A sub-microliter SFE coupled to the SFC-PTR ionization apparatus was designed.

Compared to SFE-PTR, a 16-fold sensitivity improvement was achieved by the SFC.

A split injection drastically improves theoretical plate number.

The SFC-PTR MS has an extremely high sensitivity for deprotonated FFAs.

The application of proton transfer ionization reaction mass spectrometry (PTR MS) combined with microscale supercritical fluid extraction (SFE) and supercritical fluid chromatography (SFC) aiming to quantitate single-cell fatty acid analysis levels was investigated. Using a microscale extraction vessel, the obtained low limits of quantitation (LLOQs) of arachidonic acid and arachidic acid were 1.2 and 2.7 fmol, respectively, by using less than 1 µL of sample on stainless steel frit. A series of phthalate, vitamin K1, and α-tocopherol were also tested, and the LLOQ was less than one femtomole for phthalate and 35 and 13 fmol for vitamin K1 and α-tocopherol, respectively. A microliter portion of SFE extracts was introduced into the SFC column by split injection, improving the reproducibility of the chromatography and separation efficiency. The method in the present study has great potential to quantitate lipophilic molecules on the nanogram scale of a sample without complex preparation procedures.

Supercritical fluid extraction

Supercritical fluid chromatography

Trace-level fatty acid analysis

Proton-transfer-reaction

Time-of-flight mass spectrometer

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