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MB Sample ID: SA308461

Local Sample ID:ID_85
Subject ID:SU002957
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Gender:Male and female

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Subject:

Subject ID:SU002957
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Gender:Male and female

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
ID_85SA308461FL037130D1Time-point
ID_85SA308461FL037130MPTreatment
ID_85SA308461FL037130MSex

Collection:

Collection ID:CO002950
Collection Summary:The COVID-19 infected participants in this study were drawn from the MetCOVID study, a parallel, double blind, randomized, placebo-controlled Phase IIb clinical trial, that assessed the efficacy of MP in treating hospitalized patients with suspected SARS-CoV-2 infection. Individuals with COVID-19 were admitted at the Hospital e Pronto-Socorro Delphina Rinaldi Abdel Aziz, in Manaus, Western Brazilian Amazon. The hospital was the largest public reference unit for the treatment of severe COVID-19 cases in the city. After confirmation of SARS-CoV-2 infection by RT-qPCR testing of nasopharyngeal swabs, blood was collected from patients and the separated plasma was stored at -80oC. The plasma samples were collected at the day of inclusion (D1) before treatment, and day five (D5), day seven (D7), day eleven (D11), and day fourteen (D14) after the start of treatment for metabolomic analysis.
Sample Type:Blood (plasma)

Treatment:

Treatment ID:TR002966
Treatment Summary:The participants were administered with either intravenous sodium succinate MP (0.5 mg/kg) or placebo (saline solution) twice daily for 5 days. As per hospital protocol, all patients meeting acute respiratory distress syndrome criteria also preemptively received intravenous ceftriaxone (1g twice daily for 7 days) plus azithromycin (500 mg once a day for 5 days) or clarithromycin (500 mg twice daily for 7 days), starting on Day 1. At inclusion, adult patients were excluded if they had a history of hypersensitivity to MP, were living with human immunodeficiency virus or AIDS, had a history of chronic use of corticosteroids or immunosuppressive agents, were pregnant or breastfeeding, or had decompensated cirrhosis or chronic renal failure.

Sample Preparation:

Sampleprep ID:SP002963
Sampleprep Summary:Metabolites were extracted from plasma samples using acetonitrile (2:1, v/v). Stable isotopes caffeine-¹³C3 and progesterone-d9 were used as internal standards.

Combined analysis:

Analysis ID AN004663
Analysis type MS
Chromatography type Reversed phase
Chromatography system Agilent 1220 Infinity
Column Agilent Zorbax Eclipse Plus C18 (150 x 4.6mm,3.5um)
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive Orbitrap
Ion Mode POSITIVE
Units intesity

Chromatography:

Chromatography ID:CH003509
Chromatography Summary:The binary mobile phases were water 0.5% formic acid with 5 mM of ammonium formate (A), and acetonitrile (B). Their gradient elution started with 20% (B) for 5 min, then linearly increased to 100% (B) in 30 min and kept constant for 8 min in 100% (B). The eluent was restored to the initial conditions in 4 minutes to re-equilibrate the column and held for the remaining 8 minutes. The flow rate was kept at 0.5 mL min-1. The injection volume for analysis was 3 μL, and the column temperature was set at 35 °C.
Instrument Name:Agilent 1220 Infinity
Column Name:Agilent Zorbax Eclipse Plus C18 (150 x 4.6mm,3.5um)
Column Temperature:35
Flow Gradient:gradient elution started with 20% (B) for 5 min, then linearly increased to 100% (B) in 30 min and kept constant for 8 min in 100% (B). The eluent was restored to the initial conditions in 4 minutes to re-equilibrate the column and held for the remaining 8 minutes.
Flow Rate:0.5 mL/min
Solvent A:100% water; 0.5% formic acid; 5 mM of ammonium formate
Solvent B:100% acetonitrile
Chromatography Type:Reversed phase

MS:

MS ID:MS004410
Analysis ID:AN004663
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:The electrospray ionization was operating with the following settings: spray voltage 3.5 kV; capillary temperature: 269 °C; S-lens RF level 50 V; sheath gas flow rate at 53 L min-1; aux gas flow rate at 14 L min-1; sweep gas flow rate 3 L min-1. The high-resolution mass-spectrometry was obtained under full MS/dd-MS2 mode. The mass range in the full MS scanning experiments was m/z 80-1200. The max IT was set at 200 ms, and AGC target was set at 1 x 106. For fragmentation acquisition, the top 5 (TopN, 5, loop count 5) most abundant precursors were sequentially transferred into the C-Trap (AGC target 1 x 105; max IT 50 ms) for collision. The collision energy for target analytes was 20, 30 and 35 eV. Resolving power was set at 140,000 and 70,000 for full MS and dd-MS2 acquisitions, respectively.
Ion Mode:POSITIVE
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