Summary of study ST000456

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 PR000352. The data can be accessed directly via it's Project DOI: 10.21228/M8JK5M This work is supported by NIH grant, U2C- DK119886.

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Study IDST000456
Study TitleMetabolomic changes during active immunization with anti-METH vaccines.
Study SummaryA targeted metabolomics analysis was performed on serum samples from rats immunized with a methamphetamine-like hapten covalently bound to keyhole limpet hemocyanin (KLH) monomers, and controls. The subjects were approximately 300 g adult male Sprague Dawley rats. RTI International RCMRC received pre-immune serum from RI 11-03A (n=12) which is a matched control serum that was collected before starting immunizations, and week 17 bleeds from RI 11-03A (n=12), RI 11-03B (n=8), and RI 11-03C (n=12). The A-C nomenclature denotes A) complete antigen with adjuvant, B) complete antigen without adjuvant and C) only KLH carrier protein and adjuvant without a METH-like hapten. The rats’ serum samples were extracted and prepared for a combined flow injection (FIA) and LC-MS/MS assay (Biocrates AbsoluteIDQ® p180 kit). The Biocrates AbsoluteIDQ® p180 kit (Biocrates Life Sciences AG, Innsbruck, Austria) can quantitatively measure ~188 biologically relevant metabolites from five analyte groups (acylcarnitines, amino acids, biogenic amines, glycerophospho- and sphingolipids, and hexoses) on a triple quadrupole mass spectrometer.
Institute
University of North Carolina
DepartmentDiscovery-Sciences-Technology (DST)
LaboratorySumner Lab
Last NameSumner
First NameSusan
AddressEastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition Research Institute, 500 Laureate Way, Kannapolis, NC, 28081
Emailsusan_sumner @unc.edu
Phone704-250-5066
Submit Date2016-08-31
Num Groups4
Total Subjects44
Raw Data AvailableYes
Raw Data File Type(s).wiff
Analysis Type DetailLC-MS
Release Date2017-10-03
Release Version1
Susan Sumner Susan Sumner
https://dx.doi.org/10.21228/M8JK5M
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000352
Project DOI:doi: 10.21228/M8JK5M
Project Title:Metabolomic changes during active immunization with anti-METH vaccines.
Project Summary:Metabolomics studies will be conducted on serum samples from rats immunized with a methamphetamine conjugate vaccine. The Subjects were approximately 300 g adult male Sprague Dawley rats. The following samples were sent to RTI International RCMRC: Pre-immune serum from RI 11-03A (n=12) which is a matched control serum that was collected before starting immunizations, and week 17 bleeds from RI 11-03A (n=12), RI 11-03B (n=8), and RI 11-03C (n=12). The A-C nomenclature denotes A) complete antigen with adjuvant, B) complete antigen without adjuvant and C) only KLH carrier protein and adjuvant without a METH-like hapten.
Institute:University of Arkansas
Department:Department of Pharmacology and Toxicology
Last Name:Owens
First Name:S. Michael
Address:4301 W. Markham St., #611 Little Rock, AR 72205 USA
Email:mowens@uams.edu
Phone:501-686-5487

Subject:

Subject ID:SU000477
Subject Type:Animal
Subject Species:Rattus norvegicus
Taxonomy ID:10116
Species Group:Mammal

Factors:

Subject type: Animal; Subject species: Rattus norvegicus (Factor headings shown in green)

mb_sample_id local_sample_id MCV Adjuvant Sample Collection
SA023136MO_38Act KLH RIBI Immunized (17 wks)
SA023137MO_44Act KLH RIBI Immunized (17 wks)
SA023138MO_41Act KLH RIBI Immunized (17 wks)
SA023139MO_42Act KLH RIBI Immunized (17 wks)
SA023140MO_34Act KLH RIBI Immunized (17 wks)
SA023141MO_43Act KLH RIBI Immunized (17 wks)
SA023142MO_33Act KLH RIBI Immunized (17 wks)
SA023143MO_39Act KLH RIBI Immunized (17 wks)
SA023144MO_40Act KLH RIBI Immunized (17 wks)
SA023145MO_36Act KLH RIBI Immunized (17 wks)
SA023146MO_35Act KLH RIBI Immunized (17 wks)
SA023147MO_37Act KLH RIBI Immunized (17 wks)
SA023112P17A_2- - -
SA023113PBS_03- - -
SA023114p180 Cal7- - -
SA023115p180 Cal6- - -
SA023116p180-MetaDis QC1- - -
SA023117P0_1- - -
SA023118P17C_1- - -
SA023119P17B_1- - -
SA023120p180 Cal4- - -
SA023121p180 Cal3- - -
SA023122TP_1- - -
SA023123p180 Cal5- - -
SA023124p180 Cal2- - -
SA023125p180 Cal1- - -
SA023126p180-MetaDis QC3- - -
SA023127P17A_1- - -
SA023128PBS_01- - -
SA023129p180-MetaDis QC2- - -
SA023130TP_2- - -
SA023131P17B_2- - -
SA023132TP_3- - -
SA023133P17C_2- - -
SA023134PBS_02- - -
SA023135P0_2- - -
SA023148MO_28SSM09 None Immunized (17 wks)
SA023149MO_26SSM09 None Immunized (17 wks)
SA023150MO_32SSM09 None Immunized (17 wks)
SA023151MO_29SSM09 None Immunized (17 wks)
SA023152MO_27SSM09 None Immunized (17 wks)
SA023153MO_25SSM09 None Immunized (17 wks)
SA023154MO_31SSM09 None Immunized (17 wks)
SA023155MO_30SSM09 None Immunized (17 wks)
SA023156MO_21SSM09 RIBI Immunized (17 wks)
SA023157MO_17SSM09 RIBI Immunized (17 wks)
SA023158MO_20SSM09 RIBI Immunized (17 wks)
SA023159MO_16SSM09 RIBI Immunized (17 wks)
SA023160MO_14SSM09 RIBI Immunized (17 wks)
SA023161MO_19SSM09 RIBI Immunized (17 wks)
SA023162MO_24SSM09 RIBI Immunized (17 wks)
SA023163MO_22SSM09 RIBI Immunized (17 wks)
SA023164MO_18SSM09 RIBI Immunized (17 wks)
SA023165MO_13SSM09 RIBI Immunized (17 wks)
SA023166MO_23SSM09 RIBI Immunized (17 wks)
SA023167MO_15SSM09 RIBI Immunized (17 wks)
SA023168MO_1SSM09 RIBI Pre-Immune (0 wks)
SA023169MO_10SSM09 RIBI Pre-Immune (0 wks)
SA023170MO_8SSM09 RIBI Pre-Immune (0 wks)
SA023171MO_12SSM09 RIBI Pre-Immune (0 wks)
SA023172MO_11SSM09 RIBI Pre-Immune (0 wks)
SA023173MO_9SSM09 RIBI Pre-Immune (0 wks)
SA023174MO_5SSM09 RIBI Pre-Immune (0 wks)
SA023175MO_2SSM09 RIBI Pre-Immune (0 wks)
SA023176MO_3SSM09 RIBI Pre-Immune (0 wks)
SA023177MO_7SSM09 RIBI Pre-Immune (0 wks)
SA023178MO_4SSM09 RIBI Pre-Immune (0 wks)
SA023179MO_6SSM09 RIBI Pre-Immune (0 wks)
Showing results 1 to 68 of 68

Collection:

Collection ID:CO000471
Collection Summary:Serum Samples from Rats Immunized with a Methamphetamine Conjugate Vaccine
Sample Type:Blood

Treatment:

Treatment ID:TR000491
Treatment Summary:Treatment A - complete antigen with adjuvant

Sample Preparation:

Sampleprep ID:SP000484
Sampleprep Summary:Study Phenotypic Pools Thawed serum samples were vortexed and centrifuged at 5,000 rcf for 4 minutes. The following phenotypic pools were created by combining 10 µL from each phenotypic sample into a 2 mL LoBind eppendorf tube: 1. Pre-immune serum from RI 11-03A (n=12) which is a matched control serum that was collected before starting immunizations. The final volume of this phenotypic pool was 120 µL which was vortexed for 30 seconds. Two phenotypic pools (0wk-03A_Pool1 and 0wk-03A_Pool2) of 25 µL each were created. 2. Week 17 bleeds from RI 11-03A (n=12). The final volume of this phenotypic pool was 120 µL which was vortexed for 30 seconds. Two phenotypic pools (17wk-03A_Pool1 and 17wk-03A_Pool1) of 25 µL each were created. 3. RI 11-03B (n=8).The final volume of this phenotypic pool was 80 µL which was vortexed for 30 seconds. Two phenotypic pools (17wk-03B_Pool1 and 17wk-03B_Pool2) of 25 µL each were created. 4. and RI 11-03C (n=12). The final volume of this phenotypic pool was 120 µL which was vortexed for 30 seconds. Two phenotypic pools (17wk-03C_Pool1and 17wk-03C_Pool2) of 25 µL each were created. Whole Study Pooled QC Whole study pooled QC samples were prepared by combining 25 µL of each phenotypic pool sample created above into a 2 mL LoBind eppendorf tube. This QC pooled sample was then vortexed for 30 sec. Then, three whole study pooled QC samples of 25 µL each were aliquoted into 2 mL LoBind eppendorf tubes. Biocrates Plate Preparation: All samples (Phenotypic pools, whole study pooled QC and study samples) were vortexed for 10 minutes at 5,000 rpm. Samples were then centrifuged at 16,000 rcf for 4 minutes and at 4 °C. A Biocrates p180 kit was prepared following the AbsoluteIDQ™ p180 Kit metabolomics procedure. Briefly, an internal standard mix was added to 95 of the 96 wells. Next, zero samples, QC standards and calibration standards were added to their corresponding wells. The study samples, phenotypic and whole study pooled QC Samples (20 µL) were then added to the appropriate wells and dried for 30 minutes under nitrogen flow. The plate was derivatized using a 5% phenylisothiocyanate (PITC) solution in (1:1:1) ethanol:pyridine:water (v/v/v) and, then, incubated for 20 minutes at room temperature followed by a drying step under nitrogen flow. An extraction solvent (5 mM ammonium acetate in methanol) was added to all wells. The plate was then shaken and centrifuged. After centrifugation, 150 µL was removed and transferred to a second 96-well plate (LCMS plate). This second plate was diluted with 150 µL of HPLC grade water for a subsequent LCMS (MRM analysis) for measuring amino acids and biogenic amines. All wells in the original plate were diluted with 400 µL of flow injection analysis (FIA) Running Solvent for a FIA-MS (MRM analysis) for measuring lipids, acylcarnitines, and hexose.

Combined analysis:

Analysis ID AN000713 AN000714 AN000715
Analysis type MS MS MS
Chromatography type Reversed phase Reversed phase Reversed phase
Chromatography system Agilent 1100 Agilent 1100 Agilent 1100
Column Agilent Eclipse XDB-C18 (100 x 3.0mm,3.5um) Agilent Eclipse XDB-C18 (100 x 3.0mm,3.5um) Agilent Eclipse XDB-C18 (100 x 3.0mm,3.5um)
MS Type ESI ESI ESI
MS instrument type Triple quadrupole Triple quadrupole Triple quadrupole
MS instrument name ABI Sciex API 4000 QTrap ABI Sciex API 4000 QTrap ABI Sciex API 4000 QTrap
Ion Mode POSITIVE POSITIVE NEGATIVE
Units Intensity Intensity Intensity

Chromatography:

Chromatography ID:CH000514
Instrument Name:Agilent 1100
Column Name:Agilent Eclipse XDB-C18 (100 x 3.0mm,3.5um)
Column Temperature:50 degrees C
Flow Rate:0.5mL/min
Chromatography Type:Reversed phase

MS:

MS ID:MS000633
Analysis ID:AN000713
Instrument Name:ABI Sciex API 4000 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS000634
Analysis ID:AN000714
Instrument Name:ABI Sciex API 4000 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS000635
Analysis ID:AN000715
Instrument Name:ABI Sciex API 4000 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:NEGATIVE
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