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MB Sample ID: SA170817
| Local Sample ID: | 1024035500 |
| Subject ID: | SU001913 |
| Subject Type: | Mammal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001913 |
| Subject Type: | Mammal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 1024035500 | SA170817 | FL020568 | Serum | Tissue |
| 1024035500 | SA170817 | FL020568 | RYGB_AB | Sample treatment |
Collection:
| Collection ID: | CO001906 |
| Collection Summary: | Animals were sacrificed, blood and cecum tissue were removed, snap frozen in liquid nitrogen and stored at -80°C until further analysis |
| Sample Type: | Cecum |
Treatment:
| Treatment ID: | TR001926 |
| Treatment Summary: | RYGB surgery was performed on age-matched (8-10-weeks old) male Wistar IGS rats. After four weeks of recovery, animals were put on an antibiotic cocktail consisting of ampicillin (1 g/l; Ratiopharm), vancomycin (0.5 g/l; Ratiopharm), neomycin (1 g/l; Bela-pharm), and metronidazole (1 g/l; CP-Pharma), provided freshly every day via drinking water. The parallel FMT group received fecal RYGB microbiota transplantation once per week, but were otherwise handled like DIO littermates. For fecal transplantation experiments, 100 mg of fresh stool from bodyweight-stabilized RYGB donors was re-suspended in 1 ml of PBS, homogenized carefully and administered via oral gavage with 200 μl of the suspension. |
Sample Preparation:
| Sampleprep ID: | SP001919 |
| Sampleprep Summary: | Sample preparation for MetIDQ p180 Kit measurement Solvents: Acetonitril (Merck KGaA, Darmstadt, Germany hypergrade for LC-MS) Water MiliQ, Extracting agent - ACN / H2O (1:1) Equipment 4 steel balls size M Eppendorf Tubes 2mL Tissue slicer (Rettberg, Germany) Centrifuge (Sigma) Work steps Approximately 100mg of caecum sample and 4 steel balls of size M into each tube. Per mg of sample 5µL of extracting agent was added. Shake the samples for 10 minutes at 30 Hz in the tissue slicer and centrifuge for 2 minutes at 14000 rpm. 10 µL supernatant was used for the targeted analytics. Blood serum samples 10 µL were used for analysis Kit reparation The analysis was performed using the validated MetIDQ p180 Kit and described in Siskos et al. [1]. Data processing was carried out with the provided quantitation method Kit (Biocrates Life Sciences AG, Innsbruck, Austria). 1 Siskos AP, Jain P, Römisch-Margl W, Bennett M, Achaintre D, Asad Y, et al. Interlaboratory Reproducibility of a Targeted Metabolomics Platform for Analysis of Human Serum and Plasma. Anal Chem 2017;89:656-65. |
Combined analysis:
| Analysis ID | AN002978 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | UPLC (Waters Acquity, Waters Corporation, Milford, USA) |
| Column | Agilent Zorbax Eclipse XDB C18 (100 x 3.0mm,3.5um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | ABI Sciex 5500 QTrap |
| Ion Mode | POSITIVE |
| Units | µM |
Chromatography:
| Chromatography ID: | CH002207 |
| Chromatography Summary: | LC - Instrument Parameters AbsoluteIDQ® p180 Kit (Biocrates Life Science AG, Innsbruck, Austria) System: UPLC (Waters Acquity, Waters Corporation, Milford, USA) Column: Agilent, Zorbax Eclipse XDB C18, 3.0 x 100 mm, 3.5 μM, Agilent Waldbronn, Germany Precolumn: Security Guard, Phenomenex, C18, 4 x 3 mm; Phenomenex, Aschaffenburg, Germany Materials: Water MiliQ, Methanol (Merck KGaA, Darmstadt, Germany, hypergrade for LC-MS) Acetonitril (Merck KGaA, Darmstadt, Germany hypergrade for LC-MS) Ammonium Acetate (Honeywell - Fluka, Seelze, Germany) Formic Acid (Honeywell, Fluka, Seelze, Germany) Running Solvent: 5mM ammonium acetat in methanol LC: A: 2mL Formic acid in MilliQ water B: 1mL Formic Acid in Acetonitrile Gradient Table LC Time (min) Flow Rate (mL/min) %A %B Curve Initial 0.500 100.0 0.0 Initial 0.50 0.500 100.0 0.0 6 4.00 0.500 30.0 70.0 6 5.30 0.500 30.0 70.0 6 5.40 0.500 100.0 0.0 6 7.30 0.500 100.0 0.0 6 |
| Methods Filename: | LC_parameters-metabolites.pdf |
| Instrument Name: | UPLC (Waters Acquity, Waters Corporation, Milford, USA) |
| Column Name: | Agilent Zorbax Eclipse XDB C18 (100 x 3.0mm,3.5um) |
| Flow Gradient: | FIA Time (min) Flow Rate (mL/min) %A %B Curve Initial 0.030 0.0 100.0 Initial 1.60 0.030 0.0 100.0 6 2.40 0.200 0.0 100.0 6 2.80 0.200 0.0 100.0 6 3.00 0.030 0.0 100.0 6 |
| Flow Rate: | 0.03ml/min |
| Solvent A: | 100% methanol; 5mM ammonium acetate |
| Solvent B: | 50% methanol/50% water; 5mM ammonium acetate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002768 |
| Analysis ID: | AN002978 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Analyst version 1.6 Software from SCIEX. For Validation METIDQ Software version Boron from Biocrates |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | MS_parameters_1.pdf |
