## 2 Materials and Methods
### Extractions of Bioactive Compounds from Apple Juice and Apple By-Product Samples  
Samples of three whole organic-farmed apples of the Starking variety, grown in Zagora of Pelion Mountain in Greece, were commercially purchased and then washed and pressed in a juicer, from which apple juice was produced; apple by-products (apple pomace) were also co-obtained from this processing. The extraction of total lipids (TLs) and their separation into Total Amphiphilic Compounds (TACs) and Total Lipophilic Compounds (TLCs) was performed by a modification of the Bligh and Dyer extraction [5] and Kapoulas and Galanos [6] countercurrent partitioning methodology, according to [PERSON] et al. [7]. Briefly, the juices from Greek Organic Starking apples, just after they were produced, were transferred to separatory funnels, and appropriate volumes of chloroform and methanol were added for the overall solution to reach a chloroform/methanol/water ratio of 1/2/0.8 (\(v/v/v\)). After thorough shaking, specific volumes of chloroform and water were added to obtain a biphasic chloroform/methanol/water ratio of 1/1/0.9. After a new shaking, the biphasic system was developed, from which the lower chloroform-based organic phase of the juice TL extracts was collected in a round-bottom flask. In the meantime, each apple pomace sample was further mechanically homogenized using a blender in the presence of a mixture of the same organic solvents (chloroform/methanol/water, at a ratio of \(1/2/0.8;v/v/v\)), and then subjected to filtration from the precipitated residues with a filter paper by pumping in a Buchner filtering apparatus under vacuum conditions. The filtrates were transferred to separatory funnels and additional amounts of chloroform and water were added to obtain a two-phase chloroform/methanol/water ratio of \(1/1/0.9\)\(v/v\) after vigorous shaking. Again, the lower chloroform-based organic phases of the apple pomace TL extracts were collected in a round-bottom flask. Solvents of all TL extracts were completely evaporated from the samples in several steps by using flash rotary evaporation (Rotavapor(r) R-300 Rotary Evaporator Standalone, BUCHI, DPTH Campus, Kavala, Greece) under a vacuum in different pressures for each solvent. The final extracts of TL and TLC had an oily consistency; the TLs were easily dissolved in chloroform/methanol 1:1 (\(v/v\)); the TLCs in petroleum ether, while the TAC extracts were a more complex mixture of a semi-oily dried paste texture that was also easily dissolved in chloroform/methanol 1:1 (\(v/v\)). The extracts were transferred to small pre-weighed glass vials using small amounts of chloroform/methanol \(1/1\) (\(v/v\)), where all remaining solvents were further evaporated under a stream of nitrogen. The obtained juice and by-product extracts were weighed and stored under a nitrogen atmosphere at \(-20\)\({}^{\circ}\)C for further analysis. As mentioned in an earlier paper by [PERSON] et al. [7], a counter-current partitioning methodology based on previously equilibrated petroleum ether (for acquiring TLCs in the upper phase of the developed biphasic system) and 87% ethanol in water (for obtaining TACs in the lower phase of the developed biphasic system) was used to extract the total amphiphilic (TAC) and lipophilic components (TLC) from these TL extracts. Small volumes (1-3 mL) of petroleum ether for TLC and a \(1/1\) (\(v/v\)) solution of chloroform/methanol for TAC were used to evaporate the solvents from the separated TACs and TLCs once more in a rotary flash evaporator at 37 \({}^{\circ}\)C under a vacuum, before being transferred to small, pre-weighed glass tubes. Under a nitrogen stream, the solvents used to transfer the extracted lipid samples to each tube were evaporated, and all the acquired TLs, TACs and TLCs from each sample were weighed to determine the extraction yield and eventually stored at \(-20\)\({}^{\circ}\)C (under nitrogen) for additional analysis. Just prior to this additional analysis, all dried TAC and TLC extracts were redissolved in 1 mL of ethanol and octane, respectively.