Summary of Study ST000608

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench,, 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.


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 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.
Pacific Northwest National Laboratory
Last NameBaker
First NameErin
Address902 Battelle Boulevard, Richland, WA, 99354, USA
Submit Date2017-05-13
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailGC/LC-MS
Release Date2017-07-10
Release Version1
Erin Baker Erin Baker application/zip

Select appropriate tab below to view additional metadata details:

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.