#METABOLOMICS WORKBENCH tcavallo_20170104_085013 DATATRACK_ID:812 STUDY_ID:ST000533 ANALYSIS_ID:AN000812 PROJECT_ID:PR000392 VERSION 1 CREATED_ON January 24, 2017, 1:21 pm #PROJECT PR:PROJECT_TITLE CHEAR Plasma Dilution 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 RTI International PR:DEPARTMENT Discovery-Sciences-Technology (DST) 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 Dilution Reference Material Proficiency Test NMR (Part I) 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 Discovery-Sciences-Technology (DST) ST:LABORATORY RTI CHEAR ANALYTICAL HUB/UARC 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 - D_1 SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=320 SUBJECT_SAMPLE_FACTORS - D_2 SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=420 SUBJECT_SAMPLE_FACTORS - D_3 SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=520 SUBJECT_SAMPLE_FACTORS - D_4 SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=620 SUBJECT_SAMPLE_FACTORS - D_5 SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=720 SUBJECT_SAMPLE_FACTORS - D_6 SAMPLE TYPE:Replicate CHEAR ID=CHE000054; Aliquot ID=820 #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 Plasma Reference Material aliquots were thawed on ice for SP:SAMPLEPREP_SUMMARY sample preparation, 400 L of the thawed plasma samples were transferred to SP:SAMPLEPREP_SUMMARY labeled tubes on ice where they were mixed with 300 L of NMR Master Mix SP:SAMPLEPREP_SUMMARY solution containing formate (2.3 mM), 0.9% saline, and 0.02% NaN3 (in D2O). SP:SAMPLEPREP_SUMMARY Sample tubes were vortexed for 2 min on a multi-tube vortexer and centrifuged at SP:SAMPLEPREP_SUMMARY 16,000 rcf for 5 min. A 600 L of each sample supernatant was transferred into SP:SAMPLEPREP_SUMMARY a pre-labeled 5mm NMR tubes for data acquisition on a 700 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 Topshim NM:PULSE_SEQUENCE cpmgpr1d NM:WATER_SUPPRESSION yes NM:PULSE_WIDTH 12.3us NM:POWER_LEVEL 25.119W NM:RECEIVER_GAIN 4 NM:OFFSET_FREQUENCY 3299.00 Hz NM:CHEMICAL_SHIFT_REF_CPD Formate 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 0.5Hz NM:ZERO_FILLING yes NM:APODIZATION Lorentzian NM:BASELINE_CORRECTION_METHOD polynomial NM:CHEMICAL_SHIFT_REF_STD Formate #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS uM NMR_METABOLITE_DATA_START Samples D_1 D_2 D_3 D_4 D_5 D_6 Factors SAMPLE TYPE:Replicate SAMPLE TYPE:Replicate SAMPLE TYPE:Replicate SAMPLE TYPE:Replicate SAMPLE TYPE:Replicate SAMPLE TYPE:Replicate 2-Hydroxybutyrate 53.2 46.8 47.5 46.9 48.7 50.8 3-Methyl-2-oxovalerate 5.9 8.1 8 6.4 6.4 7.6 Acetate 46.9 38.1 38.7 39.3 39.3 39.7 Acetone 34.8 26.3 26.7 26.1 34.7 27.2 Alanine 171.5 165.9 159.9 158.9 159.2 164.7 Betaine 37 37.4 35.4 36.6 36.7 37.3 Citrate 772.3 761.3 732.2 690.1 757.5 746.3 Cysteine 167.3 192.1 237.3 234.7 203.9 226.1 Dimethylamine 158.4 165.1 164.5 161.4 164.9 170.8 Ethanol 462 496.5 442.9 435.3 413.9 462.6 Glucose 2524 2536 2550.4 2594 2633.9 2564.9 Glutamate 115.5 141.5 93.3 87 92.3 93.3 Glutamine 143.8 110.8 152.3 114.6 115.1 109.9 Glycerol 138.8 132.7 117.9 123.6 117.9 102.9 Glycine 146.1 118.1 118.8 119.1 117.5 120 Isobutyrate 1.3 1.5 1.8 2 2 2 Isoleucine 36.8 33.7 33.4 33.6 34.6 34.9 Lactate 1145.6 1109.1 1119.2 1110 1113.6 1124.6 Leucine 85.9 74.3 77.2 75.6 79.2 79.8 Lysine 88.5 76.4 73.8 71 79.1 69.4 Malate 104.6 55.2 57.7 69.1 74.2 69.6 Mannose 25.1 25.3 33.5 43.9 36.3 43.3 Methanol 78.2 77 83.4 87.2 88.8 89 Methionine 20 18.1 12.9 21.2 22.8 19.2 O-Phosphocholine 92.5 96.4 97 86 86 94.6 Phenylalanine 10.8 9 11.2 9.8 10.3 11.2 Proline 109.3 110.5 96.6 105.2 109.6 91.9 Propionate 13.4 12.6 12.1 9.6 9.8 11.1 Propylene glycol 11.6 13.5 14.9 13.6 12.8 14.2 Pyruvate 28.3 24.1 23.9 25.4 27.4 23.2 Succinate 6.1 6.2 7.1 5.6 5.8 6.5 Tyrosine 25.6 23.1 24.8 24.1 23.6 24.9 Valine 96 90.3 90.1 90.7 91.2 91.8 sn-Glycero-3-phosphocholine 37.7 35 34.5 34.2 36.6 37.4 τ-Methylhistidine 21.7 19.8 21.2 22.2 21.4 23.8 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID 2-Hydroxybutyrate 3-Methyl-2-oxovalerate Acetate Acetone Alanine Betaine Citrate Cysteine Dimethylamine Ethanol Glucose Glutamate Glutamine Glycerol Glycine Isobutyrate Isoleucine Lactate Leucine Lysine Malate Mannose Methanol Methionine O-Phosphocholine Phenylalanine Proline Propionate Propylene glycol Pyruvate Succinate Tyrosine Valine sn-Glycero-3-phosphocholine τ-Methylhistidine METABOLITES_END #END