*2.2. Milling*

Leaves were separated from stems prior to the milling process. The milling was done using a Mixer Mill MM 400 (Retsch, Arzberg, Germany) which utilizes a metal ball inside a stainless-steel grinding jar that vibrates at a high horizontal speed to perform the grinding. Leaves were loaded in jars, sealed and securely positioned on the mixer instrument with a locking mechanism. Leaves were ground into a fine powder for 30 s at a vibrational frequency of 30 Hz. Milling was done on the same day of the experiment and powder used immediately for solvent extraction.

### *2.3. Solvent Extraction*

Pressurized liquid extraction method was performed using an accelerated solvent extraction Dionex 350 instrument (Dionex, Sunnyvale, CA, USA). Twenty grams of each milled sample was mixed with 10 g of diatomaceous earth (Thermo-Fisher Scientific, Waltham, MA, USA) and placed in a 100 mL stainless steel extraction cell with a paper filter installed at the bottom end of the cell. Extraction settings were at 60 ◦C with five cycles under a nitrogen pressure of 1000 psi. Four extraction solvents, ethanol (≥98%) hexane (≥98.5%), methanol (≥98%) and water were used for extraction. Each extract was collected into an amber glass bottle, filtered with No. 1 Whatman filter, transferred into a centrifuge tube and placed in a miVac DUO centrifugal vacuum concentrator (Genevac Ltd., Gardiner, NY, USA) to evaporate the organic solvent at 45 ◦C. Dried extracts were weighed and stored at 4 ◦C until further use.
