Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA014166

Local Sample ID:	L_12_287
Subject ID:	SU000334
Subject Type:	Animal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57/BL-6
Gender:	Male
Species Group:	Mammal

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Sample Preparation:

Sampleprep ID:	SP000342
Sampleprep Summary:	Aliquots of liver sample were shipped to the NIH RTI-RCMRC on dry ice and immediately stored at -70 °C after being logged in for metabolomics analysis. Frozen tissue samples were transferred to labeled tubes containing stainless steel homogenization beads on dry ice to confirm weights. A total of 83 study samples were thawed on ice for sample preparation. Total tissue contents were extracted during homogenization with a 50:50 acetonitrile:water solution, to generate 2.0-2.5 mg/µl sample homogenates. Samples were centrifuged and an 80 mg equivalent volume (400/320 µl) of homogenized liver supernatants were transferred to new 2.0 mL tubes for the experimental samples. Analytical quality control (QC) phenotypic pool samples were generated by transferring a 50/40 µL aliquot of each supernatant from each respective phenotypic group based on HS-exposure or Controls into different 5 mL tubes. The two phenotypic QC pooled samples were vortexed to mix and a total study QC pool was generated by transferring 650 µL aliquots of each phenotypic pool sample into a new 5 mL tube for mixing. All pooled samples were aliquoted into labeled 2.0 mL tubes like the experimental samples, three for each QC group for an additional 9 samples. The samples were frozen for 2 hr at -70 °C and lyophilized to dryness overnight. Samples were reconstituted in 700 µl of D2O master mix containing 0.2 M Phosphate buffer, pH 7.4 + Chenomx ISTD with 6 mM Imidazole, 9:1 v/v. The tubes were vortexed for 2 min on a multi-tube vortexer and centrifuged at 16,000 rcf for 4 min. A 600 µl aliquot of the supernatant was transferred into pre-labeled 5 mm (4”) NMR tubes for data acquisition on a 700 MHz spectrometer.

Sampleprep ID:

SP000342

Sampleprep Summary:

Aliquots of liver sample were shipped to the NIH RTI-RCMRC on dry ice and immediately stored at -70 °C after being logged in for metabolomics analysis. Frozen tissue samples were transferred to labeled tubes containing stainless steel homogenization beads on dry ice to confirm weights. A total of 83 study samples were thawed on ice for sample preparation. Total tissue contents were extracted during homogenization with a 50:50 acetonitrile:water solution, to generate 2.0-2.5 mg/µl sample homogenates. Samples were centrifuged and an 80 mg equivalent volume (400/320 µl) of homogenized liver supernatants were transferred to new 2.0 mL tubes for the experimental samples. Analytical quality control (QC) phenotypic pool samples were generated by transferring a 50/40 µL aliquot of each supernatant from each respective phenotypic group based on HS-exposure or Controls into different 5 mL tubes. The two phenotypic QC pooled samples were vortexed to mix and a total study QC pool was generated by transferring 650 µL aliquots of each phenotypic pool sample into a new 5 mL tube for mixing. All pooled samples were aliquoted into labeled 2.0 mL tubes like the experimental samples, three for each QC group for an additional 9 samples. The samples were frozen for 2 hr at -70 °C and lyophilized to dryness overnight. Samples were reconstituted in 700 µl of D2O master mix containing 0.2 M Phosphate buffer, pH 7.4 + Chenomx ISTD with 6 mM Imidazole, 9:1 v/v. The tubes were vortexed for 2 min on a multi-tube vortexer and centrifuged at 16,000 rcf for 4 min. A 600 µl aliquot of the supernatant was transferred into pre-labeled 5 mm (4”) NMR tubes for data acquisition on a 700 MHz spectrometer.