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MB Sample ID: SA138193
| Local Sample ID: | 33 |
| Subject ID: | SU001710 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001710 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 33 | SA138193 | FL017573 | MCAD-/- | Mouse_Genotype |
| 33 | SA138193 | FL017573 | Cs | Treatment |
Collection:
| Collection ID: | CO001703 |
| Collection Summary: | MCAD-/- mice were re-derived as germ-free (GF) for the purpose of this study. MCAD-/- mice or MCAD+/+ controls were maintained in sterile gnotobiotic isolators or cages with HEPA-filtered air supply. Urine from GF MCAD-/- and MCAD+/+ mice was collected under sterile conditions. Seven days subsequent, mice were mono-colonized with wild-type C. sporogenes (Cs) by gavage of 200 µL saturated overnight culture. Urine was collected one week after colonization. C. sporogenes colonization was verified with serial dilution plating. Urine samples were immediately frozen on dry ice and stored under -80 oC until analysis. |
| Sample Type: | Urine |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001723 |
| Treatment Summary: | MCAD-/- mice were re-derived as germ-free (GF) for the purpose of this study. MCAD-/- mice or MCAD+/+ controls were maintained in sterile gnotobiotic isolators or cages with HEPA-filtered air supply. Urine from GF MCAD-/- and MCAD+/+ mice was collected under sterile conditions. Seven days subsequent, mice were mono-colonized with wild-type C. sporogenes (Cs) by gavage of 200 µL saturated overnight culture. Urine was collected one week after colonization. C. sporogenes colonization was verified with serial dilution plating. |
Sample Preparation:
| Sampleprep ID: | SP001716 |
| Sampleprep Summary: | . Mouse urine samples (2.5 μL) were diluted with LC-MS grade water (5 μL) and mixed with internal standard (7.5 μL, d3-creatinine,1 mM; d9-phenylpropionic acid, 0.2 mM; 2,2-d2-hippuric acid, 0.2 mM), then 60 μL of extraction solvent containing reference standards (d7-glucose, d3-methionine, L-4-hydroxyphenyl-d4-alanine, d5-hippuric acid, d5-tryptophan, d3-leucine, di-n-octyl phthalate-3,4,5,6-d4, d19-decanoic acid, d15-octanoic acid, d27-tetradecanoic acid, 2-flurophenylglycine, d9-carnitine in methanol) was added to precipitate proteins. The solution was vortexed, then the plate was centrifuged at 15,000 x g for 5 min at 4 C. Supernatant (60 μL) was transferred to a new plate and 20 μL of each was diluted with 60 μL LC-MS grade water. Three procedure blanks were included using the same preparation method with LC-MS water (2.5 μL) instead of urine sample. Quality controls (QC) were pooled from each LC-MS ready sample. |
Combined analysis:
| Analysis ID | AN002669 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1290 Infinity II UPLC |
| Column | Waters BEH C18 (100 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6545XT QTOF |
| Ion Mode | UNSPECIFIED |
| Units | peak height normalized with creatinine |
Chromatography:
| Chromatography ID: | CH001964 |
| Instrument Name: | Agilent 1290 Infinity II UPLC |
| Column Name: | Waters BEH C18 (100 x 2.1mm,1.7um) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002468 |
| Analysis ID: | AN002669 |
| Instrument Name: | Agilent 6545XT QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Acquisition was done in All-Ions fragmentation mode using collision energies of 0, 20, and 40 eV. Suspect screening was performed with MS-DIAL (version 4.24) and MS-CleanR. Peak height was normalized with creatinine |
| Ion Mode: | UNSPECIFIED |
