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MB Sample ID: SA080591
| Local Sample ID: | M12D6 |
| Subject ID: | SU001230 |
| Subject Type: | Invertebrate |
| Subject Species: | Metacarcinus magister |
| Taxonomy ID: | 29965 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001230 |
| Subject Type: | Invertebrate |
| Subject Species: | Metacarcinus magister |
| Taxonomy ID: | 29965 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| M12D6 | SA080591 | FL012334 | High | ph treatment |
| M12D6 | SA080591 | FL012334 | Low | DO treatment |
Collection:
| Collection ID: | CO001224 |
| Collection Summary: | After each crab completed the treatment for the predetermined length of time, crabs were frozen whole specimen at -80°C and shipped to the West Coast Metabolomics Center. |
| Sample Type: | Whole Animal |
Treatment:
| Treatment ID: | TR001245 |
| Treatment Summary: | 15 crabs were placed into 1 of 4 treatment groups: High pH:High Dissolved Oxygen, High pH:Low Dissolved Oxygen, Low pH:Low Dissolved Oxygen, Low pH: High Dissolved Oxygen. Crabs were subjected to their treatment group for 30-33 days. |
Sample Preparation:
| Sampleprep ID: | SP001238 |
| Sampleprep Summary: | 15mg of whole crab was placed into a 1.5mL ependorph tube. 2 x 3mm grinding beads were added to each sample. 225 µL of cold MeOH with quality controls was added to each samples. Batches of samples were ground with GenoGrinder for 30 seconds at 1500 rpm. 750µL of methyl tert-butyl Ether (MTBE) was added to each sample. Samples were vortexed for 10 seconds and then shaken at 4°C for 5 minutes using an Orbital Mixer. 188 uL of LC-MS grade water was added to each sample. Vortex for 10 seconds and then centrifuged for 2 minutes at 14,000 rcf. 2 x 350µL aliquots were removed from the top, organic layer, one submitted for analysis and the other stored as backup in -20°C. 2 x 125µL aliquots were removed from the bottom, polar layer, one submitted for analysis, the other stored at -20°C for backup. |
| Sample Resuspension: | After drying, samples were resuspended in 110 µL of 9:1 methanol:toulene with 50 nM CUDA as an internal standard. Samples were vortexed for 10 seconds, then sonicated in room temperature water for 5 minutes, then centrifuged at 16,100 rcf for 2 minutes. 50 µL of the supernatant was removed and placed in an amber vial for lipidomics analysis. |
Combined analysis:
| Analysis ID | AN001926 | AN001927 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Agilent 1290 UPLC | Agilent 1290 UPLC |
| Column | Waters Acquity CSH C18 2.1x10 0mm 1.7m | Waters Acquity CSH C18 2.1x10 0mm 1.7m |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Agilent 6530 QTOF | Agilent 6530 QTOF |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Peak Height | Peak Height |
Chromatography:
| Chromatography ID: | CH001398 |
| Instrument Name: | Agilent 1290 UPLC |
| Column Name: | Waters Acquity CSH C18 2.1x10 0mm 1.7m |
| Column Pressure: | 500-1000bar |
| Column Temperature: | 65°C |
| Flow Rate: | 600µL/min |
| Sample Injection: | 1.67µL |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS001782 |
| Analysis ID: | AN001926 |
| Instrument Name: | Agilent 6530 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | N/A |
| Ion Mode: | POSITIVE |
| MS ID: | MS001783 |
| Analysis ID: | AN001927 |
| Instrument Name: | Agilent 6530 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | N/A |
| Ion Mode: | NEGATIVE |
