Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA025356

Local Sample ID:	A35
Subject ID:	SU000508
Subject Type:	Animal
Subject Species:	Megalobrama amblycephala
Taxonomy ID:	75352
Species Group:	Fish

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Subject:

Subject ID:	SU000508
Subject Type:	Animal
Subject Species:	Megalobrama amblycephala
Taxonomy ID:	75352
Species Group:	Fish

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
A35	SA025356	FL005987	Control	Treatment

Collection:

Collection ID:	CO000502
Collection Summary:	Blood samples were obtained following the standard protocol, using sodium heparin as anticoagulant. Liver tissue samples were weighed and put in tube then immediately flash-frozen with liquid nitrogen and stored at −80 °C till used for metabolomics analysis.
Collection Protocol Filename:	Sample collection and NMR protocol.pdf
Sample Type:	Plasma, Liver

Treatment:

Treatment ID:	TR000522
Treatment Summary:	Three tanks were randomly assigned to each diet group (3 replicates x 4 diet-groups): control (C; containing 8.0% fat and 26.7% carbohydrates), high fat diet (HFD; containing 12.2% fat and 27.8% carbohydrates), high carbohydrate diet (HCD; containing 8.3% fat and 34.1% carbohydrates) and high fat high carbohydrate diet (HFHCD; containing 10.4% fat and 30.9% carbohydrates.

Sample Preparation:

Sampleprep ID:	SP000515
Sampleprep Summary:	Each plasma sample (170 μL) was mixed with 340 μL phosphate buffer (45 mM, pH 7.47, 50% D2O) containing 0.9% NaCl in a 5 mm NMR tube and used directly for the NMR analysis. Liver tissues (about 50 mg) were homogenized in cold methanol and water (v/v=2:1) using a Qiagen Tissue-Lyser (Retsch GmBH, Germany). The supernatant (550uL) was collected after centrifugation (16 099 g, 4°C, 10 min). The extraction was repeated thrice using the same procedure. The supernatants collected from the above procedure were mixed together and freeze-dried after the removal of methanol in vacuo. Then the residue was dissolved in 600 μL of phosphate buffer (0.15 M, pH 7.38, 80% D2O) containing 0.001% TSP and 0.01% NaN3. The supernatant (550 μL) was transferred into a 5 mm NMR tube.

Sampleprep ID:

SP000515

Sampleprep Summary:

Each plasma sample (170 μL) was mixed with 340 μL phosphate buffer (45 mM, pH 7.47, 50% D2O) containing 0.9% NaCl in a 5 mm NMR tube and used directly for the NMR analysis. Liver tissues (about 50 mg) were homogenized in cold methanol and water (v/v=2:1) using a Qiagen Tissue-Lyser (Retsch GmBH, Germany). The supernatant (550uL) was collected after centrifugation (16 099 g, 4°C, 10 min). The extraction was repeated thrice using the same procedure. The supernatants collected from the above procedure were mixed together and freeze-dried after the removal of methanol in vacuo. Then the residue was dissolved in 600 μL of phosphate buffer (0.15 M, pH 7.38, 80% D2O) containing 0.001% TSP and 0.01% NaN3. The supernatant (550 μL) was transferred into a 5 mm NMR tube.

Analysis:

MB Sample ID:	SA025356
Analysis ID:	AN000753
Analysis Type:	NMR
Num Factors:	4

NMR:

NMR ID:	NM000087
Analysis ID:	AN000753
Instrument Name:	Bruker AVance III
Instrument Type:	FT-NMR
NMR Experiment Type:	1D 1H
Field Frequency Lock:	Deuterium
Standard Concentration:	0.058mM
Spectrometer Frequency:	600 MHz
NMR Probe:	cryo, inverse
NMR Solvent:	D2O
NMR Tube Size:	5mm x 7 in
Shimming Method:	topshim