Summary of Study ST001976

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench,, where it has been assigned Project ID PR001255. The data can be accessed directly via it's Project DOI: 10.21228/M8GT41 This work is supported by NIH grant, U2C- DK119886.


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 IDST001976
Study TitleAnti-oxidation metabolism measurement in mammalian cells and tissues by quantitative LC/MS method (III)
Study SummaryAnti-oxidative metabolism measurement in mouse CSF by quantitative LC/MS method of mouse CSF at 24H of MTX treatment, for either control GFP or SOD3-overexpressing ChP mice.
Boston Children's Hospital, Harvard Medical School
Last NamePetrova
First NameBoryana
Address300 Longwood Ave
Submit Date2021-09-14
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2022-08-29
Release Version1
Boryana Petrova Boryana Petrova application/zip

Select appropriate tab below to view additional metadata details:


Project ID:PR001255
Project DOI:doi: 10.21228/M8GT41
Project Title:Gene-therapy enhances CSF’s anti-oxidative activity to mitigate chemotherapy side effects
Project Type:Anti-oxidative Metabolism Measurement in CSF of MTX-treated mice under gene therapy by quantitative LC/MS method
Project Summary:In order to test the protective activity of SOD3 levels on the redox state of the CSF of MTX-treated mice, targeted metabolomics on CSF was employed at different timepoints after treatment, on mice of both sexes, at various levels of impacted hSOD3 expression.
Institute:Boston Childrens Hospital
Laboratory:Naama Kanarek
Last Name:Petrova
First Name:Boryana
Address:300 Longwood Av, Boston, MA, 2115, USA


Subject ID:SU002056
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Gender:Male and female


Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Factor
SA185342pooled sample QCa-
SA185343pooled sample QC 1/10 dilution b-
SA185344pooled sample QC 1/10 dilution a-
SA185345pooled sample QCb-
SA185346pooled sample QCc-
SA185347pooled sample QC 1/3 dilution-
SA185348pooled sample QCf-
SA185349pooled sample QCd-
SA185350blank end-
SA185351pooled sample QCg-
SA185352blank start 3-
SA185353blank start 2-
SA185354blank start 1-
SA185355blank start 4-
SA185356mouse CSF 124h GFP 1
SA185357mouse CSF 224h GFP 2
SA185358mouse CSF 324h GFP 3
SA185359mouse CSF 424h GFP 4
SA185360mouse CSF 524h GFP 5
SA185361mouse CSF 624h GFP 6
SA185362mouse CSF 724h GFP 7
SA185363mouse CSF 824h MTX 1
SA185364mouse CSF 924h MTX 2
SA185365mouse CSF 1024h MTX 3
SA185366mouse CSF 1124h MTX 4
SA185367mouse CSF 1224h MTX 5
SA185368mouse CSF 1324h MTX 6
SA185369mouse CSF 1424h MTX 7
SA185377mouse CSF 1524h SOD3-1
SA185378mouse CSF 1624h SOD3-2
SA185379mouse CSF 1724h SOD3-3
SA185380mouse CSF 1824h SOD3-4
SA185381mouse CSF 1924h SOD3-5
SA185370mouse CSF 2024h SOD3+MTX-1
SA185371mouse CSF 2124h SOD3+MTX-2
SA185372mouse CSF 2224h SOD3+MTX-3
SA185373mouse CSF 2324h SOD3+MTX-4
SA185374mouse CSF 2424h SOD3+MTX-5
SA185375mouse CSF 2524h SOD3+MTX-6
SA185376mouse CSF 2624h SOD3+MTX-7
Showing results 1 to 40 of 40


Collection ID:CO002049
Collection Summary:All animal studies were performed under the protocol approved by the Institutional Animal Care and Use Committee (IACUC) of Boston Children’s Hospital (BCH). CD-1 mice (timed pregnant and 6-8 weeks) and Sprague Dawley rats (6-8 weeks) were purchased (Charles River Laboratories). Male and female animals were used equally for all studies. All animals were housed in a 12h light-dark cycle with ad libitum access to food and water. CSF was collected from the cisterna magna and centrifuged at 10,000g for 10 min. at 4 °C to remove any tissue debris (38). CSF samples were flash frozen and stored at −80°C until use.
Sample Type:CSF


Treatment ID:TR002068
Treatment Summary:Control AAV-GFP, MTX, AAV-SOD3 + buffers from Study Design. The pAAV-IRES-hrGFP vector was purchased from Agilent Technologies (240075) and pcDNA3.1-myc HisA (-)/ human SOD3 (myc-his tag at C-terminal) (36). Virus production and purification were performed by the Viral Core at BCH according to standard procedures (37). CD-1 mice (6-8 weeks) or Sprague Dawley rats (8 weeks) were given a single intravenous injection of 75 mg/kg MTX (Fisher Scientific, AAJ66364MD) dissolved in 0.9 % NaCl (Mountainside Medical, 0409-4888-10). MTX was freshly prepared for each experiment. MTX in mouse serum or CSF was assessed by MTX ELISA according to the manufacturer's instructions (Enzo Life Sciences, 142-0001).

Sample Preparation:

Sampleprep ID:SP002062
Sampleprep Summary:CSF was collected from the cisterna magna and centrifuged at 10,000g for 10 min. at 4 °C to remove any tissue debris (38). CSF samples were flash frozen and stored at −80°C until use. For characterization by mass spectrometry, CSF was acquired 4h, 24h, and 48h following a single 75 mg/kg MTX injection from 6-8 mice and flash frozen for further analysis. Per condition, 3 μl of CSF were extracted by brief sonication in 240 μl 100% methanol, supplemented with isotopically labeled internal standards (17 amino acids and reduced glutathione, Cambridge Isotope Laboratories, MSK-A2-1.2 and CNLM-6245-10) and 60 μl 20 mM Ellman’s reagent in water (Sigma-Aldrich, D8130). After centrifugation for 10min. at maximum speed on a benchtop centrifuge (Eppendorf) the cleared supernatant was dried using a nitrogen dryer and reconstituted in 30 µl water by brief sonication. Extracted metabolites were spun again and cleared supernatant was transferred in LC-MS microvials. A small amount of each sample was pooled and serially diluted 3 and 10-fold to be used as quality controls throughout the batch run. Two microliters (equivalent to 0.2 ul of CSF) of reconstituted sample were injected into a ZIC-pHILIC 150 × 2.1 mm (5 µm particle size) column (EMD Millipore) operated on a Dionex UltiMate 3000 UPLC system (Thermo Fisher Scientific). Chromatographic separation was achieved using the following conditions: buffer A was acetonitrile; buffer B was 20 mM ammonium carbonate, 0.1% ammonium hydroxide. Gradient conditions were: linear gradient from 20% to 80% B; 20–20.5 min: from 80% to 20% B; 20.5–28 min: hold at 20% B. The column oven and autosampler tray were held at 25 °C and 4 °C, respectively. The MS data acquisition was on a QExactive benchtop orbitrap mass spectrometer equipped with an Ion Max source and a HESI II probe and was performed in a range of m/z= 70–1000, with the resolution set at 70,000, the AGC target at 1x106, and the maximum injection time (Max IT) at 20 msec. For tSIM scans, the resolution was set at 70,000, the AGC target was 1x105, and the max IT was 100 msec. Relative quantitation of polar metabolites was performed with TraceFinder 4.1 (Thermo Fisher Scientific) using a 5 ppm mass tolerance and referencing an in-house library of chemical standards. Pooled samples and fractional dilutions were prepared as quality controls and only those metabolites were taken for further analysis, for which the correlation between the dilution factor and the peak area was >0.95 (high confidence metabolites). Normalization for biological material amounts was based on the total integrated peak area values of high-confidence metabolites within an experimental batch after normalizing to the averaged factor from all mean-centered chromatographic peak areas of isotopically labeled amino acids internal standards (Cambridge Isotope Laboratories). The data were Log transformed and Pareto scaled for MetaboAnalyst-based statistical or pathway analysis (41). We profiled 200 metabolites, 85 of which were detected in CSF and passed our quality control protocol.

Combined analysis:

Analysis ID AN003224
Analysis type MS
Chromatography type HILIC
Chromatography system Thermo Q Exactive Orbitrap
Column EMD Millipore ZIC-HILIC (100 x 2.1mm,3.5um)
MS instrument type Orbitrap
MS instrument name Thermo Exactive Plus Orbitrap
Units ppm


Chromatography ID:CH002378
Instrument Name:Thermo Q Exactive Orbitrap
Column Name:EMD Millipore ZIC-HILIC (100 x 2.1mm,3.5um)
Chromatography Type:HILIC


MS ID:MS002998
Analysis ID:AN003224
Instrument Name:Thermo Exactive Plus Orbitrap
Instrument Type:Orbitrap
MS Comments:Xcalibur + Tracefinder