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MB Sample ID: SA187667
| Local Sample ID: | d02 |
| Subject ID: | SU002085 |
| Subject Type: | Other abiotic sample |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002085 |
| Subject Type: | Other abiotic sample |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| d02 | SA187667 | FL023138 | B | Reactor |
| d02 | SA187667 | FL023138 | - | Day |
Collection:
| Collection ID: | CO002078 |
| Collection Summary: | Sludge samples from two reactors were collected at days 0, 80, 177, 218, 281, 353, and 462. Sludge was collected in 50-mL sterile Falcon tubes and immediately transported on dry ice to INRAE, where they were frozen at -80 °C. |
| Sample Type: | Sewage sludge |
Treatment:
| Treatment ID: | TR002097 |
| Treatment Summary: | During the time of the experiment, both reactors (3,000 m3 each) worked at half capacity due to a constraint in the availability of substrate to treat. On average, the full-scale digesters were operated at a mean hydraulic retention time (HRT) of 34 days and a mean organic loading rate of 0.82 kg of volatile solids m-3 day-1. For the first four months of study, the two reactors operated at 35 °C. Reactor A was kept at this temperature during all the experiment. For digester B, the temperature was decreased abruptly to 30 °C on the 123th day and maintained at this level for 2 HRTs until the 190th day. Then, the temperature was modified to 25 °C and maintained for 3 HRTs (between the 191st and the 373th days). Finally, the heating of reactor B was increased from day 374 onwards to reach the initial temperature of 35 °C. |
Sample Preparation:
| Sampleprep ID: | SP002091 |
| Sampleprep Summary: | For every sample, four aliquots with 0.75 mL of sludge were prepared in 2-mL Eppendorf tubes. Then, 1 mL of phosphate buffer saline was added into each tube and the sludge volumes were washed by centrifuging them at 15500 rcf for 3 min at 4 °C. This process was repeated once. The samples were freeze-dried overnight and, for every sample, 20 mg of dried sludge were transferred in new Eppendorf tubes. Then, the dried sludge powders were resuspended in 1 mL of a cold ddH2O: MeOH mixture (1:1) containing 2.5 mM PIPES. The samples were sonicated for 6 min, and submerged in liquid nitrogen. After one minute, the samples were thawed in ice. The cold-shock step was repeated twice. Afterwards, the samples were agitated for 15 min at 420 rpm in an orbital shaker. Following this, the samples were centrifuged at 15500 rcf for 3 min at 4 °C and the supernatants were collected. The supernatants were immediately filtered with 0.45 μM Nylon filters and freeze-dried overnight. The dried extracts were left at -80°C until the HPLC-MS analyses. |
Combined analysis:
| Analysis ID | AN003267 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Accela 1250 |
| Column | Macherey Nagel Nucleodur HILIC (100 x 2 mm,1.8um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo LTQ XL |
| Ion Mode | POSITIVE |
| Units | Peak areas |
Chromatography:
| Chromatography ID: | CH002411 |
| Instrument Name: | Thermo Accela 1250 |
| Column Name: | Macherey Nagel Nucleodur HILIC (100 x 2 mm,1.8um) |
| Flow Gradient: | 95% B: 3 min, 95% - 60% B: 3 - 14 min, 60% - 95 % B: 14 -24 min, 95% B: 24 - 29 min |
| Flow Rate: | 0.4 mL/min |
| Sample Injection: | 10 µL |
| Solvent A: | 100% water; 50 mM ammonium acetate, pH 5 |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003039 |
| Analysis ID: | AN003267 |
| Instrument Name: | Thermo LTQ XL |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | XCMS online for assignment METLIN, HMDB |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 320 °C |
