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MB Sample ID: SA207547
| Local Sample ID: | L8_MNAM |
| Subject ID: | SU002249 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002249 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| L8_MNAM | SA207547 | FL025449 | WT | Genotype |
| L8_MNAM | SA207547 | FL025449 | Sham | Sample group |
| L8_MNAM | SA207547 | FL025449 | female | Sex |
| L8_MNAM | SA207547 | FL025449 | MNAM treatment | Treatment |
Collection:
| Collection ID: | CO002242 |
| Collection Summary: | Mouse livers were crushed in liquid-nitrogen and homogenized. |
| Sample Type: | Liver |
Treatment:
| Treatment ID: | TR002261 |
| Treatment Summary: | Daily 250 mg/kg MNAM injection for 12 days in the absence (sham) and presence (4T1cancers). |
Sample Preparation:
| Sampleprep ID: | SP002255 |
| Sampleprep Summary: | Metabolites were extracted from the frozen livers (approximately 5 mg) using the Bligh and Dyer’s method with some modifications. Briefly, each sample was mixed with 1 mL of cold methanol containing 10-camphorsulfonic acid (1.5 nmol) and piperazine-1,4-bis (2-ethanesulfonic acid) (PIPES, 1.5 nmol) as internal standards for mass spectrometry-based metabolomic analysis. The samples were vigorously mixed by vortexing for 1 min followed by 5 min of sonication. The extracts were then centrifuged at 16,000 × g for 5 min at 4 °C, and the resultant supernatant (400 uL) was collected. After mixing 400 uL of supernatant with 400 uL of chloroform and 320 uL of water, the aqueous and organic layers were separated by vortexing and subsequent centrifugation at 16,000 × g and 4 °C for 5 min. The aqueous (upper) layer (500 uL) was transferred into a clean tube. After the aqueous layer extracts were evaporated under vacuum, the dried extracts were stored at −80 °C until the analysis of hydrophilic metabolites. Prior to analysis, the dried aqueous layer was reconstituted in 50 uL of water. |
Combined analysis:
| Analysis ID | AN003544 | AN003545 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Ion exchange | Reversed phase |
| Chromatography system | Dionex ICS-5000+ HPIC system | Shimadzu Nexera X2 |
| Column | Dionex IonPac AS11-HC (250 x 2mm,4um) | Discovery HS (150 x 2.1mm,3um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | peak area | peak area |
Chromatography:
| Chromatography ID: | CH002618 |
| Chromatography Summary: | Anionic polar metabolites (i.e., organic acids, sugar phosphates, nucleotides, etc.) were analyzed via IC/HRMS/MS. |
| Instrument Name: | Dionex ICS-5000+ HPIC system |
| Column Name: | Dionex IonPac AS11-HC (250 x 2mm,4um) |
| Chromatography Type: | Ion exchange |
| Chromatography ID: | CH002619 |
| Chromatography Summary: | Cationic polar metabolites (i.e., amino acids, bases, nucleosides, NAM, SAM, MNAM, SAH, me2PY, me4PY, etc) were analyzed via PFPP-LC/HRMS/MS. |
| Instrument Name: | Shimadzu Nexera X2 |
| Column Name: | Discovery HS (150 x 2.1mm,3um) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003302 |
| Analysis ID: | AN003544 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | NEGATIVE |
| MS ID: | MS003303 |
| Analysis ID: | AN003545 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | POSITIVE |
