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MB Sample ID: SA340990

Local Sample ID:06_Fasnall_MCF7_Tum_Gluc_1h_F_1072
Subject ID:SU003269
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:NOD Scid Gamma (NSG)
Age Or Age Range:~4.5 months
Weight Or Weight Range:24.3 +/- 1.9 g (mean +/- SD)
Gender:Female
Animal Housing:Wistar Animal Facility
Species Group:Rodents (order Rodentia)

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Subject:

Subject ID:SU003269
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:NOD Scid Gamma (NSG)
Age Or Age Range:~4.5 months
Weight Or Weight Range:24.3 +/- 1.9 g (mean +/- SD)
Gender:Female
Animal Housing:Wistar Animal Facility
Species Group:Rodents (order Rodentia)

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
06_Fasnall_MCF7_Tum_Gluc_1h_F_1072SA340990FL040339Xenograft tumorsSample source
06_Fasnall_MCF7_Tum_Gluc_1h_F_1072SA340990FL0403391 g/kg [U-13C6] D-glucose in PBSIsotopic Tracer
06_Fasnall_MCF7_Tum_Gluc_1h_F_1072SA340990FL04033910 mg/kg FasnallDrug Treatment

Collection:

Collection ID:CO003262
Collection Summary:Tissue metabolite extracts were prepared from ~50 mg snap-frozen tissue samples. Each sample was weighed while frozen. Frozen samples were ground at the temperature of liquid nitrogen by Retch Cryomill. Then, 80% methanol was added to each tube at a ratio of 1 ml per 50 mg of tissue.
Collection Protocol Filename:DM_metabolomics_samples.txt
Sample Type:Tumor cells
Collection Method:80% methanol extraction
Volumeoramount Collected:~1 ml
Storage Conditions:-80℃
Collection Vials:1.5 ml plastic centrifuge tubes
Storage Vials:1.5 ml plastic centrifuge tubes

Treatment:

Treatment ID:TR003278
Treatment Summary:Animals received 1 g/kg [U-13C6] D-glucose bolus intraperitoneally with a 10 mg/kg dose of Fasnall.
Treatment Compound:Fasnall
Treatment Route:Intraperitoneal injection
Treatment Dose:10 mg/kg
Treatment Dosevolume:50 ul
Treatment Doseduration:1 h
Treatment Vehicle:1:1 DMSO/PBS
Animal Anesthesia:None
Animal Fasting:None
Animal Endp Euthanasia:CO2
Animal Endp Tissue Coll List:50 mg tumor samples

Sample Preparation:

Sampleprep ID:SP003276
Sampleprep Summary:Tissue metabolite extracts were prepared from ~50 mg snap-frozen tissue samples. Each sample was weighed while frozen. Frozen samples were ground at the temperature of liquid nitrogen by Retch Cryomill. Then, 80% methanol was added to each tube at a ratio of 1 ml per 50 mg of tissue. All samples were centrifuged at 18,000 g and 4 °C for 20 min. The supernatant was then transferred to new Eppendorf tubes and centrifuged again with the same parameters. After centrifugation, the extracts were transferred to glass LC-MS vials.
Sampleprep Protocol Filename:DM_metabolomics_samples.txt
Extraction Method:80% methanol
Extract Enrichment:None
Extract Cleanup:None
Extract Storage:-80℃
Sample Resuspension:None
Sample Derivatization:None
Sample Spiking:None
Subcellular Location:Intracellular metabolites

Combined analysis:

Analysis ID AN005171 AN005172
Analysis type MS MS
Chromatography type HILIC HILIC
Chromatography system Thermo Dionex Ultimate 3000 RS Thermo Dionex Ultimate 3000 RS
Column Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive HF-X Orbitrap Thermo Q Exactive HF-X Orbitrap
Ion Mode NEGATIVE POSITIVE
Units AU AU

Chromatography:

Chromatography ID:CH003912
Instrument Name:Thermo Dionex Ultimate 3000 RS
Column Name:Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um)
Column Temperature:40
Flow Gradient:A linear solvent gradient of a total duration 22.5 min was starting with 0.2 ml/min flow rate of 80% solvent B, 12.5 min – 30%, 15 min – 30%, 15.2 min – 80%, 20 min – 80%, 21 min – flow rate 0.3 ml/min, 22 min – flow rate 0.3 ml/min, 22.1 min – flow rate 0.2 ml/min.
Flow Rate:0.2 ml/min
Solvent A:100% Water; 0.01% ammonium hydroxide; 20 mM ammonium bicarbonate
Solvent B:100% Acetonitrile
Chromatography Type:HILIC

MS:

MS ID:MS004906
Analysis ID:AN005171
Instrument Name:Thermo Q Exactive HF-X Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:The HESI ion source voltage was set to the following parameters: 3,600 V in both polarity modes, sheath gas 30, auxiliary gas 5, spare gas 0, probe heater 200 °C, capillary temperature 325 °C, S-Lens RF level 65. The mass spectrometer was set to acquire data in the polarity-switching mode averaging two microscans with 60,000 resolution, automatic gain control (AGC) target 5e6, scan range 72-1080 m/z, and maximum orbitrap injection time (IT) 200 ms. Data were converted into the mzXML format by ProteoWizard and analyzed in MAVEN.
Ion Mode:NEGATIVE
Ion Source Temperature:200
Ion Spray Voltage:3600
Ionization:Both
Source Temperature:200
Automatic Gain Control:5e6
Dataformat:RAW
  
MS ID:MS004907
Analysis ID:AN005172
Instrument Name:Thermo Q Exactive HF-X Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:The HESI ion source voltage was set to the following parameters: 3,600 V in both polarity modes, sheath gas 30, auxiliary gas 5, spare gas 0, probe heater 200 °C, capillary temperature 325 °C, S-Lens RF level 65. The mass spectrometer was set to acquire data in the polarity-switching mode averaging two microscans with 60,000 resolution, automatic gain control (AGC) target 5e6, scan range 72-1080 m/z, and maximum orbitrap injection time (IT) 200 ms. Data were converted into the mzXML format by ProteoWizard and analyzed in MAVEN.
Ion Mode:POSITIVE
Ion Source Temperature:200
Ion Spray Voltage:3600
Ionization:Both
Source Temperature:200
Automatic Gain Control:5e6
Dataformat:RAW
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