Supercritical fluid chromatography-tandem mass spectrometry of oxygen heterocyclic compounds in Citrus essential oils

Oxygen heterocyclic compounds are secondary metabolites mainly present in the non-volatile fraction of cold-pressed Citrus essential oils. Under this denomination are included coumarins, furocoumarins, and polymethoxyflavones. These compounds possess numerous beneficial properties for human health, but the ingestion of large amounts of coumarins is often related to toxic effects, whereas the phototoxicity caused by furocoumarins and UVA exposure has been well known for a long time. This research has been aimed at the validation of an analytical approach, based on supercritical fluid chromatography coupled to tandem mass spectrometry, for the analysis of OHCs in Citrus essential oils. Among eight columns tested, packed with different stationary phases, the pentafluorophenyl allowed the best baseline separation in 8 min and by using less than 10% of methanol. Calibration curves of twenty-eight standards (coumarins, furocoumarins, polymethoxyflavones) were constructed on spiked lemon distilled essential oil and the method was validated according to the EURACHEM guidelines, by calculating linearity, limit of detection (LoD), limit of quantification (LoQ), accuracy, intra-day, and inter-day precision. Specifically, recoveries were in the 80.0-118.6% range, regression coefficients were between 0.9904 and 0.9998, the LoDs were in the 0.0004-0.0470 mg kg(-1) range, the LoQs were in the 0.0014-0.1536 mg kg(-1) range, and coefficients of variation were between 0.3 and 2.6% (intra-day) and 1.1 and 7.4% (inter-day). The quantitative profiles of thirteen cold-pressed Citrus essential oils were determined.

Automated summary

Oxygen heterocyclic compounds are beneficial secondary metabolites present in cold-pressed Citrus essential oils. An analytical approach based on supercritical fluid chromatography coupled to tandem mass spectrometry was validated for the analysis of these compounds. The method showed good separation and sensitivity, with accurate quantification. The study provides quantitative profiles of different Citrus essential oils, highlighting their content of oxygen heterocyclic compounds.