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MB Sample ID: SA001579
| Local Sample ID: | 131221ajcsa34_1 |
| Subject ID: | SU000042 |
| Subject Type: | Insect |
| Subject Species: | Centroptilum triangulifer;Atherix;Megaloptera |
| Taxonomy ID: | 2078957;124299;50553 |
| Species Group: | Insect |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000042 |
| Subject Type: | Insect |
| Subject Species: | Centroptilum triangulifer;Atherix;Megaloptera |
| Taxonomy ID: | 2078957;124299;50553 |
| Species Group: | Insect |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 131221ajcsa34_1 | SA001579 | FL000532 | Mayfly | species |
| 131221ajcsa34_1 | SA001579 | FL000532 | Mayfly Heat Treated | Treatment |
| 131221ajcsa34_1 | SA001579 | FL000532 | no wing pads | wing pads |
Collection:
| Collection ID: | CO000025 |
| Collection Summary: | whole insects were collected |
| Collection Protocol Comments: | Original culture material was obtained from the Stroud Water Research Center (Avondale, PA). Larvae were reared at 22C and fed a periphytic diet. When larvae reached a suitable size, they were separated from their food source for 12 hours to evacuate gut contents. Larvae were then divided into two treatment groups a control group maintained at the culturing temperature of 22C, and a thermal challenge group. The thermal challenge group was subjected to a temperature increase at a rate of 1C per hour. This rate of temperature change is commonly observed in temperate streams. When the treatment temperature reached 30C, larvae from both control and treatment groups were flash frozen in liquid nitrogen in groups of 12-13 larvae. Each grouping of 12-13 larvae comprised a composite replicate for the metabolic profiling analysis. A total of 3 replicates each for the control and thermal treatments were used in this study. Some larvae had developed dark wing pads during the experiment. This is a signal that the larvae were very close to emerging from the aquatic larval phase to a winged sub-adult phase. Enough of these larvae were present in the control treatment to be removed from the control cohort and considered as an independent sample. |
| Sample Type: | whole insect |
| Volumeoramount Collected: | 50 mg |
| Storage Conditions: | -80C |
Treatment:
| Treatment ID: | TR000043 |
| Treatment Summary: | Metabolomics profile of mayfly larvae exposed to heat |
| Treatment Protocol Comments: | Heat Treated | Control | ControlX |
| Treatment: | Abiotic |
| Animal Acclimation Duration: | exposed to heat (22C to 30C ramp); fasted for 8 hours | control (fasted for 8 hours) | control (fasted for 6 hours) |
Sample Preparation:
| Sampleprep ID: | SP000038 |
| Sampleprep Summary: | Insect samples homogenized in 50% acetonitrile at a concentration of 50 mg/mL. Sample dried and aliquot taken for NMR. Sample returned and extract processed for GC/MS analysis. Samples were derivatized using 2 step derivatization method (methoximation with methoxy amine followed by TMS derivatization with MSTFA). |
| Sampleprep Protocol Filename: | RTI_DB_INSECT_Metabolomics_Procedure_March12_2014.docx |
| Processing Method: | Homogenization and Solvent Removal with SpeedVac |
| Processing Storage Conditions: | 4C |
| Extraction Method: | 50% Acetonitrile |
| Extract Concentration Dilution: | dried on SpeedVac |
| Extract Storage: | -80C |
| Sample Resuspension: | resuspended in D2O + Chenomix ISTD solution |
| Sample Derivatization: | Methoximation with methoxy amine at 40 mg/ml in pyridine followed by TMS derivatization with MSTFA |
| Sample Spiking: | FAMES (C8-C30) added to MSTFA as retention time markers |
Combined analysis:
| Analysis ID | AN000045 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Leco Pegasus IV GC |
| Column | Restek Rtx-5Sil (30m x 0.25mm,0.25um) |
| MS Type | EI |
| MS instrument type | GC x GC-TOF |
| MS instrument name | Leco Pegasus IV TOF |
| Ion Mode | POSITIVE |
| Units | peak area |
Chromatography:
| Chromatography ID: | CH000026 |
| Chromatography Summary: | GC/MS analysis is based on Oliver Fiehn methods |
| Instrument Name: | Leco Pegasus IV GC |
| Column Name: | Restek Rtx-5Sil (30m x 0.25mm,0.25um) |
| Column Pressure: | 10.4 psi |
| Column Temperature: | 50C(0.5 min)-330C(@20C/min)-330C(5 min) |
| Flow Gradient: | constant flow |
| Flow Rate: | 1.0 mL/min |
| Injection Temperature: | 250C |
| Retention Index: | FAMEs markers (C8-C30) |
| Retention Time: | FAMEs markers (C8-C30) |
| Analytical Time: | 20 minutes |
| Oven Temperature: | 50C(0.5 min)-330C(@20C/min)-330C(5 min) |
| Transferline Temperature: | 280C |
| Sample Syringe Size: | 10 L |
| Chromatography Type: | GC |
MS:
| MS ID: | MS000040 |
| Analysis ID: | AN000045 |
| Instrument Name: | Leco Pegasus IV TOF |
| Instrument Type: | GC x GC-TOF |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 280C (transfer line) |
| Helium Flow: | 1.0 ml/min |
| Ion Source Temperature: | 250C |
| Ionization Energy: | 70 eV |
| Source Temperature: | 250C |
| Dataformat: | netcdf |
| Resolution Setting: | 1200 |
| Scan Range Moverz: | 50-800 |
| Scanning: | 20 spectra/sec |
