Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA126727

Local Sample ID:	138_Old_Liver_40a
Subject ID:	SU001581
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6JOlaHsd
Age Or Age Range:	6 months (young) and 20-24 months (old)
Gender:	Male
Animal Animal Supplier:	Envigo
Animal Housing:	Animal rooms were maintained at 21°C-24°C and 35%-75% relative humidity
Animal Light Cycle:	12/12-h (7am to 7pm) dark-light cycle
Animal Feed:	Altromin 1324 (chow)
Animal Water:	Ad lib

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Sample Preparation:

Sampleprep ID:	SP001589
Sampleprep Summary:	To maximize accuracy and to take into account tissue inhomogeneity, metabolites were extracted from 3 separate liver pieces for each mouse, and the average metabolite value of the 3 technical replicates was calculated at analysis. In detail, frozen liver pieces weighting ~30mg were transferred into soft tissue homogenizing CK 14 tubes containing 1.4 mm ceramic beads (Bertin corp.) prefilled with 1 ml of cold (-20°C) metabolites extraction solvent (Methanol: Acetonitrile:H2O, 50:30:20) and kept on ice. Samples were homogenized using precellys 24 tissue homogenizer (3X20sec at 6000 rpm, with a 30sec gap between each of the three cycles, Bertin Technologies) cooled to 2°C. Homogenized extracts were centrifuged in the Precellys tubes at 18,000g for 15 minutes at 4°C, supernatants were collected in microcentrifuge tubes and centrifuged again at 18,000g for 10 minutes at 4°C. The supernatants were transferred to glass HPLC vials and kept at -75 °C prior to LC-MS analysis.

Sampleprep ID:

SP001589

Sampleprep Summary:

To maximize accuracy and to take into account tissue inhomogeneity, metabolites were extracted from 3 separate liver pieces for each mouse, and the average metabolite value of the 3 technical replicates was calculated at analysis. In detail, frozen liver pieces weighting ~30mg were transferred into soft tissue homogenizing CK 14 tubes containing 1.4 mm ceramic beads (Bertin corp.) prefilled with 1 ml of cold (-20°C) metabolites extraction solvent (Methanol: Acetonitrile:H2O, 50:30:20) and kept on ice. Samples were homogenized using precellys 24 tissue homogenizer (3X20sec at 6000 rpm, with a 30sec gap between each of the three cycles, Bertin Technologies) cooled to 2°C. Homogenized extracts were centrifuged in the Precellys tubes at 18,000g for 15 minutes at 4°C, supernatants were collected in microcentrifuge tubes and centrifuged again at 18,000g for 10 minutes at 4°C. The supernatants were transferred to glass HPLC vials and kept at -75 °C prior to LC-MS analysis.