Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA221006

Local Sample ID:	urine_44319
Subject ID:	SU002384
Subject Type:	Mammal
Subject Species:	Rattus norvegicus
Taxonomy ID:	10116

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Sample Preparation:

Sampleprep ID:	SP002390
Sampleprep Summary:	Samples of plasma (255 μl) were mixed with 255 μl of phosphate D2O buffer solution (NaH2PO4 and K2HPO4, 60 mM, pH 7.4). After centrifugation at 10000 × g at 4°C for 10 min to remove the precipitates, the supernatants were transferred to 5 mm NMR tubes and analyzed by NMR. Samples of urine (455 μl) were mixed with 55 μl of D2O buffer solution (NaH2PO4 and K2HPO4, 1.5 M, including 0.1% TSP (sodium 3-(trimethylsilyl) propionate-2,2,3,3-d4), pH 7.4) to minimize any gross variation in the pH of the urine samples. The mixture was left to stand for 10 min and centrifuged at 10000 × g at 4°C for 10 min to remove the precipitates. The supernatants were transferred to 5 mm NMR tubes and analyzed by NMR. The polar metabolites in the rat tissue were extracted according to the protocol established in our previous work. In brief, pre-weighed brain, kidney, liver, lung, or spleen samples (100 mg) were homogenized in 400 μl of CH3OH and 85 μl of H2O at 4°C. The homogenates were transferred into a 2.5-ml tube, combined with 400 μl of CHCl3 and 200 μl of H2O, and then kept in a vortex for 60 s. After 10-min partitioning on ice, the samples were centrifuged for 5 min (10000 × g, 4°C). The upper supernatants were transferred into 1.5-ml tubes and lyophilized to remove CH3OH and H2O. The extracts were reconstituted in 0.5 ml D2O containing 1 mM TSP, then transferred into 5-mm NMR tubes and analyzed by NMR spectroscopy.

Sampleprep ID:

SP002390

Sampleprep Summary:

Samples of plasma (255 μl) were mixed with 255 μl of phosphate D2O buffer solution (NaH2PO4 and K2HPO4, 60 mM, pH 7.4). After centrifugation at 10000 × g at 4°C for 10 min to remove the precipitates, the supernatants were transferred to 5 mm NMR tubes and analyzed by NMR. Samples of urine (455 μl) were mixed with 55 μl of D2O buffer solution (NaH2PO4 and K2HPO4, 1.5 M, including 0.1% TSP (sodium 3-(trimethylsilyl) propionate-2,2,3,3-d4), pH 7.4) to minimize any gross variation in the pH of the urine samples. The mixture was left to stand for 10 min and centrifuged at 10000 × g at 4°C for 10 min to remove the precipitates. The supernatants were transferred to 5 mm NMR tubes and analyzed by NMR. The polar metabolites in the rat tissue were extracted according to the protocol established in our previous work. In brief, pre-weighed brain, kidney, liver, lung, or spleen samples (100 mg) were homogenized in 400 μl of CH3OH and 85 μl of H2O at 4°C. The homogenates were transferred into a 2.5-ml tube, combined with 400 μl of CHCl3 and 200 μl of H2O, and then kept in a vortex for 60 s. After 10-min partitioning on ice, the samples were centrifuged for 5 min (10000 × g, 4°C). The upper supernatants were transferred into 1.5-ml tubes and lyophilized to remove CH3OH and H2O. The extracts were reconstituted in 0.5 ml D2O containing 1 mM TSP, then transferred into 5-mm NMR tubes and analyzed by NMR spectroscopy.