Parastrephia quadrangularis (tola) is a native plant of the Chilean Andean Altiplano that is traditionally used for its anti-inflammatory properties. In this study, the aerial parts of the plant were analysed to determine their fatty acid (FA) profile and to identify bioactive compounds using gas chromatography-mass spectrometry (GC-MS). Both conventional extraction methods and Supercritical Fluid Extraction (SFE) were employed, using a 23 factorial design with centre-point replicates. The variables included temperature (30-60 °C), pressure (15-45 MPa), and ethanol as a cosolvent (0-30% v/v). Extraction kinetics were evaluated using a linear spline model under central conditions (45 °C, 30 MPa, 15% ethanol). Response variables included extraction yield, Total Phenolic Content (TPC), antioxidant activity measured by Trolox Equivalent Antioxidant Capacity (TEAC), and FA composition. A factorial design identified pressure and ethanol concentration as key drivers of phenolic content and antioxidant activity, as supported by confocal autofluorescence microscopy. Multi-response optimisation based on the desirability function was applied to simultaneously maximise all response variables, yielding predicted optimal extraction conditions at 60 °C, 45 MPa, and 30% v/v ethanol for P. quadrangularis. The FA profile highlighted polyunsaturated FAs such as oleic, linoleic, and linolenic acids, as well as saturated FAs including palmitic and lignoceric acids, and short-chain non-volatile FAs. GC-MS analysis revealed metabolites potentially responsible for the plant's traditionally reported therapeutic effects. Overall, these results highlight ethanol-based SFE as a sustainable strategy for recovering phenolic compounds and antioxidant-related fractions from ancestral medicinal plants.
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