Summary of study ST000608

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 PR000445. The data can be accessed directly via it's Project DOI: 10.21228/M8J60X This work is supported by NIH grant, U2C- DK119886.

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Study IDST000608
Study TitleComparing identified and statistically significant lipids and polar metabolites in 15-year old serum and dried blood spot samples for longitudinal studies
Study SummaryThe use of dried blood spots (DBS) has many advantages over traditional plasma and serum samples such as the smaller blood volume required, storage at room temperature, and ability to sample in remote locations. However, understanding the robustness of different analytes in DBS samples is essential, especially in older samples collected for longitudinal studies. Here we analyzed the stability of polar metabolites and lipids in DBS samples collected in 2000-2001 and stored at room temperature. The identified and statistically significant molecules were then compared to matched serum samples stored at –80°C to determine if the DBS samples could be effectively used in a longitudinal study following metabolic disease. A total of 400 polar metabolites and lipids were identified in the serum and DBS samples using gas chromatograph/mass spectrometry (GC/MS), liquid chromatography (LC)/MS, and LC/ion mobility spectrometry-MS (LC/IMS-MS). The identified polar metabolites overlapped well between the sample types, though only one statistically significant metabolite was conserved in a case-control study of older diabetic males with low amounts of high-density lipoproteins and high body mass indices, triacylglycerides and glucose levels when compared to non-diabetic patients with normal levels, indicating that degradation in the DBS samples affects polar metabolite quantitation. Differences in the lipid identifications indicated that some oxidation occurs in the DBS samples. However, 36 statistically significant lipids correlated in both sample types. The difference in the number of statistically significant polar metabolites and lipids indicated that the lipids did not degrade to as great of a degree as the polar metabolites in the DBS samples and lipid quantitation was still possible.
Institute
Pacific Northwest National Laboratory
Last NameBaker
First NameErin
Address902 Battelle Boulevard, Richland, WA, 99354, USA
Emailerin.baker@pnnl.gov
Phone509-371-6219
Submit Date2017-05-13
Raw Data AvailableYes
Raw Data File Type(s).raw
Analysis Type DetailGC/LC-MS
Release Date2017-07-10
Release Version1
Erin Baker Erin Baker
https://dx.doi.org/10.21228/M8J60X
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000445
Project DOI:doi: 10.21228/M8J60X
Project Title:Comparing identified and statistically significant lipids and polar metabolites in 15-year old serum and dried blood spot samples for longitudinal studies
Project Summary:The use of dried blood spots (DBS) has many advantages over traditional plasma and serum samples such as the smaller blood volume required, storage at room temperature, and ability to sample in remote locations. However, understanding the robustness of different analytes in DBS samples is essential, especially in older samples collected for longitudinal studies. Here we analyzed the stability of polar metabolites and lipids in DBS samples collected in 2000-2001 and stored at room temperature. The identified and statistically significant molecules were then compared to matched serum samples stored at –80°C to determine if the DBS samples could be effectively used in a longitudinal study following metabolic disease. A total of 400 polar metabolites and lipids were identified in the serum and DBS samples using gas chromatograph/mass spectrometry (GC/MS), liquid chromatography (LC)/MS, and LC/ion mobility spectrometry-MS (LC/IMS-MS). The identified polar metabolites overlapped well between the sample types, though only one statistically significant metabolite was conserved in a case-control study of older diabetic males with low amounts of high-density lipoproteins and high body mass indices, triacylglycerides and glucose levels when compared to non-diabetic patients with normal levels, indicating that degradation in the DBS samples affects polar metabolite quantitation. Differences in the lipid identifications indicated that some oxidation occurs in the DBS samples. However, 36 statistically significant lipids correlated in both sample types. The difference in the number of statistically significant polar metabolites and lipids indicated that the lipids did not degrade to as great of a degree as the polar metabolites in the DBS samples and lipid quantitation was still possible.
Institute:Pacific Northwest National Laboratory
Last Name:Baker
First Name:Erin
Address:902 Battelle Boulevard, Richland, WA, 99354, USA
Email:erin.baker@pnnl.gov
Phone:(509) 371-6219
Funding Source:Portions of this research were supported by grants from the National Institute of Environmental Health Sciences of the NIH (R01 ES022190), NIH Eunice Kennedy Shriver National Institute of Child Health and Human Development (R21 HD084788), National Institute of General Medical Sciences (P41 GM103493), and the Laboratory Directed Research and Development Program at Pacific Northwest National Laboratory. This research utilized capabilities developed by the Pan-omics program (funded by the U.S. Department of Energy Office of Biological and Environmental Research Genome Sciences Program) and by the National Institute of Allergy and Infectious Diseases under grant U19 AI106772. The Osteoporotic Fractures in Men (MrOS) Study in the US was supported by the National Institute on Aging (NIA), the National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS), the National Center for Advancing Translational Sciences (NCATS), and NIH Roadmap for Medical Research under the following grant numbers: U01 AG027810, U01 AG042124, U01 AG042139, U01 AG042140, U01 AG042143, U01 AG042145, U01 AG042168, U01 AR066160, and UL1 TR000128. This work was performed in the W. R. Wiley Environmental Molecular Sciences Laboratory (EMSL), a DOE national scientific user facility at the Pacific Northwest National Laboratory (PNNL). PNNL is operated by Battelle for the DOE under contract DE-AC05-76RL0 1830.
Publications:Comparing identified and statistically significant lipids and polar metabolites in 15-year old serum and dried blood spot samples for longitudinal studies

Subject:

Subject ID:SU000631
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Human

Factors:

Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Sample_Type Factor
SA033526OHSUblotter_PO6976_1blotter case
SA033527OHSUblotter_PO6927_3blotter case
SA033528OHSUblotter_PO6927_2blotter case
SA033529OHSUblotter_PO6976_2blotter case
SA033530OHSUblotter_PO6976_3blotter case
SA033531OHSUblotter_PO7324_3blotter case
SA033532OHSUblotter_PO7324_1blotter case
SA033533OHSUblotter_PO6927_1blotter case
SA033534OHSUblotter_PO7324_2blotter case
SA033535OHSUblotter_PO6716_2blotter case
SA033536OHSUblotter_PO6716_3blotter case
SA033537OHSUblotter_PO6764_2blotter case
SA033538OHSUblotter_PO6764_3blotter case
SA033539OHSUblotter_PO6764_1blotter case
SA033540OHSUblotter_PO6716_1blotter case
SA033541OHSUblotter_PO6573_3blotter control
SA033542OHSUblotter_PO7355_2blotter control
SA033543OHSUblotter_PO6573_2blotter control
SA033544OHSUblotter_PO6882_2blotter control
SA033545OHSUblotter_PO7355_3blotter control
SA033546OHSUblotter_PO7117_3blotter control
SA033547OHSUblotter_PO7355_1blotter control
SA033548OHSUblotter_PO6573_1blotter control
SA033549OHSUblotter_PO6935_1blotter control
SA033550OHSUblotter_PO6882_1blotter control
SA033551OHSUblotter_PO7117_2blotter control
SA033552OHSUblotter_PO6935_2blotter control
SA033553OHSUblotter_PO6882_3blotter control
SA033554OHSUblotter_PO6935_3blotter control
SA033555OHSUblotter_PO7117_1blotter control
SA033556OHSUserum_PO6976_3serum case
SA033557OHSUserum_PO6927_1serum case
SA033558OHSUserum_PO6927_3serum case
SA033559OHSUserum_PO6927_2serum case
SA033560OHSUserum_PO7324_1serum case
SA033561OHSUserum_PO7324_3serum case
SA033562OHSUserum_PO6976_2serum case
SA033563OHSUserum_PO6976_1serum case
SA033564OHSUserum_PO7324_2serum case
SA033565OHSUserum_PO6764_2serum case
SA033566OHSUserum_PO6716_2serum case
SA033567OHSUserum_PO6716_1serum case
SA033568OHSUserum_PO6764_1serum case
SA033569OHSUserum_PO6716_3serum case
SA033570OHSUserum_PO6764_3serum case
SA033571OHSUserum_PO7117_2serum control
SA033572OHSUserum_PO7355_3serum control
SA033573OHSUserum_PO7117_3serum control
SA033574OHSUserum_PO7355_1serum control
SA033575OHSUserum_PO7355_2serum control
SA033576OHSUserum_PO6573_1serum control
SA033577OHSUserum_PO6882_1serum control
SA033578OHSUserum_PO6573_3serum control
SA033579OHSUserum_PO6573_2serum control
SA033580OHSUserum_PO6882_2serum control
SA033581OHSUserum_PO6882_3serum control
SA033582OHSUserum_PO6935_3serum control
SA033583OHSUserum_PO6935_2serum control
SA033584OHSUserum_PO6935_1serum control
SA033585OHSUserum_PO7117_1serum control
Showing results 1 to 60 of 60

Collection:

Collection ID:CO000625
Collection Summary:Matched DBS and serum samples were simultaneously collected in 2000–2001 from 10 overnight fasted male participants having an average age of 75.5 ± 6.15 years. The institutional review boards at the participating institutions approved the protocol and the study participants provided informed consent. Case participants included 5 diabetic men with high BMIs, TGs and glucose levels, and low amounts of HDL. The 5 control participants had normal metabolic profiles (normal BMIs, TGs, HDL and glucose levels).
Sample Type:Blood
Storage Conditions:The DBS were stored at room temperature in the dark until analysis, while the serum was stored at –80°C.

Treatment:

Treatment ID:TR000645
Treatment Summary:Samples collected in 2000–2001 were stored until June 2015.

Sample Preparation:

Sampleprep ID:SP000638
Sampleprep Summary:For the serum samples, 10 mL of whole blood was drawn and centrifuged so the serum could be extracted. For the DBS samples, 500 μL of blood was collected and 125 μL was blotted in four areas on a Whatman FTA Classic Card. Polar metabolites and lipid extracts were derived from the same sample type (DBS or serum) using a modified Folch extraction in June 2015 and analyzed with MS shortly after. The DBS punch from each patient was transferred into to a 2.0 mL tube where 50 μL of water and then 1200 μL of –20°C 2:1 chloroform/methanol were added. Each sample was vortexed for 30 s then transferred into a shaker at 22°C for 60 min at 600 rpm. The samples were vortexed again for 30 s and then 250 μL of water was added to induce a phase separation. The sample was gently inverted several times, placed at room temperature for 5 min and then centrifuged at 10,000 g for 5 min at 4°C and put on ice to maintain the clear phase separation. Finally, 400 μL of the top polar layer was removed, dried in a Speedvac, and stored at –80°C for analysis of polar metabolites, while 700 μL of the bottom nonpolar layer was removed, dried in a Speedvac, and stored at –20°C in 250 μL of 2:1 chloroform/methanol for lipid analyses. Serum lipids were extracted using a similar procedure except 25 μL of serum was used and then 600 μL of –20°C 2:1 chloroform/methanol was added. After vortexing and shaking, 125 μL of water was added to induce a phase separation and 200 μL of the top polar layer and 350 μL of the bottom nonpolar lipid layer were removed and stored as outlined above. Prior to analysis, the total lipid extracts were dried down and then reconstituted in 70 μL and 100 μL of MeOH for the DBS and serum samples, respectively. To generate pooled case and control samples for LC/MS/MS and LC/IMS-MS analyses, 5 μL aliquots from each reconstituted DBS and serum sample were removed and combined.

Combined analysis:

Analysis ID AN000929 AN000930 AN000931
Analysis type MS MS MS
Chromatography type Reversed phase Reversed phase GC
Chromatography system Waters NanoAcquity Waters NanoAcquity Agilent 7890A
Column Waters Acquity HSS T3 (150 x 1mm, 1.8um) Waters Acquity HSS T3 (150 x 1mm, 1.8um) Agilent HP5-MS (30m × 0.25mm, 0.25 um)
MS Type ESI ESI EI
MS instrument type Orbitrap Orbitrap Single quadrupole
MS instrument name Thermo Velos Pro Orbitrap Thermo Velos Pro Orbitrap Agilent 7890A
Ion Mode POSITIVE NEGATIVE POSITIVE
Units Peak Height Peak Peak Area

Chromatography:

Chromatography ID:CH000663
Instrument Name:Waters NanoAcquity
Column Name:Waters Acquity HSS T3 (150 x 1mm, 1.8um)
Chromatography Type:Reversed phase
  
Chromatography ID:CH000664
Instrument Name:Agilent 7890A
Column Name:Agilent HP5-MS (30m × 0.25mm, 0.25 um)
Chromatography Type:GC

MS:

MS ID:MS000825
Analysis ID:AN000929
Instrument Name:Thermo Velos Pro Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS000826
Analysis ID:AN000930
Instrument Name:Thermo Velos Pro Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
Ion Mode:NEGATIVE
  
MS ID:MS000827
Analysis ID:AN000931
Instrument Name:Agilent 7890A
Instrument Type:Single quadrupole
MS Type:EI
Ion Mode:POSITIVE
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