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MB Sample ID: SA342884
| Local Sample ID: | CNS_RPLC_POS_WT_4 |
| Subject ID: | SU003281 |
| Subject Type: | Insect |
| Subject Species: | Drosophila melanogaster |
| Taxonomy ID: | 7227 |
| Species Group: | Insects |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003281 |
| Subject Type: | Insect |
| Subject Species: | Drosophila melanogaster |
| Taxonomy ID: | 7227 |
| Species Group: | Insects |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| CNS_RPLC_POS_WT_4 | SA342884 | FL040442 | Wild-type | Genotype |
| CNS_RPLC_POS_WT_4 | SA342884 | FL040442 | CNS | Sample source |
Collection:
| Collection ID: | CO003274 |
| Collection Summary: | Whole larvae and dissected CNS |
| Sample Type: | Larvae, CNS |
Treatment:
| Treatment ID: | TR003290 |
| Treatment Summary: | Untargeted lipidomics analysis was conducted on whole larva and dissected CNS of wild type and ifc-/- mutants at the late-third instar stage. Five replicates were prepared for each set of experiments. For whole larvae, at least 15 larvae of each genotype were used for each replicate. For the dissected CNS, at least 50 wild type and 60 ifc-/- CNS were used per replicate. Immediately following collection or dissection, larvae and the dissected CNS were flash frozen in liquid nitrogen and placed at -80°C. |
Sample Preparation:
| Sampleprep ID: | SP003288 |
| Sampleprep Summary: | Whole larvae and CNS samples were extracted utilizing a tissue homogenizer, employing a solvent mixture of acetonitrile, methanol, and water in a 2:2:1 ratio, using 40 µL per mg of wet weight. The samples were then stored at -20°C overnight. Following centrifugation at 14,000 x g for 10 minutes at 4°C, the supernatant was transferred into an LC/MS vial and subsequently stored at -80°C until LC/MS analysis. |
Combined analysis:
| Analysis ID | AN005187 | AN005188 | AN005189 | AN005190 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | HILIC | HILIC | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish Flex UHPLC Systems | Thermo Vanquish Flex UHPLC Systems | Agilent 1290 Infinity II | Agilent 1290 Infinity II |
| Column | HILICON iHILIC-(P) Classic (100 x 2.1mm,5um) | HILICON iHILIC-(P) Classic (100 x 2.1mm,5um) | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.7um) | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.7um) |
| MS Type | ESI | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap | QTOF | Orbitrap |
| MS instrument name | Thermo Orbitrap ID-X Tribrid | Thermo Orbitrap ID-X Tribrid | Agilent 6545 QTOF | Thermo Orbitrap ID-X Tribrid |
| Ion Mode | NEGATIVE | NEGATIVE | POSITIVE | POSITIVE |
| Units | peak area | peak area | peak area | peak area |
Chromatography:
| Chromatography ID: | CH003924 |
| Instrument Name: | Thermo Vanquish Flex UHPLC Systems |
| Column Name: | HILICON iHILIC-(P) Classic (100 x 2.1mm,5um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0–1 min: 90% B, 1–12 min: 90-35% B, 12–12.5 min: 35-25% B, 12.5–14.5 min: 25% B |
| Flow Rate: | 250 uL/min |
| Solvent A: | water/acetonitrile (95/5); 20 mM ammonium bicarbonate; 0.1% ammonium hydroxide; 2.5 μM medronic acid |
| Solvent B: | acetonitrile/water (95/5) |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH003925 |
| Instrument Name: | Agilent 1290 Infinity II |
| Column Name: | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.7um) |
| Column Temperature: | 60 |
| Flow Gradient: | 0-2min: 30% B, 2-17 min: 30%-75% B, 17-20 min: 75%-85% B, 20-23 min: 85%-100% B, 23-26 min: 100% B |
| Flow Rate: | 250 uL/min |
| Solvent A: | acetronitrile/water (60/40); 10mM ammonium formate; 2.5 uM medronic acid; 0.1% formic acid |
| Solvent B: | 2-propanol/acetonitrile (90/10); 10mM ammonium formate; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004920 |
| Analysis ID: | AN005187 |
| Instrument Name: | Thermo Orbitrap ID-X Tribrid |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Data were collected with the following MS source settings: spray voltage, -2.8 kV; sheath gas, 50; auxiliary gas, 10; sweep gas, 1; ion transfer tube temperature, 300°C; vaporizer temperature, 200°C; mass range, 67 – 1000 Da; resolution, 120,000; maximum injection time, 200 ms; isolation window, 1.6 Da. XCMS and Skyline software were used for data processing |
| Ion Mode: | NEGATIVE |
| MS ID: | MS004921 |
| Analysis ID: | AN005188 |
| Instrument Name: | Thermo Orbitrap ID-X Tribrid |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Data were collected with the following MS source settings: spray voltage, -2.8 kV; sheath gas, 50; auxiliary gas, 10; sweep gas, 1; ion transfer tube temperature, 300°C; vaporizer temperature, 200°C; mass range, 67 – 1000 Da; resolution, 120,000; maximum injection time, 200 ms; isolation window, 1.6 Da. XCMS and Skyline software were used for data processing |
| Ion Mode: | NEGATIVE |
| MS ID: | MS004922 |
| Analysis ID: | AN005189 |
| Instrument Name: | Agilent 6545 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Data were collected with the following MS source settings: gas temp, 250°C; drying gas, 11 l/min; nebulizer, 35 psi; sheath gas temp, 300°C; sheath gas flow 12 l/min; capillary voltage, 3 kV; nozzle voltage, 500 V; isolation window, 1.3 Da. XCMS and Skyline software were used for data processing |
| Ion Mode: | POSITIVE |
| MS ID: | MS004923 |
| Analysis ID: | AN005190 |
| Instrument Name: | Thermo Orbitrap ID-X Tribrid |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Data were collected with the following MS source settings: spray voltage, -3 kV; sheath gas, 50; auxiliary gas, 10; sweep gas, 1; ion transfer tube temperature, 300°C; vaporizer temperature, 350°C; mass range, 100 – 1500 Da; resolution, 120,000; maximum injection time, 150 ms; isolation window, 1.6 Da. XCMS and Skyline software were used for data processing |
| Ion Mode: | POSITIVE |
