#METABOLOMICS WORKBENCH kuppal2_20170217_133440_mwtab.txt DATATRACK_ID:848 STUDY_ID:ST000596 ANALYSIS_ID:AN000914 PROJECT_ID:PR000435 VERSION 1 CREATED_ON April 19, 2017, 2:07 pm #PROJECT PR:PROJECT_TITLE Characterization of CHEAR reference material PlasmaRef_20160726 PR:PROJECT_TYPE Untargeted HRMS profiling PR:PROJECT_SUMMARY Results from untargeted LC-HRMS analyses of pooled CHEAR reference materials. PR:PROJECT_SUMMARY PlasmaRef_20160726 was analyzed with additional NIST reference materials: SRM PR:PROJECT_SUMMARY 1950, SRM 1957 and SRM 1958. PR:INSTITUTE Emory University School of Medicine PR:DEPARTMENT Division of Pulmonary, Allergy and Critical Care Medicine PR:LABORATORY Clinical Biomarkers Laboratory PR:LAST_NAME Walker PR:FIRST_NAME Douglas PR:ADDRESS 625 Michael St. Ste 225, Atlanta, GA, 30322, USA PR:EMAIL douglas.walker@emory.edu PR:PHONE 4047275984 PR:FUNDING_SOURCE NIEHS ES026560 #STUDY ST:STUDY_TITLE Emory University high-resolution metabolomic profiling CHEAR pooled reference ST:STUDY_TITLE materials ST:STUDY_TYPE Untargeted HRMS for cross-laboratory characterization of common reference ST:STUDY_TYPE material. ST:STUDY_SUMMARY Proper QA/QC is an essential component of any analytical procedure. The ST:STUDY_SUMMARY variability in platforms and approaches used by the untargeted cores requires ST:STUDY_SUMMARY that each lab develop specific QA/QC protocols, making cross-laboratory ST:STUDY_SUMMARY assessments challenging. To address the difference in measures across the ST:STUDY_SUMMARY untargeted cores, the untargeted quality assurance working group (UQAWG) has ST:STUDY_SUMMARY identified the importance of using the same biological reference sample across ST:STUDY_SUMMARY the different laboratories. To reach this goal, Dr. Jones and Mr. Walker of ST:STUDY_SUMMARY Emory University volunteered to take the lead in identifying vendors, requesting ST:STUDY_SUMMARY quotes, purchasing the pooled material, delivering the specified volumes to each ST:STUDY_SUMMARY of the lab hubs and storing additional material on-site at Emory University. ST:STUDY_SUMMARY Based upon discussion with the UQAWG, it was decided that each participating ST:STUDY_SUMMARY laboratory (five total) will receive 500 mL of plasma and 2000 mL of urine, ST:STUDY_SUMMARY providing sufficient volume for daily analysis over the next five years. ST:STUDY_SUMMARY Initially, NIST standard reference materials were considered; however, limited ST:STUDY_SUMMARY supply and cost makes the use of NIST materials impractical. Vendors, including ST:STUDY_SUMMARY BioreclamationIVT, Lee Biosolutions Inc and Gemini Bio-Products were contacted ST:STUDY_SUMMARY to request quotes for preparing pooled plasma and urine meeting the following ST:STUDY_SUMMARY requirements: K2EDTA recovered whole blood (plasma), equal distribution ST:STUDY_SUMMARY male/female, equal distribution of Black/Hispanic/Caucasian donors and be ST:STUDY_SUMMARY collected from greater than 40 donors. In addition, we requested all blood ST:STUDY_SUMMARY samples be collected in FDA inspected facilities, provide collection details, ST:STUDY_SUMMARY provide general demographics for each donor and be delivered in 500 mL aliquots. ST:STUDY_SUMMARY BioreclamationIVT, who also provided the biological samples for the NIH ST:STUDY_SUMMARY metabolomics ring trial, was selected as the most appropriate vendor due to the ST:STUDY_SUMMARY ability to provide the requested materials, lead-time to deliver and price. To ST:STUDY_SUMMARY date, both pools have been purchased. The urine pool was received on July 21, ST:STUDY_SUMMARY 2016 and the plasma pool will be received July 26, 2016. Upon receipt, both ST:STUDY_SUMMARY pools will be stored at -80°C. Summary demographics were provided by ST:STUDY_SUMMARY BioreclamationIVT, the urine pool was created by combining urine samples from 60 ST:STUDY_SUMMARY males and 60 females with an average age of 38 (range 18-60) and equal ST:STUDY_SUMMARY distribution of races. The plasma pool was created by combining recovered plasma ST:STUDY_SUMMARY obtained from 50 males and 50 females with an average age of 38.4 (range 19-72) ST:STUDY_SUMMARY and equal distribution of races. Both urine and plasma pools will be delivered ST:STUDY_SUMMARY to the lab hubs frozen in units of 500 mL. Upon receipt, plasma and urine pools ST:STUDY_SUMMARY should be aliquoted into volumes that provide sufficient material so that ST:STUDY_SUMMARY freeze-thaw of the pooled material is minimized. For unambiguous identification, ST:STUDY_SUMMARY the urine pool has been named UrineRef_20160721 and the plasma pool has been ST:STUDY_SUMMARY named PlasmaRef_20160726. The purpose of providing each untargeted lab hub with ST:STUDY_SUMMARY the pooled material is two-fold. Through analysis of UrineRef_20160721 (if study ST:STUDY_SUMMARY samples are urine) or PlasmaRef_20160726 (if study samples are plasma) in each ST:STUDY_SUMMARY batch of samples analyzed for CHEAR, it will be possible to have a common ST:STUDY_SUMMARY reference across the different lab hubs. Thus, the ability to compare analyses ST:STUDY_SUMMARY on different platforms will be greatly improved. The use of a consistent ST:STUDY_SUMMARY reference material in each batch will also facilitate reference standardization, ST:STUDY_SUMMARY a concept developed for retrospective quantification of high-resolution ST:STUDY_SUMMARY metabolomics (Go, Walker et al. 2015). Using reference standardization, ST:STUDY_SUMMARY quantification post-data acquisition is possible by referencing the pooled ST:STUDY_SUMMARY material analyzed with each series of samples. The pooled material can be ST:STUDY_SUMMARY characterized and analytes quantified using traditional analytical chemistry ST:STUDY_SUMMARY techniques, such as quantification by methods of addition or standardization ST:STUDY_SUMMARY with a certified reference material, such as NIST SRM 1950. Known concentrations ST:STUDY_SUMMARY within the pooled material can be used to determine an analyte response factor ST:STUDY_SUMMARY and calculate sample concentrations based on single-point calibration. The ST:STUDY_SUMMARY benefit of this approach is that targeted quantification is only required in the ST:STUDY_SUMMARY pooled sample (i.e. UrineRef_20160721 or PlasmaRef_20160726), chemicals selected ST:STUDY_SUMMARY for quantification do not need to be selected a priori, and population wide ST:STUDY_SUMMARY estimates of plasma chemical concentrations can be determined without having to ST:STUDY_SUMMARY re-analyze samples using a targeted approach. Current Clinical Biomarker ST:STUDY_SUMMARY Laboratory protocol requires three analytical replicates of pooled reference ST:STUDY_SUMMARY material per day (three technical replicates per analytical replicate per column ST:STUDY_SUMMARY configuration, or 18 injections/instrument-day), resulting in analyses of the ST:STUDY_SUMMARY reference material every 10 samples. Results from untargeted LC-HRMS analyses of ST:STUDY_SUMMARY pooled CHEAR reference materials. PlasmaRef_20160726 was analyzed with ST:STUDY_SUMMARY additional NIST reference materials: SRM 1950, SRM 1957 and SRM 1958. ST:STUDY_SUMMARY UrineRef_20160721 was analyzed with NIST reference materials SRM 3672 and 3673. ST:INSTITUTE Emory University School of Medicine ST:DEPARTMENT Division of Pulmonary, Allergy and Critical Care Medicine ST:LABORATORY Clincal Biomarkers Laboratory ST:LAST_NAME Walker ST:FIRST_NAME Douglas ST:ADDRESS 615 Michael St. Ste 225, Atlanta, GA, 30322, USA ST:EMAIL douglas.walker@emory.edu ST:PHONE 4047275984 ST:NUM_GROUPS 2 ST:STUDY_COMMENTS Only pooled material was analyzed. #SUBJECT SU:SUBJECT_TYPE Human plasma 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 - Pla-q3June2014_1a Type:Plasma Description=In-house QC pool; Sample_type=Quality control SUBJECT_SAMPLE_FACTORS - Pla-q3June2014_1b Type:Plasma Description=In-house QC pool; Sample_type=Quality control SUBJECT_SAMPLE_FACTORS - NIST_1957_01 Type:Serum Description=NIST SRM 1957: Organic Contaminants in Non-Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1958_01 Type:Serum Description=NIST SRM 1958: Organic Contaminants in Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - ChearPlasma_01 Type:Plasma Description=PlasmaRef_20160726; Sample_type=CHEAR Sample SUBJECT_SAMPLE_FACTORS - NIST_1950_01 Type:Plasma Description=NIST SRM 1950: Metabolites in Frozen Human Plasma; Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1957_02 Type:Serum Description=NIST SRM 1957: Organic Contaminants in Non-Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1958_02 Type:Serum Description=NIST SRM 1958: Organic Contaminants in Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - ChearPlasma_02 Type:Plasma Description=PlasmaRef_20160727; Sample_type=CHEAR Sample SUBJECT_SAMPLE_FACTORS - NIST_1950_02 Type:Plasma Description=NIST SRM 1950: Metabolites in Frozen Human Plasma; Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1957_03 Type:Serum Description=NIST SRM 1957: Organic Contaminants in Non-Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1958_03 Type:Serum Description=NIST SRM 1958: Organic Contaminants in Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - ChearPlasma_03 Type:Plasma Description=PlasmaRef_20160728; Sample_type=CHEAR Sample SUBJECT_SAMPLE_FACTORS - NIST_1950_03 Type:Plasma Description=NIST SRM 1950: Metabolites in Frozen Human Plasma; Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1957_04 Type:Serum Description=NIST SRM 1957: Organic Contaminants in Non-Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1958_04 Type:Serum Description=NIST SRM 1958: Organic Contaminants in Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - ChearPlasma_04 Type:Plasma Description=PlasmaRef_20160729; Sample_type=CHEAR Sample SUBJECT_SAMPLE_FACTORS - NIST_1950_04 Type:Plasma Description=NIST SRM 1950: Metabolites in Frozen Human Plasma; Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1957_05 Type:Serum Description=NIST SRM 1957: Organic Contaminants in Non-Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1958_05 Type:Serum Description=NIST SRM 1958: Organic Contaminants in Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - ChearPlasma_05 Type:Plasma Description=PlasmaRef_20160730; Sample_type=CHEAR Sample SUBJECT_SAMPLE_FACTORS - NIST_1950_05 Type:Plasma Description=NIST SRM 1950: Metabolites in Frozen Human Plasma; Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - Pla-q3June2014_1c Type:Plasma Description=In-house QC pool; Sample_type=Quality control SUBJECT_SAMPLE_FACTORS - Pla-q3June2014_1d Type:Plasma Description=In-house QC pool; Sample_type=Quality control SUBJECT_SAMPLE_FACTORS - NIST_1957_06 Type:Serum Description=NIST SRM 1957: Organic Contaminants in Non-Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1958_06 Type:Serum Description=NIST SRM 1958: Organic Contaminants in Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - ChearPlasma_06 Type:Plasma Description=PlasmaRef_20160731; Sample_type=CHEAR Sample SUBJECT_SAMPLE_FACTORS - NIST_1950_06 Type:Plasma Description=NIST SRM 1950: Metabolites in Frozen Human Plasma; Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1957_07 Type:Serum Description=NIST SRM 1957: Organic Contaminants in Non-Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1958_07 Type:Serum Description=NIST SRM 1958: Organic Contaminants in Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - ChearPlasma_07 Type:Plasma Description=PlasmaRef_20160732; Sample_type=CHEAR Sample SUBJECT_SAMPLE_FACTORS - NIST_1950_07 Type:Plasma Description=NIST SRM 1950: Metabolites in Frozen Human Plasma; Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1957_08 Type:Serum Description=NIST SRM 1957: Organic Contaminants in Non-Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1958_08 Type:Serum Description=NIST SRM 1958: Organic Contaminants in Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - ChearPlasma_08 Type:Plasma Description=PlasmaRef_20160733; Sample_type=CHEAR Sample SUBJECT_SAMPLE_FACTORS - NIST_1950_08 Type:Plasma Description=NIST SRM 1950: Metabolites in Frozen Human Plasma; Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1957_09 Type:Serum Description=NIST SRM 1957: Organic Contaminants in Non-Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1958_09 Type:Serum Description=NIST SRM 1958: Organic Contaminants in Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - ChearPlasma_09 Type:Plasma Description=PlasmaRef_20160734; Sample_type=CHEAR Sample SUBJECT_SAMPLE_FACTORS - NIST_1950_09 Type:Plasma Description=NIST SRM 1950: Metabolites in Frozen Human Plasma; Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1957_10 Type:Serum Description=NIST SRM 1957: Organic Contaminants in Non-Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - NIST_1958_10 Type:Serum Description=NIST SRM 1958: Organic Contaminants in Fortified Human Serum (Freeze-Dried); Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - ChearPlasma_10 Type:Plasma Description=PlasmaRef_20160735; Sample_type=CHEAR Sample SUBJECT_SAMPLE_FACTORS - NIST_1950_10 Type:Plasma Description=NIST SRM 1950: Metabolites in Frozen Human Plasma; Sample_type=NIST Reference Material SUBJECT_SAMPLE_FACTORS - Pla-q3June2014_1e Type:Plasma Description=In-house QC pool; Sample_type=Quality control SUBJECT_SAMPLE_FACTORS - Pla-q3June2014_1f Type:Plasma Description=In-house QC pool; Sample_type=Quality control #COLLECTION CO:COLLECTION_SUMMARY The plasma pool was received by Emory University on July 26, 2016 and stored at CO:COLLECTION_SUMMARY -80°C. Summary demographics were provided by BioreclamationIVT. The plasma pool CO:COLLECTION_SUMMARY was created by combining recovered EDTA plasma obtained from 50 males and 50 CO:COLLECTION_SUMMARY females with an average age of 38.4 (range 19-72) and equal distribution of CO:COLLECTION_SUMMARY races. Prior to analysis, the plasma pool was aliquoted into 0.5 mL volumes t CO:COLLECTION_SUMMARY For unambiguous identification, the plasma pool has been named CO:COLLECTION_SUMMARY PlasmaRef_20160726. CO:SAMPLE_TYPE Human plasma CO:ADDITIVES EDTA CO:BLOOD_SERUM_OR_PLASMA Plasma #TREATMENT TR:TREATMENT_SUMMARY Samples were delivered in individual units of 500 mL. Aliquots of 0.5 mL were TR:TREATMENT_SUMMARY prepared after receiving the material from BioreclamationIVT and stored at -80C #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Samples were prepared for metabolomics analysis using established methods SP:SAMPLEPREP_SUMMARY (Johnson et al. (2010). Analyst; Go et al. (2015). Tox Sci). Prior to analysis, SP:SAMPLEPREP_SUMMARY plasma aliquots were removed from storage at -80°C and thawed on ice. Each SP:SAMPLEPREP_SUMMARY cryotube was then vortexed briefly to ensure homogeneity, and 50 μL was SP:SAMPLEPREP_SUMMARY transferred to a clean microfuge tube. Immediately after, the plasma was treated SP:SAMPLEPREP_SUMMARY with 100 μL of ice-cold LC-MS grade acetonitrile (Sigma Aldrich) containing 2.5 SP:SAMPLEPREP_SUMMARY μL of internal standard solution with eight stable isotopic chemicals selected SP:SAMPLEPREP_SUMMARY to cover a range of chemical properties. Following addition of acetonitrile, SP:SAMPLEPREP_SUMMARY plasma was equilibrated for 30 min on ice, upon which precipitated proteins were SP:SAMPLEPREP_SUMMARY removed by centrifuge (16.1 ×g at 4°C for 10 min). The resulting supernatant SP:SAMPLEPREP_SUMMARY (100 μL) was removed, added to a low volume autosampler vial and maintained at SP:SAMPLEPREP_SUMMARY 4°C until analysis (<22 h). SP:SAMPLEPREP_PROTOCOL_ID HRM_SP_082016_01 SP:SAMPLEPREP_PROTOCOL_FILENAME EmoryUniversity_HRM_SP_082016_01.pdf SP:SAMPLEPREP_PROTOCOL_COMMENTS Date effective: 30 July 2016 SP:EXTRACTION_METHOD 2:1 acetonitrile: sample followed by vortexing and centrifugation SP:SAMPLE_SPIKING 2.5 uL [13C6]-D-glucose, [15N,13C5]-L-methionine, [13C5]-L-glutamic acid, SP:SAMPLE_SPIKING [15N]-L-tyrosine, [3,3-13C2]-cystine, [trimethyl-13C3]-caffeine, [U-13C5, SP:SAMPLE_SPIKING U-15N2]-L-glutamine, [15N]-indole #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY The C18 column is operated parallel to the HILIC column for simultaneous CH:CHROMATOGRAPHY_SUMMARY analytical separation and column flushing through the use of a dual head HPLC CH:CHROMATOGRAPHY_SUMMARY pump equipped with 10-port and 6-port switching valves. During operation of the CH:CHROMATOGRAPHY_SUMMARY C18 method, the MS is operated in negative ion mode and 10 μL of sample is CH:CHROMATOGRAPHY_SUMMARY injected onto the C18 column while the HILIC column is flushing with wash CH:CHROMATOGRAPHY_SUMMARY solution. Flow rate is maintained at 0.4 mL/min until 1.5 min, increased to 0.5 CH:CHROMATOGRAPHY_SUMMARY mL/min at 2 min and held for 3 min. Solvent A is 100% LC-MS grade water, solvent CH:CHROMATOGRAPHY_SUMMARY B is 100% LC-MS grade acetonitrile and solvent C is 10mM ammonium acetate in CH:CHROMATOGRAPHY_SUMMARY LC-MS grade water. Initial mobile phase conditions are 60% A, 35% B, 5% C hold CH:CHROMATOGRAPHY_SUMMARY for 0.5 min, with linear gradient to 0% A, 95% B, 5% C at 1.5 min, hold for 3.5 CH:CHROMATOGRAPHY_SUMMARY min, resulting in a total analytical run time of 5 min. During the flushing CH:CHROMATOGRAPHY_SUMMARY phase (HILIC analytical separation), the C18 column is equilibrated with a wash CH:CHROMATOGRAPHY_SUMMARY solution of 0% A, 95% B, 5% C until 2.5 min, followed by an equilibration CH:CHROMATOGRAPHY_SUMMARY solution of 60% A, 35% B, 5% C for 2.5 min. CH:CHROMATOGRAPHY_TYPE Reversed phase MS:INSTRUMENT_NAME Thermo Fusion Orbitrap CH:COLUMN_NAME Thermo Higgins C18 (50 x 2.1mm, 3um) CH:COLUMN_NAME with Thermo Accucore C18 guard column CH:FLOW_GRADIENT A= water, B= acetontrile, C= 10mM ammonium acetate in water; 60% A, 35% B, 5% C CH:FLOW_GRADIENT hold for 0.5 min, linear gradient to 0% A, 95% B, 5% C at 1.5 min, hold for 3 CH:FLOW_GRADIENT min CH:FLOW_RATE 0.4 mL/min for 1.5 min; linear increase to 0.5 mL/min at 2 min held for 3 min CH:COLUMN_TEMPERATURE 40C CH:METHODS_FILENAME 20160728_c18neg_HILICposwash_FullScan_5min CH:SOLVENT_A LC-MS grade water CH:SOLVENT_B LC-MS grade acetonitrile CH:METHODS_ID 10mM ammonium acetate in LC-MS grade water CH:SAMPLE_INJECTION 10 uL CH:ANALYTICAL_TIME 5 min CH:SAMPLE_LOOP_SIZE 15 uL CH:SAMPLE_SYRINGE_SIZE 25 uL CH:CHROMATOGRAPHY_COMMENTS Triplicate injections for each chromatography mode #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Clinical Biomarkers Laboratory AN:OPERATOR_NAME Vilinh Tran AN:DETECTOR_TYPE Obitrap AN:SOFTWARE_VERSION Xcalibur 3.0 Rev. 3 AN:ACQUISITION_DATE January 2015 AN:ANALYSIS_PROTOCOL_FILE EmoryUniversity_HRM_FusionMS_082016_01.pdf AN:DATA_FORMAT Profile #MS MS:INSTRUMENT_NAME Thermo Fusion Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:CAPILLARY_TEMPERATURE 300C MS:COLLISION_GAS N2 MS:DRY_GAS_FLOW 45 MS:DRY_GAS_TEMP 250C MS:MASS_ACCURACY < 3ppm MS:SPRAY_VOLTAGE -4000 MS:ACTIVATION_PARAMETER 5e5 MS:ACTIVATION_TIME 118ms MS:INTERFACE_VOLTAGE S-Lens RF level= 69 MS:RESOLUTION_SETTING 60,000 MS:SCANNING_RANGE 85-1275 MS:MS_RESULTS_FILE ST000596_AN000914_Results.txt #END