Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA019416

Local Sample ID:	Case_70057
Subject ID:	SU000426
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	female
Species Group:	Human

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Sample Preparation:

Sampleprep ID:	SP000433
Sampleprep Summary:	Sample Preparation Prior to Biocrates p180 Kit Plate Analysis: Serum samples were thawed on ice and pooled QC samples were prepared. Samples (study and pooled QC samples) were then diluted, vortexed, and transferred into properly labeled 2 mL Lo-Bind eppendorf tubes and dried on a lyophilizer overnight. The residue was reconstituted with 30 µL of PBS buffer. Samples were then vortexed and then centrifuged at room temperature and at 16,000 rcf for 4.0 minutes. All study samples and pooled QC samples were splitted into two AbsoluteIDQ p180 Kit-96 wells plates (Plate 1502 and 1503). Biocrates Plate Preparation: The Biocrates p180 kit-96 wells plates were prepared following the AbsoluteIDQ p180 Kit metabolomics procedure. Briefly, an internal standard mix was added to 95 of the 96 wells. Next, zero samples, QC standards and calibration standards were added to their corresponding wells. The study samples and pooled QC Samples were then added to the appropriate wells and dried for 30 minutes under nitrogen flow. The plate was derivatized using a 5% Phenylisothiocyanate (PITC) solution in (1:1:1) Ethanol:Pyridine:Water and, then, incubated for 20 minutes followed by a drying step under nitrogen flow. An extraction solvent (5 mM ammonium acetate in methanol) was added to all wells. The plate was then vortexed and filtrated by centrifugation. After centrifugation, 150 µL was removed and transferred to a second 96-well plate (LC-MS/MS plate). This second plate was diluted with 150 µL of HPLC grade water for a subsequent LC-MS/MS assay (analysis of amino acids and biogenic amines). All the wells in the original plate was diluted with 400 µL of flow injection analysis (FIA) Running Solvent for a FIA-MS analysis (lipids, acylcarnitines, and hexose analysis).

Sampleprep ID:

SP000433

Sampleprep Summary:

Sample Preparation Prior to Biocrates p180 Kit Plate Analysis: Serum samples were thawed on ice and pooled QC samples were prepared. Samples (study and pooled QC samples) were then diluted, vortexed, and transferred into properly labeled 2 mL Lo-Bind eppendorf tubes and dried on a lyophilizer overnight. The residue was reconstituted with 30 µL of PBS buffer. Samples were then vortexed and then centrifuged at room temperature and at 16,000 rcf for 4.0 minutes. All study samples and pooled QC samples were splitted into two AbsoluteIDQ p180 Kit-96 wells plates (Plate 1502 and 1503). Biocrates Plate Preparation: The Biocrates p180 kit-96 wells plates were prepared following the AbsoluteIDQ p180 Kit metabolomics procedure. Briefly, an internal standard mix was added to 95 of the 96 wells. Next, zero samples, QC standards and calibration standards were added to their corresponding wells. The study samples and pooled QC Samples were then added to the appropriate wells and dried for 30 minutes under nitrogen flow. The plate was derivatized using a 5% Phenylisothiocyanate (PITC) solution in (1:1:1) Ethanol:Pyridine:Water and, then, incubated for 20 minutes followed by a drying step under nitrogen flow. An extraction solvent (5 mM ammonium acetate in methanol) was added to all wells. The plate was then vortexed and filtrated by centrifugation. After centrifugation, 150 µL was removed and transferred to a second 96-well plate (LC-MS/MS plate). This second plate was diluted with 150 µL of HPLC grade water for a subsequent LC-MS/MS assay (analysis of amino acids and biogenic amines). All the wells in the original plate was diluted with 400 µL of flow injection analysis (FIA) Running Solvent for a FIA-MS analysis (lipids, acylcarnitines, and hexose analysis).