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MB Sample ID: SA126357
| Local Sample ID: | Spike_600_3 |
| Subject ID: | SU001572 |
| Subject Type: | Bacteria |
| Subject Species: | Fecal Bacteria |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001572 |
| Subject Type: | Bacteria |
| Subject Species: | Fecal Bacteria |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Spike_600_3 | SA126357 | FL015533 | 600 | Sulfonate after Dilution (ng/mL) |
Collection:
| Collection ID: | CO001567 |
| 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: | TR001587 |
| Treatment Summary: | Fecal slurries were s spiked with cysteate, sulfoquinovose (SQ), 2,3-dihydroxypropane-1-sulfonate (DHPS), taurine and isethionate . |
Sample Preparation:
| Sampleprep ID: | SP001580 |
| 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 | AN002481 |
|---|---|
| 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: | CH001818 |
| 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: | MS002301 |
| Analysis ID: | AN002481 |
| 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 |
