Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA151896

Local Sample ID:	P16_t0
Subject ID:	SU001733
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	Male and female

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Sample Preparation:

Sampleprep ID:	SP001739
Sampleprep Summary:	After thawing the samples, 150 µL of cold acetonitrile (0.1% formic acid, v/v) was added to 50 µL of serum, vortex and kept at -20ºC for 30 min for protein precipitation. Then, 25 µl of cleaned supernatant were transferred to a 96 well-plate for UPLC-MS analysis. Finally, 125 µL of H2O (0.1% formic acid, v/v), and 2 µL of internal standard (IS) mix solution, containing reserpine, leucine enkephaline, caffeine-d9 and phenylalanine-d5 in H20:CH3OH (1:1, 0.1% v/v formic acid) at 20 µM were added to each sample. Blank samples were prepared by replacing serum by ultrapure H20 in order to identify potential artefacts from the tube, reagents and other materials. A quality control (QC) was prepared by mixing 10 µL from each prepared sample. Samples were randomly injected in the chromatographic system (UPLC-ToF-MS) injection a QC every 6 serum samples in order to avoid intra-batch variability, as well as to enhance quality and reproducibility. Blank analysis was performed at the beginning and at end of the sequence. Sample stability and analytical drift were investigated through the internal standard intensities.

Sampleprep ID:

SP001739

Sampleprep Summary:

After thawing the samples, 150 µL of cold acetonitrile (0.1% formic acid, v/v) was added to 50 µL of serum, vortex and kept at -20ºC for 30 min for protein precipitation. Then, 25 µl of cleaned supernatant were transferred to a 96 well-plate for UPLC-MS analysis. Finally, 125 µL of H2O (0.1% formic acid, v/v), and 2 µL of internal standard (IS) mix solution, containing reserpine, leucine enkephaline, caffeine-d9 and phenylalanine-d5 in H20:CH3OH (1:1, 0.1% v/v formic acid) at 20 µM were added to each sample. Blank samples were prepared by replacing serum by ultrapure H20 in order to identify potential artefacts from the tube, reagents and other materials. A quality control (QC) was prepared by mixing 10 µL from each prepared sample. Samples were randomly injected in the chromatographic system (UPLC-ToF-MS) injection a QC every 6 serum samples in order to avoid intra-batch variability, as well as to enhance quality and reproducibility. Blank analysis was performed at the beginning and at end of the sequence. Sample stability and analytical drift were investigated through the internal standard intensities.