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.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

Perform statistical analysis  |  Show all samples  |  Show named metabolites  |  Download named metabolite data  
Download mwTab file (text)   |  Download mwTab file(JSON)   |  Download data files (Contains raw data)
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

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR000761
Project DOI:doi: 10.21228/M89Q32
Project Title:Changes in the Canine Plasma Lipidome after Short- and Long-Term Excess Glucocorticoid Exposure
Project Summary:Glucocorticoids (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
Department:Singapore Lipidomics Incubator (SLING);Vetsuisse Faculty, University of Zurich
Laboratory:Singapore Lipidomics Incubator (SLING)
Last Name:Burla
First Name:Bo
Address:28 Medical Drive, Singapore 117456, Singapore
Email:bo.burla@nus.edu.sg
Phone:+6565166683

Subject:

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
Species Group:Mammals

Factors:

Subject type: Mammal; Subject species: Canis lupus familiaris (Factor headings shown in green)

mb_sample_id local_sample_id TreatmentGroup TreatmentDuration SamplingTimePoint
SA078299Prednisolone-d0-P5Prednisolone 0d before
SA078300Prednisolone-d0-P6Prednisolone 0d before
SA078301Prednisolone-d0-P1Prednisolone 0d before
SA078302Prednisolone-d0-P4Prednisolone 0d before
SA078303Prednisolone-d0-P7Prednisolone 0d before
SA078304Prednisolone-d0-P8Prednisolone 0d before
SA078305Prednisolone-d0-P2Prednisolone 0d before
SA078306Prednisolone-d0-P3Prednisolone 0d before
SA078307Prednisolone-d4-P4Prednisolone 4d after
SA078308Prednisolone-d4-P1Prednisolone 4d after
SA078309Prednisolone-d4-P7Prednisolone 4d after
SA078310Prednisolone-d4-P8Prednisolone 4d after
SA078311Prednisolone-d4-P2Prednisolone 4d after
SA078312Prednisolone-d4-P3Prednisolone 4d after
SA078313Prednisolone-d4-P6Prednisolone 4d after
SA078314Prednisolone-d4-P5Prednisolone 4d after
SA078315Tetracosactide-w00-T6Tetracosactide 00w before
SA078316Tetracosactide-w00-T5Tetracosactide 00w before
SA078317Tetracosactide-w00-T4Tetracosactide 00w before
SA078318Tetracosactide-w00-T2Tetracosactide 00w before
SA078319Tetracosactide-w00-T1Tetracosactide 00w before
SA078320Tetracosactide-w00-T3Tetracosactide 00w before
SA078321Tetracosactide-w25-T5Tetracosactide 25w after
SA078322Tetracosactide-w25-T6Tetracosactide 25w after
SA078323Tetracosactide-w25-T1Tetracosactide 25w after
SA078324Tetracosactide-w25-T2Tetracosactide 25w after
SA078325Tetracosactide-w25-T3Tetracosactide 25w after
SA078326Tetracosactide-w25-T4Tetracosactide 25w after
Showing results 1 to 28 of 28

Collection:

Collection ID:CO001198
Collection Summary:Blood was collected from the jugular vein after overnight fasting right before start and at the end of the treatments (12 h after the last prednisolone administration and 25 weeks after start of tetracosactide infusion, respectively). Plasma was prepared lithium heparin anticoagulated blood (Vacuette Greiner Bio-One, Germany) that centrifuged immediately after collection (1,862 g, 10 min, 4°C). Plasma samples were stored at −80°C until lipidomic analysis.
Collection Protocol Comments:Blood collected after overnight fasting
Sample Type:Blood (plasma)
Collection Method:Venepuncture
Collection Location:Jugular vein
Collection Frequency:Before and after treatment
Storage Conditions:-80℃
Collection Vials:VACUETTE® lithium heparin tubes (Greiner Bio-One, Germany)
Collection Tube Temp:Room temperature
Additives:Lithium heparin

Treatment:

Treatment ID:TR001219
Treatment Summary:In the short-term prednisolone group, dogs were treated with 50 mg prednisolone (Streuli Pharma AG, Switzerland) orally twice daily for 3 consecutive days. For the long-term tetracosactide treatment, ALZET osmotic pumps (Durect Corporation, USA) subcutaneously delivering tetracosactide (synthetic ACTH; Bachem AG, Switzerland) were implanted into the dorsolateral neck. New pumps were implanted every 4 weeks. Total infusion time was 25 weeks, with a starting dose of 1.3–1.95 µg/kg/d tetracosactide, which was gradually increased to a final dose of 6–10 µg/kg/d.

Sample Preparation:

Sampleprep ID:SP001212
Sampleprep Summary:Plasma lipids were extracted using a single-phase butanol/methanol extraction method (Alshehry et al, Metabolites 2015 with modifications). After thawing on ice, a 10 µL aliquot of each plasma sample was transferred into a 2 mL polypropylene tube (Eppendorf, Germany). Subsequently, 1 µL BHT (2,6-di-tert-butyl-4-methylphenol, 10 mmol/L in ethanol) and 90 µL of 1-butanol:methanol (1:1, v/v) containing internal standards was added to each sample. Following internal standards were added at 50 ng/mL final concentration in the extraction solvent: ceramide d18:1/17:0 (Avanti 860517P), Glucosylceramide (Matreya 1533), Lactosylceramide d18:1/16:0 D3 (Matreya 1534), LPC 20:0 (Avanti 855777P), LPE 14:0 (Avanti 856735P), PI 12:0/13:0 (Avanti LM-1500), PE 14:0/14:0 (Avanti 850745P), PS 14:0/14:0 (Avanti 840033P), SM d18:1/12:0 (Avanti 860583P), and at 100 ng/mL: DG 12:0_12:0 (Avanti 800812P), PC 14:0/14:0 (Avanti 850345P), TAG 16:0_16:0_16:0 d5 (CDN Isotopes D5815). Samples were then vortexed (30 sec) and sonicated in an ultrasound water bath for 30 min (20°C). After centrifugation (14,000 g, 10 min, 4°C), 90 µL of supernatant were transferred into 1.5 mL polypropylene tubes (Sarstedt, Germany) and dried under a nitrogen stream at 37°C. Dried extracts were stored at −80°C until LC-MS analysis. Dried samples were reconstituted with 90 µL 1-butanol:methanol (1:1, v/v) and sonication in an ultrasound water for 10 min. After centrifugation for 10 min at 20,800 g (4°C), supernatants (80 µL) were transferred into autosampler vials with glass inserts (Agilent Technologies, USA). For the analysis of sphingosine-1-phosphate (S1P), lipid extracts were derivatized according to the method described by Narayanaswamy et al. (Anal. Chem., 2014). To 50 µL lipid extract (see above), 50 µL 13C2D2–S1P d18:1 internal standard solution (Toronto Research Chemicals, Canada; 20 ng/mL in 1-butanol:methanol 1:1 [v/v]) was added. Derivatization was performed by adding 20 µL TMS-diazomethane (2 mol/L in hexanes; Acros Organics, Thermo Fisher Scientific, USA) and subsequent incubation for 20 min at 25°C and 700 rpm (Thermomixer, Eppendorf, Germany). To stop the reaction and inactivate TMS, 1 µL acetic acid 100% (glacial) was added. After centrifugation for 10 min at 20,800 g (7°C), supernatants were transferred into autosampler vials for LC-MS analysis. Process Quality Control (PQC) samples were generated by pooling equal volumes of plasma samples within an experimental group. For Blank samples, plasma was omitted. PQC and Blank were processed together with the study plasma samples with the same procedures.
Extraction Method:Liquid–liquid extraction using butanol:methanol
Extract Storage:-80℃

Chromatography:

Chromatography ID:CH001351
Instrument Name:Agilent 1290 Infinity
Column Name:Agilent Zorbax RRHD Eclipse Plus C18 (50 x 2.1mm,1.8um,95 Å)
Column Temperature:40
Flow Gradient:20% B: 2 min, 20% - 60% B: 2 - 7 min, 60% - 100% B: 7 - 9 min, 20% B: 9 - 10.8 min
Flow Rate:0.4 mL/min
Sample Injection:2 µL
Solvent A:60% acetonitrile/40% water; 10 mM ammonium formate
Solvent B:90% isopropanol/10% acetonitrile; 10 mM ammonium formate
Chromatography Type:Reversed phase
  
Chromatography ID:CH001352
Instrument Name:Agilent 1290 Infinity
Column Name:Agilent Zorbax RRHD Eclipse Plus C18 (50 x 2.1mm,1.8um,95 Å)
Column Temperature:40
Flow Gradient:20% B: 2 min, 20% - 60% B: 2 - 7 min, 60% - 100% B: 7 - 9 min, 20% B: 9 - 10.8 min
Flow Rate:0.4 mL/min
Sample Injection:1 µL
Solvent A:60% acetonitrile/40% water; 10 mM ammonium formate
Solvent B:90% isopropanol/10% acetonitrile; 10 mM ammonium formate
Chromatography Type:Reversed phase
  
Chromatography ID:CH001353
Instrument Name:Agilent 1290 Infinity
Column Name:Waters Acquity BEH HILIC (100 x 2.1mm,1.7um,130 Å)
Column Temperature:60
Flow Gradient:99.9% B: 0 - 5 min; 40% B: 5 - 5.5 min; 10% B: 5.5 - 6.6 min and 99.9% B: 6.6 - 9.6 min (total run time 9.6 min)
Flow Rate:0.4 mL/min
Sample Injection:2 µL
Solvent A:50% water/50% acetonitrile; 25 mM ammonium formate, pH 4.6
Solvent B:95% acetonitrile/5% water; 25 mM ammonium formate, pH 4.6
Chromatography Type:HILIC
  
Chromatography ID:CH001354
Instrument Name:Agilent 1100
Column Name:Agilent Zorbax Eclipse XDB C18 (150 x 3.0mm,1.8um)
Column Temperature:25
Flow Gradient:Isocratic (25 min)
Flow Rate:0.128 mL/min
Sample Injection:10 µL
Solvent A:50% chloroform/50% methanol; 2 mM ammonium acetate
Chromatography Type:Reversed phase

Analysis:

Analysis ID:AN001870
Laboratory Name:Singapore Lipidomics Incubator
Analysis Type:MS
Software Version:. MassHunter Data Acquisition B.06
Data Format:.d
Chromatography ID:CH001351
Num Factors:4
Num Metabolites:157
Units:µmol/L
  
Analysis ID:AN001871
Laboratory Name:Singapore Lipidomics Incubator
Analysis Type:MS
Software Version:. MassHunter Data Acquisition B.06
Data Format:.d
Chromatography ID:CH001352
Num Factors:4
Num Metabolites:66
Units:µmol/L
  
Analysis ID:AN001872
Laboratory Name:Singapore Lipidomics Incubator
Analysis Type:MS
Software Version:. MassHunter Data Acquisition B.06
Data Format:.d
Chromatography ID:CH001353
Num Factors:4
Num Metabolites:6
Units:µmol/L
  
Analysis ID:AN001873
Laboratory Name:Singapore Lipidomics Incubator
Analysis Type:MS
Software Version:Analyst 1.6.2
Data Format:.wiff
Chromatography ID:CH001354
Num Factors:4
Num Metabolites:33
Units:µmol/L
  logo