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MB Sample ID: SA217520
Local Sample ID: | Smp_S515mMS_B |
Subject ID: | SU002352 |
Subject Type: | Water sample |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002352 |
Subject Type: | Water sample |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Smp_S515mMS_B | SA217520 | FL026414 | NA | Sampling_Date |
Smp_S515mMS_B | SA217520 | FL026414 | NA | Latitude |
Smp_S515mMS_B | SA217520 | FL026414 | NA | Longitude |
Smp_S515mMS_B | SA217520 | FL026414 | NA | Station |
Smp_S515mMS_B | SA217520 | FL026414 | NA | Cast |
Smp_S515mMS_B | SA217520 | FL026414 | NA | Depth |
Smp_S515mMS_B | SA217520 | FL026414 | NA | sample_vol_filtered_L |
Collection:
Collection ID: | CO002345 |
Collection Summary: | Samples for particulate metabolites were collected from 5 different stations surrounding the lava flowing into the ocean off the island of Hawaii, all from a depth of 5m. At each sampling location, single, duplicate, or triplicate filters were collected using either niskins attached to a conductivity, temperature, depth array (CTD) or the underway intake. Samples (10 L) 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. In addition to our samples, we filtered MilliQ water through a 0.2 μm PTFE filter and extracted this filter alongside samples as a methodological blank. |
Sample Type: | Phytoplankton |
Collection Method: | CTD Niskin arrays |
Collection Location: | Hawai'i |
Volumeoramount Collected: | 10L |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR002364 |
Treatment Summary: | Samples (10 L) 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. |
Sample Preparation:
Sampleprep ID: | SP002358 |
Sampleprep Summary: | Each sample was extracted using a modified Bligh-Dyer extraction. Briefly, filters were cut up and 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 ~2 mL of cold dichloromethane was added to the combined aqueous layer. Tubes were shaken and centrifuged at 4,300 rpm for 2 minutes at 4°C. The aqueous layer was removed to a new glass vial and dried under N2 gas. The remaining organic layer in the bead beating tubes was transferred into the glass centrifuge tube and the bead beating tube was rinsed two more times with cold organic solvent. The combined organic rinses were centrifuged, transferred to a new glass vial, and dried under N2 gas. Dried aqueous fractions were re-dissolved in 380 µL of water. Dried organic fractions were re-dissolved in 400 µL of 90:10 methanol:toluene. 20 µL of isotope-labeled injection standards in water were added to the aqueous fractions. Blank filters were extracted alongside samples as methodological blanks. |
Combined analysis:
Analysis ID | AN003701 | AN003702 | AN003703 |
---|---|---|---|
Analysis type | MS | MS | MS |
Chromatography type | HILIC | HILIC | GC |
Chromatography system | Q Exactive™ Plus Hybrid Quadrupole-Orbitrap | Q Exactive™ Plus Hybrid Quadrupole-Orbitrap | Q Exactive™ Plus Hybrid Quadrupole-Orbitrap |
Column | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | Waters Acquity UPLC HSS Cyano (100 x 2.1mm,1.8um) |
MS Type | ESI | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
Ion Mode | POSITIVE | NEGATIVE | POSITIVE |
Units | Normalized peak area | Normalized peak area | Normalized peak area |
Chromatography:
Chromatography ID: | CH002742 |
Chromatography Summary: | See attached summary |
Methods Filename: | Ingalls_Metabolomics_LC_HOT-LAVA.txt |
Instrument Name: | Q Exactive™ Plus Hybrid Quadrupole-Orbitrap |
Column Name: | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
Column Temperature: | 30 |
Flow Rate: | 0.15 ml/min |
Solvent A: | 85% acetonitrile/15% water; 10 mM ammonium carbonate |
Solvent B: | 15% acetonitrile/85% water; 10 mM ammonium carbonate |
Chromatography Type: | HILIC |
Chromatography ID: | CH002743 |
Chromatography Summary: | See attached summary |
Methods Filename: | Ingalls_Metabolomics_LC_HOT-LAVA.txt |
Instrument Name: | Q Exactive™ Plus Hybrid Quadrupole-Orbitrap |
Column Name: | Waters Acquity UPLC HSS Cyano (100 x 2.1mm,1.8um) |
Chromatography Type: | GC |
MS:
MS ID: | MS003451 |
Analysis ID: | AN003701 |
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_HOT-LAVA.txt |
MS ID: | MS003452 |
Analysis ID: | AN003702 |
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_HOT-LAVA.txt |
MS ID: | MS003453 |
Analysis ID: | AN003703 |
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_HOT-LAVA.txt |