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MB Sample ID: SA207111
| Local Sample ID: | 180227_Smp_Cr_C |
| Subject ID: | SU002245 |
| Subject Type: | Cultured cells |
| Subject Species: | Marine Plankton |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002245 |
| Subject Type: | Cultured cells |
| Subject Species: | Marine Plankton |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 180227_Smp_Cr_C | SA207111 | FL025369 | C | Replicate |
Collection:
| Collection ID: | CO002238 |
| Collection Summary: | Cultured marine plankton cells grown to mid-to-late exponential phase were filtered onto 0.2-micron durapore filters and extracted for metabolites according to extraction protocol. |
| Sample Type: | Plankton cells |
Treatment:
| Treatment ID: | TR002257 |
| Treatment Summary: | Marine plankton were cultured according to standard protocols. |
Sample Preparation:
| Sampleprep ID: | SP002251 |
| Sampleprep Summary: | Marine plankton cells were extracted using the described protocols. 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 beadbeating 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: | SP_Ingalls_extraction_protocol.pdf |
Combined analysis:
| Analysis ID | AN003535 | AN003536 | AN003537 | AN003538 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | HILIC | HILIC | HILIC | 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) | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | Waters Acquity UPLC HSS Cyano (100 x 2.1 mm,1.8um) |
| MS Type | ESI | ESI | ESI | ESI |
| MS instrument type | Triple quadrupole | Orbitrap | Orbitrap | Orbitrap |
| MS instrument name | Waters Xevo-TQ-S | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | UNSPECIFIED | POSITIVE | NEGATIVE | POSITIVE |
| Units | presence (1), absence (0), or background level (NA) | presence (1), absence (0), or background level (NA) | presence (1), absence (0), or background level (NA) | presence (1), absence (0), or background level (NA) |
Chromatography:
| Chromatography ID: | CH002611 |
| Chromatography Summary: | For HILIC chromatography, a SeQuant ZIC-pHILIC column (5 µm particle size, 2.1 mm x 150 mm, from Millipore) was used with 10 mM ammonium carbonate in 85:15 acetonitrile to water (Solvent A) and 10 mM ammonium carbonate in 60:40 water to acetonitrile (Solvent B) at a flow rate of 0.15 mL/min. |
| Methods Filename: | CH_Ingalls_LC_protocol.pdf |
| Instrument Name: | Waters Acquity I-Class |
| 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: | 40% acetonitrile/60% water; 10 mM ammonium carbonate |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH002612 |
| Chromatography Summary: | For reversed phase (RP) chromatography, a Waters Acquity UPLC HSS Cyano column (1.8 µm particle size, 2.1 mm x 100 mm) equipped with a Waters Acquity UPLC HSS Cyano guard column (1.8 µm particle size, 2.1 mm x 5 mm) was used with 0.1% formic acid in water (Solvent A) and 0.1% formic acid in acetonitrile (Solvent B) at a flow rate of 0.4 mL/ min. |
| Methods Filename: | CH_Ingalls_LC_protocol.pdf |
| Instrument Name: | Waters Acquity I-Class |
| Column Name: | Waters Acquity UPLC HSS Cyano (100 x 2.1 mm,1.8um) |
| Column Temperature: | 35 |
| Flow Rate: | 0.4 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003293 |
| Analysis ID: | AN003535 |
| Instrument Name: | Waters Xevo-TQ-S |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | MS acquisition according to attached protocol. |
| Ion Mode: | UNSPECIFIED |
| Analysis Protocol File: | Ingalls_Metabolomics_MS_Parameters_2015.txt |
| MS ID: | MS003294 |
| Analysis ID: | AN003536 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | MS acquisition according to attached protocol. |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | Ingalls_Metabolomics_MS_Parameters_2015.txt |
| MS ID: | MS003295 |
| Analysis ID: | AN003537 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | MS acquisition according to attached protocol. |
| Ion Mode: | NEGATIVE |
| Analysis Protocol File: | Ingalls_Metabolomics_MS_Parameters_2015.txt |
| MS ID: | MS003296 |
| Analysis ID: | AN003538 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | MS acquisition according to attached protocol. |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | Ingalls_Metabolomics_MS_Parameters_2015.txt |
