Supercritical fluid extraction as a preparation method for mass spectrometry of dried blood spots

•The application of dried blood spots (DBS) with mass spectrometry (MS) is expanding.•Supercritical fluid extraction (SFE) was applied as a preparation method.•Metabolites with a wide range of polarity in DBS were extracted at once.•Under optimized conditions, over 160 metabolites were analyzed stably.•These results clearly show the usefulness of SFE in MS of DBS.

The potential of supercritical fluid extraction (SFE) as a preparation method for mass spectrometry of dried blood spots (DBS) was examined. SFE is generally used for the extraction of hydrophobic compounds, but hydrophilic metabolites such as amino acids, amines, and nucleic-acid-related metabolites could be extracted by adding a low level of methanol as a modifier. Under the optimized conditions, over 200 metabolites were detected from a dried serum spot, of which over 160 metabolites could be analyzed stably (RSD <20%). These results show that SFE is an effective extraction method of metabolites with a wide range of polarity in DBS.