Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA317923

Local Sample ID:	C4
Subject ID:	SU003040
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Select appropriate tab below to view additional metadata details:

Sample Preparation:

Sampleprep ID:	SP003046
Sampleprep Summary:	Briefly, equal confluency of cells in 6-well plate with 3 biological replicates were rinsed by 1 mL HPLC-grade water rapidly and immediately quenched by directly added by 1 mL liquid nitrogen to the plate. Then, 1 mL ice-cold 90% MC buffer (MC, 9: 1 MeOH: CHCl3. 90% MC, 9 : 1 MC: water) with 5 µM methionine sulfone as internal signal was added to each well of plate and the cells were scraped and transferred to1.5-mL tubes. Extracts were centrifuged at 4 ℃ for 5 min at 16,000 xg and the supernatants were transferred to new clean 1.5-mL tubes and equal volume of each sample was pooled for the QC sample. Finally, the extracts were dried using vacuum concentrator system and store at -80 ℃ until analysis. For targeted analysis, the dried extracts were resuspended by 100 µL 60% acetonitrile and centrifuged at 4 ℃ for 15 min at 15,000 xg after 30 min incubation at 4 ℃. Only the supernatant was transferred to autosampler vials for MS analysis.

Sampleprep ID:

SP003046

Sampleprep Summary:

Briefly, equal confluency of cells in 6-well plate with 3 biological replicates were rinsed by 1 mL HPLC-grade water rapidly and immediately quenched by directly added by 1 mL liquid nitrogen to the plate. Then, 1 mL ice-cold 90% MC buffer (MC, 9: 1 MeOH: CHCl3. 90% MC, 9 : 1 MC: water) with 5 µM methionine sulfone as internal signal was added to each well of plate and the cells were scraped and transferred to1.5-mL tubes. Extracts were centrifuged at 4 ℃ for 5 min at 16,000 xg and the supernatants were transferred to new clean 1.5-mL tubes and equal volume of each sample was pooled for the QC sample. Finally, the extracts were dried using vacuum concentrator system and store at -80 ℃ until analysis. For targeted analysis, the dried extracts were resuspended by 100 µL 60% acetonitrile and centrifuged at 4 ℃ for 15 min at 15,000 xg after 30 min incubation at 4 ℃. Only the supernatant was transferred to autosampler vials for MS analysis.