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MB Sample ID: SA098591
| Local Sample ID: | S43R3 |
| Subject ID: | SU001430 |
| Subject Type: | Other |
| Subject Species: | Ilyonectria mors-panacis;Ilyonectria rufa;Ilyonectria robusta;Ilyonectria torresensis;Ilyonectria estremocensis;Neonectria obtusispora |
| Taxonomy ID: | 1079131;1079113;1079257;1199091;1079267;78417 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001430 |
| Subject Type: | Other |
| Subject Species: | Ilyonectria mors-panacis;Ilyonectria rufa;Ilyonectria robusta;Ilyonectria torresensis;Ilyonectria estremocensis;Neonectria obtusispora |
| Taxonomy ID: | 1079131;1079113;1079257;1199091;1079267;78417 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| S43R3 | SA098591 | FL013886 | 1400 | Time |
| S43R3 | SA098591 | FL013886 | S69C001 | Station and Cast |
Collection:
| Collection ID: | CO001425 |
| Collection Summary: | Samples for particulate metabolites were collected from 15 m water depth by niskin bottles attached to a conductivity, temperature, depth array (CTD). Metabolite samples were collected in triplicate at each time point by filtering approximately 3.5 L of seawater onto 47 mm 0.2 micron Omnipore filters using peristaltic pumps, polycarbonate filter holders, and Masterflex PharMed BPT tubing (Cole-Parmer). Filters were frozen in liquid nitrogen immediately after filtration and stored in a -80 C freezer until extraction. |
| Sample Type: | Suspended Marine Particulate Matter |
Treatment:
| Treatment ID: | TR001445 |
| Treatment Summary: | No treatments - this was a study of the natural marine microbial population in the surface ocean at approximately 24.5 degrees N, 156.5 degrees W. |
Sample Preparation:
| Sampleprep ID: | SP001438 |
| Sampleprep Summary: | Each sample was extracted using a modified Bligh-Dyer extraction. Briefly, filters were cut up and split between two bead beating tubes containing a mixture of 100 µm and 400 µm silica beads. Heavy isotope-labeled internal standards were added along with 750 µL of cold aqueous solved (50:50 methanol:water) and 750 µL of cold organic solvent (dichloromethane). The samples were shaken on a FastPrep-24 Homogenizer for 30 seconds and chilled in a -20 °C freezer repeatedly for three cycles of bead-beating and a total of 30 minutes of chilling. The organic and aqueous layers were separated by spinning samples in a microcentrifuge at 5,000 rpm for 90 seconds at 4 °C. The aqueous layer was removed to a new glass centrifuge tube. The remaining organic fraction was rinsed three more times with additions of 750 µL of 50:50 methanol:water. All aqueous rinses were combined for each sample and dried down under N2 gas. The remaining organic layer was transferred into a clean glass centrifuge tube and the remaining bead beating tube was rinsed two more times with cold organic solvent. The combined organic rinses were centrifuged, transferred to a new tube, and dried under N2 gas. Dried aqueous fractions were re-dissolved in 380 µL of water. Dried organic fractions were re-dissolved in 380 µL of 1:1 water:acetonitrile. 20 µL of isotope-labeled injection standards in water were added to both fractions. Blank filters were extracted alongside samples as methodological blanks. |
| Sampleprep Protocol Filename: | Ingalls_Metabolomics_Sample_Processing_2015.txt |
| Processing Storage Conditions: | On ice |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN002255 | AN002256 | AN002257 | AN002258 | AN002259 |
|---|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | HILIC | HILIC | HILIC |
| Chromatography system | Waters Acquity I-Class | Waters Acquity I-Class | Waters Acquity I-Class | Waters Acquity I-Class | Waters Acquity I-Class |
| Column | Waters Acquity UPLC HSS Cyano (CN) ( 50 x 2.1mm,1.8um) | Waters Acquity UPLC HSS Cyano (CN) ( 50 x 2.1mm,1.8um) | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| MS Type | ESI | ESI | ESI | ESI | ESI |
| MS instrument type | Triple quadrupole | Orbitrap | Triple quadrupole | Triple quadrupole | Orbitrap |
| MS instrument name | Waters Xevo-TQ-S | Thermo Q Exactive HF hybrid Orbitrap | Waters Xevo-TQ-S | Waters Xevo-TQ-S | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | POSITIVE | POSITIVE | POSITIVE | NEGATIVE | POSITIVE |
| Units | Normalized Peak Area Per L Seawater Filtered | Normalized Peak Area Per L Seawater Filtered | Normalized Peak Area Per L Seawater Filtered | Normalized Peak Area Per L Seawater Filtered | Normalized Peak Area per L of SW filtered |
Chromatography:
| Chromatography ID: | CH001657 |
| Chromatography Summary: | RP |
| Methods Filename: | Ingalls_Metabolomics_MS_Parameters_2015.pdf |
| Instrument Name: | Waters Acquity I-Class |
| Column Name: | Waters Acquity UPLC HSS Cyano (CN) ( 50 x 2.1mm,1.8um) |
| Column Temperature: | 35 C |
| Flow Gradient: | 5% B for 2 minutes, ramped to 100% B over 16 minutes, held at 100% B for 2 minutes, and equilibrated at 5% B for 5 minutes |
| Flow Rate: | 0.4 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Analytical Time: | 20 min |
| Preconditioning: | equilibrated at the starting conditions for at least 10 minutes; several water blanks were run before |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH001658 |
| Chromatography Summary: | HILIC |
| Methods Filename: | Ingalls_Metabolomics_MS_Parameters_2015.pdf |
| Chromatography Comments: | In marine samples, a major salt peak elutes at approximately 23 minutes, To improve the performance of the HILIC column, we maintained the same injection volume, kept the instrument running water blanks between samples as necessary. |
| Instrument Name: | Waters Acquity I-Class |
| Column Name: | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| Column Temperature: | 30 C |
| Flow Gradient: | 100% A for 2 minutes, ramped to 100% B over 18 minutes, held at 100% B for 5 minutes, and equilibrated at 100% A for 25 minutes |
| Flow Rate: | 0.15 mL/min |
| Solvent A: | 85% acetonitrile/15% water; 10 mM ammonium carbonate |
| Solvent B: | 40% acetonitrile/60% water; 10 mM ammonium carbonate |
| Analytical Time: | 25 min |
| Preconditioning: | , the column was equilibrated at the starting conditions for at least 30 minutes |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS002099 |
| Analysis ID: | AN002255 |
| Instrument Name: | Waters Xevo-TQ-S |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | See File |
| Ion Mode: | POSITIVE |
| Acquisition Parameters File: | Ingalls_Metabolomics_MS_Parameters_2015.txt |
| MS ID: | MS002100 |
| Analysis ID: | AN002256 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | See File |
| Ion Mode: | POSITIVE |
| Acquisition Parameters File: | Ingalls_Metabolomics_MS_Parameters_2015.txt |
| MS ID: | MS002101 |
| Analysis ID: | AN002257 |
| Instrument Name: | Waters Xevo-TQ-S |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | See File |
| Ion Mode: | POSITIVE |
| Acquisition Parameters File: | Ingalls_Metabolomics_MS_Parameters_2015.txt |
| MS ID: | MS002102 |
| Analysis ID: | AN002258 |
| Instrument Name: | Waters Xevo-TQ-S |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | See File |
| Ion Mode: | NEGATIVE |
| Acquisition Parameters File: | Ingalls_Metabolomics_MS_Parameters_2015.txt |
| MS ID: | MS002103 |
| Analysis ID: | AN002259 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | See File |
| Ion Mode: | POSITIVE |
| Acquisition Parameters File: | Ingalls_Metabolomics_MS_Parameters_2015.txt |
