"None"}
},
{
"Subject ID":"QC",
"Sample ID":"Inj078_CSH_QC_08.d",
"Factors":{"Source":"Plasma","Species":"Human","Treatment":"None"}
},
{
"Subject ID":"QC",
"Sample ID":"Inj089_CSH_QC_09.d",
"Factors":{"Source":"Plasma","Species":"Human","Treatment":"None"}
},
{
"Subject ID":"QC",
"Sample ID":"Inj096_CSH_QC_10.d",
"Factors":{"Source":"Plasma","Species":"Human","Treatment":"None"}
},
{
"Subject ID":"HFS59",
"Sample ID":"Inj012_CSH_Kid_HFS59.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"HFS70",
"Sample ID":"Inj013_CSH_Kid_HFS70.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"CF90",
"Sample ID":"Inj014_CSH_Kid_CF90.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"VHF97",
"Sample ID":"Inj015_CSH_Kid_VHF97.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"CF54",
"Sample ID":"Inj016_CSH_Kid_CF54.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"CF78",
"Sample ID":"Inj017_CSH_Kid_CF78.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"CF49",
"Sample ID":"Inj018_CSH_Kid_CF49.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"HFS82",
"Sample ID":"Inj019_CSH_Kid_HFS82.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"HFS1",
"Sample ID":"Inj020_CSH_Kid_HFS1.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"VHF45",
"Sample ID":"Inj021_CSH_Kid_VHF45.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"VHF92",
"Sample ID":"Inj023_CSH_Kid_VHF92.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"VHF2",
"Sample ID":"Inj024_CSH_Kid_VHF2.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"VHF80",
"Sample ID":"Inj025_CSH_Kid_VHF80.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"CF57",
"Sample ID":"Inj026_CSH_Kid_CF57.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"VHF64",
"Sample ID":"Inj027_CSH_Kid_VHF64.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"CF82",
"Sample ID":"Inj028_CSH_Kid_CF82.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"CF70",
"Sample ID":"Inj029_CSH_Kid_CF70.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"VHF86",
"Sample ID":"Inj030_CSH_Kid_VHF86.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"CF26",
"Sample ID":"Inj031_CSH_Kid_CF26.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"HFS69",
"Sample ID":"Inj032_CSH_Kid_HFS69.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"HFS87",
"Sample ID":"Inj034_CSH_Kid_HFS87.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"VHF31",
"Sample ID":"Inj035_CSH_Kid_VHF31.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"HFS55",
"Sample ID":"Inj036_CSH_Kid_HFS55.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"VHF70",
"Sample ID":"Inj037_CSH_Kid_VHF70.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"CF11",
"Sample ID":"Inj038_CSH_Kid_CF11.d",
"Factors":{"Source":"Kidney","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"HFS55",
"Sample ID":"Inj068_CSH_Liv_HFS55.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"HFS1",
"Sample ID":"Inj069_CSH_Liv_HFS1.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"VHF64",
"Sample ID":"Inj070_CSH_Liv_VHF64.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"HFS82",
"Sample ID":"Inj071_CSH_Liv_HFS82.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"CF78",
"Sample ID":"Inj072_CSH_Liv_CF78.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"VHF2",
"Sample ID":"Inj073_CSH_Liv_VHF2.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"VHF86",
"Sample ID":"Inj074_CSH_Liv_VHF86.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"VHF31",
"Sample ID":"Inj075_CSH_Liv_VHF31.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"VHF45",
"Sample ID":"Inj076_CSH_Liv_VHF45.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"HFS59",
"Sample ID":"Inj077_CSH_Liv_HFS59.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"CF57",
"Sample ID":"Inj079_CSH_Liv_CF57.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"VHF92",
"Sample ID":"Inj080_CSH_Liv_VHF92.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"VHF97",
"Sample ID":"Inj081_CSH_Liv_VHF97.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"HFS69",
"Sample ID":"Inj082_CSH_Liv_HFS69.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"HFS87",
"Sample ID":"Inj083_CSH_Liv_HFS87.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"CF49",
"Sample ID":"Inj084_CSH_Liv_CF49.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"CF70",
"Sample ID":"Inj085_CSH_Liv_CF70.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"VHF70",
"Sample ID":"Inj086_CSH_Liv_VHF70.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"VHF80",
"Sample ID":"Inj087_CSH_Liv_VHF80.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"CF54",
"Sample ID":"Inj088_CSH_Liv_CF54.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"CF11",
"Sample ID":"Inj090_CSH_Liv_CF11.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"CF82",
"Sample ID":"Inj091_CSH_Liv_CF82.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"CF90",
"Sample ID":"Inj092_CSH_Liv_CF90.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"HFS92",
"Sample ID":"Inj093_CSH_Liv_HFS92.d",
"Factors":{"Source":"Liver","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"HFS55",
"Sample ID":"Inj040_CSH_Lung_HFS55.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"CF54",
"Sample ID":"Inj041_CSH_Lung_CF54.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"HFS82",
"Sample ID":"Inj042_CSH_Lung_HFS82.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"VHF2",
"Sample ID":"Inj043_CSH_Lung_VHF2.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"CF90",
"Sample ID":"Inj044_CSH_Lung_CF90.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"CF82",
"Sample ID":"Inj045_CSH_Lung_CF82.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"VHF80",
"Sample ID":"Inj046_CSH_Lung_VHF80.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"VHF31",
"Sample ID":"Inj047_CSH_Lung_VHF31.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"HFS87",
"Sample ID":"Inj048_CSH_Lung_HFS87.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"VHF45",
"Sample ID":"Inj049_CSH_Lung_VHF45.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"VHF70",
"Sample ID":"Inj051_CSH_Lung_VHF70.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"VHF86",
"Sample ID":"Inj052_CSH_Lung_VHF86.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"CF70",
"Sample ID":"Inj053_CSH_Lung_CF70.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"CF49",
"Sample ID":"Inj054_CSH_Lung_CF49.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"VHF92",
"Sample ID":"Inj055_CSH_Lung_VHF92.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"HFS59",
"Sample ID":"Inj056_CSH_Lung_HFS59.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"HFS92",
"Sample ID":"Inj057_CSH_Lung_HFS92.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"CF78",
"Sample ID":"Inj058_CSH_Lung_CF78.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"HFS69",
"Sample ID":"Inj059_CSH_Lung_HFS69.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"CF57",
"Sample ID":"Inj060_CSH_Lung_CF57.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"CF11",
"Sample ID":"Inj062_CSH_Lung_CF11.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Control Diet"}
},
{
"Subject ID":"VHF64",
"Sample ID":"Inj063_CSH_Lung_VHF64.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Very High Fat Diet"}
},
{
"Subject ID":"HFS70",
"Sample ID":"Inj064_CSH_Lung_HFS70.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"HFS1",
"Sample ID":"Inj065_CSH_Lung_HFS1.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"High Fat Diet"}
},
{
"Subject ID":"CF26",
"Sample ID":"Inj066_CSH_Lung_CF26.d",
"Factors":{"Source":"Lung","Species":"Mouse","Treatment":"Control Diet"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"C57BL/6 mice, 6-7 weeks of age upon diet assignment were fed for 150 days and weighed daily. Mice were euthanized with an overdose of pentobarbital IP and lungs were flash frozen. 6 mg of lung tissue was extracted for GC-TOFMS and HILIC-QTOFMS analysis.","SAMPLE_TYPE":"Tissue"},

"TREATMENT":{"TREATMENT_SUMMARY":"Male C57BL/6N mice (6-7 weeks of age) were provided ad libitum access to one of three diets for 150 days: low fat (10% kcals) control (CTRL) chow, high fat (45% kcals) with sugar (HFS) chow or very high (60% kcals) fat (VHF) chow. Body weight and food intake were measured daily.","TREATMENT_DOSEDURATION":"150 Days"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"1. Weigh 50 mg tissue sample in to a 25 ml conical polypropylene centrifuge tube. 2. Add 2.5mL extraction solvent to the tissue sample and homogenize for 45 seconds ensuring that sample resembles a powder. In between samples, clean the homogenizer in solutions of methanol, acetone, water, and the extraction solvent. 3. Centrifuge the samples at 2500 rpm. for 5 minutes. Aliquot 2 X 500?l supernatant, one for analysis and one for a backup sample. Store backup aliquot in the -20°C freezer. 4. Evaporate one 500?l aliquot of the sample in the Labconco Centrivap cold trap concentrator to complete dryness 5. The dried aliquot is then re-suspended with 500l 50% acetonitrile (degassed as given) 6. Centrifuge for 2 min at 14000 rcf using the centrifuge Eppendorf 5415. 7. Remove supernatant to a new Eppendorff tube. 8. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. 9. Submit to derivatization.","SAMPLEPREP_PROTOCOL_FILENAME":"SOP_Extraction_of_Mammalian_Tissue_Samples.pdf","SAMPLEPREP_PROTOCOL_COMMENTS":"This study combined 2 different injection volumes for certain samples when processing positive mode data. The reason for this was because in certain samples the TG's were overloaded with the 1uL injections, but everything else was at a normal peak height. Therefore samples were run at a 0.1uL injection volume and took the data from 9-12.5 minutes because that's when TG's elute. The samples that were injected at 0.1uL were used for times 9 min-12.5 min and the 1uL samples were used for times 0-8.99min."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Data Dictionary Fiehn laboratory_CSH QTOF lipidomics_05-29-2014.pdf","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent 6550","COLUMN_NAME":"Waters Acquity CSH C18 (100 x 2.1mm, 1.7um) 1.7um Pre-Column","FLOW_GRADIENT":"15% B to 99% B","FLOW_RATE":"0.6 mL/min","COLUMN_TEMPERATURE":"65 C","SOLVENT_A":"60:40 Acetonitrile:Water +10mM Ammonium Acetate +10mM Acetic Acid Formate +10mM Formic Acid","SOLVENT_B":"9:1 Isopropanol:Acetonitrile +10mM Ammonium Acetate +10mM Acetic Acid","COLUMN_PRESSURE":"450-850 bar","INJECTION_TEMPERATURE":"4 C","INTERNAL_STANDARD":"See data dictionary","RETENTION_TIME":"See data dictionary","SAMPLE_INJECTION":"5.0 uL","ANALYTICAL_TIME":"13 min","CAPILLARY_VOLTAGE":"3500","TIME_PROGRAM":"15 min","WEAK_WASH_SOLVENT_NAME":"Isopropanol","STRONG_WASH_SOLVENT_NAME":"Isopropanol","TARGET_SAMPLE_TEMPERATURE":"Autosampler temp 4 C","RANDOMIZATION_ORDER":"Excel"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"WCMC Metabolomics Core","DETECTOR_TYPE":"TOF MCP","SOFTWARE_VERSION":"Masshunter","DATA_FORMAT":".d"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Agilent 6550 QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","CAPILLARY_VOLTAGE":"3500","COLLISION_ENERGY":"25 eV","COLLISION_GAS":"Nitrogen","DRY_GAS_FLOW":"13L/min","DRY_GAS_TEMP":"200 C","FRAGMENT_VOLTAGE":"175","FRAGMENTATION_METHOD":"Auto MSMS","ION_SOURCE_TEMPERATURE":"325 C","ION_SPRAY_VOLTAGE":"1000","IONIZATION":"Neg","PRECURSOR_TYPE":"Intact Molecule","REAGENT_GAS":"Nitrogen","SOURCE_TEMPERATURE":"325 C","DATAFORMAT":".d","DESOLVATION_GAS_FLOW":"11 L/min","DESOLVATION_TEMPERATURE":"350 C","NEBULIZER":"35 psig","OCTPOLE_VOLTAGE":"750","RESOLUTION_SETTING":"Exteded Dyamic Range","SCAN_RANGE_MOVERZ":"60-1700 Da","SCANNING_CYCLE":"2 Hz","SCANNING_RANGE":"60-1700 Da","SKIMMER_VOLTAGE":"65","MS_RESULTS_FILE":"ST000604_AN000925_Results.txt UNITS:Counts"}

}