Metabolite annotations were assigned based on accurate mass and the m/z difference between the C12 and C13 peaks detected for a given feature. Molecular formulas were mapped to internal libraries in the ClusterFinder software to generate putative annotations.
"],"repository":["MetaboLights"],"study_status":["Public"],"ptm_modification":[""],"instrument_platform":["Liquid Chromatography MS - negative - reverse-phase","Liquid Chromatography MS - positive - reverse-phase"],"chromatography_protocol":["Lab 1
LC system: Thermo Scientific Vanquish HPLC
Mobile Phase A: Water + 0.1% formic acid
Mobile Phase B: Acetonitrile
Column: Avantor ACE EXCEL 2 C18-PFP (2.1 x 100 mm, 2 um)
Flow Rate: 0.35 mL/min
Injection volume: 6 uL
Lab 2
LC system: Thermo Scientific Vanquish HPLC
Mobile Phase A: Water + 0.1% formic acid
Mobile Phase B: Methanol + 0.1% formic acid
Column: Acquity HSS T3 (2.1 x 150 mm, 1.8 um)
Flow Rate: 0.3 mL/min
Injection volume: 5 uL
Lab 3
LC system: Thermo Scientific Vanquish HPLC
Mobile Phase A: Water + 0.1% formic acid + 10 mM ammonium acetate
Mobile Phase B: Methanol + 0.1% formic acid + 10 mM ammonium acetate
Column: Phenomenex Kinetex C18 (2.1 x 150 mm, 2.6 um)
Flow Rate: 0.2 mL/min
Injection volume: 5 uL
Lab 4
LC system: SCIEX Exion UPLC
Mobile Phase A: Water + 0.1% formic acid
Mobile Phase B: Acetonitrile + 0.1% formic acid
Column: Phenomenex Luna Polar Omega (2.1 x 100 mm, 1.6 um)
Flow Rate: 0.5 mL/min
Injection volume: 10 uL
Lab 5
LC system: Agilent 1290 Infinity II
Mobile Phase A: Water + 0.1% formic acid
Mobile Phase B: Acetonitrile + 0.1% formic acid
Column: Acquity HSS T3 (2.1 x 150 mm, 1.8 um)
Flow Rate: 0.3 mL/min
Injection volume: 5 uL
Lab 6
LC system: Waters nanoAcquity UPLC
Mobile Phase A: Water + 0.1% formic acid
Mobile Phase B: Acetonitrile + 0.1% formic acid
Column: Waters (2.1 x 150 mm, 1.8 um)
Flow Rate: 0.07 mL/min
Injection volume: 5 uL
Lab 7
LC system: SCIEX Exion LC 3.0
Mobile Phase A: POS - Water + 0.2% formic acid | NEG – Water + 0.1% formic acid + 10 mM ammonium formate
Mobile Phase B: POS - Acetonitrile + 0.2% formic acid | NEG – Acetonitrile
Column: Phenomenex Luna Polar Omega (2.1 x 100 mm, 1.6 um)
Flow Rate: 0.45 mL/min
Injection volume: 10 uL
Lab 8
LC system: Agilent 1290 UPLC
Mobile Phase A: 95/5 Water/Acetonitrile + 0.1% formic acid
Mobile Phase B: 95/5 Acetonitrile/Water + 0.1% formic acid
Column: Waters Acquity CHS C18 (2.1 x 100 mm, 1.7 um)
Flow Rate: 0.4 mL/min
Injection volume: 5 uL
"],"publication":["Metabolomic profiles reproducibly differentiate clinically relevant, heterogeneous sample mixtures: a multi-lab “round robin” study."],"submitter_name":["HALEY CHATELAINE"],"submitter_affiliation":["National Center for Advancing Translational Sciences - National Institutes of Health"],"organism_part":["mixture"],"technology_type":["mass spectrometry assay"],"disease":[""],"extraction_protocol":["Meat homogenates were protein-normalized, and metabolites were extracted, centrally. Aliquots of extracts were distributed to all labs and maintained at -80 C until analysis.
"],"organism":["Gallus gallus; Sus scrofa; Bos taurus","Bos taurus","Sus scrofa","Gallus gallus"],"full_dataset_link":["https://www.ebi.ac.uk/metabolights/MTBLS15676"],"author":["Haley Chatelaine. haley.chatelaine@nih.gov.","Chris Beecher. chris@iroatech.com.","Ewy Mathe. National Center for Advancing Translational Sciences. ewy.mathe@nih.gov."],"data_transformation_protocol":["Data was transformed to mzXML using ProteoWizard. The mzXML files were input to ClusterFinder software for peak picking using the LTRS standards, which generated a library that was then applied for targeted peak picking on the samples. The picked and grouped peaks were output in a matrix as Clean Data. The Clean Data was then corrected for ion suppression to create the Suppression Corrected Data. Suppression correction involved dividing C12 peaks by the corresponding C13 standard peak and multiplying the resulting ratio by the mean integral of the C13 peak. The Suppression Corrected data was then normalized (SC Normalized Data) using a 'Dual MSTUS' (multiply each Suppression Corrected feature by the sum of the C12 peaks divided by the sum of the C13 peaks). A separate, normalized-only data set (Clean Normalized Data) by applying a MSTUS algorithm to the Clean Data.
"],"study_factor":["Relative concentration","Sample group"],"submitter_email":["haley.chatelaine@nih.gov"],"sample_collection_protocol":["Samples comprised homogenized, protein-normalized beef, chicken, and pork extracts. Seven sample solutions were prepared that comprised all possible equal mixtures of these three extract types: 1) Pure sample chicken (a), pork (b) or beef (c); 2) Dual mixtures of extracts (beef + pork (bc), beef + chicken (ac), pork + chicken (ab)); 3) Mix of all extracts (abc). Concentrations of pure of mixtures of extracts were varied as follows, resulting in 16 samples: Pure samples as 0.5, 1, and 1.5 concentration each (N=18); Dual mixtures at 0.3 and 0.5 concentration each (N=12); Mix of all extracts at 0.3 concentration each (N=2).
"],"omics_type":["Metabolomics"],"study_design":["pooled quality control sample","Agilent 6546 LC/Q-TOF","untargeted analysis","ZenoTOF 7600","ring trial design","isotope ratio mass spectrometry","Bos taurus","Agilent 1290 Infinity UHPLC","experimental sample","Thermo Scientific Vanquish UHPLC System","Thermo Scientific Q Exactive","Thermo Scientific Orbitrap Fusion Lumos Tribrid","SCIEX ExionLC AD","Waters Xevo G2-XS QTof","mixture","Gallus gallus; Sus scrofa; Bos taurus","Shimadzu Nexera UHPLC system","untargeted metabolites","SCIEX 5600 TripleToF","Waters nanoACQUITY UPLC system","Sus scrofa","Gallus gallus"],"curator_keywords":["pooled quality control sample","Agilent 6546 LC/Q-TOF","untargeted analysis","ZenoTOF 7600","ring trial design","isotope ratio mass spectrometry","Bos taurus","Agilent 1290 Infinity UHPLC","experimental sample","Thermo Scientific Vanquish UHPLC System","Thermo Scientific Q Exactive","Thermo Scientific Orbitrap Fusion Lumos Tribrid","SCIEX ExionLC AD","Waters Xevo G2-XS QTof","mixture","Gallus gallus; Sus scrofa; Bos taurus","Shimadzu Nexera UHPLC system","untargeted metabolites","SCIEX 5600 TripleToF","Waters nanoACQUITY UPLC system","Sus scrofa","Gallus gallus"],"mass_spectrometry_protocol":["Lab 1
MS instrument: Thermo Scientific Q Exactive Orbitrap
Drying gas temperature: 350
Sheath gas temperature:
Voltage: 3500 V
m/z range: 70-1000
Scan rate: 6 scan/second
Resolution: 35,000
Lab 2
MS instrument: Thermo Scientific Orbitrap Fusion Lumos
Drying gas temperature:
Sheath gas temperature: 350
Voltage: 3500 V
m/z range: 70-1200
Scan rate: 4 Hz
Resolution: 120,000
Lab 3
MS instrument: Thermo Scientific Orbitrap Fusion Lumos
Drying gas temperature: 250
Sheath gas temperature:
Voltage: 3500 V
m/z range: 70-1400
Scan rate: 20 spectra/second
Resolution: 240,000
Lab 4
MS instrument: SCIEX 5600 Triple TOF
Drying gas temperature: 400
Sheath gas temperature: 40
Voltage: 40 V
m/z range: 50-1000
Scan rate: 8 scans/second
Resolution: 30,000
Lab 5
MS instrument: Agilent 6546 QTOF
Drying gas temperature: 150
Sheath gas temperature: 250
Voltage: 2000 V
m/z range: 50-1700
Scan rate: 5 spectra/second
Resolution:
Lab 6
MS instrument: Waters Xevo G2-XS QTOF
Drying gas temperature: 20
Sheath gas temperature: 120
Voltage: 3000 V
m/z range: 50-1200
Scan rate: 2 spectra/second
Resolution:
Lab 7
MS instrument: ZenoTOF 7600
Drying gas temperature: 600
Sheath gas temperature:
Voltage: POS = 5500 V | NEG = 4500 V
m/z range: 50-1000
Scan rate: Total scan time: 0.839 sec (0.1 s TOF + 25 SWATH win x 0.02s)
Resolution: <0.02 Da
Lab 8
MS instrument: Agilent 6546 QTOF
Drying gas temperature: 300
Sheath gas temperature: 350
Voltage: 3000 V
m/z range: 100-1700
Scan rate: 1 spectrum/second
Resolution:
"],"additional_accession":[]},"is_claimable":false,"name":"Metabolomic profiles generated using the IROA TruQuant platform result in reproducible differentiation of sample mixtures in a multi-lab “round robin” study","description":"The clinical utility of metabolomics hinges on reproducibly detecting differences between groups across analytical platforms. No multi-lab studies, to date, have attempted to evaluate the ability to deconvolve heterogeneous mixtures, which reflect mixtures of cell types present in clinical samples, using metabolomics profiles. We evaluated the reproducibility of the TruQuant metabolomics approach in eight labs in the US and UK. Each lab received identical aliquots of 32 samples, containing known mixtures of beef, pork, and chicken extracts of varying homogeneity levels. Metabolomic profiles were generated for each mixture in duplicate using each lab’s LC-MS methods. Anonymized, processed data were analyzed centrally. Despite differences in the number of features detected (560 – 1727 features), all labs correctly differentiated sample mixture types via unsupervised approaches. Pearson correlations of suppression corrected metabolite abundances, compared between lab pairs, were strong (mean = 0.87 ± 0.12 across all labs). Overall, this “round-robin” study design and data show that metabolomic analyses with appropriate study designs and controls enable reproducible conclusions across labs and platforms.
","dates":{"publication":"2026-09-28","submission":"2026-09-15"},"accession":"MTBLS15676","cross_references":{}}