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  2. In curation
  3. In review
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MTBLS247: Positional Enrichment by Proton Analysis (PEPA): a One-Dimensional 1H-NMR Approach for 13C Stable Isotope Tracer Studies in Metabolomics

Abstract

A novel metabolomics approach for NMR-based stable isotope tracer studies called PEPA is presented, and its performance validated using human cancer cells. PEPA detects the position of carbon label in isotopically enriched metabolites and quantifies fractional enrichment by indirect determination of 13C-satellite peaks using 1D-1H-NMR spectra. In comparison with 13C-NMR, TOCSY and HSQC, PEPA improves sensitivity, accelerates the elucidation of 13C positions in labeled metabolites and the quantification of the percentage of stable isotope enrichment. Altogether, PEPA provides a novel framework for extending the high-throughput of 1H-NMR metabolic profiling to stable isotope tracing in metabolomics, facilitating and complementing the information derived from 2D-NMR experiments and expanding the range of isotopically enriched metabolites detected in cellular extracts. The study also contains data, subjected to this technique, pertaining to exonuclease domain like 2 (EXD2), a nuclear encoded gene that we have found is targeted to the mitochondria and is critical for normal metabolism.

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 Authors: Maria Vinaixa

  Release date: 23-Feb-2017

 Status: Public

Organism(s)

Homo sapiens

  Study Design

PEPA

CHMO:nuclear magnetic resonance spectroscopy

untargeted metabolites

tracer

PR:exonuclease 3'-5' domain-containing protein 2

  Experimental Factors

Substrate

Labeling

 Publications
[1]  Positional Enrichment by Proton Analysis (PEPA): a One- Dimensional 1H-NMR Approach for 13C Stable Isotope Tracer Studies in Metabolomics
[2]  EXD2 governs stem cell homeostasis and lifespan by regulating mitochondrial translation and cellular respiration
Protocol Description
Sample collection U2OS-cells human osteosarcoma cell line were cultured in Dulbeccos modified Eagle Medium (DMEM, Invitrogen 41966) supplemented with 10% (v/v) fetal bovine serum (FBS, Sigma F7524-500ML) and 100 U/ml penicillin/streptomycin (Reactiva 01030311B000). The cells (in duplicated batches) were cultured in a humidified atmosphere at 5% CO2 , 37 ºC and grown for 8-9 days in polypropylene bottles; the medium was changed once daily initially and twice daily near the end of the growth period. Then, U2OS-cells were seeded on 15-cm tissue culture plates in triplicates. During 6 hr, cells were supplemented with DMEM powder without glucose and glutamine (Sigma) supplemented with 5 mM glucose and 2 mM L-glutamine that was unlabeled or [U-13C]-Glucose or [U-13C]-Glutamine labeled (Cambridge Isotope Laboratories). After 6 hr, culture medium was removed and cells were collected by trypsinization and 106 cells were pelleted and snap-frozen for further metabolites extraction.
Extraction Metabolites were extracted into the extraction solvent by adding 2 ml of a cold mixture of chloroform/ methanol (2:1 v/v). The resulting suspension was bath-sonicated for 3 min, and 2 ml of cold water was added. Then, 1 ml of chloroform/methanol (2:1 v/v) was added to the samples and bath-sonicated for 3 min. Cell lysates were centrifuged (5000 x g, 15 min at 4 °C) and the aqueous phase was carefully transferred into a new tube. The sample was frozen, lyophilized and stored at 80 °C until further NMR analysis.
NMR sample For NMR measurements, the hydrophilic extracts were reconstituted in 600 µl of D2O phosphate buffer (PBS 0.05 mM, pH 7.4, 99.5% D2O) containing 0.73 mM of deuterared trisilylpropionic acid (d6-TSP). Samples were then vortexed, homogenized for 5 min, and centrifuged for 15 min at 14000 x g at 4 °C. Finally, clear redissolved samples were tranferred into 5 mm NMR tubes.
NMR spectroscopy NMR spectra were recorded at 300 K on an Avance III 600 spectrometer (Bruker, Germany) operating at a proton frequency of 600.20 MHz and a carbon frequency of 150.93 MHz using a 5 mm CPTCI triple resonance (1H, 13C, 31P) gradient cryoprobe.
NMR assay One-dimensional 1H pulse experiments were performed using the nuclear Overhauser Effect Spectroscopy (NOESY) presaturation sequence (RD-90-t1-90-tm-90-ACQ) to suppress the residual water peak with mixing time (tm) of 100 ms. Solvent presaturation with irradiaton power of 50 Hz was applied during recycling delay (RD = 5 s) and mixing time. The 90 pulse length was calibrated for each sample and varied from 8.06 to 10.26 µs. The spectral width was 12 kHz (20 ppm), and a total of 256 transients were collected into 64 k data points for each 1H spectrum. The exponential line broadening applied before Fourier transformation was 0.3 Hz.

One-dimensional 13C-NMR spectra were recorded using Inverse Gate decoupled 13C pulse experiment and WALTZ-16 scheme proton presaturation and a relaxation delay of 10 s. The 90 pulse length was 14.75 µs. The spectral width was 36 kHz (240 ppm), and a total of 6144 transients were collected into 64 k data points for each 13C spectrum. The exponential line broadening applied before Fourier transformation was 1 Hz.

Two-dimensional TOCSY spectra were recorded using MLEV17 spin lock with 63 ms of duration, B1 field strength of 8,33 kHz, and acquisition times of 0.28 s in t2 and 0.071 s in t1. The recycling delay was set to 5 s, and 64 scans for each transient were collected into 2 k of data points in t2 and 1024 transients were acquired in t1. Data were zero-filled to 4 k points, apodized with a shifted Gaussian and a 1 Hz line broadening exponential function in both dimensions prior to Fourier transformation.

Two-dimensional gradient 1H-13C-HSQC spectra were recorded using an acquisition time of 0.128 s in t2 and 0.0084 s in t1 and a recycle time of 1.5 s, with the 13C GARP decoupling set to 100 ppm, and the evolution delay set for 145 Hz corresponding approximately to an average value of JCH coupling constants. The data were apodized with an unshifted Gaussian and a 1 Hz line broadening exponential in both dimensions prior to Fourier transformation.
Data transformation The frequency domain spectra were phased, baseline-corrected and referenced to TSP signal ( = 0 ppm) using TopSpin software (version 2.1, Bruker). Dolphin 1D[1] was used to profile metabolites identified in the 1H-NMR spectra. Poorly resolved and partially overlapped resonances are resolved using line shape fitting and consequently, their area can be properly quantified. Profiled areas were row-wise normalized using ERETIC signal. Area of each profiled metabolite in unlabeled experiments were compared with the same area in experiments using isotopically enriched substrates using t-test (p<0.05 were considered for statistical significance). Data analysis was performed using R version 3.2.0.

Ref:
[1] Gómez J, Brezmes J, Mallol R, Rodríguez MA, Vinaixa M, Salek RM, Correig X, Cañellas N. Dolphin: a tool for automatic targeted metabolite profiling using 1D and 2D (1)H-NMR data. Anal Bioanal Chem. 2014 Dec;406(30):7967-76. doi:10.1007/s00216-014-8225-6. PMID:25370160
Metabolite identification 1H-NMR resonances were assigned using different database search engines BBioref AMIX database from Bruker, Chenomx, HMDB[1] and Complex Mixture Analysis by NMR (COLMAR)[2]. Both heteronuclear 2D-1H-13C-HSQC, 1H-13C-HMBC and homonuclear 2D 1H-1H-COSY, 1H-1H-TOCSY were used for structural confirmation.

Ref:
[1] Wishart DS, Jewison T, Guo AC, Wilson M, Knox C, Liu Y, Djoumbou Y, Mandal R, Aziat F, Dong E, Bouatra S, Sinelnikov I, Arndt D, Xia J, Liu P, Yallou F, Bjorndahl T, Perez-Pineiro R, Eisner R, Allen F, Neveu V, Greiner R, Scalbert A. HMDB 3.0--The Human Metabolome Database in 2013. Nucleic Acids Res. 2013 Jan;41(Database issue):D801-7. doi:10.1093/nar/gks1065. PMID:23161693
[2] Bingol K, Li DW, Bruschweiler-Li L, Cabrera OA, Megraw T, Zhang F, Brüschweiler R. Unified and isomer-specific NMR metabolomics database for the accurate analysis of (13)C-(1)H HSQC spectra. ACS Chem Biol. 2015 Feb 20;10(2):452-9. doi:10.1021/cb5006382. PMID:25333826
Source Name Organism Organism part Protocol REF Sample Name Substrate Labeling
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_12CGln_107 Glutamine Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_12CGln_108 Glutamine Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_12CGln_109 Glutamine Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_12C_10 Glucose Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_12C_11 Glucose Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_12C_12 Glucose Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_13CGln_122 Glutamine [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_13CGln_123 Glutamine [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_13CGln_124 Glutamine [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_13C_22 Glucose [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_13C_23 Glucose [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_13C_24 Glucose [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_12C_01 Glucose Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_12C_02 Glucose Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_12C_03 Glucose Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_12CGln_101 Glutamine Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_12CGln_102 Glutamine Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_12CGln_103 Glutamine Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_13C_13 Glucose [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_13C_14 Glucose [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_13C_15 Glucose [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_13CGln_116 Glutamine [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_13CGln_117 Glutamine [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh22_AQ_13CGln_118 Glutamine [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_12C_10_TOCSY Glucose Unlabeled
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_13CGln_122_HSQC Glutamine [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_13CGln_TOCSY Glutamine [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_13C_23_HSQC Glucose [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection sh00_AQ_13C_23_TOCSY Glucose [U-13C]
U-2 OS Homo sapiens U2-OS cell Sample collection Sh00_AQ_12CGln_109_TOCSY Glutamine Unlabeled

Assay  1

Assay file name: a_13C_EXD2_NMR_spectroscopy.txt
Technology: NMR spectroscopy
Platform: Bruker

Instrumentation

Sample Name Protocol REF Extraction Method Extract Name Protocol REF NMR tube type Solvent Sample pH Temperature Unit Labeled Extract Name Label Protocol REF Instrument NMR Probe Number of transients Pulse sequence name Magnetic field strength Unit Acquisition Parameter Data File Protocol REF NMR Assay Name Free Induction Decay Data File Protocol REF Normalization Name Derived Spectral Data File Protocol REF Data Transformation Name Metabolite Assignment File
Sh00_AQ_12CGln_107 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12CGln_107 Sh00_AQ_12CGln_107.rar Data transformation Metabolite identification
Sh00_AQ_12CGln_108 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12CGln_108 Sh00_AQ_12CGln_108.rar Data transformation Metabolite identification
Sh00_AQ_12CGln_109 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12CGln_109 Sh00_AQ_12CGln_109.rar Data transformation Metabolite identification
Sh00_AQ_12C_10 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12C_10 Sh00_AQ_12C_10.rar Data transformation Metabolite identification
Sh00_AQ_12C_11 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12C_11 Sh00_AQ_12C_11.rar Data transformation Metabolite identification
Sh00_AQ_12C_12 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12C_12 Sh00_AQ_12C_12.rar Data transformation Metabolite identification
Sh00_AQ_13CGln_122 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13CGln_122 Sh00_AQ_13CGln_122.rar Data transformation Metabolite identification
Sh00_AQ_13CGln_123 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13CGln_123 Sh00_AQ_13CGln_123.rar Data transformation Metabolite identification
Sh00_AQ_13CGln_124 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13CGln_124 Sh00_AQ_13CGln_124.rar Data transformation Metabolite identification
Sh00_AQ_13C_22 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13C_22 Sh00_AQ_13C_22.rar Data transformation Metabolite identification
Sh00_AQ_13C_23 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13C_23 Sh00_AQ_13C_23.rar Data transformation Metabolite identification
Sh00_AQ_13C_24 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13C_24 Sh00_AQ_13C_24.rar Data transformation Metabolite identification
Sh22_AQ_12C_01 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_12C_01 Sh22_AQ_12C_01.rar Data transformation Metabolite identification
Sh22_AQ_12C_02 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_12C_02 Sh22_AQ_12C_02.rar Data transformation Metabolite identification
Sh22_AQ_12C_03 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_12C_03 Sh22_AQ_12C_03.rar Data transformation Metabolite identification
Sh22_AQ_12CGln_101 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_12CGln_101 Sh22_AQ_12CGln_101.rar Data transformation Metabolite identification
Sh22_AQ_12CGln_102 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_12CGln_102 Sh22_AQ_12CGln_102.rar Data transformation Metabolite identification
Sh22_AQ_12CGln_103 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_12CGln_103 Sh22_AQ_12CGln_103.rar Data transformation Metabolite identification
Sh22_AQ_13C_13 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_13C_13 Sh22_AQ_13C_13.rar Data transformation Metabolite identification
Sh22_AQ_13C_14 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_13C_14 Sh22_AQ_13C_14.rar Data transformation Metabolite identification
Sh22_AQ_13C_15 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_13C_15 Sh22_AQ_13C_15.rar Data transformation Metabolite identification
Sh22_AQ_13CGln_116 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_13CGln_116 Sh22_AQ_13CGln_116.rar Data transformation Metabolite identification
Sh22_AQ_13CGln_117 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_13CGln_117 Sh22_AQ_13CGln_117.rar Data transformation Metabolite identification
Sh22_AQ_13CGln_118 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh22_AQ_13CGln_118 Sh22_AQ_13CGln_118.rar Data transformation Metabolite identification

Assay  2

Assay file name: a_TOCSY_PEPA_NMR_spectroscopy.txt
Technology: NMR spectroscopy
Platform: Bruker

Instrumentation

Sample Name Protocol REF Extraction Method Extract Name Protocol REF NMR tube type Solvent Sample pH Temperature Unit Labeled Extract Name Label Protocol REF Instrument NMR Probe Number of transients Pulse sequence name Magnetic field strength Unit Acquisition Parameter Data File Protocol REF NMR Assay Name Free Induction Decay Data File Protocol REF Normalization Name Derived Spectral Data File Protocol REF Data Transformation Name Metabolite Assignment File
Sh00_AQ_12C_10_TOCSY Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 1024 2D TOCSY with presaturation using MLEV (mlevphpr) 14.1 tesla NMR assay Sh00_AQ_12C_10_TOCSY Sh00_AQ_12C_10_TOCSY.rar Data transformation Metabolite identification
Sh00_AQ_13CGln_TOCSY Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 1024 2D TOCSY with presaturation using MLEV (mlevphpr) 14.1 tesla NMR assay Sh00_AQ_13CGln_TOCSY Sh00_AQ_13CGln_TOCSY.rar Data transformation Metabolite identification
sh00_AQ_13C_23_TOCSY Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 1024 2D TOCSY with presaturation using MLEV (mlevphpr) 14.1 tesla NMR assay sh00_AQ_13C_23_TOCSY sh00_AQ_13C_23_TOCSY.rar Data transformation Metabolite identification
Sh00_AQ_12CGln_109_TOCSY Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 1024 2D TOCSY with presaturation using MLEV (mlevphpr) 14.1 tesla NMR assay Sh00_AQ_12CGln_109_TOCSY Sh00_AQ_12CGln_109_TOCSY.rar Data transformation Metabolite identification

Assay  3

Assay file name: a_NOESY_EXD2_NMR_spectroscopy.txt
Technology: NMR spectroscopy
Platform: Bruker

Instrumentation

Sample Name Protocol REF Extraction Method Extract Name Protocol REF NMR tube type Solvent Sample pH Temperature Unit Labeled Extract Name Label Protocol REF Instrument NMR Probe Number of transients Pulse sequence name Magnetic field strength Unit Acquisition Parameter Data File Protocol REF NMR Assay Name Free Induction Decay Data File Protocol REF Normalization Name Derived Spectral Data File Protocol REF Data Transformation Name Metabolite Assignment File
Sh00_AQ_12CGln_107 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12CGln_107 Sh00_AQ_12CGln_107.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_12CGln_108 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12CGln_108 Sh00_AQ_12CGln_108.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_12CGln_109 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12CGln_109 Sh00_AQ_12CGln_109.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_12C_10 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12C_10 Sh00_AQ_12C_10.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_12C_11 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12C_11 Sh00_AQ_12C_11.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_12C_12 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12C_12 Sh00_AQ_12C_12.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13CGln_122 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13CGln_122 Sh00_AQ_13CGln_122.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13CGln_123 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13CGln_123 Sh00_AQ_13CGln_123.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13CGln_124 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13CGln_124 Sh00_AQ_13CGln_124.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13C_22 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13C_22 Sh00_AQ_13C_22.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13C_23 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13C_23 Sh00_AQ_13C_23.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13C_24 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13C_24 Sh00_AQ_13C_24.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_12C_01 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_12C_01 Sh22_AQ_12C_01.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_12C_02 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_12C_02 Sh22_AQ_12C_02.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_12C_03 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_12C_03 Sh22_AQ_12C_03.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_12CGln_101 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_12CGln_101 Sh22_AQ_12CGln_101.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_12CGln_102 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_12CGln_102 Sh22_AQ_12CGln_102.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_12CGln_103 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_12CGln_103 Sh22_AQ_12CGln_103.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_13C_13 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_13C_13 Sh22_AQ_13C_13.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_13C_14 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_13C_14 Sh22_AQ_13C_14.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_13C_15 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_13C_15 Sh22_AQ_13C_15.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_13CGln_116 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_13CGln_116 Sh22_AQ_13CGln_116.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_13CGln_117 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_13CGln_117 Sh22_AQ_13CGln_117.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_13CGln_118 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh22_AQ_13CGln_118 Sh22_AQ_13CGln_118.rar Data transformation Metabolite identification m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv

Assay  4

Assay file name: a_TOCSY_EXD2_NMR_spectroscopy.txt
Technology: NMR spectroscopy
Platform: Bruker

Instrumentation

Sample Name Protocol REF Extraction Method Extract Name Protocol REF NMR tube type Solvent Sample pH Temperature Unit Labeled Extract Name Label Protocol REF Instrument NMR Probe Number of transients Pulse sequence name Magnetic field strength Unit Acquisition Parameter Data File Protocol REF NMR Assay Name Free Induction Decay Data File Protocol REF Normalization Name Derived Spectral Data File Protocol REF Data Transformation Name Metabolite Assignment File
Sh00_AQ_12C_10_TOCSY Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 1024 2D TOCSY with presaturation using MLEV (mlevphpr) 14.1 tesla NMR assay Sh00_AQ_12C_10_TOCSY Sh00_AQ_12C_10_TOCSY.rar Data transformation Metabolite identification
Sh00_AQ_13CGln_TOCSY Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 1024 2D TOCSY with presaturation using MLEV (mlevphpr) 14.1 tesla NMR assay Sh00_AQ_13CGln_TOCSY Sh00_AQ_13CGln_TOCSY.rar Data transformation Metabolite identification
sh00_AQ_13C_23_TOCSY Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 1024 2D TOCSY with presaturation using MLEV (mlevphpr) 14.1 tesla NMR assay sh00_AQ_13C_23_TOCSY sh00_AQ_13C_23_TOCSY.rar Data transformation Metabolite identification
Sh00_AQ_12CGln_109_TOCSY Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 1024 2D TOCSY with presaturation using MLEV (mlevphpr) 14.1 tesla NMR assay Sh00_AQ_12CGln_109_TOCSY Sh00_AQ_12CGln_109_TOCSY.rar Data transformation Metabolite identification

Assay  5

Assay file name: a_13C_PEPA_NMR_spectroscopy.txt
Technology: NMR spectroscopy
Platform: Bruker

Instrumentation

Sample Name Protocol REF Extraction Method Extract Name Protocol REF NMR tube type Solvent Sample pH Temperature Unit Labeled Extract Name Label Protocol REF Instrument NMR Probe Number of transients Pulse sequence name Magnetic field strength Unit Acquisition Parameter Data File Protocol REF NMR Assay Name Free Induction Decay Data File Protocol REF Normalization Name Derived Spectral Data File Protocol REF Data Transformation Name Metabolite Assignment File
Sh00_AQ_12C_10 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12C_10 Sh00_AQ_12C_10.rar Data transformation Metabolite identification
Sh00_AQ_12C_11 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12C_11 Sh00_AQ_12C_11.rar Data transformation Metabolite identification
Sh00_AQ_12C_12 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12C_12 Sh00_AQ_12C_12.rar Data transformation Metabolite identification
Sh00_AQ_13C_22 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13C_22 Sh00_AQ_13C_22.rar Data transformation Metabolite identification
Sh00_AQ_13C_23 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13C_23 Sh00_AQ_13C_23.rar Data transformation Metabolite identification
Sh00_AQ_13C_24 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13C_24 Sh00_AQ_13C_24.rar Data transformation Metabolite identification
Sh00_AQ_12CGln_107 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12CGln_107 Sh00_AQ_12CGln_107.rar Data transformation Metabolite identification
Sh00_AQ_12CGln_108 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12CGln_108 Sh00_AQ_12CGln_108.rar Data transformation Metabolite identification
Sh00_AQ_12CGln_109 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_12CGln_109 Sh00_AQ_12CGln_109.rar Data transformation Metabolite identification
Sh00_AQ_13CGln_122 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13CGln_122 Sh00_AQ_13CGln_122.rar Data transformation Metabolite identification
Sh00_AQ_13CGln_123 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13CGln_123 Sh00_AQ_13CGln_123.rar Data transformation Metabolite identification
Sh00_AQ_13CGln_124 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin carbon-13 atom NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 6144 1D 13C with power-gated decoupling (zgpg) 14.1 tesla NMR assay Sh00_AQ_13CGln_124 Sh00_AQ_13CGln_124.rar Data transformation Metabolite identification

Assay  6

Assay file name: a_HSQC_PEPA_NMR_spectroscopy.txt
Technology: NMR spectroscopy
Platform: Bruker

Instrumentation

Sample Name Protocol REF Extraction Method Extract Name Protocol REF NMR tube type Solvent Sample pH Temperature Unit Labeled Extract Name Label Protocol REF Instrument NMR Probe Number of transients Pulse sequence name Magnetic field strength Unit Acquisition Parameter Data File Protocol REF NMR Assay Name Free Induction Decay Data File Protocol REF Normalization Name Derived Spectral Data File Protocol REF Data Transformation Name Metabolite Assignment File
Sh00_AQ_12C_10 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 512 2D HSQC with presaturation (hsqcphpr) 14.1 tesla NMR assay Sh00_AQ_12C_10 Sh00_AQ_12C_10.rar Data transformation Metabolite identification
Sh00_AQ_12C_11 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 512 2D HSQC with presaturation (hsqcphpr) 14.1 tesla NMR assay Sh00_AQ_12C_11 Sh00_AQ_12C_11.rar Data transformation Metabolite identification

Assay  7

Assay file name: a_NOESY_PEPA_NMR_spectroscopy.txt
Technology: NMR spectroscopy
Platform: Bruker

Instrumentation

Sample Name Protocol REF Extraction Method Extract Name Protocol REF NMR tube type Solvent Sample pH Temperature Unit Labeled Extract Name Label Protocol REF Instrument NMR Probe Number of transients Pulse sequence name Magnetic field strength Unit Acquisition Parameter Data File Protocol REF NMR Assay Name Free Induction Decay Data File Protocol REF Normalization Name Derived Spectral Data File Protocol REF Data Transformation Name Metabolite Assignment File
Sh00_AQ_12C_10 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12C_10 Sh00_AQ_12C_10.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_12C_11 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12C_11 Sh00_AQ_12C_11.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_12C_12 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12C_12 Sh00_AQ_12C_12.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13C_22 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13C_22 Sh00_AQ_13C_22.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13C_23 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13C_23 Sh00_AQ_13C_23.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13C_24 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13C_24 Sh00_AQ_13C_24.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_12CGln_107 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12CGln_107 Sh00_AQ_12CGln_107.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_12CGln_108 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12CGln_108 Sh00_AQ_12CGln_108.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_12CGln_109 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_12CGln_109 Sh00_AQ_12CGln_109.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13CGln_122 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13CGln_122 Sh00_AQ_13CGln_122.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13CGln_123 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13CGln_123 Sh00_AQ_13CGln_123.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
Sh00_AQ_13CGln_124 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 256 1D NOESY with presaturation (noesypr1d) 14.1 tesla NMR assay Sh00_AQ_13CGln_124 Sh00_AQ_13CGln_124.rar Data transformation Metabolite identification m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv

Assay  8

Assay file name: a_HSQC_EXD2_NMR_spectroscopy.txt
Technology: NMR spectroscopy
Platform: Bruker

Instrumentation

Sample Name Protocol REF Extraction Method Extract Name Protocol REF NMR tube type Solvent Sample pH Temperature Unit Labeled Extract Name Label Protocol REF Instrument NMR Probe Number of transients Pulse sequence name Magnetic field strength Unit Acquisition Parameter Data File Protocol REF NMR Assay Name Free Induction Decay Data File Protocol REF Normalization Name Derived Spectral Data File Protocol REF Data Transformation Name Metabolite Assignment File
Sh00_AQ_12C_10 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 512 2D HSQC with presaturation (hsqcphpr) 14.1 tesla NMR assay Sh00_AQ_12C_10 Sh00_AQ_12C_10.rar Data transformation Metabolite identification
Sh00_AQ_12C_11 Extraction NMR sample 5 mm standard 0.05 mM phosphate buffered D2O 7.4 300 kelvin hydrogen molecular entity NMR spectroscopy Bruker AVANCE III 600 MHz spectrometer 5 mm CPTCI 1H/13C/31P Z-GRD cryoprobe 512 2D HSQC with presaturation (hsqcphpr) 14.1 tesla NMR assay Sh00_AQ_12C_11 Sh00_AQ_12C_11.rar Data transformation Metabolite identification
   Download study (FTP)  |     Download metadata    

Aspera Download Details:

List of study files   Subset

File
audit
Dolphin_Glc_metabolights.csv
Sh00_AQ_12CGln_107.rar
Sh00_AQ_12CGln_108.rar
Dolphin_Gln_metabolights.csv
Sh00_AQ_12C_10.rar
Sh00_AQ_12CGln_109_TOCSY.rar
a_pepa_metabolite_profiling_NMR_spectroscopy.txt
Sh00_AQ_12CGln_109.rar
Sh00_AQ_12C_10_TOCSY.rar
Sh00_AQ_12C_11.rar
Sh00_AQ_12C_12.rar
Sh00_AQ_13CGln_122.rar
Sh00_AQ_13CGln_122_HSQC.rar
Sh00_AQ_13CGln_123.rar
Sh00_AQ_13CGln_124.rar
Sh00_AQ_13CGln_TOCSY.rar
Sh00_AQ_13C_22.rar
Sh00_AQ_13C_23.rar
Sh00_AQ_13C_23_HSQC.rar
Sh00_AQ_13C_24.rar
i_Investigation.txt
s_PEPA.txt
sh00_AQ_13C_23_TOCSY.rar
a_exd2_metabolite_profiling_NMR_spectroscopy.txt
m_exd2_metabolite_profiling_NMR_spectroscopy_v2_maf.tsv
m_pepa_metabolite_profiling_NMR_spectroscopy_v2_maf.tsv
Sh22_AQ_12C_03.rar
Sh22_AQ_12CGln_101.rar
Sh22_AQ_12C_02.rar
Sh22_AQ_12C_01.rar
s_EXD2.txt
Sh22_AQ_12CGln_102.rar
Sh22_AQ_12CGln_103.rar
Sh22_AQ_13C_13.rar
Sh22_AQ_13C_14.rar
Sh22_AQ_13C_15.rar
Sh22_AQ_13CGln_116.rar
Sh22_AQ_13CGln_117.rar
Sh22_AQ_13CGln_118.rar
metexplore_mapping.json
a_13C_EXD2_NMR_spectroscopy.txt
a_13C_PEPA_NMR_spectroscopy.txt
a_HSQC_EXD2_NMR_spectroscopy.txt
a_HSQC_PEPA_NMR_spectroscopy.txt
a_NOESY_EXD2_NMR_spectroscopy.txt
a_NOESY_NMR_spectroscopy.txt
a_NOESY_PEPA_NMR_spectroscopy.txt
a_TOCSY_NMR_spectroscopy.txt
a_TOCSY_EXD2_NMR_spectroscopy.txt
a_TOCSY_PEPA_NMR_spectroscopy.txt
m_NOESY_EXD2_NMR_spectroscopy_v2_maf.tsv
m_NOESY_PEPA_NMR_spectroscopy_v2_maf.tsv
s_mtbls247.txt

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Validations marked with (*) have been allowed by the MetaboLights Curators.
Click here for the detailed description of Validations.
Condition Status Description Requirement Group Message
PASSES Study Title MANDATORY STUDY OK
PASSES Study Description MANDATORY STUDY OK
PASSES Study text successfully parsed OPTIONAL STUDY OK
PASSES Study Contact(s) have listed email MANDATORY CONTACT OK
PASSES Sample(s) MANDATORY SAMPLES OK
PASSES Sample Name consistency check MANDATORY ASSAYS OK
* PASSES Publication PubMed ID and Publication DOI are missing OPTIONAL PUBLICATION Please provide Pubmed ID and/or DOI
PASSES Minimal Experimental protocol MANDATORY PROTOCOLS OK
PASSES Comprehensive Experimental protocol OPTIONAL PROTOCOLS OK
PASSES Extraction protocol description MANDATORY PROTOCOLS OK
PASSES Data transformation protocol description MANDATORY PROTOCOLS OK
PASSES Metabolite Identification protocol description MANDATORY PROTOCOLS OK
PASSES NMR sample protocol description MANDATORY PROTOCOLS OK
PASSES NMR spectroscopy protocol description MANDATORY PROTOCOLS OK
PASSES NMR assay protocol description MANDATORY PROTOCOLS OK
PASSES Sample Collection protocol description MANDATORY PROTOCOLS OK
PASSES Protocols text successfully parsed OPTIONAL PROTOCOLS OK
PASSES Organism name MANDATORY ORGANISM OK
PASSES Organism part MANDATORY ORGANISM OK
PASSES Study Factors MANDATORY FACTORS OK
PASSES Assay platform information OPTIONAL ASSAYS OK
PASSES Assay has raw files referenced MANDATORY FILES OK
PASSES Assay referenced raw files detection in filesystem MANDATORY FILES OK
PASSES Raw files in the Assay(s) have the correct format MANDATORY FILES OK
PASSES Assay(s) MANDATORY ASSAYS OK
* PASSES All Assays have Metabolite Assignment File (MAF) referenced OPTIONAL FILES Metabolite identification protocol is described but the following Assay(s) have no Metabolite Assignment File (MAF) reference: Assay 1, Assay 2, Assay 4, Assay 5, Assay 6, Assay 8
PASSES At least one assay has Metabolite Assignment Files (MAFs) referenced MANDATORY FILES OK
PASSES Metabolite Assignment File (MAF) is present in Study folder MANDATORY FILES OK
PASSES Metabolite Assignment File (MAF) has correct format MANDATORY FILES OK
PASSES Metabolite Identification File (MAF) content MANDATORY FILES OK
PASSES ISA-Tab investigation file check MANDATORY ISATAB OK

Pathways - Assay  1



MetExplore Pathways Mapping

Name DB Identifier Mapped Metabolite(s)

Pathways - Assay  2



MetExplore Pathways Mapping

Name DB Identifier Mapped Metabolite(s)

Pathways - Assay  3



MetExplore Pathways Mapping

Name DB Identifier Mapped Metabolite(s)

Pathways - Assay  4



MetExplore Pathways Mapping

Name DB Identifier Mapped Metabolite(s)

Pathways - Assay  5



MetExplore Pathways Mapping

Name DB Identifier Mapped Metabolite(s)

Pathways - Assay  6



MetExplore Pathways Mapping

Name DB Identifier Mapped Metabolite(s)

Pathways - Assay  7



MetExplore Pathways Mapping

Name DB Identifier Mapped Metabolite(s)

Pathways - Assay  8



MetExplore Pathways Mapping

Name DB Identifier Mapped Metabolite(s)