{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003273","ANALYSIS_ID":"AN005359","PROJECT_ID":"PR002030","VERSION":"1","CREATED_ON":"June 18, 2024, 10:04 pm"},

"PROJECT":{"PROJECT_TITLE":"Metabolomic changes in mouse tissues following modulation of serine through the diet","PROJECT_SUMMARY":"We analyzed metabolites in the retina, choroid/RPE, and plasma from WT and PHGDH heterozygous mice that were fed either a control or serine/glycine-deprived diet. Additionally, we also analyzed metabolites from the retina, choroid/RPE, paw skin, and plasma from mice that were previously on either a control or serine/glycine-deprived diet and then switched back to a control or serine-supplemented diet.","INSTITUTE":"Salk Institute for Biological Studies","DEPARTMENT":"Molecular and Cell Biology Laboratory","LABORATORY":"Metallo Lab","LAST_NAME":"Lim","FIRST_NAME":"Esther","ADDRESS":"10010 N Torrey Pines Rd, La Jolla, California, 92037, USA","EMAIL":"ewlim2024@gmail.com","PHONE":"(858) 453-4100","FUNDING_SOURCE":"R01CA234245"},

"STUDY":{"STUDY_TITLE":"Impact of serine supplementation following treatment with serine/glycine-depleted diet","STUDY_SUMMARY":"We analyzed metabolites in the retina, choroid/RPE, plasma, and paw skin from mice that were previously on control or serine/glycine-depleted diet and then switched over to a control or serine-supplemented diet.","INSTITUTE":"Salk Institute for Biological Studies","DEPARTMENT":"Molecular and Cell Biology Laboratory","LABORATORY":"Metallo Lab","LAST_NAME":"Lim","FIRST_NAME":"Esther","ADDRESS":"10010 N Torrey Pines Rd","EMAIL":"ewlim2024@gmail.com","PHONE":"(858) 453-4100","NUM_GROUPS":"4","TOTAL_SUBJECTS":"20","NUM_MALES":"20"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"C57BL/6J","AGE_OR_AGE_RANGE":"12-16 months","WEIGHT_OR_WEIGHT_RANGE":"25-45 g","GENDER":"Male","ANIMAL_ANIMAL_SUPPLIER":"The Scripps Research Institute Vivarium","ANIMAL_HOUSING":"5/cage","ANIMAL_LIGHT_CYCLE":"12h dark/12 h light","ANIMAL_FEED":"Special diets; see group labeling","ANIMAL_WATER":"Ad libitum"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20B2",
"Factors":{"Treatment":"Plasma  control","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20B2.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20B3",
"Factors":{"Treatment":"Plasma  control","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20B3.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20B4",
"Factors":{"Treatment":"Plasma  control","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20B4.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20C2",
"Factors":{"Treatment":"Plasma  control","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20C2.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20C3",
"Factors":{"Treatment":"Plasma  control","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20C3.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20D1",
"Factors":{"Treatment":"Plasma  -SG diet","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20D1.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20D2",
"Factors":{"Treatment":"Plasma  -SG diet","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20D2.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20D3",
"Factors":{"Treatment":"Plasma  -SG diet","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20D3.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20D4",
"Factors":{"Treatment":"Plasma  -SG diet","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20D4.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20D5",
"Factors":{"Treatment":"Plasma  -SG diet","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20D5.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20H1",
"Factors":{"Treatment":"Plasma  High serine rescue","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20H1.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20H2",
"Factors":{"Treatment":"Plasma  High serine rescue","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20H2.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20H3",
"Factors":{"Treatment":"Plasma  High serine rescue","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20H3.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20H4",
"Factors":{"Treatment":"Plasma  High serine rescue","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20H4.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20H5",
"Factors":{"Treatment":"Plasma  High serine rescue","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20H5.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20J1",
"Factors":{"Treatment":"Plasma  Control rescue","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20J1.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20J2",
"Factors":{"Treatment":"Plasma  Control rescue","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20J2.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20J3",
"Factors":{"Treatment":"Plasma  Control rescue","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20J3.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20J4",
"Factors":{"Treatment":"Plasma  Control rescue","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20J4.d"}
},
{
"Subject ID":"-",
"Sample ID":"LMRI_DietRev_plasma_20J5",
"Factors":{"Treatment":"Plasma  Control rescue","Sample source":"Mouse plasma"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"LMRI_DietRev_plasma_20J5.d"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Blood samples collected into EDTA-coated microvette tubes. EDTA microvettes were spun at 2,000g at 4 °C for 5 min to obtain plasma, and samples stored at −80 °C until analysis.","SAMPLE_TYPE":"Blood (plasma)"},

"TREATMENT":{"TREATMENT_SUMMARY":"Mice were fed control or serine/glycine-depleted (-SG) diet for 12 months. After 12 months, a subset of the mice that were on the -SG diet were switched back to either the control or supplemented diet for 4 months."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"For sphingolipid extraction from retina and choroid/RPE, frozen tissue was homogenized with a ball mill (Retsch Mixer Mill MM 400) at 30 Hz for 3 min in 500 μl of −20°C methanol, 400 μl of ice-cold saline, and 100 μl of ice-cold MilliQ water, spiked with deuterated internal standards as described earlier. The mixture was then transferred into a 2-ml Eppendorf tube containing 1 ml of chloroform. The tubes were vortexed for 5 min and centrifuged at 16,000g at 4°C for 5 min. The lower organic phase was collected, and 2 μl of formic acid was added to the remaining polar phase, which was re-extracted with 1 ml of chloroform. Combined organic phases were dried and resuspended in 50 μl of 0.2% formic acid and 1 mM ammonium formate in methanol. Last, the tubes were sonicated in a bath sonicator for 10 min and spun at 16,000g for 10 min at 4°C. For plasma, 50 μl of plasma was extracted with 500 μl of −20°C methanol, 400 μl of saline, and 100 μl of water spiked with deuterated internal standards. Chloroform (1 ml) was then added to the tubes. The tubes were vortexed for 5 min and centrifuged at 16,000g at 4°C for 5 min. The lower organic phase was collected, and 2 μl of formic acid was added to the remaining polar phase, which was reextracted with 1 ml of chloroform. Combined organic phases were dried and resuspended in 100 μl of 0.2% formic acid and 1 mM ammonium formate in methanol. Next, the tubes were sonicated in a bath sonicator for 10 min and spun at 16,000g for 10 min at 4°C","PROCESSING_STORAGE_CONDITIONS":"Room temperature","EXTRACT_STORAGE":"4℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"The gradient elution program consisted of the following profile: 0 min, 82% B; 3 min, 82% B; 4 min, 90% B, 18 min, 99% B; 25 min, 99%, 27 min, 82% B, 30 min, 82% B.","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent 6460","COLUMN_NAME":"Spectra 3µm C8SR column (3 μm particle, 3.0 × 150 mm; Peeke Scientific)","SOLVENT_A":"100% water; 0.2% formic acid; 2 mM ammonium formate","SOLVENT_B":"100% methanol; 0.2% formic acid; 1 mM ammonium formate","FLOW_GRADIENT":"0 min, 82% B; 3 min, 82% B; 4 min, 90% B; 18 min, 99% B; 25 min, 99%; 27 min, 82% B; and 30 min, 82% B","FLOW_RATE":"0.5 mL/min","COLUMN_TEMPERATURE":"40"},

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

"MS":{"INSTRUMENT_NAME":"Agilent 6460 QQQ","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Software: Agilent MassHunter Sphingolipid species were analyzed by selective reaction monitoring of the transition from precursor to product ions at associated optimized collision energies, and fragmentor voltages are provided elsewhere (https://doi.org/10.1007/978-1-60761-322-0_22). The m/z values of the precursor and product ions are provided in the following format: Precursor ion -> Product ion."},

"MS_METABOLITE_DATA":{
"Units":"Normalized ion count/L",

"Data":[{"Metabolite":"Cer d18:1/14:0","LMRI_DietRev_plasma_20B2":"0.001540888","LMRI_DietRev_plasma_20B3":"0.003277925","LMRI_DietRev_plasma_20B4":"0.002735089","LMRI_DietRev_plasma_20C2":"0.003494261","LMRI_DietRev_plasma_20C3":"0.003018156","LMRI_DietRev_plasma_20D1":"0.001241107","LMRI_DietRev_plasma_20D2":"0.00124779","LMRI_DietRev_plasma_20D3":"0.000804783","LMRI_DietRev_plasma_20D4":"0.000656254","LMRI_DietRev_plasma_20D5":"0.001497828","LMRI_DietRev_plasma_20H1":"0.000974917","LMRI_DietRev_plasma_20H2":"0.001586607","LMRI_DietRev_plasma_20H3":"0.000834861","LMRI_DietRev_plasma_20H4":"0.000572454","LMRI_DietRev_plasma_20H5":"0.002266307","LMRI_DietRev_plasma_20J1":"0.001652809","LMRI_DietRev_plasma_20J2":"0.000453495","LMRI_DietRev_plasma_20J3":"0.002596889","LMRI_DietRev_plasma_20J4":"0.001828832","LMRI_DietRev_plasma_20J5":"0.000905584"},{"Metabolite":"Cer d18:1/16:0","LMRI_DietRev_plasma_20B2":"0.207970529","LMRI_DietRev_plasma_20B3":"0.251007658","LMRI_DietRev_plasma_20B4":"0.137984966","LMRI_DietRev_plasma_20C2":"0.136540859","LMRI_DietRev_plasma_20C3":"0.112199419","LMRI_DietRev_plasma_20D1":"0.097065861","LMRI_DietRev_plasma_20D2":"0.111473306","LMRI_DietRev_plasma_20D3":"0.047708182","LMRI_DietRev_plasma_20D4":"0.052910772","LMRI_DietRev_plasma_20D5":"0.0567471","LMRI_DietRev_plasma_20H1":"0.065242","LMRI_DietRev_plasma_20H2":"0.111055882","LMRI_DietRev_plasma_20H3":"0.055606243","LMRI_DietRev_plasma_20H4":"0.101735089","LMRI_DietRev_plasma_20H5":"0.088870868","LMRI_DietRev_plasma_20J1":"0.109348651","LMRI_DietRev_plasma_20J2":"0.027500393","LMRI_DietRev_plasma_20J3":"0.096254647","LMRI_DietRev_plasma_20J4":"0.114089603","LMRI_DietRev_plasma_20J5":"0.093283231"},{"Metabolite":"Cer d18:1/16:1","LMRI_DietRev_plasma_20B2":"0.000131152","LMRI_DietRev_plasma_20B3":"9.675E-05","LMRI_DietRev_plasma_20B4":"0.001597623","LMRI_DietRev_plasma_20C2":"0.000211415","LMRI_DietRev_plasma_20C3":"0.001166319","LMRI_DietRev_plasma_20D1":"0.000455524","LMRI_DietRev_plasma_20D2":"0.000291762","LMRI_DietRev_plasma_20D3":"0.001081502","LMRI_DietRev_plasma_20D4":"0.000387356","LMRI_DietRev_plasma_20D5":"0.001318047","LMRI_DietRev_plasma_20H1":"0.000137904","LMRI_DietRev_plasma_20H2":"0.000218868","LMRI_DietRev_plasma_20H3":"6.26302E-05","LMRI_DietRev_plasma_20H4":"0.000344443","LMRI_DietRev_plasma_20H5":"5.76239E-05","LMRI_DietRev_plasma_20J1":"0.000394463","LMRI_DietRev_plasma_20J2":"2.02824E-05","LMRI_DietRev_plasma_20J3":"0.000424029","LMRI_DietRev_plasma_20J4":"0.000198003","LMRI_DietRev_plasma_20J5":"0.000193948"},{"Metabolite":"Cer d18:1/18:0","LMRI_DietRev_plasma_20B2":"0.014423106","LMRI_DietRev_plasma_20B3":"0.032108611","LMRI_DietRev_plasma_20B4":"0.035541504","LMRI_DietRev_plasma_20C2":"0.020171491","LMRI_DietRev_plasma_20C3":"0.008877141","LMRI_DietRev_plasma_20D1":"0.015351664","LMRI_DietRev_plasma_20D2":"0.012102002","LMRI_DietRev_plasma_20D3":"0.003331723","LMRI_DietRev_plasma_20D4":"0.004624577","LMRI_DietRev_plasma_20D5":"0.004092113","LMRI_DietRev_plasma_20H1":"0.005883424","LMRI_DietRev_plasma_20H2":"0.015370857","LMRI_DietRev_plasma_20H3":"0.005989328","LMRI_DietRev_plasma_20H4":"0.009990453","LMRI_DietRev_plasma_20H5":"0.007921886","LMRI_DietRev_plasma_20J1":"0.008282782","LMRI_DietRev_plasma_20J2":"0.003255072","LMRI_DietRev_plasma_20J3":"0.010100734","LMRI_DietRev_plasma_20J4":"0.007257727","LMRI_DietRev_plasma_20J5":"0.006054432"},{"Metabolite":"Cer d18:1/20:0","LMRI_DietRev_plasma_20B2":"0.018232017","LMRI_DietRev_plasma_20B3":"0.07983698","LMRI_DietRev_plasma_20B4":"0.088509504","LMRI_DietRev_plasma_20C2":"0.05682402","LMRI_DietRev_plasma_20C3":"0.033133396","LMRI_DietRev_plasma_20D1":"0.039098803","LMRI_DietRev_plasma_20D2":"0.045929391","LMRI_DietRev_plasma_20D3":"0.019695384","LMRI_DietRev_plasma_20D4":"0.004426753","LMRI_DietRev_plasma_20D5":"0.029397362","LMRI_DietRev_plasma_20H1":"0.030389089","LMRI_DietRev_plasma_20H2":"0.038695386","LMRI_DietRev_plasma_20H3":"0.019477999","LMRI_DietRev_plasma_20H4":"0.038584042","LMRI_DietRev_plasma_20H5":"0.043029621","LMRI_DietRev_plasma_20J1":"0.035615963","LMRI_DietRev_plasma_20J2":"0.004292077","LMRI_DietRev_plasma_20J3":"0.03260917","LMRI_DietRev_plasma_20J4":"0.03419628","LMRI_DietRev_plasma_20J5":"0.035995477"},{"Metabolite":"Cer d18:1/22:0","LMRI_DietRev_plasma_20B2":"0.406595686","LMRI_DietRev_plasma_20B3":"0.816508369","LMRI_DietRev_plasma_20B4":"1.152331471","LMRI_DietRev_plasma_20C2":"0.793351941","LMRI_DietRev_plasma_20C3":"0.629769526","LMRI_DietRev_plasma_20D1":"0.571711377","LMRI_DietRev_plasma_20D2":"0.707528791","LMRI_DietRev_plasma_20D3":"0.495312733","LMRI_DietRev_plasma_20D4":"0.136218304","LMRI_DietRev_plasma_20D5":"0.935587636","LMRI_DietRev_plasma_20H1":"0.622267633","LMRI_DietRev_plasma_20H2":"0.782749385","LMRI_DietRev_plasma_20H3":"0.566082635","LMRI_DietRev_plasma_20H4":"0.808920255","LMRI_DietRev_plasma_20H5":"0.830618127","LMRI_DietRev_plasma_20J1":"0.941094569","LMRI_DietRev_plasma_20J2":"0.04681344","LMRI_DietRev_plasma_20J3":"0.744509373","LMRI_DietRev_plasma_20J4":"0.884041604","LMRI_DietRev_plasma_20J5":"0.904377959"},{"Metabolite":"Cer d18:1/22:1","LMRI_DietRev_plasma_20B2":"0.017038835","LMRI_DietRev_plasma_20B3":"0.064213915","LMRI_DietRev_plasma_20B4":"0.075149449","LMRI_DietRev_plasma_20C2":"0.047589429","LMRI_DietRev_plasma_20C3":"0.046737083","LMRI_DietRev_plasma_20D1":"0.024406544","LMRI_DietRev_plasma_20D2":"0.034917763","LMRI_DietRev_plasma_20D3":"0.029701653","LMRI_DietRev_plasma_20D4":"0.004037028","LMRI_DietRev_plasma_20D5":"0.050200764","LMRI_DietRev_plasma_20H1":"0.03884476","LMRI_DietRev_plasma_20H2":"0.05518269","LMRI_DietRev_plasma_20H3":"0.041791894","LMRI_DietRev_plasma_20H4":"0.049002218","LMRI_DietRev_plasma_20H5":"0.061367384","LMRI_DietRev_plasma_20J1":"0.048443968","LMRI_DietRev_plasma_20J2":"0.001958556","LMRI_DietRev_plasma_20J3":"0.056162562","LMRI_DietRev_plasma_20J4":"0.070772923","LMRI_DietRev_plasma_20J5":"0.06933246"},{"Metabolite":"Cer d18:1/23:0","LMRI_DietRev_plasma_20B2":"0.299132442","LMRI_DietRev_plasma_20B3":"0.348984031","LMRI_DietRev_plasma_20B4":"0.371906572","LMRI_DietRev_plasma_20C2":"0.360419633","LMRI_DietRev_plasma_20C3":"0.260219525","LMRI_DietRev_plasma_20D1":"0.189225748","LMRI_DietRev_plasma_20D2":"0.186445873","LMRI_DietRev_plasma_20D3":"0.124055544","LMRI_DietRev_plasma_20D4":"0.121267204","LMRI_DietRev_plasma_20D5":"0.161080819","LMRI_DietRev_plasma_20H1":"0.189338155","LMRI_DietRev_plasma_20H2":"0.319721647","LMRI_DietRev_plasma_20H3":"0.245471013","LMRI_DietRev_plasma_20H4":"0.25162827","LMRI_DietRev_plasma_20H5":"0.300087191","LMRI_DietRev_plasma_20J1":"0.313047935","LMRI_DietRev_plasma_20J2":"0.025874157","LMRI_DietRev_plasma_20J3":"0.2416308","LMRI_DietRev_plasma_20J4":"0.251593298","LMRI_DietRev_plasma_20J5":"0.244724404"},{"Metabolite":"Cer d18:1/24:0","LMRI_DietRev_plasma_20B2":"0.76814448","LMRI_DietRev_plasma_20B3":"0.775191138","LMRI_DietRev_plasma_20B4":"0.842270305","LMRI_DietRev_plasma_20C2":"0.755229658","LMRI_DietRev_plasma_20C3":"0.536932582","LMRI_DietRev_plasma_20D1":"0.46172973","LMRI_DietRev_plasma_20D2":"0.38028078","LMRI_DietRev_plasma_20D3":"0.39052124","LMRI_DietRev_plasma_20D4":"0.284786976","LMRI_DietRev_plasma_20D5":"0.587166444","LMRI_DietRev_plasma_20H1":"0.461214605","LMRI_DietRev_plasma_20H2":"0.732148917","LMRI_DietRev_plasma_20H3":"0.498043667","LMRI_DietRev_plasma_20H4":"0.589594397","LMRI_DietRev_plasma_20H5":"0.672779137","LMRI_DietRev_plasma_20J1":"0.616757127","LMRI_DietRev_plasma_20J2":"0.069076282","LMRI_DietRev_plasma_20J3":"0.527907929","LMRI_DietRev_plasma_20J4":"0.600130538","LMRI_DietRev_plasma_20J5":"0.672209311"},{"Metabolite":"Cer 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}

}