Ethyl acetate

Three NeuroPs (4(RS)-4F_4t-NeuroP, 4-epi-4-F_3t-NeuroP, 4-F_4t-NeuroP) and three F_2t-dihomo-IsoPss (17-epi-17-F_2t-dihomo-IsoP, 17-F_2t-dihomo-IsoP, Ent-7(RS)-7F_2t-dihomo-IsoP) were synthesized by Durand’s team at the Institut des Biomolecules Max Mosseron (IBMM) (Montpellier, France). Table 1 shows the different NeuroPs and F_2t-dihomo-IsPs molecules tested in this study.
The enzyme β-glucuronidase, type H2 from Helix pomatia and BIS-TRIS (bis-(2hydroxyethyl)-amino-tris(hydroxymethyl)-tris(hydroxymethyl)-methane) were obtained from Sigma-Aldrich (St Louis, MO, USA). All LC-MS-grade solvents were obtained from J.T. Baker (Phillipsburg, NJ, USA). Hydrochloric acid, hexane, trichloroacetic acid, and ethyl acetate were obtained from Panreac (Caste3llar del Vallés, Barcelona, Spain). Strata X-AW solid-phase extraction (SPE) cartridges, 100 mg per 3 mL, were purchased from Phenomenex (Torrance, CA, USA). Water was treated in a Milli-Q water purification system from Millipore (Bedford, MA, USA).

L-cysteine, γ-benzyl-L-glutamate ((Bzl)Glu), ɛ-Z-L-lysine ((Z)Lys), L-leucine (Leu), triphosgene, α-pinene, trifluoromethanesulfonic acid (TFMSA), trifluoroacetic acid (TFA), 2-(p-tolyl)-pyridine (N^Ctpy) and other reagents were purchased from Sigma–Aldrich (Germany) and used as received. 1,4-Dioxane, n-hexane were purchased from Vecton Ltd. (Russia) and distilled prior to use. Dimethyl sulfoxide (DMSO), also purchased from Vecton Ltd. (Russia), was dried under molecular sieves 4 Ǻ and distilled under vacuum. Ethyl acetate purchased from Panreac (Spain) was distilled before application. All solvents were purified and distilled using standard procedures.
HEK 293 (human embryonic kidney cells), HELF (human embryonic lung fibroblasts) and U937 (human lymphoblast lung cells) cell lines were purchased from BioloT (Russia). The cells were cultured in Dulbecco’s Modified Eagle Medium (BioloT, Russia) and grown in Dulbecco’s Modified Eagle’s Medium containing 10% (v/v) heat-inactivated fetal bovine serum (FBS, HyClone Laboratories, UT, USA), 1% L-glutamine, 1% sodium pyruvate, 50 U/mL penicillin and 50 μg/mL streptomycin (BioloT).

In this study, Comercializadora Tint Café S.A.S, Villeta, Colombia, supplied coffee pulp samples of both colors (red and yellow). β-Carotene of 97.0% (w/w) purity was used from Sigma-Aldrich (product number 7235-40-7). The chemicals 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-Azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), and anhydrous caffeine (C₈H₁₀N₄O₂), were also used from Sigma-Aldrich. Absolute ethanol (C₂H₅OH, 99.8% v/v), ethyl acetate (C₄H₈O₂, 99.5% v/v), acetone (C₃H₆O, 99.5% v/v), hydrochloric acid (HCl, 37.0% v/v), and sodium hydroxide (NaOH, 98.0% w/w) were obtained from Panreac (Gmbh). For the determination of total dietary fiber (TDF), insoluble dietary fiber (IDF), and soluble dietary fiber (SDF) heat-stable α-amylase, protease, and amyloglycosidase (Sigma® Life Science) were utilized for enzymatic digestion. Anhydrous disodium hydrogen phosphate (Na₂HPO₄, 99.0% w/w) and anhydrous sodium dihydrogen phosphate (NaH₂PO₄, 99.0% w/w) from Panreac were used for preparing the phosphate buffer in TDF, IDF, and SDF quantification. For the extract dilution a Don Olio (Colombia) sunflower oil was used. As wall materials, arabic gum from CIACOMEQ S.A.S. and maltodextrin from Cimpa S.A.S. were used.