Protocol detail

Untargeted Metabolomics Analysis of Plasma

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Untargeted metabolomics data were performed as described (17 (link)). Metabolites were extracted from plasma samples using acetonitrile (2:1, vol/vol). Stable isotopes caffeine-¹³C3, tyrosine-¹⁵N, and progesterone-d⁹ were used as internal standards. LC-MS/MS analysis was performed with an HPLC-UV, 1220 Infinity (Agilent Technologies, Santa Clara, CA, USA) coupled with Q Exactive hybrid Quadrupole-Orbitrap high-resolution mass spectrometer (Thermo Fisher, Waltham, MA, USA). Reverse phase C18 chromatography was performed with Zorbax Eclipse Plus C18 column (4.6 × 150 mm², 3.5 µm Agilent) and positive electrospray ionization.

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Mwangi V.I., Netto R.L., Borba M.G., Santos G.F., Lima G.S., Machado L.S., Yakubu M.N., Val F.F., Sampaio V.S., Sartim M.A., Koolen H.H., Costa A.G., Toméi M.C., Guimarães T.P., Chaves A.R., Vaz B.G., Lacerda M.V., Monteiro W.M., Gardinassi L.G, & Melo G.C. (2023). Methylprednisolone therapy induces differential metabolic trajectories in severe COVID-19 patients. mSystems, 8(6), e00726-23.

Publication 2023

Q Exactive hybrid Quadrupole-Orbitrap high-resolution mass spectrometer Zorbax Eclipse Plus C18 column

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Variable analysis

independent variables

Extraction of metabolites from plasma samples using acetonitrile (2:1, vol/vol)

dependent variables

Metabolite levels measured by LC-MS/MS analysis

control variables

Stable isotopes caffeine-¹³C3, tyrosine-¹⁵N, and progesterone-d⁹ used as internal standards
Reverse phase C18 chromatography performed with Zorbax Eclipse Plus C18 column (4.6 × 150 mm², 3.5 µm Agilent)
Positive electrospray ionization used

controls

Positive control: None specified
Negative control: None specified

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