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MB Sample ID: SA126539
| Local Sample ID: | LPS-4 |
| Subject ID: | SU001577 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001577 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| LPS-4 | SA126539 | FL015558 | Model | Treatment |
Collection:
| Collection ID: | CO001572 |
| Collection Summary: | Samples of blood were gathered in heparinized tubes and then spun down at 3000 rpm at 4 °C for 10 min to obtain supernatant of plasma samples for subsequent preparation. Heart, lung, liver, spleen and kidney samples were homogenized in a five-fold volume of normal saline, respectively, and spun down at 12,000 rpm at 4 °C for 20 min to acquire supernatants for further preparation. |
| Collection Protocol Filename: | shanpingwang_Collection_Protocol.docx |
| Sample Type: | Kidney |
Treatment:
| Treatment ID: | TR001592 |
| Treatment Summary: | After being acclimatized for 1 week, the mice were separated, at random, into four groups: 1) saline + saline; 2) saline + LPS; 3) STV-Na+ LPS; and 4) dexamethasone (Dex) + LPS. Mice were intraperitoneally administered saline (0.1 mL/10 g) or STV-Na (5, 10, 20 mg/kg) and Dex (10 mg/kg) two times per day every 12 h for 3 consecutive days, and one hour after the first intraperitoneal injection on day 3, saline (0.1 mL/g body weight) or LPS from E. coli (0111: B4, 20 mg/kg) was intraperitoneally administered. |
| Treatment Protocol Filename: | shanpingwang_Treatment_Protocol.docx |
Sample Preparation:
| Sampleprep ID: | SP001585 |
| Sampleprep Summary: | A total of 160 µL of MTBE solution (methyl-T-butyl-ether: methanol: water, 6/3/1, v/v/v) was applied to 40 µL of the plasma or tissue homogenate supernatant, vortexed for 30 min at 4°C and spun at 12,000 rpm for 30 min. Two extract components were produced: an organic hydrophobic layer and a hydrophilic layer. These two extracts were vacuum-dried and dissolved in 0.1% (v/v) formic acid in water (45 µL), followed by analysis. The pooled quality control (QC) samples including whole plasma and tissues were utilized for monitoring data acquisition performance throughout the analysis. Finally, 6 duplicate QC samples were prepared and injected at the start of the sequence, and after each of the six tissue samples was inserted, the QC samples were added to determine system stability. |
| Sampleprep Protocol Filename: | shanpingwang_Sampleprep_protocol.docx |
Combined analysis:
| Analysis ID | AN002490 | AN002491 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
| Column | Waters Acquity BEH C18 (50 x 2.1mm,1.7um) | Waters Acquity BEH C18 (50 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Bruker TIMS TOF | |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Intensity | Intensity |
Chromatography:
| Chromatography ID: | CH001823 |
| Chromatography Summary: | Chromatographic separations were conducted utilizing a Waters BEH C18, 2.1 mm×50 mm 1.7 µm particle column with a Dionex Ultimate 3000 UHPLC system from Thermo Fisher Scientific (CA, USA). The mobile phase encompassed water with 0.1% v/v formic acid (A) and acetonitrile with 0.1% v/v formic acid (B). Columns were kept at 40°C and eluted using a linear gradient: 2-30% B at 0-4 min, 30-40% B at 4-5 min, 40% B at 5-8 min, 40-60% B at 8-10 min, 60-100% B at 10-17 min, 100% B at 17-19 min, 100-2% B at 19-19.1 min, and 2% B at 19.1-25 min. To increase the amount of metabolites and save experimental time, a new sampling method was used to detect both the organic phase and the aqueous phase extracts (Qiuhui Xuan et al., 2018; Shanping Wang et al., 2019): 5 µL of organic phase extracts were first loaded without running the elution gradient, which lasted for one minute at the initial mobile phase, and then 5 µL of the aqueous phase extracts were added to the same column in order to start running the elution gradient using a 0.4 mL/min flow rate. |
| Methods Filename: | shanpingwang_Chromatography_Methods.docx |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters Acquity BEH C18 (50 x 2.1mm,1.7um) |
| Column Temperature: | 40 |
| Flow Rate: | 0.4 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002310 |
| Analysis ID: | AN002490 |
| Instrument Name: | Bruker TIMS TOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | MSMS Progenesis QI 2.1 software EZinfo 3.0 software |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | shanpingwang_Analysis_Protocol.docx |
| MS ID: | MS002311 |
| Analysis ID: | AN002491 |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | MSMS Progenesis QI 2.1 software EZinfo 3.0 software |
| Ion Mode: | NEGATIVE |
| Analysis Protocol File: | shanpingwang_Analysis_Protocol.docx |
