To develop a protocol for extracting metabolites and lipids from a single dried blood spot for dual omics analyses.
Approach:
Extraction Method: A modified Folch extraction is used to separate metabolites and lipids from a 10 µL dried blood spot using a 2:1 mixture of chloroform and methanol, followed by sonication, water addition, and centrifugation.
Analysis Technique: The extracts are analyzed using reversed-phase liquid chromatography-tandem mass spectrometry, specifically with a Q Exactive Orbitrap mass spectrometer.
Data Processing: Two open-source tools, Mzmine and LipidMatch, are utilized for metabolite and lipid data processing, respectively.
Key Findings:
The method allows for the extraction of both metabolites and lipids from a single dried blood spot.
Results are semiquantitative, enabling comparison of peak heights or areas between groups.
The protocol includes controls to assess contamination, analytical variation, and instrument performance.
Limitations:
Absolute concentrations cannot be determined without additional standards and method validation.
Variability in lipid feature identification was observed between extraction trials.
Conclusion:
This protocol provides a streamlined approach for dual omics analyses from a single dried blood spot, facilitating research in small sample contexts.
