Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA204429

Local Sample ID:	ybtP-0-6S
Subject ID:	SU002213
Subject Type:	Bacteria
Subject Species:	Escherichia coli
Taxonomy ID:	562

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Subject:

Subject ID:	SU002213
Subject Type:	Bacteria
Subject Species:	Escherichia coli
Taxonomy ID:	562

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
ybtP-0-6S	SA204429	FL025109	standard growth conditions	Treatment

Collection:

Collection ID:	CO002206
Collection Summary:	After 18h of culture, the sample supernatant was isolated.Then, 2μL 0.1M ferric chloride was mixed with 2 mL of cell supernatant. After incubating at room temperature for 15 minutes, the precipitate was removed by centrifugation at 20000 × g for 15 min at 4 °C. The supernatant was added to an SPE plate (Waters, Oasis HLB) and washed with 0.5 mL 5% methanol, and then eluted with 0.5 mL 100% methanol to obtain the siderophores.
Sample Type:	Bacterial cells

Treatment:

Treatment ID:	TR002225
Treatment Summary:	M63 medium (1.36% monopotassium phosphate, 0.2% ammonium sulfate, 0.024% magnesium sulfate, 0.001% calcium chloride, and 0.0015% nicotinic acid) was used to form siderophores. The E. coli strain was incubated in LB-agar plate for 12 hours, one colony was isolated to LB broth for further 4 hours incubation, then diluted the solution into M63 medium at a ratio of 1:100 and the cultures were incubated for another18 h at 37°C, 200rpm to culture E. coli.

Treatment ID:

TR002225

Treatment Summary:

M63 medium (1.36% monopotassium phosphate, 0.2% ammonium sulfate, 0.024% magnesium sulfate, 0.001% calcium chloride, and 0.0015% nicotinic acid) was used to form siderophores. The E. coli strain was incubated in LB-agar plate for 12 hours, one colony was isolated to LB broth for further 4 hours incubation, then diluted the solution into M63 medium at a ratio of 1:100 and the cultures were incubated for another18 h at 37°C, 200rpm to culture E. coli.

Sample Preparation:

Sampleprep ID:	SP002219
Sampleprep Summary:	Siderophores were extracted as previously described. Briefly, 12μL 0.1M ferric chloride was mixed with 2 mL of cell supernatant. After incubating at room temperature for 15 minutes, the precipitate was removed by centrifugation at 20000 × g for 15 min at 4 °C. The supernatant was added to an SPE plate (Waters, Oasis HLB) and washed with 0.5 mL 5% methanol, and then eluted with 0.5 mL 100% methanol to obtain the siderophores. LC/MS analysis was performed using 5μL aliquots.

Sampleprep ID:

SP002219

Sampleprep Summary:

Siderophores were extracted as previously described. Briefly, 12μL 0.1M ferric chloride was mixed with 2 mL of cell supernatant. After incubating at room temperature for 15 minutes, the precipitate was removed by centrifugation at 20000 × g for 15 min at 4 °C. The supernatant was added to an SPE plate (Waters, Oasis HLB) and washed with 0.5 mL 5% methanol, and then eluted with 0.5 mL 100% methanol to obtain the siderophores. LC/MS analysis was performed using 5μL aliquots.

Combined analysis:

Analysis ID	AN003481
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Agilent 1290 Infinity
Column	Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um)
MS Type	ESI
MS instrument type	QTOF
MS instrument name	Agilent 6560 Ion Mobility
Ion Mode	POSITIVE
Units	peak area

Chromatography:

Chromatography ID:	CH002570
Instrument Name:	Agilent 1290 Infinity
Column Name:	Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS003242
Analysis ID:	AN003481
Instrument Name:	Agilent 6560 Ion Mobility
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	Agilent MassHunter Workstation Data Acquisition Agilent MassHunter QualitativeAnalysis B.07.00 Agilent MassHunter Quantitative Analysis (for QTOF)
Ion Mode:	POSITIVE