Summary of study ST001140

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000761. The data can be accessed directly via it's Project DOI: 10.21228/M89Q32 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001140
Study TitleChanges in the Canine Plasma Lipidome after Short- and Long-Term Excess Glucocorticoid Exposure
Study SummaryGlucocorticoids (GCs) are widely used in veterinary and human medicine. Chromic endogenous or iatrogenic GC overexposure impairs metabolic function and can result in diverse side-effects, including Cushing’s syndrome. This study examines the effects of experimentally induced short-term and long-term GC excess (induced by prednisolone and tetracosactide, respectively) on the plasma lipidome of Beale dogs. Both, long- and short-term GC resulted in significant changes of the plasma lipidome.
Institute
National University of Singapore;University of Zurich
DepartmentSingapore Lipidomics Incubator (SLING);Vetsuisse Faculty, University of Zurich
LaboratorySingapore Lipidomics Incubator (SLING), National University of Singapore
Last NameBurla
First NameBo
Address28 Medical Drive, Singapore 117456, Singapore
Emailbo.burla@nus.edu.sg
Phone+6565166683
Submit Date2019-01-19
Num Groups2
Total Subjects14
Num Males9
Num Females5
Raw Data AvailableYes
Raw Data File Type(s).d
Analysis Type DetailLC-MS
Release Date2019-03-06
Release Version1
Bo Burla Bo Burla
https://dx.doi.org/10.21228/M89Q32
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Combined analysis:

Analysis ID AN001870 AN001871 AN001872 AN001873
Analysis type MS MS MS MS
Chromatography type Reversed phase Reversed phase HILIC Normal phase
Chromatography system Agilent 1290 Infinity Agilent 1290 Infinity Agilent 1290 Infinity Agilent 1100
Column Agilent Zorbax RRHD Eclipse Plus C18 (50 x 2.1 mm, 1.8 µm, 95 Å) Agilent Zorbax RRHD Eclipse Plus C18 (50 x 2.1 mm, 1.8 µm, 95 Å) Waters Acquity BEH HILIC (100 x 2.1mm,1.7 µm, 130 Å) Agilent Zorbax Eclipse XDB-C18 Silica (150 x 3mm, 1.8 μm, 80 Å)
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
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