Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA209203

Local Sample ID:	2020-08-17_Zang-43_Box2Redo-22_NEG
Subject ID:	SU002264
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Age Or Age Range:	10-week and 24-month
Gender:	Male

Select appropriate tab below to view additional metadata details:

Sample Preparation:

Sampleprep ID:	SP002270
Sampleprep Summary:	Sample preparation: aqueous metabolites were extracted using a methanol-based protein precipitation method as described previously. Briefly, heart tissue samples were homogenized in cold water using zirconium oxide beads, methanol was added, and samples were vortexed and then stored for 30 minutes at -20˚C. Afterwards, samples were first sonicated in an ice bath for 10 minutes, centrifuged for 15 min at 18,000g and 4˚C, and then a fixed volume of supernatant was collected from each sample. Lastly, recovered supernatants were dried on a SpeedVac and reconstituted for LC-MS analysis. Protein pallets that were left over from the sample prep were saved for BCA assay. (2) LC-MS analysis: samples were analyzed on a duplex-LC-MS system composed of two Shimadzu UPLC pumps, CTC Analytics PAL HTC-xt temperature-controlled auto-sampler and AB Sciex 6500+ Triple Quadrupole MS equipped with ESI ionization source. UPLC pumps were connected to the auto-sampler in parallel and were able to perform two chromatography separations independently from each other. Each sample was injected twice on two identical analytical columns (Waters XBridge BEH Amide XP) performing separations in hydrophilic interaction liquid chromatography (HILIC) mode. While one column was performing separation and MS data acquisition in ESI(+) ionization mode, the other column was getting equilibrated for sample injection, chromatography separation and MS data acquisition in ESI(-) mode. Each chromatography separation was 18 minutes (total analysis time per sample was 36 minutes).

Sampleprep ID:

SP002270

Sampleprep Summary:

Sample preparation: aqueous metabolites were extracted using a methanol-based protein precipitation method as described previously. Briefly, heart tissue samples were homogenized in cold water using zirconium oxide beads, methanol was added, and samples were vortexed and then stored for 30 minutes at -20˚C. Afterwards, samples were first sonicated in an ice bath for 10 minutes, centrifuged for 15 min at 18,000g and 4˚C, and then a fixed volume of supernatant was collected from each sample. Lastly, recovered supernatants were dried on a SpeedVac and reconstituted for LC-MS analysis. Protein pallets that were left over from the sample prep were saved for BCA assay. (2) LC-MS analysis: samples were analyzed on a duplex-LC-MS system composed of two Shimadzu UPLC pumps, CTC Analytics PAL HTC-xt temperature-controlled auto-sampler and AB Sciex 6500+ Triple Quadrupole MS equipped with ESI ionization source. UPLC pumps were connected to the auto-sampler in parallel and were able to perform two chromatography separations independently from each other. Each sample was injected twice on two identical analytical columns (Waters XBridge BEH Amide XP) performing separations in hydrophilic interaction liquid chromatography (HILIC) mode. While one column was performing separation and MS data acquisition in ESI(+) ionization mode, the other column was getting equilibrated for sample injection, chromatography separation and MS data acquisition in ESI(-) mode. Each chromatography separation was 18 minutes (total analysis time per sample was 36 minutes).