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MB Sample ID: SA218443
| Local Sample ID: | 1024019764 |
| Subject ID: | SU002365 |
| Subject Type: | Fish |
| Subject Species: | Oncorhynchus mykiss |
| Species Group: | Fish |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002365 |
| Subject Type: | Fish |
| Subject Species: | Oncorhynchus mykiss |
| Species Group: | Fish |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 1024019764 | SA218443 | FL026468 | 6_weeks | Sample time |
| 1024019764 | SA218443 | FL026468 | 5% ß-glucan | Treatment |
Collection:
| Collection ID: | CO002358 |
| Collection Summary: | For the metabolite and microbiota analyses, 10 fish from the control group were collected on day 0 and 10 fish from each group (control, 0.1, 1.0, 5.0% β-glucan) at week 6. Fish were euthanized by immersion in an overdose (200 mg∙L-1) of MS-222 (cat no. A5040, Sigma-Aldrich). The tail was cut, and blood was sampled from the vena caudalis using Na-heparinised 25 mL and 50 mL capillary pipettes (Hirschmann Laborgeräte, Germany). Blood samples were centrifuged at 3,000 g for 10 min at 4 °C, and serum was isolated and stored at -80 °C. The intestine was aseptically sampled and immediately immersed in RNAlater® (Sigma-Aldrich) and transferred to -20 °C. |
| Sample Type: | Blood (serum) |
| Storage Conditions: | -20℃ |
Treatment:
| Treatment ID: | TR002377 |
| Treatment Summary: | A commercial pelleted (1.5 mm) trout feed (INICIO 917, BioMar A/S) based on a mixture of fish and plant protein and containing 47% crude protein, 20% crude lipid, 18% carbohydrates (NFE), 1.2% fibre, 8.5% ash, and 1.1% phosphorus was used for preparation of the diets. A β-1,3;1,6-glucan (purity: 81.6%; mean particle size: 37.7 µm; supplementary table S2) purified from yeast (Saccharomyces cerevisiae) (Biorigin, Brazil) was supplemented to this diet at varying doses: 0 g, 1 g, 10 g, and 50 g β-glucan were added to 1 kg of INICIO 917, respectively, during continuous mixing and sealed to the pellets by spraying with 30 mL organic, cold-pressed rapeseed oil (Gyldenmark), resulting in a control diet without β-glucan and three experimental diets of 0.1%, 1.0%, and 5.0% Wt/Wt β-glucan. |
Sample Preparation:
| Sampleprep ID: | SP002371 |
| 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 100 mg of sample and 4 steel balls of size M into each tube. Per mg of sample 5 µL 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. 10µL of supernatant was used for the targeted analytics. For blood sample analysis 10 µL plasma was taken. 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. |
| Sampleprep Protocol Filename: | Sample_preparation.pdf |
Combined analysis:
| Analysis ID | AN003723 |
|---|---|
| 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: | CH002757 |
| 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 FIA: B: Mix running solvent and MiliQ water 1:1 Gradient Table 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 |
| Methods Filename: | LC_parameters_amino_acids.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: | MS003471 |
| Analysis ID: | AN003723 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | MS - Instrument Parameter AbsoluteIDQ® p180 Kit (Biocrates Life Science AG, Innsbruck, Austria) MS -Q-Trap 5500 Sciex Ion Source: Turbo Spray Curtian Gas: 20 CAD Gas: Medium Ion Spray Voltage: 5500 V Temperature: 500°C Ion Source Gas 1: 40psi Ion Source Gas 2: 50 psi |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | MS_amino_acids.pdf |
