#METABOLOMICS WORKBENCH tcavallo_20170119_084207 DATATRACK_ID:823 STUDY_ID:ST000537 ANALYSIS_ID:AN000816 PROJECT_ID:PR000393 VERSION 1 CREATED_ON January 24, 2017, 1:47 pm #PROJECT PR:PROJECT_TITLE CHEAR Plasma Extraction Reference Material Proficiency Test NMR PR:PROJECT_TYPE CHEAR-UT-1 PR:PROJECT_SUMMARY CHEAR Reference Material Plasma was provided by Emory University. The material PR:PROJECT_SUMMARY was prepared and analyzed using the NMR metabolomics workflow of the Eastern PR:PROJECT_SUMMARY Regional Metabolomics Resource Core (protocols available in metabolomics PR:PROJECT_SUMMARY workbench). Data of six samples were acquired of the sample reference material PR:PROJECT_SUMMARY prepared in replicate. PR:INSTITUTE NIEHS PR:DEPARTMENT CHEAR - Children's Health Exposure Analysis Resource PR:LABORATORY RTI CHEAR Analytical Hub - Untargeted Analysis Resource Core PR:LAST_NAME Fennell PR:FIRST_NAME Timothy PR:ADDRESS 3040 E Cornwallis Road, Research Triangle Park, NC 27709 PR:EMAIL fennell@rti.org PR:PHONE 919-485-2781 #STUDY ST:STUDY_TITLE CHEAR Plasma Extraction Reference Material Proficiency Test NMR (Part II) ST:STUDY_TYPE Metabolomics ST:STUDY_SUMMARY CHEAR Reference Material Plasma was provided by Emory University. The material ST:STUDY_SUMMARY was prepared and analyzed using the NMR metabolomics workflow of the Eastern ST:STUDY_SUMMARY Regional Metabolomics Resource Core (protocols available in metabolomics ST:STUDY_SUMMARY workbench). Data of six samples were acquired of the sample reference material ST:STUDY_SUMMARY prepared in replicate. ST:INSTITUTE RTI International ST:DEPARTMENT Analytical Chemistry & Pharmaceuticals ST:LABORATORY RTI CHEAR Analytical Hub - Untargeted Analysis Resource Core ST:LAST_NAME Fennell ST:FIRST_NAME Timothy ST:ADDRESS 3040 E Cornwallis Road, Research Triangle Park, NC 27709 ST:EMAIL fennell@rti.org ST:PHONE 919-485-2781 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - E_1 Sample Type:Replicate CHEAR ID=CHE000054; Aliquot ID=310 SUBJECT_SAMPLE_FACTORS - E_2 Sample Type:Replicate CHEAR ID=CHE000054; Aliquot ID=410 SUBJECT_SAMPLE_FACTORS - E_3 Sample Type:Replicate CHEAR ID=CHE000054; Aliquot ID=510 SUBJECT_SAMPLE_FACTORS - E_4 Sample Type:Replicate CHEAR ID=CHE000054; Aliquot ID=610 SUBJECT_SAMPLE_FACTORS - E_5 Sample Type:Replicate CHEAR ID=CHE000054; Aliquot ID=710 SUBJECT_SAMPLE_FACTORS - E_6 Sample Type:Replicate CHEAR ID=CHE000054; Aliquot ID=810 #COLLECTION CO:COLLECTION_SUMMARY CHEAR Reference Material Plasma was provided by Emory University. #TREATMENT TR:TREATMENT_SUMMARY None. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY A total of 6 CHEAR Reference Plasma aliquots were thawed on ice for sample SP:SAMPLEPREP_SUMMARY preparation, 400 uL of the thawed plasma samples were transferred to labeled SP:SAMPLEPREP_SUMMARY Eppendorf tubes on ice. Methanol (1200 uL) was added to each tube, sample tubes SP:SAMPLEPREP_SUMMARY were vortexed for 2 min on a multi-tube vortexer and centrifuged at 16,000 rcf SP:SAMPLEPREP_SUMMARY for 5 min. A 1000 µl aliquot of the supernatant was transferred into new SP:SAMPLEPREP_SUMMARY pre-labeled 2.0 mL Eppendorf tubes and lyophilized to complete dryness SP:SAMPLEPREP_SUMMARY overnight. Samples were reconstituted with 700 uL of NMR Master Mix solution SP:SAMPLEPREP_SUMMARY containing Chenomx ISTD: DSS-d6, 0.2% NaN3 and 0.2M phosphate buffer (in D2O) at SP:SAMPLEPREP_SUMMARY 7.4 pH. The tubes were vortexed for 4 min on a multi-tube vortexer and SP:SAMPLEPREP_SUMMARY centrifuged at 16,000 rcf for 5 min. A 600 uL aliquot of each sample supernatant SP:SAMPLEPREP_SUMMARY was transferred into a pre-labeled 5mm NMR tubes for data acquisition on a 700 SP:SAMPLEPREP_SUMMARY MHz spectrometer. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE - CH:INSTRUMENT_NAME - CH:COLUMN_NAME - #ANALYSIS AN:ANALYSIS_TYPE NMR #NMR NM:INSTRUMENT_NAME Bruker NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 1D-1H NM:FIELD_FREQUENCY_LOCK Deuterium NM:STANDARD_CONCENTRATION 1mM NM:SPECTROMETER_FREQUENCY 700 MHz NM:NMR_PROBE 5mm ATMA Cryoprobe NM:NMR_SOLVENT D2O NM:NMR_TUBE_SIZE 5mm NM:SHIMMING_METHOD top shim NM:PULSE_SEQUENCE noesypr1d NM:WATER_SUPPRESSION yes NM:PULSE_WIDTH 12.6s NM:POWER_LEVEL 25.119 W NM:RECEIVER_GAIN 4.5 NM:OFFSET_FREQUENCY 3299.5 Hz NM:CHEMICAL_SHIFT_REF_CPD DSS NM:TEMPERATURE 300K NM:NUMBER_OF_SCANS 128 NM:DUMMY_SCANS 4 NM:ACQUISITION_TIME 3.9s NM:RELAXATION_DELAY 2s NM:SPECTRAL_WIDTH 12.02ppm NM:NUM_DATA_POINTS_ACQUIRED 65536 NM:REAL_DATA_POINTS 32768 NM:LINE_BROADENING 0.5 Hz NM:ZERO_FILLING yes NM:APODIZATION Lorentzian NM:BASELINE_CORRECTION_METHOD Polynomial NM:CHEMICAL_SHIFT_REF_STD DSS-d6 #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS uM NMR_METABOLITE_DATA_START Samples E_1 E_2 E_3 E_4 E_5 E_6 Factors Sample Type:Replicate Sample Type:Replicate Sample Type:Replicate Sample Type:Replicate Sample Type:Replicate Sample Type:Replicate Acetate 23156.3 22507.3 26619.8 25692.3 24549.5 25378.3 Alanine 119.2 113.3 107.4 101.8 112.1 125 Citrate 620 504.3 534.9 481.6 513.3 671.3 Dimethylamine 112.9 141.9 88.1 69.8 110.1 121.8 Formate 8.4 8.9 10.6 13.4 9.6 12.1 Glucose 1761.8 1646.6 1507.1 1480.7 1632.7 1777.2 Glutamate 93.2 84.4 80.5 73.8 87.9 94 Glutamine 91.6 88.4 100.3 79.2 83.2 105 Glycine 94.1 78.8 81.3 72.3 83.1 88.7 Histidine 21.6 17.7 22.8 17.5 21.8 25.2 Isoleucine 23.4 22 21 20 20.2 24.8 Lactate 891.9 854 725.2 713.2 836.9 797.1 Leucine 40.8 38.6 35.3 31.6 37.4 39.4 Malonate 720.5 653.2 569.3 599.2 743 846.6 Methanol 5744.3 4384.3 2410.6 2281.1 4809 5155.3 Methylamine 923.9 1241.8 806.4 595.9 881.5 1115.8 Phenylalanine 13.9 16.9 11 13.4 15.6 19.3 Pyruvate 11.3 8.5 10.2 9.4 11.2 15.1 Succinate 5.4 5.6 5.3 4.7 5.8 4.7 Threonine 49.7 51.5 47 40.6 45.7 68.8 Thymine 1255.5 1368 1451.3 1280.5 1298.4 1380.9 Trimethylamine N-oxide 332.9 255 239.3 283.7 399.3 304.6 Tyrosine 28.5 27.9 26 27.4 25.6 23.3 Valine 75.4 66.4 64.3 57.5 62.9 70.9 methyl sulfone 924.6 951.7 634.4 762.5 938.9 1233.8 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID Acetate Alanine Citrate Dimethylamine Formate Glucose Glutamate Glutamine Glycine Histidine Isoleucine Lactate Leucine Malonate Methanol Methylamine Phenylalanine Pyruvate Succinate Threonine Thymine Trimethylamine N-oxide Tyrosine Valine methyl sulfone METABOLITES_END #END