{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001333","ANALYSIS_ID":"AN002222","VERSION":"1","CREATED_ON":"March 30, 2020, 10:57 am"},

"PROJECT":{"PROJECT_TITLE":"Huntington’s Disease Genotype Suppresses Global Manganese-Responsive Processes","PROJECT_TYPE":"Untargeted Metabolomics analysis","PROJECT_SUMMARY":"Global untargeted metabolomics studies performed in the striatum tissue, the brain region most sensitive to neurodegeneration in Huntington’s Disease, to investigate global Mn-dependent and Mn-responsive biology following various Mn exposures in a mouse model of HD.","INSTITUTE":"Vanderbilt University","DEPARTMENT":"Chemistry","LABORATORY":"Center for Innovative Technology","LAST_NAME":"Codreanu","FIRST_NAME":"Simona","ADDRESS":"1234 Stevenson Center Lane","EMAIL":"simona.codreanu@vanderbilt.edu","PHONE":"6158758422"},

"STUDY":{"STUDY_TITLE":"Huntington’s Disease Genotype Suppresses Global Manganese-Responsive Processes","STUDY_TYPE":"Untargeted metabolomics analysis","STUDY_SUMMARY":"Global untargeted metabolomics studies performed in the striatum tissue, the brain region most sensitive to neurodegeneration in Huntington’s Disease, to investigate global Mn-dependent and Mn-responsive biology following various Mn exposures in a mouse model of HD.","INSTITUTE":"Vanderbilt University","DEPARTMENT":"Chemistry","LABORATORY":"Center for Innovative Technology","LAST_NAME":"Codreanu","FIRST_NAME":"Simona","ADDRESS":"1234 Stevenson Center Lane","EMAIL":"simona.codreanu@vanderbilt.edu","PHONE":"6158758422"},

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"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__136_1911_A_M_WT_0"}
},
{
"Subject ID":"1912",
"Sample ID":"96_1912_A_M_WT_50",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__96_1912_A_M_WT_50"}
},
{
"Subject ID":"1913",
"Sample ID":"20_1913_A_M_WT_0",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__20_1913_A_M_WT_0"}
},
{
"Subject ID":"1914",
"Sample ID":"46_1914_C_M_WT_0",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__46_1914_C_M_WT_0"}
},
{
"Subject ID":"1915",
"Sample ID":"70_1915_C_M_128_50",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__70_1915_C_M_128_50"}
},
{
"Subject ID":"1916",
"Sample ID":"43_1916_A_M_WT_50",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__43_1916_A_M_WT_50"}
},
{
"Subject ID":"1917",
"Sample ID":"71_1917_A_M_128_0",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__71_1917_A_M_128_0"}
},
{
"Subject ID":"1918",
"Sample ID":"115_1918_A_F_WT_50",
"Factors":{"Sex":"F","Genotype":"WT","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__115_1918_A_F_WT_50"}
},
{
"Subject ID":"1919",
"Sample ID":"88_1919_C_F_128_0",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__88_1919_C_F_128_0"}
},
{
"Subject ID":"1920",
"Sample ID":"62_1920_C_F_128_5",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"5"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__62_1920_C_F_128_5"}
},
{
"Subject ID":"1921",
"Sample ID":"10_1921_C_F_128_15",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"15"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__10_1921_C_F_128_15"}
},
{
"Subject ID":"1922",
"Sample ID":"122_1922_A_F_WT_0",
"Factors":{"Sex":"F","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__122_1922_A_F_WT_0"}
},
{
"Subject ID":"1923",
"Sample ID":"60_1923_C_F_128_50",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__60_1923_C_F_128_50"}
},
{
"Subject ID":"1924",
"Sample ID":"27_1924_C_M_WT_0",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__27_1924_C_M_WT_0"}
},
{
"Subject ID":"1928",
"Sample ID":"134_1928_A_F_128_0",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__134_1928_A_F_128_0"}
},
{
"Subject ID":"1929",
"Sample ID":"25_1929_A_F_WT_50",
"Factors":{"Sex":"F","Genotype":"WT","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__25_1929_A_F_WT_50"}
},
{
"Subject ID":"1930",
"Sample ID":"65_1930_C_M_128_15",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"15"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__65_1930_C_M_128_15"}
},
{
"Subject ID":"1931",
"Sample ID":"6_1931_C_M_WT_15",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"15"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__6_1931_C_M_WT_15"}
},
{
"Subject ID":"1933",
"Sample ID":"119_1933_C_F_WT_0",
"Factors":{"Sex":"F","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__119_1933_C_F_WT_0"}
},
{
"Subject ID":"1935",
"Sample ID":"74_1935_A_M_WT_0",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__74_1935_A_M_WT_0"}
},
{
"Subject ID":"1936",
"Sample ID":"29_1936_A_M_WT_50",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__29_1936_A_M_WT_50"}
},
{
"Subject ID":"1937",
"Sample ID":"92_1937_A_F_128_50",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__92_1937_A_F_128_50"}
},
{
"Subject ID":"1938",
"Sample ID":"97_1938_A_M_128_50",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__97_1938_A_M_128_50"}
},
{
"Subject ID":"1939",
"Sample ID":"105_1939_A_F_128_50",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__105_1939_A_F_128_50"}
},
{
"Subject ID":"1943",
"Sample ID":"121_1943_A_F_WT_50",
"Factors":{"Sex":"F","Genotype":"WT","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__121_1943_A_F_WT_50"}
},
{
"Subject ID":"1944",
"Sample ID":"31_1944_A_M_WT_0",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__31_1944_A_M_WT_0"}
},
{
"Subject ID":"1945",
"Sample ID":"137_1945_A_M_128_0",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__137_1945_A_M_128_0"}
},
{
"Subject ID":"1946",
"Sample ID":"138_1946_A_M_WT_50",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__138_1946_A_M_WT_50"}
},
{
"Subject ID":"1948",
"Sample ID":"139_1948_A_M_WT_0",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__139_1948_A_M_WT_0"}
},
{
"Subject ID":"1949",
"Sample ID":"140_1949_A_F_128_0",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__140_1949_A_F_128_0"}
},
{
"Subject ID":"1950",
"Sample ID":"141_1950_A_F_128_50",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__141_1950_A_F_128_50"}
},
{
"Subject ID":"1951",
"Sample ID":"48_1951_A_F_WT_0",
"Factors":{"Sex":"F","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__48_1951_A_F_WT_0"}
},
{
"Subject ID":"1953",
"Sample ID":"103_1953_A_M_128_50",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__103_1953_A_M_128_50"}
},
{
"Subject ID":"1954",
"Sample ID":"126_1954_C_F_128_0",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__126_1954_C_F_128_0"}
},
{
"Subject ID":"1955",
"Sample ID":"52_1955_C_F_WT_0",
"Factors":{"Sex":"F","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__106_1956_C_M_WT_0"}
},
{
"Subject ID":"1957",
"Sample ID":"35_1957_C_M_WT_5",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"5"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__35_1957_C_M_WT_5"}
},
{
"Subject ID":"1958",
"Sample ID":"8_1958_C_M_128_5",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"5"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__8_1958_C_M_128_5"}
},
{
"Subject ID":"1959",
"Sample ID":"118_1959_C_M_WT_15",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"15"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__118_1959_C_M_WT_15"}
},
{
"Subject ID":"1960",
"Sample ID":"63_1960_C_M_WT_50",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__63_1960_C_M_WT_50"}
},
{
"Subject ID":"1961",
"Sample ID":"120_1961_A_M_WT_0",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__120_1961_A_M_WT_0"}
},
{
"Subject ID":"1963",
"Sample ID":"40_1963_C_F_128_5",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"5"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__40_1963_C_F_128_5"}
},
{
"Subject ID":"1964",
"Sample ID":"9_1964_C_F_128_15",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"15"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__9_1964_C_F_128_15"}
},
{
"Subject ID":"1965",
"Sample ID":"36_1965_C_F_128_50",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__36_1965_C_F_128_50"}
},
{
"Subject ID":"1966",
"Sample ID":"14_1966_A_M_128_0",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__14_1966_A_M_128_0"}
},
{
"Subject ID":"1967",
"Sample ID":"15_1967_A_M_128_50",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__15_1967_A_M_128_50"}
},
{
"Subject ID":"1970",
"Sample ID":"81_1970_A_M_WT_50",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__81_1970_A_M_WT_50"}
},
{
"Subject ID":"1971",
"Sample ID":"13_1971_A_M_WT_50",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__13_1971_A_M_WT_50"}
},
{
"Subject ID":"1972",
"Sample ID":"98_1972_A_M_128_0",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__98_1972_A_M_128_0"}
},
{
"Subject ID":"1973",
"Sample ID":"51_1973_A_M_WT_0",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__51_1973_A_M_WT_0"}
},
{
"Subject ID":"1974",
"Sample ID":"12_1974_A_M_128_50",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__12_1974_A_M_128_50"}
},
{
"Subject ID":"1976",
"Sample ID":"18_1976_A_F_128_0",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__18_1976_A_F_128_0"}
},
{
"Subject ID":"1978",
"Sample ID":"67_1978_A_M_128_50",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__67_1978_A_M_128_50"}
},
{
"Subject ID":"1979",
"Sample ID":"83_1979_A_M_WT_50",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__83_1979_A_M_WT_50"}
},
{
"Subject ID":"1980",
"Sample ID":"109_1980_A_M_128_0",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__109_1980_A_M_128_0"}
},
{
"Subject ID":"1981",
"Sample ID":"53_1981_A_M_WT_0",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__53_1981_A_M_WT_0"}
},
{
"Subject ID":"1982",
"Sample ID":"2_1982_A_M_128_50",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__2_1982_A_M_128_50"}
},
{
"Subject ID":"1983",
"Sample ID":"17_1983_A_M_128_0",
"Factors":{"Sex":"M","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__17_1983_A_M_128_0"}
},
{
"Subject ID":"1984",
"Sample ID":"11_1984_A_F_128_0",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__11_1984_A_F_128_0"}
},
{
"Subject ID":"1985",
"Sample ID":"19_1985_A_F_128_50",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__19_1985_A_F_128_50"}
},
{
"Subject ID":"1986",
"Sample ID":"16_1986_A_F_128_0",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__16_1986_A_F_128_0"}
},
{
"Subject ID":"1987",
"Sample ID":"34_1987_A_F_128_50",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"50"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__34_1987_A_F_128_50"}
},
{
"Subject ID":"1890",
"Sample ID":"44_1890_A_F_128_0",
"Factors":{"Sex":"F","Genotype":"128","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__44_1890_A_F_128_0"}
},
{
"Subject ID":"1956",
"Sample ID":"106_1956_C_M_WT_0",
"Factors":{"Sex":"M","Genotype":"WT","Treatment Dosage":"-"},
"Additional sample data":{"RAW_FILE_NAME":"SC_20180814_HILICp_PRM_AP__106_1956_C_M_WT_0"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"For the single injection exposure, mice were sacrificed either 0, 1, 4, 12 or 24 hours after the injection to examine acute changes to Mn. For 1-week and 20-week Mn exposures, mice were sacrificed 24 hours after the final injection. For all studies, mice were sacrificed by cervical dislocation without anesthesia. Mice were decapitated with sharp scissors and the brain removed. The striatum was dissected out under a dissecting microscope, flash-frozen in liquid nitrogen and stored at -80C until later analysis.","SAMPLE_TYPE":"Brain"},

"TREATMENT":{"TREATMENT_SUMMARY":"This study utilized three Mn exposure paradigms: 1) a single subcutaneous exposure of either a 1% solution of MnCl2•4(H2O) in filtered MilliQ water at 50 mg/kg body weight or vehicle (filtered water), 2) a 1-week, and 3) 20-week chronic exposure. The 1-week exposure is a previously published week-long exposure protocol (intermittent injections given on days 0, 3 and 6, with sacrifice on day 7) known to elevate brain Mn levels1. The 20-week exposure is a twice weekly injection (Mondays and Thursdays) based on the estimated half-life of 3.5 days for Mn administered via this route and pilot work suggesting the absence of observable toxicity with this exposure paradigm (including no weight loss or physical signs of distress).","TREATMENT_DOSE":"0, 5, 15, 50 mg/kg body weight","TREATMENT_COMPOUND":"Mn"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Previously frozen striatal tissue was lysed in 400µL ice-cold lysing buffer (1:1:2, ACN:MeOH:Ammonium Bicarbonate (0.1M, pH 8.0) (LC-MS grade). Individual samples were sonicated using a probe tip sonicator, 10 pulses, at 30% power and cooled down on ice between samples. A BCA protein assay was used to determine the protein concentration for each individual sample, and adjusted to a total amount of protein of 200µg in 200 µL of lysis buffer. Isotopically labeled standards, Creatinine-D3 and Lysine-D4, were added to each sample to assess sample processing steps (metabolite extraction and reconstitution). Following lysis and standard addition, protein precipitation was performed by adding 800µL of ice-cold methanol (4x by volume). Samples were incubated at -80°C overnight. Following incubation, samples were centrifuged at 10,000 rpm for 10 min to eliminate proteins. The supernatants containing metabolites were dried via speed-vacuum. Dried metabolite extracts were stored frozen at -80C until ready to use. Prior to mass spectrometry analysis, extracts were reconstituted in 100 μl of acetonitrile/ water (80:20, v/v) and centrifuged for 5 min at 15K rpm to remove insoluble material. Quality control samples were prepared by pooling equal volumes of each sample. Isotopically labeled standards, Valine-D8 and Inosine-4N15, were added to each sample to determine MS instrument reproducibility.","SAMPLEPREP_PROTOCOL_FILENAME":"Codreags00_20200327_075634_PR_MS_Metabolomics_Protocol.pdf","PROCESSING_STORAGE_CONDITIONS":"-80℃","EXTRACTION_METHOD":"Following lysis and standard addition, protein precipitation was performed by adding 800µL of ice-cold methanol (4x by volume). Samples were incubated at -80°C overnight. Following incubation, samples were centrifuged at 10,000 rpm for 10 min to eliminate proteins. The supernatants containing metabolites were dried via speed-vacuum.","EXTRACT_STORAGE":"-80℃","SAMPLE_RESUSPENSION":"100 μl of acetonitrile/ water (80:20, v/v) and centrifuged for 5 min at 15K rpm to remove insoluble material","SAMPLE_SPIKING":"Valine-D8 and Inosine-4N15, were added to each sample to determine MS instrument reproducibility."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Vanquish UHPLC binary system","COLUMN_NAME":"SeQuant ZIC-HILIC 3.5-µm, 2.1 mm × 100 mm column","FLOW_RATE":"0.2 mL/min","COLUMN_TEMPERATURE":"40","SOLVENT_A":"5mM Ammonium formate in 90% water, 10% acetonitrile","SOLVENT_B":"5mM Ammonium formate in 90% acetonitrile, 10% water"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","ANALYSIS_PROTOCOL_FILE":"Codreags00_20200327_075634_PR_MS_Metabolomics_Protocol.pdf"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive HF hybrid Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Ultra-performance liquid chromatography - tandem mass spectrometry (UPLC-MS/MS and UPLC-MS) raw data were imported, processed, normalized and reviewed using Progenesis QI v.2.1 (Non-linear Dynamics, Newcastle, UK).","MS_RESULTS_FILE":"ST001333_AN002222_Results.txt UNITS:peak area Has m/z:Yes Has RT:Yes RT units:Minutes"}

}