Silica gel 60 f254 aluminium sheets

All organic solvents (analytical grade) and other chemicals used in this study were supplied by either Merck (Darmstadt, Germany) or Sigma (St. Louis, USA) chemical companies. Merck 20 cm × 20 cm silica gel 60 F₂₅₄ aluminium sheets were used for thin layer chromatography (TLC). Developed TLC plates were viewed under an ultraviolet lamp (254 and 366 nm) and further visualized with 10% H₂SO₄ in MeOH followed by heating^{36 (link)}. Column chromatography (CC) was carried out using Merck silica gel 60 (0.040–0.063 mm). ¹H, ¹³C and 2D nuclear magnetic resonance (NMR) spectra were recorded using deuterated chloroform (CDCl₃) at room temperature on a Bruker Avance^III 400 MHz spectrometer.

Column chromatography (SPE) of the sponge extract was performed on Supelco Dianion^® HP20SS (Bellefonte, PA, USA) and silica gel 60 (0.040–0.063 mm) from Merck KGaA (Darmstadt, Germany). Normal phase TLC was performed on silica gel 60 F₂₅₄ aluminium sheets purchased from Merck KGaA (Germany) and visualized under UV light at 254 and 365 nm. All high-performance liquid chromatography procedures were carried out with Agilent technologies equipped with ultraviolet and refractive index detectors using a Whatman 10 μm silica (2) semi-preparative column 50 cm × 10 mm (i.d.). NMR samples were prepared in deuterated solvents and all experiments acquired on a Bruker Avance III HD 400 MHz spectrometer equipped with a 5 mm BBO probe at 298K. Chemical shifts were referenced to deuterated solvent peaks (CDCl₃ δ_H 7.25, δ_C 77.00) and reported in ppm. Homogenisation and sonication of the SLNs was accomplished using an IKA^® T18 digital Ultra Turrax^® homogenizer and Bandelin Sonoplus HD 2070, respectively. Cell studies were performed under a class II biological safety cabinet. To visualize the cells, a Nikon light microscope with 20x magnification was used together with a Leica EC digital camera.

Pharmaceutical grade propylene glycol, glycerol, ethyl acetate, hexane, petroleum ether, ethanol, and KOH were purchased from Fischer Scientific (Thermo Fisher Scientific, Canada). Plant-based CBD isolate material (> 99% purity) was purchased by an independent manufacturer (The Valens Company, Canada), under authorization from Health Canada. Reactions were performed in disposable VWR® 1 dram (4 mL) or 0.5 dram (2 mL) vials. Column chromatography was performed using SiliaFlash F60 (40–63 mm) silica (Silicycle, Canada). Thin-layer chromatography was run on silica gel 60 F₂₅₄ aluminium sheets (Merck, Canada) and visualized by UV fluorescence (254 nm), followed by KMnO₄. Sun lamp experiments used a 275 W tanning light bulb (General Electric, Canada). White light experiments used a CF1EL/MICRO/865 bulb with a colour temperature of 6500 K (Sylvania, Canada). A Royal Blue Rebel LED on a SinkPAD-II 40 mm round 7-Up base with the power supply controlled by a XANTREX LXQ 30–2 Dual Power Supply was used to supply 448-nm light.

All commercial chemicals were purchased from Merck (Darmstadt, Germany) and were used without further purification. (±)-cis-N-normetazocine was obtained from Fabbrica Italiana Sintetici. Melting points were determined in open capillary tubes with a Büchi 530 apparatus and are uncorrected. Analytical TLC was performed on silica gel 60 F₂₅₄ aluminium sheets (Merck) with a fluorescent indicator. Components were visualized by UV light (λ = 254 nm) and iodine vapour. Flash column chromatography was carried out on Merck silica gel 60 (230–400 mesh). Optical rotations were determined in MeOH solution with a Perkin-Elmer 241 polarimeter. ¹H and ¹³C NMR spectra were routinely recorded on a Varian Inova-200 spectrometer in CDCl₃ solution; chemical shifts δ are expressed in ppm with reference to tetramethylsilane as an internal standard. Elemental analyses (C, H, and N) were performed on a Carlo Erba 1106 analyzer and the analysis results were within ± 0.4% of the theoretical values. All reported compounds had a purity of at least 95%.

Ethyl acetimidate hydrochloride and 4-(dibutylamino)benzaldehyde (DBA) were purchased from Sigma-Aldrich and used without further purification. The solvents and other reagents were purchased from Sigma Aldrich, Fluka and Merck and were dried and purified by standard laboratory methods. Trimethylolpropane triacrylate (TTA, Genomer 1330) and ethoxylated-(20/3)-trimethylolpropane triacrylate (ETA, Sartomer 415) were received as a gift from Rahn and Sartomer, respectively. Thin layer chromatography analysis was performed on silica gel 60 F₂₅₄ aluminium sheets from Merck. Column chromatography was performed by preparative MPLC using a Buechi Sepacore Flash System (Buechi pump module C-605, Buechi control unit C-620, Buechi UV-Photometer C-635 with peak detection set at λ = 295 nm, Buechi fraction collector C-660). Glass and polyethylene columns were used, packed with Silicagel 60 on VWR silica gel 60 (0.040–0.063 mm particle size).
The preparation and analysis of the photosensitive compounds and formulations was conducted in an orange light lab. The windows and fluorescent lamps were covered in adhesive foils or coated so that light with a wavelength < 520 nm was cut off.