Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA202378

Local Sample ID:	BW45-27
Subject ID:	SU002194
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Sample Preparation:

Sampleprep ID:	SP002200
Sampleprep Summary:	Metabolomics analyses were performed as extensively described in previous studies (Issaian et al., Hematologica 2021). A volume of 20μl of frozen plasma was extracted in either 480μl of methanol:acetonitrile:water (5:3:2, v/v/v) (D'Alessandro et al. JCI Insight 2021). After vortexing at 4°C for 30 min, extracts were separated from the protein pellet by centrifugation for 10 min at 10,000g at 4°C and stored at −80°C until analysis. Ultra-High-Pressure Liquid Chromatography-Mass Spectrometry analyses were performed using a Vanquish UHPLC coupled online to a Q Exactive mass spectrometer (Thermo Fisher, Bremen, Germany) (Nemkov et al. Methods Mol Bio 2019). Samples were analyzed using a 5-minute gradient as described ( Nemkov et al. Methods Mol Bio 2019, Nemkov et al. JCI Insight 2020). Solvents were supplemented with 0.1% formic acid for positive mode runs and 1 mM ammonium acetate for negative mode runs. MS acquisition, data analysis and elaboration were performed as described.

Sampleprep ID:

SP002200

Sampleprep Summary:

Metabolomics analyses were performed as extensively described in previous studies (Issaian et al., Hematologica 2021). A volume of 20μl of frozen plasma was extracted in either 480μl of methanol:acetonitrile:water (5:3:2, v/v/v) (D'Alessandro et al. JCI Insight 2021). After vortexing at 4°C for 30 min, extracts were separated from the protein pellet by centrifugation for 10 min at 10,000g at 4°C and stored at −80°C until analysis. Ultra-High-Pressure Liquid Chromatography-Mass Spectrometry analyses were performed using a Vanquish UHPLC coupled online to a Q Exactive mass spectrometer (Thermo Fisher, Bremen, Germany) (Nemkov et al. Methods Mol Bio 2019). Samples were analyzed using a 5-minute gradient as described ( Nemkov et al. Methods Mol Bio 2019, Nemkov et al. JCI Insight 2020). Solvents were supplemented with 0.1% formic acid for positive mode runs and 1 mM ammonium acetate for negative mode runs. MS acquisition, data analysis and elaboration were performed as described.