Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA204459

Local Sample ID:	WT-0-6S
Subject ID:	SU002214
Subject Type:	Bacteria
Subject Species:	Escherichia coli
Taxonomy ID:	562

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Subject:

Subject ID:	SU002214
Subject Type:	Bacteria
Subject Species:	Escherichia coli
Taxonomy ID:	562

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
WT-0-6S	SA204459	FL025111	standard growth conditions	Treatment

Collection:

Collection ID:	CO002207
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:	TR002226
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 MG1655 and UTI89. In addition, add ferric chloride solution to the medium to prepare 10μM iron M63 medium, we cultured the wild UTI89 strain and MG1655 in the presence of 10μM iron. 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:

TR002226

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 MG1655 and UTI89. In addition, add ferric chloride solution to the medium to prepare 10μM iron M63 medium, we cultured the wild UTI89 strain and MG1655 in the presence of 10μM iron. 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:	SP002220
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:

SP002220

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	AN003482
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:	CH002571
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:	MS003243
Analysis ID:	AN003482
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