Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA301032

Local Sample ID:	20220202_B8_7j_M2_82_1_676
Subject ID:	SU002914
Subject Type:	Other organism
Subject Species:	Biofilm microbial community

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Subject:

Subject ID:	SU002914
Subject Type:	Other organism
Subject Species:	Biofilm microbial community

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
20220202_B8_7j_M2_82_1_676	SA301032	FL036563	7	Sampling day
20220202_B8_7j_M2_82_1_676	SA301032	FL036563	j	Experimental group
20220202_B8_7j_M2_82_1_676	SA301032	FL036563	1 x 10-6	Cobalt concentration

Collection:

Collection ID:	CO002907
Collection Summary:	Four weeks before the start of the experiment, the mature biofilms were collected by natural colonisation of glass slides (5 x 10 cm).
Sample Type:	Microbial biofilm

Treatment:

Treatment ID:	TR002923
Treatment Summary:	A total of 15 microcosms were filled with 15 L of the Gave de Pau water; three for each of the five exposure conditions: 2 x 10-9 M (background concentration used as control), 1 x 10-7, 1 x 10-6, 5 x 10-6 and 1 x10-5 M Co. Cobalt (Cobalt Standard for ICP, 1000 mg·L-1, Supelco, Germany) was added in each microcosm and let equilibrating for 24 h (D-1). The microcosms were placed in one of the artificial streams of the facility in order to maintain the water temperature at 13.5 ± 1.0°C (Table S2). After one, three and seven days of exposure (D1, D3 and D7), slides were collected from each microcosm for metabolomics.

Treatment ID:

TR002923

Treatment Summary:

A total of 15 microcosms were filled with 15 L of the Gave de Pau water; three for each of the five exposure conditions: 2 x 10-9 M (background concentration used as control), 1 x 10-7, 1 x 10-6, 5 x 10-6 and 1 x10-5 M Co. Cobalt (Cobalt Standard for ICP, 1000 mg·L-1, Supelco, Germany) was added in each microcosm and let equilibrating for 24 h (D-1). The microcosms were placed in one of the artificial streams of the facility in order to maintain the water temperature at 13.5 ± 1.0°C (Table S2). After one, three and seven days of exposure (D1, D3 and D7), slides were collected from each microcosm for metabolomics.

Sample Preparation:

Sampleprep ID:	SP002920
Sampleprep Summary:	Briefly, freeze-dried biofilms (1 mg) were diluted in 10 µL of cold 75% methanol acidified with 0.1% formic acid and sonicated on ice during 30 s, at 80 % of the maximum intensity (SONICS Vibra Cell, Newton, CT, USA; 130 Watts, 20 kHz). The homogenates were then centrifuged at 4°C (12,000 g: 10 min). The supernatants were then collected and stored in the dark at -20°C.
Processing Storage Conditions:	-20℃

Combined analysis:

Analysis ID	AN004564
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Waters Acquity
Column	Thermo Acclaim Polar Advantage II (150 x 3mm,3um)
MS Type	ESI
MS instrument type	QTOF
MS instrument name	Bruker maXis Impact qTOF
Ion Mode	POSITIVE
Units	Dalton

Chromatography:

Chromatography ID:	CH003429
Instrument Name:	Waters Acquity
Column Name:	Thermo Acclaim Polar Advantage II (150 x 3mm,3um)
Column Temperature:	40
Flow Gradient:	5-90%
Flow Rate:	300 microliter per minute
Solvent A:	Water
Solvent B:	ACN
Chromatography Type:	Reversed phase

MS:

MS ID:	MS004310
Analysis ID:	AN004564
Instrument Name:	Bruker maXis Impact qTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	Metabolites were subsequently ionised and analysed using an electrospray ionization hybrid quadrupole time-of-flight high-resolution mass spectrometer (ESI-Qq-TOF Compact, Bruker) at 2 Hz speed on the 50–1500 m/z range on positive autoMS/MS mode between 2 and 8 Hz speed on the 50–1500 m/z range with information-dependent acquisition (IDA).
Ion Mode:	POSITIVE