Supercritical CO2 Extraction and Tandem Mass Spectrometry of the Medicinal Plant Sagan Dalya (Rhododendron adamsii)

<b>Background:</b> In Siberian folk medicine, Sagan-Dalya (<i>Rhododendron adamsii</i> Rehder) of the <i>Ericaceae</i> family is used as a tonic and restorative in the form of infusions and decoctions. Pharmacological studies have shown that alcoholic extracts of this plant enhance performance and have anti-inflammatory and immunomodulatory effects. <i>Rhododendron adamsii</i> shoots accumulate essential oil (up to 1.6%), flavonoids (1.8-3.0%), tannins (up to 6.9%), phenolic carbolic acids, β-sitosterin, oleanolic and ursolic acids, simple phenolic compounds, and coumarins. <b>Methods:</b> Supercritical carbon dioxide extraction (SC-CO₂) is the most preferred environmentally friendly and selective method for extracting these natural compounds from the plant matrix of <i>Rh. adamsii</i> due to their high thermolability. Tandem mass spectrometry was applied to detect chemical compounds. Mass-spectrometry (MS) analysis was performed on an ion trap equipped with an ESI source in negative and positive ion modes. The capture rate was one spectrum/s for MS and two spectrum/s for MS/MS. All experiments were repeated three times. A four-stage ion separation mode (MS/MS mode) was implemented. <b>Results:</b> The operative parameters and working conditions have been optimized by different pressure (100-400 bars) and temperature (31-70 °C) regimes, and CO₂ flow rate (10-25 mL/min) with 1 C₂H₅OH as a co-solvent. The extraction time varied from 60 to 90 min. The maximum global yield of biologically active substances (BAS) from <i>R. adamsii</i> leaves and stems was observed under the following extraction conditions: Pressure: 350 bar, extraction temperature: 65 °C, extraction time: 1 h; the global yield of BAS was 8.5 mg/g of plant sample; the share of the co-solvent (C₂H₅OH) was 2%. In total, forty-nine different BAS were identified in the <i>Rh. adamsii</i> SC-CO₂ extracts. <b>Conclusions:</b> The obtained results may shed new light on the scientific basis for the traditional medicinal use of <i>Rh. adamsii</i> leaf and stem extracts. The pharmacological contribution of the identified phytocannabinoids requires further detailed study. It is hypothesized that the excellent transdermal permeability of supercritical extracts may open new therapeutic approaches using transdermal formulations based on SC-CO₂ extracts of <i>Rh. adamsii</i>.