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MB Sample ID: SA254631
Local Sample ID: | A35097-120 |
Subject ID: | SU002629 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002629 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
A35097-120 | SA254631 | FL032629 | Plasma | Sample type |
A35097-120 | SA254631 | FL032629 | Arg1fl/fl | Condition |
Collection:
Collection ID: | CO002622 |
Collection Summary: | 6~8-month-old female & male Lyz2-Cre +/+; Arg1fl/fl and litter mate control Arg1fl/fl mice with murine PDAC orthotopic tumors underwent dual jugular vein & carotid artery catheterization surgery. On day 5 of post recovery, mice received a 0.28 mg/g 10 min. bolus followed by a continuous 4 hr. infusion 0.005 mg/g/min infusion of 15N2-glutamine (Cambridge Isotope Laboratory #NLM-1328-PK). Plasma samples were taken at time points: 0' 15' 30' 60' 120' 180’ 240’. Tumors and tissues were harvested at 240’ and immediately snap frozen with liquid nitrogen stored at -80°C prior to analysis. |
Sample Type: | Tumor, plasma |
Treatment:
Treatment ID: | TR002641 |
Treatment Summary: | 6~8-month-old female & male Lyz2-Cre +/+; Arg1fl/fl and litter mate control Arg1fl/fl mice with murine PDAC orthotopic tumors underwent dual jugular vein & carotid artery catheterization surgery. On day 5 of post recovery, mice received a 0.28 mg/g 10 min. bolus followed by a continuous 4 hr. infusion 0.005 mg/g/min infusion of 15N2-glutamine (Cambridge Isotope Laboratory #NLM-1328-PK). Plasma samples were taken at time points: 0' 15' 30' 60' 120' 180’ 240’. Tumors and tissues were harvested at 240’ and immediately snap frozen with liquid nitrogen stored at -80°C prior to analysis. |
Sample Preparation:
Sampleprep ID: | SP002635 |
Sampleprep Summary: | Cryogenically frozen tumor pieces were ground to a fine homogenous powder with a liquid nitrogen cooled mortar and pestle. ~30mg of tissue powder was weighed into sample tubes, and metabolites were extracted with 600µL HPLC grade methanol, 300µL HPLC grade water, and 400µL chloroform. Samples were vortexed for 10min at 4°C, centrifuged 21,000xg at 4°C for 10 min. 400µL of the aqueous top layer was removed into a new tube and dried under nitrogen. Dried tumor extracts were resuspended in 100µL HPLC grade water and LC-MS analysis was performed as described before(Sullivan et al., 2019b, 2019a). For plasma samples, we extracted polar metabolites from 5µL of sample using 45µL of a 75:25:0.1 HPLC grade acetonitrile:methanol:formic acid extraction mix. Samples in extraction mix were vortexed for 10 min at 4°C and centrifugated at 15,000x rpm for 10 min at 4°C to pellet insoluble material. 20µL of the soluble polar metabolite supernatant was moved to sample vials for analysis by LC-MS. |
Combined analysis:
Analysis ID | AN004163 |
---|---|
Analysis type | MS |
Chromatography type | HILIC |
Chromatography system | Thermo Vanquish |
Column | Waters Atlantis BEH Z-HILIC (150 x 2.1 mm, 2.5um) |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo Orbitrap IQ-X Tribrid |
Ion Mode | UNSPECIFIED |
Units | Natural abundance corrected percentages |
Chromatography:
Chromatography ID: | CH003082 |
Instrument Name: | Thermo Vanquish |
Column Name: | Waters Atlantis BEH Z-HILIC (150 x 2.1 mm, 2.5um) |
Column Temperature: | 30°C |
Flow Gradient: | 0 minute: 85% B, 0.5 minute: 85% B, 18 minutes: 20% B, 20 minutes: 20% B, 20.5 minutes: 85% B and 28 minutes: 85% B |
Flow Rate: | 0.2 mL/minute |
Solvent A: | 20 mM ammonium bicarbonate at pH 9.6, adjusted by ammonium hydroxide addition |
Solvent B: | 100% Acetonitrile |
Chromatography Type: | HILIC |
MS:
MS ID: | MS003910 |
Analysis ID: | AN004163 |
Instrument Name: | Thermo Orbitrap IQ-X Tribrid |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Data acquisition was done using the Xcalibur software (Thermo Scientific) in full-scan mode with a range of 70-1000 m/z at 60K. Metabolite identification was done by matching the retention time and MS/MS fragmentation to the reference standards. Data analysis was performed using Tracefinder 5.1 software (Thermo Scientific). |
Ion Mode: | UNSPECIFIED |