Summary of Study ST000596

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000435. The data can be accessed directly via it's Project DOI: 10.21228/M8TK53 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST000596
Study TitleEmory University high-resolution metabolomic profiling CHEAR pooled reference materials
Study TypeUntargeted HRMS for cross-laboratory characterization of common reference material.
Study SummaryProper QA/QC is an essential component of any analytical procedure. The variability in platforms and approaches used by the untargeted cores requires that each lab develop specific QA/QC protocols, making cross-laboratory assessments challenging. To address the difference in measures across the untargeted cores, the untargeted quality assurance working group (UQAWG) has identified the importance of using the same biological reference sample across the different laboratories. To reach this goal, Dr. Jones and Mr. Walker of Emory University volunteered to take the lead in identifying vendors, requesting quotes, purchasing the pooled material, delivering the specified volumes to each of the lab hubs and storing additional material on-site at Emory University. Based upon discussion with the UQAWG, it was decided that each participating laboratory (five total) will receive 500 mL of plasma and 2000 mL of urine, providing sufficient volume for daily analysis over the next five years. Initially, NIST standard reference materials were considered; however, limited supply and cost makes the use of NIST materials impractical. Vendors, including BioreclamationIVT, Lee Biosolutions Inc and Gemini Bio-Products were contacted to request quotes for preparing pooled plasma and urine meeting the following requirements: K2EDTA recovered whole blood (plasma), equal distribution male/female, equal distribution of Black/Hispanic/Caucasian donors and be collected from greater than 40 donors. In addition, we requested all blood samples be collected in FDA inspected facilities, provide collection details, provide general demographics for each donor and be delivered in 500 mL aliquots. BioreclamationIVT, who also provided the biological samples for the NIH metabolomics ring trial, was selected as the most appropriate vendor due to the ability to provide the requested materials, lead-time to deliver and price. To date, both pools have been purchased. The urine pool was received on July 21, 2016 and the plasma pool will be received July 26, 2016. Upon receipt, both pools will be stored at -80°C. Summary demographics were provided by BioreclamationIVT, the urine pool was created by combining urine samples from 60 males and 60 females with an average age of 38 (range 18-60) and equal distribution of races. The plasma pool was created by combining recovered plasma obtained from 50 males and 50 females with an average age of 38.4 (range 19-72) and equal distribution of races. Both urine and plasma pools will be delivered to the lab hubs frozen in units of 500 mL. Upon receipt, plasma and urine pools should be aliquoted into volumes that provide sufficient material so that freeze-thaw of the pooled material is minimized. For unambiguous identification, the urine pool has been named UrineRef_20160721 and the plasma pool has been named PlasmaRef_20160726. The purpose of providing each untargeted lab hub with the pooled material is two-fold. Through analysis of UrineRef_20160721 (if study samples are urine) or PlasmaRef_20160726 (if study samples are plasma) in each batch of samples analyzed for CHEAR, it will be possible to have a common reference across the different lab hubs. Thus, the ability to compare analyses on different platforms will be greatly improved. The use of a consistent reference material in each batch will also facilitate reference standardization, a concept developed for retrospective quantification of high-resolution metabolomics (Go, Walker et al. 2015). Using reference standardization, quantification post-data acquisition is possible by referencing the pooled material analyzed with each series of samples. The pooled material can be characterized and analytes quantified using traditional analytical chemistry techniques, such as quantification by methods of addition or standardization with a certified reference material, such as NIST SRM 1950. Known concentrations within the pooled material can be used to determine an analyte response factor and calculate sample concentrations based on single-point calibration. The benefit of this approach is that targeted quantification is only required in the pooled sample (i.e. UrineRef_20160721 or PlasmaRef_20160726), chemicals selected for quantification do not need to be selected a priori, and population wide estimates of plasma chemical concentrations can be determined without having to re-analyze samples using a targeted approach. Current Clinical Biomarker Laboratory protocol requires three analytical replicates of pooled reference material per day (three technical replicates per analytical replicate per column configuration, or 18 injections/instrument-day), resulting in analyses of the reference material every 10 samples. Results from untargeted LC-HRMS analyses of pooled CHEAR reference materials. PlasmaRef_20160726 was analyzed with additional NIST reference materials: SRM 1950, SRM 1957 and SRM 1958. UrineRef_20160721 was analyzed with NIST reference materials SRM 3672 and 3673.
Institute
Emory University
DepartmentSchool of Medicine, Division of Pulmonary, Allergy, Critical Care Medicine
LaboratoryClincal Biomarkers Laboratory
Last NameWalker
First NameDouglas
Address615 Michael St. Ste 225, Atlanta, GA, 30322, USA
Emaildouglas.walker@emory.edu
Phone4047275984
Submit Date2017-02-17
Num Groups2
Study CommentsOnly pooled material was analyzed.
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Chear StudyYes
Analysis Type DetailLC-MS
Release Date2024-12-31
Release Version1
Douglas Walker Douglas Walker
https://dx.doi.org/10.21228/M8TK53
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:


Project:

Project ID:PR000435
Project DOI:doi: 10.21228/M8TK53
Project Title:Characterization of CHEAR reference material PlasmaRef_20160726
Project Type:Untargeted HRMS profiling
Project Summary:Results from untargeted LC-HRMS analyses of pooled CHEAR reference materials. PlasmaRef_20160726 was analyzed with additional NIST reference materials: SRM 1950, SRM 1957 and SRM 1958.
Institute:Emory University
Department:Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine
Laboratory:Clinical Biomarkers Laboratory
Last Name:Walker
First Name:Douglas
Address:625 Michael St. Ste 225, Atlanta, GA, 30322, USA
Email:douglas.walker@emory.edu
Phone:4047275984
Funding Source:NIEHS ES026560

Subject:

Subject ID:SU000619
Subject Type:Human plasma
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Mammals

Factors:

Subject type: Human plasma; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Type
SA032704NIST_1950_06Plasma
SA032705ChearPlasma_07Plasma
SA032706NIST_1950_07Plasma
SA032707ChearPlasma_06Plasma
SA032708Pla-q3June2014_1aPlasma
SA032709NIST_1950_05Plasma
SA032710Pla-q3June2014_1cPlasma
SA032711ChearPlasma_08Plasma
SA032712NIST_1950_08Plasma
SA032713Pla-q3June2014_1ePlasma
SA032714Pla-q3June2014_1fPlasma
SA032715NIST_1950_10Plasma
SA032716ChearPlasma_10Plasma
SA032717ChearPlasma_09Plasma
SA032718NIST_1950_09Plasma
SA032719ChearPlasma_05Plasma
SA032720Pla-q3June2014_1dPlasma
SA032721NIST_1950_01Plasma
SA032722ChearPlasma_02Plasma
SA032723ChearPlasma_03Plasma
SA032724NIST_1950_02Plasma
SA032725ChearPlasma_01Plasma
SA032726NIST_1950_03Plasma
SA032727NIST_1950_04Plasma
SA032728ChearPlasma_04Plasma
SA032729Pla-q3June2014_1bPlasma
SA032730NIST_1958_09Serum
SA032731NIST_1957_09Serum
SA032732NIST_1958_02Serum
SA032733NIST_1957_10Serum
SA032734NIST_1957_01Serum
SA032735NIST_1958_10Serum
SA032736NIST_1958_01Serum
SA032737NIST_1957_02Serum
SA032738NIST_1957_08Serum
SA032739NIST_1957_06Serum
SA032740NIST_1957_04Serum
SA032741NIST_1958_04Serum
SA032742NIST_1958_05Serum
SA032743NIST_1958_06Serum
SA032744NIST_1957_07Serum
SA032745NIST_1957_05Serum
SA032746NIST_1957_03Serum
SA032747NIST_1958_03Serum
SA032748NIST_1958_07Serum
SA032749NIST_1958_08Serum
Showing results 1 to 46 of 46

Collection:

Collection ID:CO000613
Collection Summary:The plasma pool was received by Emory University on July 26, 2016 and stored at -80°C. Summary demographics were provided by BioreclamationIVT. The plasma pool was created by combining recovered EDTA plasma obtained from 50 males and 50 females with an average age of 38.4 (range 19-72) and equal distribution of races. Prior to analysis, the plasma pool was aliquoted into 0.5 mL volumes t For unambiguous identification, the plasma pool has been named PlasmaRef_20160726.
Sample Type:Human plasma
Additives:EDTA
Blood Serum Or Plasma:Plasma

Treatment:

Treatment ID:TR000633
Treatment Summary:Samples were delivered in individual units of 500 mL. Aliquots of 0.5 mL were prepared after receiving the material from BioreclamationIVT and stored at -80C

Sample Preparation:

Sampleprep ID:SP000626
Sampleprep Summary:Samples were prepared for metabolomics analysis using established methods (Johnson et al. (2010). Analyst; Go et al. (2015). Tox Sci). Prior to analysis, plasma aliquots were removed from storage at -80°C and thawed on ice. Each cryotube was then vortexed briefly to ensure homogeneity, and 50 μL was transferred to a clean microfuge tube. Immediately after, the plasma was treated with 100 μL of ice-cold LC-MS grade acetonitrile (Sigma Aldrich) containing 2.5 μL of internal standard solution with eight stable isotopic chemicals selected to cover a range of chemical properties. Following addition of acetonitrile, plasma was equilibrated for 30 min on ice, upon which precipitated proteins were removed by centrifuge (16.1 ×g at 4°C for 10 min). The resulting supernatant (100 μL) was removed, added to a low volume autosampler vial and maintained at 4°C until analysis (<22 h).
Sampleprep Protocol ID:HRM_SP_082016_01
Sampleprep Protocol Filename:EmoryUniversity_HRM_SP_082016_01.pdf
Sampleprep Protocol Comments:Date effective: 30 July 2016
Extraction Method:2:1 acetonitrile: sample followed by vortexing and centrifugation
Sample Spiking:2.5 uL [13C6]-D-glucose, [15N,13C5]-L-methionine, [13C5]-L-glutamic acid, [15N]-L-tyrosine, [3,3-13C2]-cystine, [trimethyl-13C3]-caffeine, [U-13C5, U-15N2]-L-glutamine, [15N]-indole

Combined analysis:

Analysis ID AN000913 AN000914
Analysis type MS MS
Chromatography type HILIC Reversed phase
Chromatography system Thermo Dionex Ultimate 3000 RS Thermo Dionex Ultimate 3000 RS
Column Waters XBridge Amide (50 x 2.1mm,2.5um) Thermo Higgins C18 (50 x 2.1mm,3um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Fusion Orbitrap Thermo Fusion Orbitrap
Ion Mode POSITIVE NEGATIVE
Units unspecified {"MS:MS_RESULTS_FILE ST000596_AN000914_Results.txt

Chromatography:

Chromatography ID:CH000650
Chromatography Summary:The HILIC column is operated parallel to reverse phase column for simultaneous analytical separation and column flushing through the use of a dual head HPLC pump equipped with 10-port and 6-port switching valves. During operation of HILIC separation method, the MS is operated in positive ion mode and 10 μL of sample is injected onto the HILIC column while the reverse phase column is flushing with wash solution. Flow rate is maintained at 0.35 mL/min until 1.5 min, increased to 0.4 mL/min at 4 min and held for 1 min. Solvent A is 100% LC-MS grade water, solvent B is 100% LC-MS grade acetonitrile and solvent C is 2% formic acid (v/v) in LC-MS grade water. Initial mobile phase conditions are 22.5% A, 75% B, 2.5% C hold for 1.5 min, with linear gradient to 77.5% A, 20% B, 2.5% C at 4 min, hold for 1 min, resulting in a total analytical run time of 5 min. During the flushing phase (reverse phase analytical separation), the HILIC column is equilibrated with a wash solution of 77.5% A, 20% B, 2.5% C.
Methods ID:2% formic acid in LC-MS grade water
Methods Filename:20160728_HILICpos_c18negwash_FullScan_5min
Chromatography Comments:Triplicate injections for each chromatography mode
Instrument Name:Thermo Dionex Ultimate 3000 RS
Column Name:Waters XBridge Amide (50 x 2.1mm,2.5um)
Column Temperature:40C
Flow Gradient:A= water, B= acetontrile, C= 2% formic acid in water; 22.5% A, 75% B, 2.5% C hold for 1.5 min, linear gradient to 77.5% A, 20% B, 2.5% C at 4 min, hold for 1 min
Flow Rate:0.35 mL/min for 1.5 min; linear increase to 0.4 mL/min at 4 min, hold for 1 min
Sample Injection:10 uL
Solvent A:100% water
Solvent B:100% acetonitrile
Analytical Time:5 min
Sample Loop Size:15 uL
Sample Syringe Size:25 uL
Chromatography Type:HILIC
  
Chromatography ID:CH000651
Chromatography Summary:The C18 column is operated parallel to the HILIC column for simultaneous analytical separation and column flushing through the use of a dual head HPLC pump equipped with 10-port and 6-port switching valves. During operation of the C18 method, the MS is operated in negative ion mode and 10 μL of sample is injected onto the C18 column while the HILIC column is flushing with wash solution. Flow rate is maintained at 0.4 mL/min until 1.5 min, increased to 0.5 mL/min at 2 min and held for 3 min. Solvent A is 100% LC-MS grade water, solvent B is 100% LC-MS grade acetonitrile and solvent C is 10mM ammonium acetate in LC-MS grade water. Initial mobile phase conditions are 60% A, 35% B, 5% C hold for 0.5 min, with linear gradient to 0% A, 95% B, 5% C at 1.5 min, hold for 3.5 min, resulting in a total analytical run time of 5 min. During the flushing phase (HILIC analytical separation), the C18 column is equilibrated with a wash solution of 0% A, 95% B, 5% C until 2.5 min, followed by an equilibration solution of 60% A, 35% B, 5% C for 2.5 min.
Methods ID:10mM ammonium acetate in LC-MS grade water
Methods Filename:20160728_c18neg_HILICposwash_FullScan_5min
Chromatography Comments:Triplicate injections for each chromatography mode
Instrument Name:Thermo Dionex Ultimate 3000 RS
Column Name:Thermo Higgins C18 (50 x 2.1mm,3um)
Column Temperature:40C
Flow Gradient:A= water, B= acetontrile, C= 10mM ammonium acetate in water; 60% A, 35% B, 5% C hold for 0.5 min, linear gradient to 0% A, 95% B, 5% C at 1.5 min, hold for 3 min
Flow Rate:0.4 mL/min for 1.5 min; linear increase to 0.5 mL/min at 2 min held for 3 min
Sample Injection:10 uL
Solvent A:100% water
Solvent B:100% acetonitrile
Analytical Time:5 min
Sample Loop Size:15 uL
Sample Syringe Size:25 uL
Chromatography Type:Reversed phase

MS:

MS ID:MS000812
Analysis ID:AN000913
Instrument Name:Thermo Fusion Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
Ion Mode:POSITIVE
Capillary Temperature:300C
Collision Gas:N2
Dry Gas Flow:45
Dry Gas Temp:250C
Mass Accuracy:< 3ppm
Spray Voltage:+3500
Activation Parameter:5e5
Activation Time:118ms
Interface Voltage:S-Lens RF level= 69
Resolution Setting:60,000
Scanning Range:85-1275
Analysis Protocol File:EmoryUniversity_HRM_FusionMS_082016_01.pdf
  
MS ID:MS000813
Analysis ID:AN000914
Instrument Name:Thermo Fusion Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
Ion Mode:NEGATIVE
Capillary Temperature:300C
Collision Gas:N2
Dry Gas Flow:45
Dry Gas Temp:250C
Mass Accuracy:< 3ppm
Spray Voltage:-4000
Activation Parameter:5e5
Activation Time:118ms
Interface Voltage:S-Lens RF level= 69
Resolution Setting:60,000
Scanning Range:85-1275
Analysis Protocol File:EmoryUniversity_HRM_FusionMS_082016_01.pdf
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