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MB Sample ID: SA192390
| Local Sample ID: | exp4_hilic_pos_01 |
| Subject ID: | SU002128 |
| Subject Type: | Invertebrate |
| Subject Species: | Caenorhabditis elegans |
| Taxonomy ID: | 6239 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002128 |
| Subject Type: | Invertebrate |
| Subject Species: | Caenorhabditis elegans |
| Taxonomy ID: | 6239 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| exp4_hilic_pos_01 | SA192390 | FL023629 | Sample | sample_type |
| exp4_hilic_pos_01 | SA192390 | FL023629 | 4 | experiment_number |
| exp4_hilic_pos_01 | SA192390 | FL023629 | hilic_pos | lc_ms_method |
Collection:
| Collection ID: | CO002121 |
| Collection Summary: | C. elegans were grown on large-scale culture plates (LSCPs), manually counted, aliquoted into 50,000 worm tubes, lyophilized and weighed. See detailed protocol for worm growth at: Shaver, A. O., Gouveia, G. J., Kirby, P. S., Andersen, E. C., Edison, A. S. Culture and Assay of Large-Scale Mixed-Stage Caenorhabditis elegans Populations. J. Vis. Exp. (171), e61453, doi:10.3791/61453 (2021). |
| Sample Type: | Worms |
| Volumeoramount Collected: | 50,000 worms |
| Storage Conditions: | -80℃ |
| Storage Vials: | 2.0 mL screwcap tubes |
Treatment:
| Treatment ID: | TR002140 |
| Treatment Summary: | The compared groups are different extraction protocols using a Taguchi Design of Experiments (DOE) approach. 3 different types of: solvent, volume, extraction time, and reconstitution solvent were investigated. Solvents included: IPA, MeOH, and 1:1 1-Butanol:MeOH for non-polar extractions followed by an 80:20 MeOH/H2O sequential polar extraction. Volumes of 0.5, 1.0, and 3.0 mL were used; Times: 0.5, 2.0, or 12.0 hours, and reconstitution solvents: IPA, 60/40 H2O/ACN, or 50/50 IPA/ACN (non-polar), and H2O, MeOH, or 80/20 MeOH/H2O (polar). Detailed information on the 9 different experimental conditions can be found in the attached manuscript or methods. |
Sample Preparation:
| Sampleprep ID: | SP002134 |
| Sampleprep Summary: | Samples were extracted according to the L9 orthogonal array described in the manuscript. Details are shown in the attached methods, but for full context refer to the manuscript. |
Combined analysis:
| Analysis ID | AN003330 | AN003331 | AN003332 | AN003333 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | HILIC | HILIC |
| Chromatography system | Thermo Vanquish | Thermo Vanquish | Thermo Vanquish | Thermo Vanquish |
| Column | Thermo Scientific Accucore C30 (150 x 2.1mm,2.6um) | Thermo Scientific Accucore C30 (150 x 2.1mm,2.6um) | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) |
| MS Type | ESI | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive HF-X Orbitrap | Thermo Q Exactive HF-X Orbitrap | Thermo Q Exactive HF-X Orbitrap | Thermo Q Exactive HF-X Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE | POSITIVE | NEGATIVE |
| Units | Peak height | Peak height | Peak height | Peak height |
Chromatography:
| Chromatography ID: | CH002467 |
| Chromatography Summary: | The following gradient program was used: -5.0 20% B; 0.0 min 20% B; 1.0 min 60% B; 5.0 min 70% B; 5.5 min 85% B; 8.0 min 90% B; 8.2-10.5 min 100% B; 10.7-12.0 min 20% B. A curve 5 value was set for -5.0 and 0.0 minutes, and a curve 6 for the remainder of the gradient. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Thermo Scientific Accucore C30 (150 x 2.1mm,2.6um) |
| Column Temperature: | 55 |
| Flow Rate: | 0.400 mL/min |
| Sample Injection: | 2.0 uL |
| Solvent A: | 40% water/60% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Solvent B: | 90% isopropanol/10% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH002468 |
| Chromatography Summary: | The following gradient program was used: -5.0 min 95% B; 0.0-0.5 min 95% B; 8.0-9.4 min 40% B; 9.5-11.0 min 95% B. A curve 5 value was set for -5.0 and 0.0 minutes, a curve 6 at 0.5 min, curve 7 at 8.0 min, and a curve 6 for the remainder of the gradient. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) |
| Column Temperature: | 40 |
| Flow Rate: | 0.400 mL/min |
| Sample Injection: | 2.0 uL |
| Solvent A: | 80% water/20% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003100 |
| Analysis ID: | AN003330 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Scan range 150-2000 m/z. Details in attached methods. MZmine 2.53 was used for data processing. |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 262.50 |
| Dry Gas Flow: | 50 |
| Dry Gas Temp: | 425.0 |
| Source Temperature: | 425.0 |
| Spray Voltage: | 3500 |
| Dataformat: | .raw Profile |
| MS ID: | MS003101 |
| Analysis ID: | AN003331 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Scan range 150-2000 m/z. Details in attached methods. MZmine 2.53 was used for data processing. |
| Ion Mode: | NEGATIVE |
| Capillary Temperature: | 262.50 |
| Dry Gas Flow: | 50 |
| Dry Gas Temp: | 425.0 |
| Source Temperature: | 425.0 |
| Spray Voltage: | 2500 |
| Dataformat: | .raw Profile |
| MS ID: | MS003102 |
| Analysis ID: | AN003332 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Scan range 70-1050 m/z. Details in attached methods. MZmine 2.53 was used for data processing. |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 262.50 |
| Dry Gas Flow: | 50 |
| Dry Gas Temp: | 425.0 |
| Source Temperature: | 425.0 |
| Spray Voltage: | 3500 |
| Dataformat: | .raw Profile |
| MS ID: | MS003103 |
| Analysis ID: | AN003333 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Scan range 70-1050 m/z. Details in attached methods. MZmine 2.53 was used for data processing. |
| Ion Mode: | NEGATIVE |
| Capillary Temperature: | 262.50 |
| Dry Gas Flow: | 50 |
| Dry Gas Temp: | 425.0 |
| Source Temperature: | 425.0 |
| Spray Voltage: | 2500 |
| Dataformat: | .raw Profile |
