Return to study ST001497 main page
MB Sample ID: SA126172
Local Sample ID: | Spike_002_7 |
Subject ID: | SU001571 |
Subject Type: | Bacteria |
Subject Species: | Fecal Bacteria |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001571 |
Subject Type: | Bacteria |
Subject Species: | Fecal Bacteria |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Spike_002_7 | SA126172 | FL015507 | 2 | Sulfonates after dilution (ng/mL) |
Collection:
Collection ID: | CO001566 |
Collection Summary: | One g of the human fecal sample was homogenized in anoxic phosphate-buffered saline (PBS, pH 7.0, Table 4) by vortexing with glass beads (c. 3 mm; Roth, Germany) to yield a 10% fecal suspension. The fecal suspension was further diluted to 1% in a Hungate tube containing anoxic PBS supplemented with 3.18 mM of sterile filtered Ti(III) nitrilotriacetate (Sigma-Aldrich, Germany) as reductant. Subsequently, fecal slurry aliquotes were centrifuged (14.000 x g, 4°C, 5 min) and the supernatant frozen until further processing. |
Sample Type: | Feces |
Treatment:
Treatment ID: | TR001586 |
Treatment Summary: | Fecal slurries were s spiked with cysteate, sulfoquinovose (SQ), 2,3-dihydroxypropane-1-sulfonate (DHPS), taurine and isethionate . |
Sample Preparation:
Sampleprep ID: | SP001579 |
Sampleprep Summary: | Fecal supernatants were serially diluted using 50% acetonitrile and 0.1% formic acid in water to obtain final dilution of 1:10,000. 500 µl of each sample were placed in glass vials (Wicom, Heppenheim, Germany) and stored at - 80 °C until LC-MS/MS analysis.For the analysis of sulfonates by LC-MS/MS-MRM, aliquots of 250 µl for each sample were centrifuged (14 000 x g, 4 °C, 5 min) and 50 µl of the supernatant stored at – 20 °C until further processing. Samples were thawed, centrifuged at 18 000 x g at RT for 2 min and the supernatant was diluted 1:10 000 in 50% aqueous acetonitrile. Subsequently, 500 µl of each sample were placed in glass vials (Wicom, Heppenheim, Germany) and stored at - 80 °C until LC-MS/MS analysis. |
Sampleprep Protocol Filename: | sven_haange_Sample_preparation-application_experiment.pdf |
Combined analysis:
Analysis ID | AN002480 |
---|---|
Analysis type | MS |
Chromatography type | HILIC |
Chromatography system | Dionex 3000 HPLC (Dionex,Sunnyvale, USA) |
Column | BEH amide (100 x 2.1mm,1.7um) |
MS Type | ESI |
MS instrument type | Triple quadrupole |
MS instrument name | ABI Sciex 5500 QTrap |
Ion Mode | NEGATIVE |
Units | mM |
Chromatography:
Chromatography ID: | CH001817 |
Methods Filename: | sven_haange_20201002_055740_MS-parameters.pdf sven_haange_20201002_055740_LC_parameters-sulfonates.pdf |
Chromatography Comments: | BEH amide column, 2.1 x 100 mm, 1.7 µm, Waters, Milford, USA |
Instrument Name: | Dionex 3000 HPLC (Dionex,Sunnyvale, USA) |
Column Name: | BEH amide (100 x 2.1mm,1.7um) |
Chromatography Type: | HILIC |
MS:
MS ID: | MS002300 |
Analysis ID: | AN002480 |
Instrument Name: | ABI Sciex 5500 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | As softwar used Analyst 1.6.2 (Sciex) |
Ion Mode: | NEGATIVE |
Analysis Protocol File: | sven_haange_20201002_055740_LC_parameters-sulfonates.pdf |