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MB Sample ID: SA163753
Local Sample ID: | bileacid_Exp1_21_Prevention |
Subject ID: | SU001825 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | GF 129SvEv background IL-10-deficient mice (Il10-/-); Il10-eGFP-reporter (Il10+/eGFP) mice (on a C57BL/6J background) |
Age Or Age Range: | 8-12 week old |
Animal Animal Supplier: | 8-12 week-age GF 129SvEv background IL-10-deficient mice (Il10-/-) were obtained from University of North Carolina National Gnotobiotic Rodent Resource Center; Il10-eGFP-reporter (Il10+/eGFP) mice on a C57BL/6J background mice were originally provided by Dr. C. L. Karp (Global Health, Bill & Melinda Gates Foundation, USA) |
Animal Housing: | Germ-free and gnotobiotic conditions at National Gnotobiotic Rodent Resource Center, University of North Carolina |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001825 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | GF 129SvEv background IL-10-deficient mice (Il10-/-); Il10-eGFP-reporter (Il10+/eGFP) mice (on a C57BL/6J background) |
Age Or Age Range: | 8-12 week old |
Animal Animal Supplier: | 8-12 week-age GF 129SvEv background IL-10-deficient mice (Il10-/-) were obtained from University of North Carolina National Gnotobiotic Rodent Resource Center; Il10-eGFP-reporter (Il10+/eGFP) mice on a C57BL/6J background mice were originally provided by Dr. C. L. Karp (Global Health, Bill & Melinda Gates Foundation, USA) |
Animal Housing: | Germ-free and gnotobiotic conditions at National Gnotobiotic Rodent Resource Center, University of North Carolina |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
bileacid_Exp1_21_Prevention | SA163753 | FL019468 | GUT-103 plus EER | Treatment |
Collection:
Collection ID: | CO001818 |
Collection Summary: | For samples for targeted metabolomics analysis, mouse cecal content and/or feces were collected. |
Collection Protocol ID: | University of North Carolina at Chapel Hill Institutional Animal Care and Use Committee (18-266.0-B) |
Sample Type: | Feces |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR001838 |
Treatment Summary: | For treatment 200 µl of diluted human donor stool was applied by oral gavage on day 1 to Il10-/- mice. The stool was derived from a single healthy donor, Donor-Y, and was previously found to induce moderate to severe colitis in Il10-/- mice. One gram stool was diluted 100- fold with anaerobic PBS, and vigorously mixed for 5 minutes under anaerobic conditions. For application of GUT-103 and GUT-108, 300µl resuspended strain mixture in anaerobic PBS was applied per mouse by oral gavage. GUT-103 and GUT-108 strains were grown individually, subsequently mixed to equal concentrations (cfu/ml), and provided at a dose of 2.0x10+7 cfu/strain in a total volume of 300 µl. The strain mixture was provided four times via oral gavage on days 15, 17, 22 and 25 (Il10-/- mice) or on days 1, 3, 8 and 11 (Il10+/eGFP mice). |
Sample Preparation:
Sampleprep ID: | SP001831 |
Sampleprep Summary: | Bile acids, SCFAs, and tryptophan metabolites were extracted (with careful cleanup or derivatization procedures whenever necessary) and analyzed using mass spectrometry-based platforms. |
Sampleprep Protocol Filename: | Protocols.pdf |
Processing Storage Conditions: | On ice |
Extraction Method: | Bile acid: SPE and LLE; SCFA: alkaline aqueous extraction; tryptophan metabolites: 1:1 MeOH:Water LLE |
Extract Cleanup: | For Bile acids, C18 solid-phase extraction cleanup and concentration was used. |
Extract Storage: | -80℃ |
Sample Derivatization: | For SCFAs, propyl chloroformate derivatization was performed. |
Combined analysis:
Analysis ID | AN002845 | AN002846 | AN002847 |
---|---|---|---|
Analysis type | MS | MS | MS |
Chromatography type | GC | Reversed phase | Reversed phase |
Chromatography system | Agilent 7820A | Thermo Vanquish | Thermo Vanquish |
Column | Agilent DB5-MS (30m x 0.25mm, 0.25um) | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) | Waters Acquity BEH C18 (150 x 2mm,1.7um) |
MS Type | EI | ESI | ESI |
MS instrument type | Single quadrupole | Orbitrap | Triple quadrupole |
MS instrument name | Agilent 5977B | Thermo Q Exactive Orbitrap | Thermo Quantis QQQ |
Ion Mode | POSITIVE | POSITIVE | NEGATIVE |
Units | μmol/kg | ng/g | mg/g |
Chromatography:
Chromatography ID: | CH002106 |
Chromatography Summary: | Generic DB-5ms GC capillary column for analysis of SCFA derivatives. |
Instrument Name: | Agilent 7820A |
Column Name: | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
Chromatography Type: | GC |
Chromatography ID: | CH002107 |
Chromatography Summary: | C18 LC (HSS-T3) for quantitation of tryptophan metabolites |
Instrument Name: | Thermo Vanquish |
Column Name: | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
Chromatography Type: | Reversed phase |
Chromatography ID: | CH002108 |
Chromatography Summary: | C18 LC (BEH C18) and (HSS T3) for quantitation of bile acids |
Instrument Name: | Thermo Vanquish |
Column Name: | Waters Acquity BEH C18 (150 x 2mm,1.7um) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS002638 |
Analysis ID: | AN002845 |
Instrument Name: | Agilent 5977B |
Instrument Type: | Single quadrupole |
MS Type: | EI |
MS Comments: | fullscan data with scan range of m/z 30-600; Peak integration was performed in MassHunter Qual. |
Ion Mode: | POSITIVE |
MS ID: | MS002639 |
Analysis ID: | AN002846 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Optimized PRM method scanning characteristic MS2 fragment ion for absolute quantitation. |
Ion Mode: | POSITIVE |
MS ID: | MS002640 |
Analysis ID: | AN002847 |
Instrument Name: | Thermo Quantis QQQ |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | Optimized SRM method scanning characteristic MS2 fragment ion for absolute quantitation. |
Ion Mode: | NEGATIVE |