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MB Sample ID: SA170418
| Local Sample ID: | 1024033195 |
| Subject ID: | SU001909 |
| Subject Type: | Mammal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001909 |
| Subject Type: | Mammal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 1024033195 | SA170418 | FL020337 | Serum | Tissue |
| 1024033195 | SA170418 | FL020337 | 126 | Rat_number |
| 1024033195 | SA170418 | FL020337 | DIO_8 | Rat_sample_name |
| 1024033195 | SA170418 | FL020337 | DIO | Sample treatment |
Collection:
| Collection ID: | CO001902 |
| 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: | TR001922 |
| 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 body weight-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: | SP001915 |
| Sampleprep Summary: | 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 To approximately 100 mg of sample four steel balls of size M rae added in Eppendorf tubes. Then add for each mg of sample 5 µL of extracting agent. Shake the samples for 10 minutes at 30 Hz in the tissue slicer and centrifuge for 2 minutes at 14000 rpm. The supernatant is used for targeted analytics as described in the kit manufactures manual. Kit measurement The analysis was performed using the validated Bile Acid Kit (Biocrates Life Sciences, Inssbruck, Austria) as described in Pham et al. 2017 [1]. Data processing is carried out with the provided quantitation method Kit (Biocrates Life Sciences AG, Innsbruck, Austria). 1 Pham HT, Arnhard K, Asad YJ, Deng L, Felder TK, St John-Williams L, et al. Inter-Laboratory Robustness of Next-Generation Bile Acid Study in Mice and Humans: International Ring Trial Involving 12 Laboratories. The Journal of Applied Laboratory Medicine 2019;1:129-42. |
Combined analysis:
| Analysis ID | AN002974 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | UPLC (Waters Acquity, Waters Corporation, Milford, USA) |
| Column | none |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | ABI Sciex 5500 QTrap |
| Ion Mode | NEGATIVE |
| Units | µM |
Chromatography:
| Chromatography ID: | CH002203 |
| Instrument Name: | UPLC (Waters Acquity, Waters Corporation, Milford, USA) |
| Column Name: | none |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002764 |
| Analysis ID: | AN002974 |
| 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: | NEGATIVE |
