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MB Sample ID: SA239047
| Local Sample ID: | Z22 |
| Subject ID: | SU002487 |
| Subject Type: | Fish |
| Subject Species: | Danio rerio |
| Taxonomy ID: | 7955 |
| Genotype Strain: | wildtype and tango2bwh211 |
| Age Or Age Range: | 4 weeks |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN003905 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters Acquity H-Class |
| Column | Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive HF-X Orbitrap |
| Ion Mode | POSITIVE |
| Units | peak area |
MS:
| MS ID: | MS003644 |
| Analysis ID: | AN003905 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Analysis was performed using a Thermo Q Exactive Plus coupled to a Waters Acquity H-Class LC. A 100 mm x 2.1 mm, 2.1 µm Waters BEH C18 column was used for separations. The following mobile phases were used: A- 60/40 ACN/H20 B- 90/10 IPA/ACN; both mobile phases contained 10 mM Ammonium Formate and 0.1% Formic Acid. A flow rate of 0.2 mL/minutes was used. Starting composition was 32% B, which increased to 40% B at 1 minute (held until 1.5 minutes) then 45% B at 4 minutes. This was increased to 50% B at 5 minutes, 60% B at 8 minutes, 70% B at 11 minutes, and 80% B at 14 minutes (held until 16 minutes). At 16 minutes the composition switched back to starting conditions (32% B) and was held for 4 minutes to re-equilibrate the column. Samples were analyzed in positive/negative switching ionization mode with top 5 data dependent fragmentation. Raw data were analyzed by LipidSearch 4.2. Lipids were identified by MS2 fragmentation (mass error of precursor=5 ppm, mass error of product=8 ppm). The identifications were generated individually for each sample and then aligned by grouping the samples (OxPAPC=C, HF=S1, Con=S2). Normalization was performed using EquiSplash from Avanti. Samples were normalized and biological replicates were averaged. P-value and fold change were calculated as instructed as previously described (Aguilan, Kulej, & Sidoli, 2020). P-value was set to 0.05. |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 300 |
| Capillary Voltage: | 3.50 KV |
| Collision Energy: | 25, 35, 45 V stepped collision energy |
| Fragmentation Method: | HCD |
| Ion Source Temperature: | 400 |
| Ion Spray Voltage: | 3.50 KV |
| Ionization: | HESI |
| Mass Accuracy: | 5 ppm |
| Reagent Gas: | Nitrogen |
| Source Temperature: | 400 |
| Spray Voltage: | 3.5 KV |
