#METABOLOMICS WORKBENCH tcavallo_20170105_080931 DATATRACK_ID:813 STUDY_ID:ST000534 ANALYSIS_ID:AN000813 PROJECT_ID:PR000393 VERSION 1 CREATED_ON January 24, 2017, 1:27 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 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 Extraction 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 - 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, 400uL of the thawed plasma samples were transferred to labeled SP:SAMPLEPREP_SUMMARY Eppendorf tubes on ice. Methanol (1200uL) 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 700uL 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 600uL 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 Topshim NM:PULSE_SEQUENCE noesypr1d NM:WATER_SUPPRESSION yes NM:PULSE_WIDTH 12.6us NM:POWER_LEVEL 25.119W NM:RECEIVER_GAIN 4 NM:OFFSET_FREQUENCY 3299.00 Hz NM:CHEMICAL_SHIFT_REF_CPD Formate NM:TEMPERATURE 300K #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 2-Hydroxybutyrate 26.3 20.4 26.1 20.4 26.2 25 3-Methyl-2-oxovalerate 8.3 8.7 8.8 7.4 10 10 Alanine 135.6 126.7 120.1 107.8 125.5 143.5 Citrate 725.1 619.7 602.4 577.4 610.6 722.5 Ethanol 118.7 100.2 64.3 71.4 110.6 149.7 Formate 11.9 8.4 9.3 12.7 7.5 11.7 Glucose 2077.6 1827.8 1748.6 1707.1 1817.3 2078.2 Glutamate 204.4 145.8 166.5 129.1 193.2 215.7 Glycerol 51.9 44.5 45.5 42.8 44.7 64.6 Glycine 87.3 78.6 79.5 71.1 80.8 90 Isobutyrate 4.5 3.9 3.9 2.7 4.6 6.1 Isoleucine 30.4 27.5 23.9 20.8 27.7 37.6 Lactate 962.5 929.4 877.5 799.3 884.2 1000.8 Leucine 53.7 46.2 48.3 41.2 51.9 64.8 Lysine 109.5 103.3 97.9 89.1 107.2 134.2 Malate 58.1 114 92.6 87 110.3 158.2 Methanol 5808 4456.8 2594 2281.3 4938.1 5144.3 Methylamine 6.7 6.5 5.7 5 5.6 4.4 O-Acetylcholine 18.9 15.3 17 13.5 17.9 20 O-Phosphocholine 31.1 25.7 24.6 24.2 31 36.6 Phenylalanine 26.5 21.1 18.5 22.2 20.4 25.9 Proline 88.1 82.8 73.6 67.5 50.3 104.5 Propionate 29.8 22.1 30.7 23.9 33 33 Propylene glycol 25.3 21 15.7 11.8 20.9 22.5 Pyruvate 13.6 8.6 10.3 9.4 11.2 15 Sarcosine 10.8 10.4 9 7.7 9.2 12.4 Succinate 6.3 6 6.2 5.2 5.8 6.7 Tyrosine 24.6 24 22.7 22.3 26.9 28.5 Valine 80.3 75.4 71.6 67.2 70.7 91 π-Methylhistidine 38.7 28.7 26.9 31.4 26.3 37.5 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Retention Index Moverz Quant Local ID PubChem CID KEGG ID mass spectrum 2-Hydroxybutyrate 3-Methyl-2-oxovalerate Alanine Citrate Ethanol Formate Glucose Glutamate Glycerol Glycine Isobutyrate Isoleucine Lactate Leucine Lysine Malate Methanol Methylamine O-Acetylcholine O-Phosphocholine Phenylalanine Proline Propionate Propylene glycol Pyruvate Sarcosine Succinate Tyrosine Valine π-Methylhistidine METABOLITES_END #END