Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA217817

Local Sample ID:	A02-02_58_1_1700
Subject ID:	SU002359
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	Not applicable
Cell Biosource Or Supplier:	Sigma-Aldrich (St. Louis, MO)
Cell Strain Details:	Hep3B

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Sample Preparation:

Sampleprep ID:	SP002365
Sampleprep Summary:	Metabolite extraction: For metabolomics, three biological replicates were used. An equal number of cells were utilized for each sample. The cells were centrifuged separately at 15000 rpm for 10 min to separate the cells and cell-free supernatants. The supernatants were dis-carded, and to each cell pellet (~3x 106), 1 mL of 0.1% formic acid in methanol was added, followed by vertexing for 2 min x 4, interrupted by 15 min incubation on ice. This is followed by sonication using a COPLEY probe-sonicator (QSONICA SONI-CATOR, USA) for 30 seconds with 30% amplifier in an ice bath. Subsequently, the cel-lular debris of each sample was collected by centrifugation at 15000 rpm for 10 min at 4°C. The supernatants were separated and dried at 37 ± 1 °C using EZ-2 Plus (Gene-Vac-Ipswich, UK). Finally, 200 µL of 0.1% formic acid in water was used to resuspend the dried samples, which were then vortexed for 2 min, filtered by a 0.45µm hydro-philic Nylon Syringe Filter and transferred to 200 μL (micro-inserts) in LC vials. The quality control (QC) sample was prepared by pooling the same volume (10 μL) of each sample, and all samples were placed in the autosampler at a temperature set at 4℃ and analyzed with UHPLC-QTOF-MS.

Sampleprep ID:

SP002365

Sampleprep Summary:

Metabolite extraction: For metabolomics, three biological replicates were used. An equal number of cells were utilized for each sample. The cells were centrifuged separately at 15000 rpm for 10 min to separate the cells and cell-free supernatants. The supernatants were dis-carded, and to each cell pellet (~3x 106), 1 mL of 0.1% formic acid in methanol was added, followed by vertexing for 2 min x 4, interrupted by 15 min incubation on ice. This is followed by sonication using a COPLEY probe-sonicator (QSONICA SONI-CATOR, USA) for 30 seconds with 30% amplifier in an ice bath. Subsequently, the cel-lular debris of each sample was collected by centrifugation at 15000 rpm for 10 min at 4°C. The supernatants were separated and dried at 37 ± 1 °C using EZ-2 Plus (Gene-Vac-Ipswich, UK). Finally, 200 µL of 0.1% formic acid in water was used to resuspend the dried samples, which were then vortexed for 2 min, filtered by a 0.45µm hydro-philic Nylon Syringe Filter and transferred to 200 μL (micro-inserts) in LC vials. The quality control (QC) sample was prepared by pooling the same volume (10 μL) of each sample, and all samples were placed in the autosampler at a temperature set at 4℃ and analyzed with UHPLC-QTOF-MS.