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MB Sample ID: SA178193
| Local Sample ID: | 61 |
| Subject ID: | SU002004 |
| Subject Type: | Invertebrate |
| Subject Species: | Drosophila melanogaster |
| Taxonomy ID: | 7227 |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002004 |
| Subject Type: | Invertebrate |
| Subject Species: | Drosophila melanogaster |
| Taxonomy ID: | 7227 |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 61 | SA178193 | FL022069 | Mel-56 | Line |
| 61 | SA178193 | FL022069 | females | Sex |
| 61 | SA178193 | FL022069 | melanogaster | Species |
| 61 | SA178193 | FL022069 | - | Age |
Collection:
| Collection ID: | CO001997 |
| Collection Summary: | We collected virgin males and females into vials with banana medium under light CO2 anesthesia within 8 hours of eclosion, except D. virilis, which was collected within 12 hours of eclosion. Flies were reared on banana media. For targeted analysis 3 flies from each sex/genotype combination at 5 and 31 days of age were flash frozen and stored at -80°C. For untargeted analysis five days old flies in groups of 10 flies were flash frozen in 1.5 mL tubes in liquid N2 and stored at -80°C until metabolite extraction |
| Sample Type: | Insect tissue |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002016 |
| Treatment Summary: | Sex, genotype, and age. |
Sample Preparation:
| Sampleprep ID: | SP002010 |
| Sampleprep Summary: | Metabolites were extracted by homogenizing samples in 200uL HPLC water (Sigma) using a TissueLyser (TissueLyser II, Qiagen) for 6 minutes at 25 Hz at 4°C. We then added 800uL methanol and incubated for 30 minutes on dry ice. The suspension was homogenized again for 10 minutes at 25Hz, then spun at 14,000rcf for 10 minutes at 4°C. The supernatant was transferred to a new tube and dried in a Speed-Vac at 30°C. |
Combined analysis:
| Analysis ID | AN003130 | AN003131 | AN003132 |
|---|---|---|---|
| Analysis type | MS | MS | MS |
| Chromatography type | HILIC | HILIC | HILIC |
| Chromatography system | Agilent 1260 | Agilent 1260 | Agilent 1260 |
| Column | Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um) | Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um) | Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um) |
| MS Type | ESI | ESI | ESI |
| MS instrument type | Other | QTOF | QTOF |
| MS instrument name | ABI Sciex 5500 QTrap | Agilent 6520 QTOF | Agilent 6520 QTOF |
| Ion Mode | UNSPECIFIED | POSITIVE | NEGATIVE |
| Units | Peak counts | Peak counts |
Chromatography:
| Chromatography ID: | CH002312 |
| Chromatography Summary: | Targeted LC/MS-MS 1st column. LC-MS/MS experiments were performed on an Agilent 1260 LC (Agilent Technologies, Santa Clara, CA)-AB Sciex QTrap 5500 mass spectrometer (AB Sciex, Toronto, ON, Canada) system at the University of Washington Northwest Metabolomics Research Center (UWNMRC). Each sample was injected twice, 10 µL for analysis using negative ionization mode and 2 µL for analysis using positive ionization mode. Both chromatographic separations were performed in hydrophilic interaction chromatography (HILIC) mode on two Waters XBridge BEH Amide columns (150 x 2.1 mm, 2.5 µm particle size, Waters Corporation, Milford, MA) connected in parallel. The flow rate was 0.300 mL/min, auto-sampler temperature was kept at 4 °C, and the column compartment was set at 40 °C. The mobile phase was composed of Solvents A (5 mM ammonium acetate in 90%H2O/ 10% acetonitrile + 0.2% acetic acid) and B (5 mM ammonium acetate in 90%acetonitrile/ 10% H2O + 0.2% acetic acid). After the initial 2 min isocratic elution of 90% B, the percentage of Solvent B decreased to 50% at t=5 min. The composition of Solvent B maintained at 50% for 4 min (t=9 min), and then the percentage of B gradually went back to 90%, to prepare for the next injection. Targeted data acquisition was performed in multiple-reaction-monitoring (MRM) mode. The LC-MS system was controlled by Analyst 1.5 software (AB Sciex). The extracted MRM peaks were integrated using MultiQuant 2.1 software (AB Sciex). Samples were spiked with 13C internal standards, and two types of LC-MS/MS quality control (QC) samples were run at 11 evenly-spaced intervals throughout the run to track potential drift in the assay. One QC sample was a pool of 10 fly samples, and the other was a sample of human serum. The CV for these QCs was 8.2% and 7.7% respectively |
| Instrument Name: | Agilent 1260 |
| Column Name: | Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um) |
| Column Temperature: | 40 |
| Flow Rate: | 0.300 mL/min |
| Internal Standard: | 13C |
| Solvent A: | 90% water/10% acetonitrile; 0.2% acetic acid; 5 mM ammonium acetate |
| Solvent B: | 90% acetonitrile/ 10% water; 0.2% acetic acid; 5 mM ammonium acetate |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH002313 |
| Chromatography Summary: | Targeted LC/MS-MS 1st column. LC-MS/MS experiments were performed on an Agilent 1260 LC (Agilent Technologies, Santa Clara, CA)-AB Sciex QTrap 5500 mass spectrometer (AB Sciex, Toronto, ON, Canada) system at the University of Washington Northwest Metabolomics Research Center (UWNMRC). Each sample was injected twice, 10 µL for analysis using negative ionization mode and 2 µL for analysis using positive ionization mode. Both chromatographic separations were performed in hydrophilic interaction chromatography (HILIC) mode on two Waters XBridge BEH Amide columns (150 x 2.1 mm, 2.5 µm particle size, Waters Corporation, Milford, MA) connected in parallel. The flow rate was 0.300 mL/min, auto-sampler temperature was kept at 4 °C, and the column compartment was set at 40 °C. The mobile phase was composed of Solvents A (5 mM ammonium acetate in 90%H2O/ 10% acetonitrile + 0.2% acetic acid) and B (5 mM ammonium acetate in 90%acetonitrile/ 10% H2O + 0.2% acetic acid). After the initial 2 min isocratic elution of 90% B, the percentage of Solvent B decreased to 50% at t=5 min. The composition of Solvent B maintained at 50% for 4 min (t=9 min), and then the percentage of B gradually went back to 90%, to prepare for the next injection. Targeted data acquisition was performed in multiple-reaction-monitoring (MRM) mode. The LC-MS system was controlled by Analyst 1.5 software (AB Sciex). The extracted MRM peaks were integrated using MultiQuant 2.1 software (AB Sciex). Samples were spiked with 13C internal standards, and two types of LC-MS/MS quality control (QC) samples were run at 11 evenly-spaced intervals throughout the run to track potential drift in the assay. One QC sample was a pool of 10 fly samples, and the other was a sample of human serum. The CV for these QCs was 8.2% and 7.7% respectively |
| Instrument Name: | Agilent 1260 |
| Column Name: | Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um) |
| Column Temperature: | 40 |
| Flow Rate: | 0.300 mL/min |
| Internal Standard: | 13C |
| Solvent A: | 90% water/10% acetonitrile; 0.2% acetic acid; 5 mM ammonium acetate |
| Solvent B: | 90% acetonitrile/ 10% water; 0.2% acetic acid; 5 mM ammonium acetate |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH002314 |
| Chromatography Summary: | Targeted LC/MS-MS 1st column. LC-MS/MS experiments were performed on an Agilent 1260 LC (Agilent Technologies, Santa Clara, CA)-AB Sciex QTrap 5500 mass spectrometer (AB Sciex, Toronto, ON, Canada) system at the University of Washington Northwest Metabolomics Research Center (UWNMRC). Each sample was injected twice, 10 µL for analysis using negative ionization mode and 2 µL for analysis using positive ionization mode. Both chromatographic separations were performed in hydrophilic interaction chromatography (HILIC) mode on two Waters XBridge BEH Amide columns (150 x 2.1 mm, 2.5 µm particle size, Waters Corporation, Milford, MA) connected in parallel. The flow rate was 0.300 mL/min, auto-sampler temperature was kept at 4 °C, and the column compartment was set at 40 °C. The mobile phase was composed of Solvents A (5 mM ammonium acetate in 90%H2O/ 10% acetonitrile + 0.2% acetic acid) and B (5 mM ammonium acetate in 90%acetonitrile/ 10% H2O + 0.2% acetic acid). After the initial 2 min isocratic elution of 90% B, the percentage of Solvent B decreased to 50% at t=5 min. The composition of Solvent B maintained at 50% for 4 min (t=9 min), and then the percentage of B gradually went back to 90%, to prepare for the next injection. Targeted data acquisition was performed in multiple-reaction-monitoring (MRM) mode. The LC-MS system was controlled by Analyst 1.5 software (AB Sciex). The extracted MRM peaks were integrated using MultiQuant 2.1 software (AB Sciex). Samples were spiked with 13C internal standards, and two types of LC-MS/MS quality control (QC) samples were run at 11 evenly-spaced intervals throughout the run to track potential drift in the assay. One QC sample was a pool of 10 fly samples, and the other was a sample of human serum. The CV for these QCs was 8.2% and 7.7% respectively |
| Instrument Name: | Agilent 1260 |
| Column Name: | Waters Xbridge BEH HILIC (75 x 2.1mm,2.5um) |
| Column Temperature: | 40 |
| Flow Rate: | 0.300 mL/min |
| Internal Standard: | 13C |
| Solvent A: | 90% water/10% acetonitrile; 0.2% acetic acid; 5 mM ammonium acetate |
| Solvent B: | 90% acetonitrile/ 10% water; 0.2% acetic acid; 5 mM ammonium acetate |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS002910 |
| Analysis ID: | AN003130 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Other |
| MS Type: | ESI |
| MS Comments: | Targeted data acquisition was performed in multiple-reaction-monitoring (MRM) mode. The LC-MS system was controlled by Analyst 1.5 software (AB Sciex). The extracted MRM peaks were integrated using MultiQuant 2.1 software (AB Sciex). Both positive and negative ionization modes were used. |
| Ion Mode: | UNSPECIFIED |
| MS ID: | MS002911 |
| Analysis ID: | AN003131 |
| Instrument Name: | Agilent 6520 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | The ESI voltage was 3.8 kV, and the m/z scan range was 60-1000. The Q-TOF data were extracted using Agilent MassHunter Qualitative Analysis (version B.07.00), Quantitative Analysis (version B.07.01), and Mass Profiler Professional (MPP, version B.13.00) software. The absolute intensity threshold for the LC-Q-TOF data extraction was 1000, and the mass accuracy limit was set to 10 ppm. Both positive and negative ionization modes were analyzed. |
| Ion Mode: | POSITIVE |
| MS ID: | MS002912 |
| Analysis ID: | AN003132 |
| Instrument Name: | Agilent 6520 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | The ESI voltage was 3.8 kV, and the m/z scan range was 60-1000. The Q-TOF data were extracted using Agilent MassHunter Qualitative Analysis (version B.07.00), Quantitative Analysis (version B.07.01), and Mass Profiler Professional (MPP, version B.13.00) software. The absolute intensity threshold for the LC-Q-TOF data extraction was 1000, and the mass accuracy limit was set to 10 ppm. Both positive and negative ionization modes were analyzed. |
| Ion Mode: | NEGATIVE |
