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MB Sample ID: SA255503
| Local Sample ID: | Meltwater_B |
| Subject ID: | SU002639 |
| Subject Type: | Water sample |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002639 |
| Subject Type: | Water sample |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Meltwater_B | SA255503 | FL032686 | 11/5/2018 | Sampling_Date |
| Meltwater_B | SA255503 | FL032686 | 64.78 | Latitude |
| Meltwater_B | SA255503 | FL032686 | 64.05 | Longitude |
| Meltwater_B | SA255503 | FL032686 | 1.105 | sample_vol_filtered_L |
Collection:
| Collection ID: | CO002632 |
| Collection Summary: | Samples for particulate metabolites from cruise B234P were collected from Palmer Station from November 5 to November 21 2018, all at <1 m. At each sampling date, triplicate filters were collected using polycarbonate carboys. For sea-ice cores, the bottom 5-cm sections were placed in polycarbonate tubs and allowed to melt at 4°C into filtered seawater before collection. Samples (~0.2-3L) were collected into polycarbonate carboys, filtered onto 147 mm 0.2 μm PTFE filters using peristaltic pumps, polycarbonate filter holders, and Masterflex PharMed BPT tubing (Cole-Parmer). Filters were flash frozen in liquid nitrogen and stored at -80°C until extraction. Blank PTFE filters were extracted alongside samples as methodological blanks. |
| Sample Type: | Suspended Marine Particulate Matter |
| Storage Conditions: | Described in summary |
Treatment:
| Treatment ID: | TR002651 |
| Treatment Summary: | On 12th November 2018 (sample SW_12), additional seawater was collected for incubation experiments that simulated temperature(T)-salinity(S) conditions of sea-ice meltwater (3˚C and salinity 21, designated Meltwater_T-S), ambient seawater (0˚C and salinity 35, SW_T-S), and sea ice (3˚C and salinity 52, Sea ice_T-S) in triplicate. All other samples received no treatment and were samples of the natural marine microbial population in the surface ocean and sea ice along the western Antarctic Peninsula. |
Sample Preparation:
| Sampleprep ID: | SP002645 |
| Sampleprep Summary: | Each sample was extracted using a modified Bligh-Dyer extraction. Briefly, filters were put into 15 mL teflon centrifuge tubes containing a mixture of 100 µm and 400 µm silica beads. Heavy isotope-labeled internal standards were added along with ~2 mL of cold aqueous solvent (50:50 methanol:water) and ~3 mL 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 centrifuge at 4,300 rpm for 2 minutes 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 1 to 2 mL 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. |
| Processing Storage Conditions: | On ice |
| Extraction Method: | Bligh-Dyer |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN004181 | AN004182 | AN004183 | AN004184 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | HILIC | HILIC | Reversed phase | Reversed phase |
| Chromatography system | Waters Acquity I-Class | Waters Acquity I-Class | Waters Acquity I-Class | Waters Acquity I-Class |
| Column | SeQuant ZIC- pHILIC (150 x 2.1mm,5um) | SeQuant ZIC- pHILIC (150 x 2.1mm,5um) | Waters ACQUITY UPLC CSH C18 (150 x 2.1mm,1.7um) | Waters ACQUITY UPLC HSS CN (100 x 2.1mm,1.8um) |
| MS Type | ESI | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE | POSITIVE | NEGATIVE |
| Units | Estimated metabolite carbon concentration (nmol C per L) | Estimated metabolite carbon concentration (nmol C per L) | Estimated metabolite carbon concentration per particulate carbon (nmol C per umol C) | Estimated metabolite carbon concentration (nmol C per L) |
Chromatography:
| Chromatography ID: | CH003098 |
| Chromatography Summary: | See attached summary. |
| Instrument Name: | Waters Acquity I-Class |
| Column Name: | SeQuant ZIC- pHILIC (150 x 2.1mm,5um) |
| Column Temperature: | 30 |
| Flow Gradient: | 100% B for 2 minutes, ramped to 64% A over 18 minutes, ramped up to 100% A over 1 minute, held at 100% A for 7 minutes, and equilibrated at 100% B for 22 minutes (total time is 50 minutes) |
| Flow Rate: | 0.15 mL/min |
| Solvent A: | 10mM ammonium carbonate;85% water/15% acetonitrile |
| Solvent B: | 10mM ammonium carbonate;85% acetonitrile/15% water |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH003099 |
| Chromatography Summary: | See attached summary. |
| Instrument Name: | Waters Acquity I-Class |
| Column Name: | Waters ACQUITY UPLC CSH C18 (150 x 2.1mm,1.7um) |
| Column Temperature: | 65 |
| Flow Gradient: | Initial conditions were 90% A and 10% B. The column was ramped to 80% B over 33 minutes, ramped up to 90% B over 12 minutes, held at 90% B for 1 minute, and equilibrated to 90%A and 10% B for 6 minutes (total time is 52 minutes). |
| Flow Rate: | 0.45 mL/min |
| Solvent A: | 10mM ammonium formate; 60% acetonitrile/40% water; 0.1% formic acid |
| Solvent B: | 10mM ammonium formate; 90% isopropyl alcohol/10% acetonitrile; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH003100 |
| Chromatography Summary: | See attached summary. |
| Instrument Name: | Waters Acquity I-Class |
| Column Name: | Waters ACQUITY UPLC HSS CN (100 x 2.1mm,1.8um) |
| Column Temperature: | 35 |
| Flow Gradient: | The column was held at 5% B for 2 minutes, ramped to 100% B over 18 minutes, held at 100% B for 2 minutes, and equilibrated at 5% B for 5 minutes (total run time is 25 minutes) |
| Flow Rate: | 0.4 mL/min |
| Solvent A: | 0.1% formic acid in water |
| Solvent B: | 0.1% formic acid in acetonitrile |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003928 |
| Analysis ID: | AN004181 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | See protocol. |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | Ingalls_Metabolomics_MS_PalmerStation.txt |
| MS ID: | MS003929 |
| Analysis ID: | AN004182 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | See protocol. |
| Ion Mode: | NEGATIVE |
| Analysis Protocol File: | Ingalls_Metabolomics_MS_PalmerStation.txt |
| MS ID: | MS003930 |
| Analysis ID: | AN004183 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | See protocol. |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | Ingalls_Metabolomics_MS_PalmerStation.txt |
| MS ID: | MS003931 |
| Analysis ID: | AN004184 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | See protocol. |
| Ion Mode: | NEGATIVE |
| Analysis Protocol File: | Ingalls_Metabolomics_MS_PalmerStation.txt |
