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MB Sample ID: SA078306
| Local Sample ID: | Prednisolone-d0-P3 |
| Subject ID: | SU001204 |
| Subject Type: | Mammal |
| Subject Species: | Canis lupus familiaris |
| Taxonomy ID: | 9615 |
| Genotype Strain: | Beagle |
| Age Or Age Range: | 8 to 83 months |
| Weight Or Weight Range: | 12.2 to 18.9 kg |
| Gender: | Male and female |
| Animal Animal Supplier: | In-house breeding |
| Animal Feed: | Standard pellet/kibble maintenance diet |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN001870 | AN001871 | AN001872 | AN001873 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | HILIC | Reversed phase |
| Chromatography system | Agilent 1290 Infinity | Agilent 1290 Infinity | Agilent 1290 Infinity | Agilent 1100 |
| Column | Agilent Zorbax RRHD Eclipse Plus C18 (50 x 2.1mm,1.8um,95 Å) | Agilent Zorbax RRHD Eclipse Plus C18 (50 x 2.1mm,1.8um,95 Å) | Waters Acquity BEH HILIC (100 x 2.1mm,1.7um,130 Å) | Agilent Zorbax Eclipse XDB C18 (150 x 3.0mm,1.8um) |
| MS Type | ESI | ESI | ESI | ESI |
| MS instrument type | Triple quadrupole | Triple quadrupole | Triple quadrupole | Triple quadrupole |
| MS instrument name | Agilent 6460 QQQ | Agilent 6495 QQQ | Agilent 6490 QQQ | ABI Sciex 4000 QTrap |
| Ion Mode | POSITIVE | POSITIVE | POSITIVE | POSITIVE |
| Units | µmol/L | µmol/L | µmol/L | µmol/L |
MS:
| MS ID: | MS001726 |
| Analysis ID: | AN001870 |
| Instrument Name: | Agilent 6460 QQQ |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Phospholipids, cholesteryl esters and diacylglycerols were measured with an Agilent 6460 triple quadrupole mass spectrometer in dynamic MRM mode. The ESI source settings were: polarity: positive, dry gas temperature 300°C, dry gas flow 5L/min, nebulizer pressure: 45 psi, sheath gas temperature: 250°C, sheath gas flow: 11 L/min, capillary voltage: 3500 V, noozle: 500. MRM transitions with collision energies are detailed in the attached protocol. Data were processed with Agilent MassHunter QQQ Quantitative Analysis (version B.08). For plasmalogen PE (PE-P), transitions with the fatty acid as product were used as quantifiers, and those with the head group as qualifiers. Plasmalogen PCs (PC-P), ether PCs (PC-O) and odd-chain fatty acid PCs were distinguished based on retention time (Alshehry et al., Circulation, 2016; Huynh et al, Cell Chem. Biol. 2019). Normalised peak areas were calculated by dividing the peak areas of the analyte with the corresponding class-specific internal standard. Relative abundance was obtained by multiplying the normalised peak areas with the molar concentration of the corresponding internal standard. Lipid species with a median peak area in the PQC samples below 250 or less than 5 times of the highest signal in Blank samples were excluded. Additionally, the coefficient of variation (CV) of the normalised peak area was calculated for each lipid species in the PQC samples of each experimental group. Species with a CV higher than 25% in any of the two groups were excluded from subsequent evaluation. |
| Ion Mode: | POSITIVE |
| Capillary Voltage: | 3500 V |
| Collision Gas: | Nitrogen |
| Dry Gas Flow: | 5 L/min |
| Dry Gas Temp: | 300°C |
| Fragment Voltage: | 135 V |
| Nebulizer: | 45 psi |
| MS ID: | MS001727 |
| Analysis ID: | AN001871 |
| Instrument Name: | Agilent 6495 QQQ |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Sphingolipids were measured with an Agilent 6495A triple quadrupole mass spectrometer in dynamic MRM mode. The ESI source settings were: polarity: positive, dry gas temperature 200°C, dry gas flow 12 L/min, nebulizer pressure: 25 psi, sheath gas temperature: 250°C, sheath gas flow: 12 L/min, capillary voltage: 3500 V, noozle: 500, iFunnel high pressure RF: 80, iFunnel high pressure RF: 40. MRM transitions with collision energies are detailed in the attached protocol. Data were processed with Agilent MassHunter QQQ Quantitative Analysis (version B.08). Transitions of precursor ions with water loss were used as qualifiers. Normalised peak areas were calculated by dividing the peak areas of the analyte with the corresponding class-specific internal standard. Relative abundance was obtained by multiplying the normalised peak areas with the molar concentration of the corresponding internal standard. Lipid species with a median peak area in the PQC samples below 250 or less than 5 times of the highest signal in Blank samples were excluded. Additionally, the coefficient of variation (CV) of the normalised peak area was calculated for each lipid species in the PQC samples of each experimental group. Species with a CV higher than 25% in any of the two groups were excluded from subsequent evaluation. |
| Ion Mode: | POSITIVE |
| Capillary Voltage: | 3500 V |
| Collision Gas: | Nitrogen |
| Dry Gas Flow: | 12 L/min |
| Dry Gas Temp: | 200°C |
| Fragment Voltage: | 135 V |
| Nebulizer: | 25 psi |
| MS ID: | MS001728 |
| Analysis ID: | AN001872 |
| Instrument Name: | Agilent 6490 QQQ |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Derivatized S1P species were measured with an Agilent 6490 triple quadrupole mass spectrometer in MRM mode. The ESI source settings were: polarity: positive, dry gas temperature 200°C, dry gas flow 12 L/min, nebulizer pressure: 25 psi, sheath gas temperature: 400°C, sheath gas flow: 12 L/min, capillary voltage: 3500 V, noozle: 500, iFunnel high pressure RF: 200, iFunnel high pressure RF: 110. MRM transitions with collision energies are detailed in the attached protocol. Data were processed with Agilent MassHunter QQQ Quantitative Analysis (version B.08). The m/z 60 fragments were used as quantifiers, the m/z 103 fragments as qualifiers. Normalised peak areas were calculated by dividing the peak areas of the S1P species with the internal standard. Relative abundance was obtained by multiplying the normalised peak areas with the molar concentration of the corresponding internal standard. S1P species with a median peak area in the PQC samples below 250 or less than 5 times of the highest signal in Blank samples were excluded. Additionally, the coefficient of variation (CV) of the normalised peak area was calculated for each lipid species in the PQC samples of each experimental group. Species with a CV higher than 25% in any of the two groups were excluded from subsequent evaluation. |
| Ion Mode: | POSITIVE |
| Capillary Voltage: | 3500 V |
| Collision Gas: | Nitrogen |
| Dry Gas Flow: | 12 L/min |
| Dry Gas Temp: | 200°C |
| Fragment Voltage: | 135 V |
| Nebulizer: | 25 psi |
| MS ID: | MS001729 |
| Analysis ID: | AN001873 |
| Instrument Name: | ABI Sciex 4000 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Triacylglycerol species were measured with a Sciex 4000 QTRAP mass spectrometer operated in single-ion monitoring (SIM) mode at unit resolution. The ESI source settings were: polarity: positive, electrospray voltage: 5 kV, source temperature: 250°C, drying gas: nitrogen, gas 1 flow: 40 units, gas 2 flow: 30 units and curtain gas flow: 10 units. MRM transitions with collision energies are detailed in the attached protocol. Raw data were processed with Sciex Analyst (Version 1.6.2). Normalised peak areas were calculated by dividing the peak areas of the TG species with the internal standard. Relative abundance was obtained by multiplying the normalised peak areas with the molar concentration of the internal standard. Lipid species with a median peak area in the PQC samples below 250 or less than 5 times of the highest signal in Blank samples were excluded. Additionally, the coefficient of variation (CV) of the normalised peak area was calculated for each lipid species in the PQC samples of each experimental group. Species with a CV higher than 25% in any of the two groups were excluded from subsequent evaluation. |
| Ion Mode: | POSITIVE |
| Ion Spray Voltage: | 5 kV |
| Source Temperature: | 250°C |
